Arsenic-Induced Neurocardiac Toxicity and Protective Role of Resveratrol: Histopathological and Molecular Insights

Arsenic-Induced Neurocardiac Toxicity and Protective Role of Resveratrol: Histopathological and Molecular Insights | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Arsenic-Induced Neurocardiac Toxicity and Protective Role of Resveratrol: Histopathological and Molecular Insights Saroj N/A, Kamakshi Mehta, Balpreet Kaur, Kamlesh Kumar Pandey, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6316307/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 20 May, 2025 Read the published version in Journal of Molecular Histology → Version 1 posted 8 You are reading this latest preprint version Abstract Arsenic toxicity is a global health problem chiefly targeting soft tissues of the body like the brain and heart. The major mechanism underlying arsenic-induced neurotoxicity is oxidative stress. Particularly, the neurons and cardiac myocytes show limitless susceptibility to oxidative stress. Herein, we examined the impact of prolonged arsenic exposure and resveratrol post-treatment on the cardiac and neuronal [Ventromedial hypothalamic nucleus (VMH)] morphology. Adult mice were segregated into control and experimental groups. Controls received distilled water, whereas experimental mice received oral gavage of low (2mg/kg bw) and high (4 mg/kg bw) concentrations of ATO (Arsenic trioxide) for 45 days. Mice were sacrificed on day 45 to obtain perfusion-fixed hearts and brains for histological and morphometric studies. Long-term ATO exposure resulted in a higher heart-to-body weight ratio than controls, suggesting ATO-induced hypertrophy. Microscopic observations revealed a regular arrangement of cardiac muscle fibres, branching patterns of cardiomyocytes, and fibroblasts across all the treatment groups. However, increased cardiac myocyte diameter in ventricles and substantial fibrosis in vessel walls were noticed in ATO-alone exposed hearts relative to controls. Selective vulnerability of hypothalamic neurons following ATO exposure was evident by significant alterations in morphometric parameters (reduced cell density and soma size) in the VMH nucleus of animals receiving ATO (2 and 4 mg/kg) alone. These dramatic histopathological alterations were found to be restored after ATO + Res co-treatment. We also examined the expression of ER-α in the preoptic area of the hypothalamus and indicated downregulation of ER-α due to prolonged ATO exposure. Our findings highlight Resveratrol as a potent neurocardiac protector against ATO toxicity via estrogen signaling modulation, supporting its therapeutic potential in arsenic poisoning. Arsenic trioxide Cardiotoxicity Neurotoxicity Oxidative stress Resveratrol Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 1. Introduction Exposure to heavy metals (arsenic, lead, cadmium, mercury, and copper) of hydro-geological origin may adversely affect humans (Balali-Mood et al. 2021 ). One of these toxic metalloids is Arsenic with the symbol As and an atomic number 33. A significant source of inorganic arsenic ( iAs) exposure is contaminated groundwater with higher As concentrations, thereby affecting communities that rely entirely on groundwater for drinking and cooking (Smith et al. 2000 ). According to past surveys, arsenic has been identified as the world’s second-leading water-borne cause of mortality (Straif et al. 2009 )Moreover, several Indian states lying along the GMB (Ganga-Meghna-Brahmputra) belt (West Bengal, Jharkhand, Bihar, Uttar Pradesh, Assam, Manipur, and Chhattisgarh) are reported to be adversely impacted by groundwater contamination with As levels that exceed the WHO permissible limit. Despite its reputation for toxicity, As serves paradoxically as a therapeutic agent employed since ancient eras for treating various illnesses (Paul et al. 2023 ). Arsenic trioxide (ATO), an inorganic compound of As , is currently used as an anti-neoplastic agent in treating acute promyelocytic leukemia (APL). Even though APL patients generally tolerate ATO therapy well, it can also cause toxic effects in some cases. However, its use is limited to a certain extent keeping in consideration the adverse health effects observed in APL patients (Wang et al. 2019 ). Various cardiac abnormalities such as prolonged QT interval, arrhythmias, and pericardial effusion have been observed in APL patients following ATO therapy (Ohnishi et al. 2000 ; Barbey et al. 2003 ). These cardiac complications from ATO exposure are significant because they may predict a fatal outcome in APL patients. Apart from the heart, other primary targets for iAs toxicity include the liver, lungs, kidneys, and brain. However, symptoms of iAs poisoning may take from hours to years to manifest, depending on the dose and length of exposure. With context to the brain, iAs -induced neurotoxicity leads to neuroinflammation, oxidative stress, and neurodegeneration, particularly in the hippocampus and cortex, resulting in cognitive decline and neuronal loss (Rodríguez et al. 2001 , 2003 ; Luo et al. 2012 ; Medda et al. 2020 ; Mehta et al. 2021b ). It disrupts neurotransmission by impairing dopamine, glutamate, and acetylcholine signaling, affecting learning, memory, and motor functions (Wang et al. 2012 ; Tyler and Allan 2014 ; Sun et al. 2017 ; Chandravanshi et al. 2019 ). Additionally, As increases blood-brain barrier permeability, allowing further toxin entry, and induce mitochondrial dysfunction, leading to neuronal apoptosis. These effects contribute to cognitive and behavioral deficits such as memory loss, attention impairments, anxiety, and depression. Moreover, As exposure causes epigenetic alterations, potentially leading to long-term neurodevelopmental consequences and increasing the risk of neurodegenerative diseases like Alzheimer’s and Parkinson’s (Tyler and Allan 2014 ; Medda et al. 2020 ). Our study primarily focuses on iAs -induced toxicity in the heart and Ventromedial Hypothalamic (VMH) area. Functionally, the VMH is involved in energy homeostasis, feeding behavior, and sympathetic nervous system regulation. VMH is also reported to be targeted in obesity and diabetes, which are major risk factors for cardiovascular disease (CVD). By selecting the VMH, we can investigate how central neural circuits related to metabolic regulation might influence or be influenced by cardiovascular function. Additionally, the VMH is involved in regulating the body’s stress response, which has direct implications for both metabolic and cardiovascular health. To date, numerous epidemiological studies have established a strong correlation between chronic arsenic exposure and deteriorated mental health. Nevertheless, there are scanty animal studies that could reveal the association between arsenic exposure and neurocardiac health. Mouse is an ideal species to study the in-vivo effects of drugs, toxic metals on several organs, including the nervous system. Several mechanisms have been proposed for the cardio- and neurotoxic effects of ATO including oxidative stress, apoptosis, altered DNA methylation, genotoxicity, mitochondrial damage, and calcium dysregulation (Kitchin and Conolly 2010 ; Eyvani et al. 2016 ). Additionally, As also interacts with endogenous cellular antioxidants such as glutathione (GSH) to form a transient compound called glutathione trioxide, thus depleting the cellular GSH levels and resulting in oxidative stress (Ran et al. 2020 ). Currently, clinical medicine has no effective strategies to protect against cardiotoxicity caused by inorganic arsenic ( iAs ). Given that oxidative stress is a key mechanism in iAs -induced toxicity, it is worthwhile to investigate the application of naturally occurring antioxidants in animal models of arsenic toxicity. Resveratrol (3,5,4′-trihydroxy-trans-stilbene) [ Res ] is a plant-based polyphenolic stilbene, found abundantly in grapes, all types of berries, soy, and peanuts. Because of its effective antioxidant activity, Res has been shown to protect against ATO-induced cardiotoxicity as well as neurotoxicity via upregulation of antioxidant enzymes and downregulation of apoptotic levels (Zhang et al. 2013 ; Mehta et al. 2021a ). The present study raises two possible questions: Can Arsenic Trioxide (ATO) induce toxic effects in the heart and ventromedial hypothalamic nucleus (VMH)? Given the known cardiotoxic and neurotoxic effects of ATO in Acute Promyelocytic leukemia (APL) patients and its role in generating oxidative damage, can Resveratrol supplementation alleviate these detrimental effects caused by prolonged exposure to ATO in mice, and if so, how? Hence, we intend to evaluate the potential cardio- and neuroprotective efficacy of Resveratrol, a proven antioxidant in a mouse model of ATO-induced toxicity affecting the heart and hypothalamus. 2. Methods 2.1 Animals Adult female Swiss albino mice (weight = 25–30 g) were procured from the Central Animal Facility (CAF) of All India Institute of Medical Sciences (AIIMS) after obtaining ethical clearance from the Institutional Animal Ethics Committee (IAEC). The animals were kept in cages under 12 hr light/12 hr dark cycles and temperature (20–25˚C) and humidity (50–60%) controlled rooms of CAF. They were fed routinely with a standard rodent diet ( chow ) and given free access to drinking water. Animals were acclimatized for 7 days before the start of the treatment regimen. 2.2 Study design After acclimatization, the animals were randomly divided into five groups with six animals per group: Group Ι - normal control (no treatment), Group II - vehicle control (5% Gum Acacia), Group III –Arsenic trioxide, ATO (2mg/kg b.w.) alone, Group IV- Res (40mg/kg b.w.) alone, and Group V- ATO and Res (2mg and 40 mg/kg b.w.). Both test substances were administered orally over an experimental period of 45 days. 2.3 Chemicals and reagents Arsenic trioxide (A1010), Resveratrol (R-5010), and Gum acacia (V800016) were purchased from Sigma Chemicals, St. Louis, USA. 2.4 ATO and Resveratrol dose selection The dose of ATO used in the present study was 2mg/kg body weight (bw) which is equivalent to the therapeutic dose in humans for the treatment of APL, i.e., 0.15 mg/kg per day for 60 days (Powell et al. 2011 ; Lo-Coco et al. 2013 ). The conversion of human equivalent dose (HED) to animal equivalent dose (AED) was done by applying a BSA (body surface area) based formula (Shin et al. 2010 ). To prepare a 2 mg/kg bw concentration of ATO, 20 mg of ATO was fully dissolved in 500 µl of NaOH, followed by dilution to a final volume of 20 ml with distilled water. The daily ATO dosage was determined based on the body weight of each animal. The dose for Res (40 mg/kg bw) was adopted from the previous literature, wherein an oral dose of 40 mg/kg bw has been documented to be cardio- and neuroprotective in animal models (Mukherjee et al. 2010 ; Mehta et al. 2021b , a ). Res was freshly prepared in 5% gum acacia and kept on ice until its administration through oral gavage. 2.5 Histology On Day 45, the animals were anesthetized with sodium pentobarbitone (60 mg/kg, i.p.) and underwent transcardial perfusion fixation with 4% paraformaldehyde (prepared in 0.1M PBS). The perfusion-fixed heart and brains were later processed for paraplast embedding and staining methods. The boundaries and gross histology of two AOI (Areas of Interest) were identified by different staining methods, as shown in Fig. 1 . Hematoxylin and Eosin (H-E) staining was performed to visualize the cardiac morphology (Fig. 1 C), and cresyl violet staining was done to observe the neuronal morphology in the ventromedial hypothalamic nucleus (VMH) (Fig. 1 A, B). Additionally, we performed Masson’s Trichrome staining to determine the status of connective tissue or fibrosis (if any) within the heart's ventricular walls. 2.6 Morphological observations Morphological observations were carried out on H-E and Masson’s trichrome stained transverse sections (7µm thick) of the mouse heart using a bright field Nikon E-600 microscope fitted with Nikon Digital Camera System (DS-Fil –U2). We studied various morphological features of VMH neurons and cardiac myocytes, including their branching pattern, shape, and size. The general appearance of fibroblasts and the location of intercalated discs were also observed in the heart sections of control and experimental mice. 2.7 Morphometric analysis in heart Measurement of wall thickness and nuclear size was carried out by the attached image analysis system (Nikon Imaging Software, NIS Elements AR 3.1). For determining the thickness of ventricular walls (LV, RV, and IVS), the first section was chosen randomly, and the subsequent section was every sixth section from it, and a total of six sections (6 different sites) were analyzed for further calculations. For each chamber (LV & RV), the diameter of around ten to twelve myocytes was measured at the centre of the nucleus (40X). In the same sections, the area of the nucleus was also determined. 2.8 Morphometric study in Ventromedial Hypothalamic (VMH) Nucleus The rostral-caudal limits of VMH were identified from serial brain sections, and the midpoint of the nucleus was located for analysis as previously illustrated (Garris et al. 1982 , 1985 ; Garris 1989 ). The first section was randomly chosen, followed by every seventh section, so the interval between sections considered for counting was 49 µm. This ensures that the same cell was not counted twice, given that the typical cell diameter in this region is smaller than 49 µm. A total of six sections were evaluated per animal (n = 5 animals/group) for the morphometric study. 2.8.1 Neuronal density (neurons/unit area) : A rectangular reference frame of a defined area (200×170 µm 2 ) (with solid lines on two adjacent sides and dotted lines on the other two adjacent sides) was placed on the specified region of the digital image taken at 40X (Fig. 2 ), and the neuronal cell bodies with nucleoli and lying within the VMH nucleus were considered for counting. According to the Forbidden line rule, the neurons lying along the right and upper lines of the frame were included for counting, while those lying along the left and lower lines of the reference frame were excluded. To standardize the measurements, only neurons exhibiting a well-stained nucleus with a distinct nucleolus were counted. Glia was readily distinguished from neurons by their size, nuclear shape, cytoplasm, location, and characteristic staining (Roy et al. 2005 ). 2.8.2 Soma size estimation : The Soma size of the VMH neurons was measured on digital images captured at higher magnification (100X) to have a clear and distinct observation of neuronal features such as soma, nucleus, cytoplasm, and nucleoli (Fig. 2 ). Cells showing euchromatin material and nucleoli within the nuclei and the nuclei surrounded by cytoplasm were identified as neurons (Benes et al. 2001 ; Korbo et al. 2004 ; Namavar et al. 2012 ). Sections showing the full extent (Bregma − 1.06 to -2.06 mm) of VMH nucleus per animal were analyzed for measuring the soma size of neurons in the VMH nucleus. An average of 10–15 neurons per section was considered for determining soma size estimation. Six sections per animal were considered, and an average of 450 neurons per group (n = 5/group) were analyzed to measure soma size. 2.9 Immunohistochemistry On day 46, the perfusion-fixed brains (without brainstem and cerebellum) were processed for immunohistochemistry. Briefly, the brains were cryopreserved in 15% and 30% sucrose solutions, embedded in OCT (optimum cutting temperature), and sectioned at 25µm using a cryotome (Leica CM1900). Coronal sections containing the hypothalamus were collected in PBS and treated with 3% H₂O₂ to quench peroxidase activity. Following PBS rinses, sections were blocked with 10% normal goat serum and incubated with rabbit monoclonal anti-ER alpha (estrogen receptor-α, 1:50, ab32063; Abcam) at 4°C for 48 hours. After incubation with biotinylated IgG peroxidase secondary antibody, immunostaining was visualized using a diaminobenzidine (DAB) reaction. Sections were then washed, dehydrated, cleared in xylene, cover-slipped with DPX, and scanned under a bright-field microscope to capture images. 2.10 Statistical Analysis The collected data from the control and experimental animals were analyzed by SPSS 17 (Chicago, IL, USA) statistical software, and graphs were plotted using graph pad prism (Version 9). Body weight, heart weight, and heart heart-to-body weight ratios were analyzed using one-way ANOVA followed by a post hoc LSD test for multiple comparison analysis. The data for the thickness of ventricular walls (LV, RV, IVS), the diameter of myocyte (LV and RV), and the area of the nucleus (LV and RV) were expressed as mean ± SD and analyzed by one-way analysis of variance (ANOVA) followed by the Bonferroni test for multiple comparisons among groups. Differences were considered statistically significant at p-value < 0.05. 3. Results 3.1 General physical characteristics During the experimental period, the animals were observed for their general well-being, including body weight, state of alertness, hair loss (if any), and physical activities like rearing and grooming. No significant differences were observed regarding these physical features. On day 1 of the experimental period, the body weights of the control and experimental animals were comparable (Table 1 ). On day 45, an increase in body weight was observed both in the control and the experimental groups, though the increase was not statistically significant (p = 0.64). The percentage change in body weight (Table 1 ) was maximum (12.76 ± 4.41%) in group IV (Res alone treated) followed by group III (ATO alone treated) with a value of 11.92 ± 3.48%. We herein assume that ATO at this low dose of 2mg/kg b.w. was not sufficient to alter the body weight of mice. Table 1 Descriptive statistics of Percentage (%) change in body weight, heart weight (mg), body weight (g), and heart-to-body weight ratio on Day 45 of the experimental period among control and experimental animals Animal Groups Body weight (gm) on day 1 Body weight (gm) on day 45 % age change in body weight Heart weight (mg) Heart weight/Body weight ratio NC 30.41 ± 4.60 33.31 ± 2.47 9.39 ± 3.42 285.2 ± 21.74 8.59 ± 0.81 VC 33.36 ± 1.86 33.88 ± 2.86 1.55 ± 2.04 246.28 ± 34.24 7.84 ± 2.04 ATO alone 29.6 ± 4.37 33.13 ± 2.42 11.92 ± 3.48 310.09 ± 19.13 9.37 ± 0.48 b* Res alone 31. 26 ± 1.68 35.25 ± 2.43 12.76 ± 4.41 241.08 ± 16.32 c*** 6.78 ± 0.81 c*** ATO + Res 30.51 ± 2.82 33.3 ± 3.18 9.14 ± 3.79 250.41 ± 17.73 d*** 7.58 ± 0.99 d*** Data is presented as mean ± SD; n = 6/group. Means were compared using one-way ANOVA followed by LSD test for multiple comparisons. b* VC group vs ATO alone; c*** ATO alone vs Res alone. d*** ATO alone group vs ATO + Res co-treated group. * p < 0.05; * * p < 0.01; *** p < 0.001. Abbreviations: NC- normal control, VC- vehicle control, ATO –Arsenic Trioxide, Res- Resveratrol alone, ATO + Res – ATO & Resveratrol co-treated group. Next, we observed that the heart weight was found to be higher in the ATO-alone treated animals (310.09 ± 19.13) relative to controls (Table 1 ); however, the data was not statistically significant. In Res alone and ATO + Res co-treated animals, the heart weight was significantly lower (241.08 ± 16.32; p = 0.000) and (250.41 ± 17.73; p = 0.000), respectively when compared to ATO alone treated animals. As shown in Table 1 , the heart to body weight ratio was observed to be significantly higher (9.37 ± 0.48; p = 0.03) in ATO-treated animals in comparison to vehicle controls (7.84 ± 2.04), suggesting ATO-induced hypertrophy in these animals. However, Res alone treated animals and ATO + Res co-treated animals showed significantly lower ratios (6.78 ± 0.81, p = 0.001) (7.58 ± 0.99, p = 0.01) in contrast to ATO alone treated animals, thus indicating the beneficial role of Res against this cardiotoxin. 3.2 Effects of Res on Left and right ventricular wall thickness The morphometric study on H&E-stained heart sections showed an apparent decrease in left and right ventricular thickness; however, the difference was found to be non-significant ventricular thickness on treatment with Res in the ATO-treated compared with ATO alone group (p > 0.05; non-significant) (Table 2 ). Table 2 Comparative analysis of morphometric parameters in mouse heart: Wall Thickness (LV/RV), Interventricular septum (IVS) Thickness, Cardiac Myocyte Nuclear Area and Cardiac Myocyte Diameter in Left Ventricle (LV) and Right Ventricle (RV) across the control and experimental groups. Animal Groups Wall thickness (mm)-LV Wall thickness (mm)-RV IVS Thickness (mm) Cardiac myocyte nuclear area in LV (µm 2 ) Cardiac myocyte nuclear area in RV (µm 2 ) Cardiac myocyte diameter in LV (µm) Cardiac myocyte diameter in RV (µm) NC 1.16 ± 0.08 0.55 ± 0.27 0.89 ± 0.18 35.41 ± 10.11 31.74 ± 6.04 9.64 ± 1.07 8.78 ± 1.31 VC 1.12 ± 0.10 0.66 ± 0.20 0.90 ± 0.15 38.24 ± 11.94 36.02 ± 12.91 10.32 ± 1.09 9.87 ± 1.66 ATO alone 1.18 ± 0.25 0.56 ± 0.17 0.88 ± 0.10 38.36 ± 11.14 34.83 ± 10.93 10.73 ± 1.28 a** 10.74 ± 0.22 a*** Res alone 1.06 ± 0.13 0.56 ± 0.18 0.92 ± 0.25 38.77 ± 9.41 39.52 ± 12.30 9.16 ± 1.38 c*** 9.53 ± 1.70 ATO + Res 1.02 ± 0.15 0.60 ± 0.22 0.66 ± 0.18 d*** 31.30 ± 6.20 d* 33.23 ± 7.82 8.88 ± 1.36 d*** 8.90 ± 1.27 d*** Values expressed as mean ± S.D, n = 6/group. Means were compared using one-way ANOVA followed by the Bonferroni test for multiple comparisons. a** and a*** denotes NC vs ATO alone treated group; c*** denotes ATO alone vs Res alone; d* and d*** denotes ATO alone vs ATO + Res co-treated group ( * p < 0.05, ** p < 0.01, *** p < 0.001). Abbreviations: NC- normal control, VC- vehicle control, ATO –ATO alone, Res- Resveratrol alone, ATO + Res – ATO & Resveratrol co-treated group. 3.3 Effects of Res on the Thickness of Interventricular Septum (IVS) No significant difference was observed in the thickness of IVS among control (0.89 ± 0.18) and ATO alone treated (0.88 ± 0.10, p = 1.000) animals. A significant decrease in the thickness of IVS was observed in the ATO + Res co-treated groups (0.66 ± 0.18, p = 0.000) compared with the ATO alone (0.88 ± 0.10) group, thereby indicating the beneficial role of Res on ATO-induced alterations in IVS thickness (Table 2 ). 3.4 Effects of Res on cardiac myocyte diameter and nuclear area The average values along with descriptive statistics of cardiac myocyte diameter in LV and RV walls are tabulated in Table 2 . The diameter of cardiac myocytes in LV was found to be significantly increased in ATO-alone treated animals (10.73 ± 1.28; p = 0.003) when compared to normal control. On the contrary, the animals receiving either Res alone (9.16 ± 1.38; p = 0.000) or co-treatment of ATO + Res showed a significant decrease (8.88 ± 1.36; p = 0.000) in myocyte diameter compared to ATO alone treated animals. Similarly in the RV region, animals exposed to ATO alone showed a significant increase in myocyte diameter (10.74 ± 0.22, p = 0.000) compared to control animals. ATO + Res co-treated animals revealed a significant decrease (8.90 ± 1.27, p = 0.001) in RV myocyte diameter compared to alone-treated animals (Table 2 ). ATO alone exposed animals showed an apparent increase in the cardiac myocyte nuclear area in the region of LV (38.24 ± 11.94) and RV (34.83 ± 10.93) as compared to normal control animals (LV- 35.41 ± 10.11; RV- 31.74 ± 6.04), respectively, though not statistically significant. ATO + Res co-treated group showed a significant decrease (31.30 ± 6.20, p = 0.030) in the nuclear area in the LV region as compared to ATO alone treated animals as shown in Supplementary Table 2. 3.5 General microscopic observations of cardiac tissue To study the morphological features of ventricular myocardium in the control, ATO- alone and ATO + Resveratrol co-treated mice, we examined H & E stained paraplast sections under the bright field microscope. Overall, cardiac muscle fibers were observed to be well arranged across control and experimental groups (Fig. 3– 7 ). Also, the branching patterns of cardiac myocytes along with sites of intercalated discs were evident across all the animal groups (Fig. 3– 7 ). The nuclei of the myocytes as well as fibroblasts presented a normal appearance in control as well as experimental animals (Fig. 3– 7 ). However, in Masson’s trichrome stained sections of ATO alone treated animals, substantial fibrosis in the vessel wall was noticed compared to control sections (Fig. 8 ). 3.6 Morphological observations in Ventromedial Nucleus of Hypothalamus (VMH) The ventromedial nucleus of the hypothalamus was identified as a bilateral cell group in the medial basal hypothalamus lying near the third ventricle. The outer limits of the VMH were defined by the encircling stria terminalis fibers, which separated the dense neuronal core of the nucleus from the relatively lesser neuron-concentrated area (Fig. 1 ). The cells of the ventromedial nucleus appeared to be variable in shape and size. Neuronal profiles in CV-stained sections containing VMH nuclei showed vesicular nuclei and prominent nucleoli under high-power magnification (Fig. 9 , 10). The neuronal population of the VMH nucleus showed variability based on their size and shape. Neurons of various shapes including spherical, round, oval, and triangular, and sizes, such as small, medium, and large, were observed in the VMH nucleus (Fig. 9 , 10). In most of the sections, VMH neurons presented round to oval-shaped cell bodies with round centrally placed nuclei having two or more nucleoli. The neurons were more densely packed in the ventromedial aspect of the third ventricle than in the dorsolateral aspect. Qualitatively, intensely stained small and medium-sized neurons were seen in some of the sections of ATO (2 & 4 mg/kg) alone groups in comparison to the control group (Fig. 9 , 10). Otherwise, no obvious change was observed in the morphological features of VMH neurons among experimental groups receiving ATO (2 & 4 mg/kg) alone in comparison to controls and RES supplemented. 3.7 Morphometric observations in Ventromedial Nucleus of Hypothalamus (VMH) The values for neuronal density and soma size of VMH neurons are presented in Fig. 11 . The neuronal density in VMH nucleus was found to be significantly (p = 0.0471, p = 0.0303) reduced in ATO alone treated groups (2mg/kg, 6690 ± 1410 & 4 mg/kg, 6537 ± 827) when compared to normal control (9471 ± 1337). The mean neuronal density in groups receiving RES along with ATO (ATO 2 + 40mg/kg, 9813 ± 1400 & ATO 4 + 40 mg/kg, 9480 ± 2001) was significantly ( c p=0.0174, f p=0.0295) greater than ATO (2 and 4 mg/kg) alone treated groups. The mean soma size of VMH neurons was observed to be significantly (p < 0.0001) decreased in ATO (2 & 4 mg/kg) (246 ± 12.21; 222 ± 32.89) alone exposed groups in comparison to the normal control group (418 ± 48.31). On the contrary, ATO + RES co-treated (ATO 2 + 40mg/kg = 409 ± 37.19; ATO 4 + 40mg/kg = 397 ± 26.17) groups showed a significant (p < 0.0001) increase in mean soma size of VMH neurons when compared to the mean of soma size of VMH neurons in ATO 2 and 4 mg alone treated group (409 ± 37.19; 397 ± 26.17), thereby suggesting RES-induced restoration of soma size of neurons in VMH nucleus. 3.8 Effect of ATO + RES on Estrogen Receptor-α Expression Since estrogen receptor regulation plays a crucial role in metabolism, neuroendocrine function, and cardiovascular health, we aimed to test the hypothesis that prolonged arsenic exposure disrupts estrogen signaling, potentially affecting both neuroendocrine and cardiac function. Immunohistochemical localization of ERα was observed in the coronal sections (25 µm) of the mouse brain, showing the preoptic area of the hypothalamus (bregma + 0.74 mm to -0.58 mm). Specifically, ERα immunoreactivity was observed in the cytoplasm of the neurons as well as in the neuropil among control and experimental animals (Fig. 12). Largely, the ERα + ve neurons were observed to be rounded or pyramidal in shape (Fig. 12). Qualitatively, ERα expression was less intense in the cytoplasm of the neurons in preoptic areas of the experimental groups receiving ATO (2 and 4 mg/kg bw) alone in comparison to controls and RES-supplemented groups. Hence, prolonged ATO exposure reduces ERα expression in the preoptic area of the hypothalamus, potentially disrupting estrogen signaling and neuroendocrine function, while resveratrol may offer protective effects. 4. Discussion Recently, our lab provided evidence of the neuroprotective efficacy of Resveratrol on arsenic-induced cytoarchitectural perturbations, cognitive dysfunction, and disrupted estrogen signaling in the mouse hippocampus (Mehta et al. 2021b , a ). However, the protective effects of Res on arsenic-induced cardiotoxicity remains poorly understood. Therefore, the present study was designed to determine the beneficial roles of Resveratrol (a potent antioxidant) on the morphological and morphometric features of the heart in adult mice subjected to chronic exposure to Arsenic Trioxide (ATO). We found that Resveratrol supplementation significantly alleviated cardiac tissue damage, reducing cardiac fibrosis and ventricular hypertrophy to some extent. These findings underscore resveratrol's potential as a therapeutic agent against arsenic trioxide-induced cardiotoxicity, highlighting its role in preserving cardiac integrity and function amidst environmental toxic insults. Such insights offer promising avenues for developing preventive strategies and interventions against arsenic trioxide-induced cardiac pathology. ATO is a highly potent anticancer drug being used in the treatment of APL (Emadi and Gore 2010 ). However, cardiotoxicity associated with the use of ATO limits its use to a certain extent, (Saad et al. 2010 ) although ATO has been reported to adversely affect the functions of the Liver, Kidney, and CNS as well (Zhang et al. 2013 ). ATO-induced adverse effects on the heart have been manifested as increased thickness of ventricular walls. To date, the possible mechanisms underlying ATO-induced cardiotoxicity include DNA fragmentation, ROS generation/oxidative stress, cardiac ion channel changes, and apoptosis (Zhang et al. 2013 ). Previous studies examined the effects of Res on oxidative stress and various signaling pathways in advanced stages of heart failure induced by subcutaneous administration of Isoproterenol (ISO). This study revealed that ISO administration decreased LV function and altered myocardial fibres orientation, while Res treatment restored LV function and moderated heart failure severity (Chakraborty et al. 2015 ). Likewise, another study showed the cardioprotective effects of polyphenol phytoalexin Res against doxorubicin-induced cardiotoxicity in adult female mice, thus providing in vivo evidence (Osman et al. 2013 ). In the present study, we also observed an increase in LV and RV wall thickness in H&E-stained heart sections of animals exposed to ATO alone when compared to the control group. However, the LV and RV wall thickness in animals co-treated with ATO and Res was less than in ATO alone treated groups and comparable to wall thickness in controls. These observations do suggest the beneficial role of Res against ATO-induced adverse effects on cardiac morphology. The observations of our study also revealed a significant increase in cardiac myocyte diameter (LV and RV) in ATO alone exposed animals, thereby suggestive of cardiac hypertrophy compared to normal controls and the animals receiving ATO and Res simultaneously. Prior work provided insights into the effects of Res in a post-infarction heart failure rat model, where isoproterenol was used to induce myocardial infarction and post-infarction remodeling (Riba et al. 2017 ). Riba and co-workers also showed that systolic left ventricular function was significantly increased, whereas plasma BNP levels, left ventricular wall thickness and dimensions were decreased after 8 weeks of Res treatment (15 mg/kg/day) (Riba et al. 2017 ). Likewise, attenuation of doxorubicin-induced impairment of cardiac function in aged mice was shown through the restoration of SIRT1 activity (Sin et al. 2015 ). Research has extensively documented the impact of Res on cardiac structure and function in a mouse model of heart failure induced by pressure overload (Sung et al. 2015 ). Moreover, the study also observed positive effects on cardiac energy metabolism, suggesting Res’s potential to address both functional and metabolic aspects in the context of heart failure. Res administration at a dose of 150 mg/kg/day led to improvements in diastolic function, decreased left ventricular diameters and volumes, and mitigated cardiac fibrosis, hypertrophy, and remodeling through its antifibrotic and anti-inflammatory properties (Sung et al. 2015 ). However, systolic function did not change after a 2-week-long Res supplementation. Similarly, another group found that even the low dose of Res (2.5 mg/kg/day for 28 days) was able to regress the pressure-overload-induced cardiac hypertrophy and remodeling in Sprague Dawley rats (Wojciechowski et al. 2010 ). Previous clinical studies investigated Resveratrol's potential cardioprotective effects in myocardial infarction patients with preserved ejection fraction (baseline: 54.77 ± 1.64% in the treated group). They discovered a significant improvement in diastolic function after administering 10 mg/day of Resveratrol for 3 months, yet no significant improvement was noted in systolic function (Magyar et al. 2012 ). In the current study, the morphometric analysis showed an apparent increase in the width of cardiomyocytes in the lateral wall of the left ventricle of the ATO (2mg/kg) alone treated group compared to other groups, which is suggestive of ATO-induced adverse effects on myocyte size. It has been reported that increased measurement of myocardiocytes is associated with left ventricular hypertrophy (ASHLEY 1945; Tracy 2012 ). Several clinical studies have reported that ATO treatment in APL patients is associated with various cardiac complications such as ECG abnormalities, ventricular tachycardia, pericardial effusion, myocardial infarction, and left ventricular (Unnikrishnan et al. 2004 ; Ravandi et al. 2009 ). Such studies have examined ATO-induced ventricular hypertrophy using the echocardiography (ECG) technique (Bagul et al. 2015 ). In the present study, morphometric observations carried out on H&E-stained sections (40X) showed increased nucleus size and myocyte size in ATO-alone treated animals. These changes were attenuated to some extent in animals co-treated with ATO and Res . The results of the current study align with a prior study, showing that supplementing Res at a dosage of 10 mg/kg/day for 8 weeks led to a reduction in cardiac hypertrophy in rats with diabetes. This reduction was attributed to the antioxidant effects of Res , which are mediated by SIRT1 (Riba et al. 2017 ). This highlights the fact that the biological activities of Res may be dependent on its multiple molecular targets including SIRT1. Moreover, observations (present study) from Masson’s Trichrome stained sections showed an increase in interstitial and perivascular fibrosis in the ATO alone treated group compared to the control group. A previous study also showed the accumulation of collagen fibres in transgenic mice contributing to cardiac hypertrophy, in addition, to an increase in cardiomyocyte size (Shi et al. 2014 ). Similarly, another group assessed the long-term effects of ATO therapy in Guinea pigs (Chu et al. 2012 ). Chu and co-workers observed substantial structural alterations in both Right and Left ventricles of animals treated with ATO based on analysis of Masson’s trichrome-stained heart sections compared to controls (Chu et al. 2012 ). Nevertheless, future studies are warranted to explore in-depth mechanisms of ATO-induced cardiac damage. In this study, we also examined the effects of ATO exposure on morphological features & morphometric parameters of VMH nucleus using Cresyl Violet stained paraplast sections containing region and to note any alteration in these features following co-administration of RES (ATO + RES). Given the potential link between histopathological changes in hypothalamic neurons and disrupted estrogen receptor (ER) signaling, we explored the expression patterns of ER-α within the hypothalamic nuclei. Notably, we observed a qualitative reduction in ER-α expression in the preoptic area of ATO-exposed groups (2 mg/kg and 4 mg/kg bw), suggesting a downregulation of estrogen signaling due to arsenic exposure. However, co-treatment with resveratrol (ATO + RES) appeared to mitigate this effect, partially restoring ER-α expression. These findings indicate that ATO exposure disrupts estrogen receptor regulation in the hypothalamus, potentially impacting neuroendocrine function. The partial reversal of ER-α downregulation by resveratrol suggests its neuroprotective role in counteracting arsenic-induced endocrine dysregulation, possibly through its antioxidant and anti-inflammatory mechanisms. Furthermore, these observations support the notion that inorganic arsenic (iAs) may act as an endocrine-disrupting compound (EDC) within specific brain regions. We focused on ER expression due to its highly divergent nature among steroid hormone receptors, its extensive co-regulatory interactions, and its distinct activation and deactivation phases (Stenoien et al. 2000 ; Dennis and O’Malley 2005 ; Chatterjee and Chatterji 2010a ). Moreover, our previous findings have also demonstrated that ATO disrupts estrogen signaling in the hippocampus, a brain region abundant in estrogen receptors, further reinforcing the potential endocrine-disrupting effects of arsenic (Mehta et al. 2021b ). To the best of our knowledge, we could find only one study (Ommati et al., 2020) which could explain the microscopic changes in hypothalamic tissue after iAs exposure. Recently, Transmission electron microscopy (TEM) findings revealed an increase in the number of double membrane-surrounded autophagosomes (autophagic vacuoles) in HPG tissues of As 2 O 3 -treated animals (Ommati et al., 2020). TEM findings further showed that the highest number of autophagic vacuoles was recorded in the mature hypothalamus of F1-male mice exposing to the highest dose of As 2 O 3 (20 ppm). Moreover, there are no other reports that have determined the effects of iAs exposure on the estrogen signaling in brain regions like the hippocampus. However, several in-vitro and in-vivo reports demonstrate the endocrine-disrupting effects of iAs exposure in other target tissues and cell line models (Watson and Yager 2007 ; Akram et al. 2010 ; Zhang et al. 2011 ). Endocrine disruptors can either mimic or antagonize the effects of hormones in the target tissues. The role of As as a potent EDC has been suggested by iAs -induced alteration of gene regulation by the closely related steroid hormone receptors for glucocorticoids, mineralocorticoids, progesterone, androgen, and estrogen (Bodwell et al., 2006 , 2004 ; Chatterjee and Chatterji, 2010; Kaltreider et al., 2001 ; Sun et al., 2016 ). The incidence of ATO-induced cardiotoxicity and neural defects in APL patients is relatively low, but it can be severe when it does occur. Several factors contribute to ATO-induced cardiotoxicity, including aging, pre-existing heart disease, and concurrent use of other cardiotoxic drugs. To mitigate the risk of ATO-induced cardiotoxicity and neural defects, it is important to monitor cardiac and neural functions regularly in APL patients receiving ATO treatment. This can involve performing electrocardiograms (ECGs), echocardiograms, and electroencephalograms (EEG), fMRI tests. In some cases, it may be necessary to adjust the dosage or duration of ATO treatment or to switch to an alternative antioxidative therapy. Our study presents promising implications for patient care in managing cardiotoxicity associated with Arsenic Trioxide (ATO) treatment for Acute Promyelocytic Leukemia (APL). Investigating the effects of Resveratrol (Res) on mouse hearts exposed chronically to ATO sheds light on potential interventions to mitigate ATO-induced cardiac hypertrophy and fibrosis in APL patients. Implementing Res as a supplementary treatment alongside ATO could potentially enhance oncology patient care by minimizing adverse cardiac effects. Additionally, Res might serve as a dietary supplement in arsenic-endemic areas, potentially mitigating prevailing arsenic-induced cardiotoxicity among inhabitants. A major limitation of this study is that we did not assess estrogen receptor signaling in the heart, which could provide a more comprehensive understanding of systemic estrogen regulation under ATO exposure. Since estrogen receptors play a crucial role in cardiovascular function, future studies should investigate whether ATO-induced downregulation of ER-α in the hypothalamus also occurs in the heart and whether RES offers similar protective effects. Exploring estrogen receptor-mediated pathways in both the brain and heart could help elucidate the broader endocrine and cardiovascular consequences of arsenic toxicity and the therapeutic potential of RES. 5. Conclusions Globally, millions of people are poisoned by arsenic due to its ecological prevalence and epidemiological importance. Our study revealed the significant cardiac changes in the mouse model of arsenic toxicity (ATO), notably the histological modifications. Meanwhile, resveratrol supplementation following arsenic intoxication in adult mice produced ameliorative effects against cardiac and neuronal morphology. The observed cardio- and neuroprotective effects of Resveratrol could be attributed to its potent antioxidant activity and metal-chelating action against arsenic. It could also be presumed that supplementation of Resveratrol might be an ideal approach for amelioration of ATO-induced cardio- and neurotoxicity, though countless in-depth studies are warranted in this direction to reach any substantial conclusion. Declarations Acknowledgments We duly acknowledge the technical help extended by Mr. Kirpal Singh and the financial support provided by the Department of Anatomy at the All India Institute of Medical Sciences, New Delhi, India. References Akram Z, Jalali S, Shami SA, et al (2010) Adverse effects of arsenic exposure on uterine function and structure in female rat. Experimental and Toxicologic Pathology 62:451–459. https://doi.org/10.1016/j.etp.2009.07.008 ASHLEY LM (1945) A determination of the diameters of ventricular myocardial fibers in man and other mammals. The American journal of anatomy 77:325–363. https://doi.org/10.1002/aja.1000770303 Bagul PK, Deepthi N, Sultana R, Banerjee SK (2015) Resveratrol ameliorates cardiac oxidative stress in diabetes through deacetylation of NFkB-p65 and histone 3. The Journal of nutritional biochemistry 26:1298–1307 Balali-Mood M, Naseri K, Tahergorabi Z, et al (2021) Toxic Mechanisms of Five Heavy Metals: Mercury, Lead, Chromium, Cadmium, and Arsenic. Frontiers in Pharmacology 12:1–19. https://doi.org/10.3389/fphar.2021.643972 Barbey JT, Pezzullo JC, Soignet SL (2003) Effect of arsenic trioxide on QT interval in patients with advanced malignancies. Journal of Clinical Oncology 21:3609–3615. https://doi.org/10.1200/JCO.2003.10.009 Benes FM, Vincent SL, Todtenkopf M (2001) The density of pyramidal and nonpyramidal neurons in anterior cingulate cortex of schizophrenic and bipolar subjects. Biological Psychiatry 50:395–406. https://doi.org/10.1016/S0006-3223(01)01084-8 Bodwell JE, Gosse JA, Nomikos AP, Hamilton JW (2006) Arsenic disruption of steroid receptor gene activation: Complex dose - Response effects are shared by several steroid receptors. Chemical Research in Toxicology 19:1619–1629. https://doi.org/10.1021/tx060122q Bodwell JE, Kingsley LA, Hamilton JW (2004) Arsenic at very low concentrations alters glucocorticoid receptor (GR)-mediated gene activation but not GR-mediated gene repression: Complex dose-response effects are closely correlated with levels of activated GR and require a functional GR DNA binding d. Chemical Research in Toxicology 17:1064–1076. https://doi.org/10.1021/tx0499113 Chakraborty S, Pujani M, Haque SE (2015) Combinational effect of resveratrol and atorvastatin on isoproterenol-induced cardiac hypertrophy in rats. Journal of pharmacy & bioallied sciences 7:233–238. https://doi.org/10.4103/0975-7406.160037 Chandravanshi LP, Gupta R, Shukla RK (2019) Arsenic-Induced Neurotoxicity by Dysfunctioning Cholinergic and Dopaminergic System in Brain of Developing Rats. Biological Trace Element Research 189:118–133. https://doi.org/10.1007/s12011-018-1452-5 Chatterjee A, Chatterji U (2010a) Arsenic abrogates the estrogen-signaling pathway in the rat uterus. Reproductive biology and endocrinology : RB&E 8:80. https://doi.org/10.1186/1477-7827-8-80 Chatterjee A, Chatterji U (2010b) Arsenic abrogates the estrogen-signaling pathway in the rat uterus. Reproductive Biology and Endocrinology 8:1–11. https://doi.org/10.1186/1477-7827-8-80 Chu W, Li C, Qu X, et al (2012) Arsenic-induced interstitial myocardial fibrosis reveals a new insight into drug-induced long QT syndrome. Cardiovascular research 96:90–98 Dennis AP, O’Malley BW (2005) Rush hour at the promoter: How the ubiquitin-proteasome pathway polices the traffic flow of nuclear receptor-dependent transcription. Journal of Steroid Biochemistry and Molecular Biology 93:139–151. https://doi.org/10.1016/j.jsbmb.2004.12.015 Emadi A, Gore SD (2010) Arsenic trioxide - An old drug rediscovered. Blood Reviews. https://doi.org/10.1016/j.blre.2010.04.001 Eyvani H, Moghaddaskho F, Kabuli M, et al (2016) Arsenic trioxide induces cell cycle arrest and alters DNA methylation patterns of cell cycle regulatory genes in colorectal cancer cells. Life Sciences 167:67–77. https://doi.org/10.1016/j.lfs.2016.10.020 Garris DR (1989) Morphometric analysis of obesity (ob/ob)- and diabetes (db/db)-associated hypothalamic neuronal degeneration in C57BL/KsJ mice. Brain Research 501:162–170. https://doi.org/10.1016/0006-8993(89)91037-8 Garris DR, Diani AR, Smith C, Gerritsen GC (1982) Depopulation of the ventromedial hypothalamic nucleus in the diabetic Chinese hamster. Acta Neuropathologica 56:63–66. https://doi.org/10.1007/BF00691183 Garris DR, West RL, Coleman DL (1985) Morphometric analysis of medial basal hypothalamic neuronal degeneration in diabetes (db/db) mutant C57BL/KsJ mice: Relation to age and hyperglycemia. Developmental Brain Research 20:161–168. https://doi.org/10.1016/0165-3806(85)90101-4 Kaltreider RC, Davis AM, Lariviere JP, Hamilton JW (2001) Arsenic alters the function of the glucocorticoid receptor as a transcription factor. Environmental Health Perspectives 109:245–251. https://doi.org/10.1289/ehp.01109245 Kitchin KT, Conolly R (2010) Arsenic-induced carcinogenesissoxidative stress as a possible mode of action and future research needs for more biologically based risk assessment. Chemical Research in Toxicology 23:327–335. https://doi.org/10.1021/tx900343d Korbo L, Amrein I, Lipp HP, et al (2004) No evidence for loss of hippocampal neurons in non-Alzheimer dementia patients. Acta Neurologica Scandinavica 109:132–139. https://doi.org/10.1034/j.1600-0404.2003.00182.x Lo-Coco F, Avvisati G, Vignetti M, et al (2013) Retinoic Acid and Arsenic Trioxide for Acute Promyelocytic Leukemia. New England Journal of Medicine 369:111–121. https://doi.org/10.1056/nejmoa1300874 Luo JH, Qiu ZQ, Zhang L, Shu WQ (2012) Arsenite exposure altered the expression of NMDA receptor and postsynaptic signaling proteins in rat hippocampus. Toxicology Letters 211:39–44. https://doi.org/10.1016/j.toxlet.2012.02.021 Magyar K, Halmosi R, Palfi A, et al (2012) Cardioprotection by resveratrol: A human clinical trial in patients with stable coronary artery disease. Clinical Hemorheology and Microcirculation 50:179–187. https://doi.org/10.3233/CH-2011-1424 Medda N, Patra R, Ghosh TK, Maiti S (2020) Neurotoxic Mechanism of Arsenic: Synergistic Effect of Mitochondrial Instability, Oxidative Stress, and Hormonal-Neurotransmitter Impairment. Biological Trace Element Research 198:8–15 Mehta K, Kaur B, Pandey KK, et al (2021a) Resveratrol protects against inorganic arsenic-induced oxidative damage and cytoarchitectural alterations in female mouse hippocampus. Acta Histochemica 123:151792. https://doi.org/10.1016/j.acthis.2021.151792 Mehta K, Pandey KK, Kaur B, et al (2021b) Resveratrol attenuates arsenic-induced cognitive deficits via modulation of Estrogen-NMDAR-BDNF signalling pathway in female mouse hippocampus. Psychopharmacology 238:2485–502. https://doi.org/10.1007/s00213-021-05871-2 Mukherjee S, Dudley JI, Das DK (2010) Dose-dependency of resveratrol in providing health benefits. Dose-Response 8:478–500. https://doi.org/10.2203/dose-response.09-015.Mukherjee Namavar MR, Raminfard S, Jahromi ZV, Azari H (2012) Effects of high-fat diet on the numerical density and number of neuronal cells and the volume of the mouse hypothalamus: a stereological study. Anatomy & Cell Biology 45:178. https://doi.org/10.5115/acb.2012.45.3.178 Ohnishi K, Yoshida H, Shigeno K, et al (2000) Prolongation of the QT interval and ventricular tachycardia in patients treated with arsenic trioxide for acute promyelocytic leukemia. Annals of Internal Medicine 133:881–885. https://doi.org/10.7326/0003-4819-133-11-200012050-00012 Osman AMM, Al-Harthi SE, AlArabi OM, et al (2013) Chemosensetizing and cardioprotective effects of resveratrol in doxorubicin- treated animals. Cancer Cell International 13:1. https://doi.org/10.1186/1475-2867-13-52 Paul NP, Galván AE, Yoshinaga-Sakurai K, et al (2023) Arsenic in medicine: past, present and future. BioMetals 36:283–301. https://doi.org/10.1007/s10534-022-00371-y Powell BL, Moser B, Stock W, et al (2011) Arsenic trioxide improves event-free and overall survival for adults with acute promyelocytic leukemia : North American Leukemia Intergroup Study C9710 Arsenic trioxide improves event-free and overall survival for adults with acute promyelocytic leukemia . Cancer 116:3751–3757. https://doi.org/10.1182/blood-2010-02-269621 Ran S, Liu J, Li S (2020) A Systematic Review of the Various Effect of Arsenic on Glutathione Synthesis in Vitro and in Vivo. BioMed Research International 2020:. https://doi.org/10.1155/2020/9414196 Ravandi F, Estey E, Jones D, et al (2009) Effective treatment of acute promyelocytic leukemia with all-trans-retinoic acid, arsenic trioxide, and gemtuzumab ozogamicin. Journal of Clinical Oncology 27:504 Riba A, Deres L, Sumegi B, et al (2017) Cardioprotective effect of resveratrol in a postinfarction heart failure model. Oxidative Medicine and Cellular Longevity 2017:. https://doi.org/10.1155/2017/6819281 Rodríguez VM, Carrizales L, Jiménez-Capdeville ME, et al (2001) The effects of sodium arsenite exposure on behavioral parameters in the rat. Brain Research Bulletin 55:301–308. https://doi.org/10.1016/S0361-9230(01)00477-4 Rodríguez VM, Jiménez-Capdeville ME, Giordano M (2003) The effects of arsenic exposure on the nervous system. Toxicology Letters 145:1–18. https://doi.org/10.1016/S0378-4274(03)00262-5 Roy TS, Sharma V, Seidler FJ, Slotkin TA (2005) Quantitative morphological assessment reveals neuronal and glial deficits in hippocampus after a brief subtoxic exposure to chlorpyrifos in neonatal rats. Developmental Brain Research 155:71–80. https://doi.org/10.1016/j.devbrainres.2004.12.004 Saad SY, Alkharfy KM, Arafah MM (2010) Cardiotoxic effects of arsenic trioxide/imatinib mesilate combination in rats. Journal of Pharmacy and Pharmacology 58:567–573. https://doi.org/10.1211/jpp.58.4.0017 Shi K, Zhao W, Chen Y, et al (2014) Cardiac hypertrophy associated with myeloproliferative neoplasms in JAK2V617F transgenic mice. Journal of Hematology & Oncology 7:1–8 Shin J, Seol I, Son C (2010) Interpretation of Animal Dose and Human Equivalent Dose for Drug Development. The Journal of Korean Oriental Medicine 31:1–7 Sin TK, Tam BT, Yung BY, et al (2015) Resveratrol protects against doxorubicin-induced cardiotoxicity in aged hearts through the SIRT1-USP7 axis. Journal of Physiology 593:1887–1899. https://doi.org/10.1113/jphysiol.2014.270101 Smith AH, Arroyo AP, Guha Mazumder DN, et al (2000) Arsenic-induced skin lesions among Atacameno people in Northern Chile despite good nutrition and centuries of exposure. Environmental Health Perspectives 108:617–620. https://doi.org/10.1289/ehp.00108617 Stenoien DL, Simeoni S, Sharp ZD, Mancini MA (2000) Subnuclear dynamics and transcription factor function. Journal of cellular biochemistry Supplement Suppl 35:99–106. https://doi.org/10.1002/1097-4644(2000)79:35+3.0.co;2-w Straif K, Benbrahim-Tallaa L, Baan R, et al (2009) A review of human carcinogens--part C: metals, arsenic, dusts, and fibres. The lancet oncology 10:453–454. https://doi.org/10.1016/s1470-2045(09)70134-2 Sun H, Yang Y, Shao H, et al (2017) Sodium arsenite-induced learning and memory impairment is associated with endoplasmic reticulum stress-mediated apoptosis in rat hippocampus. Frontiers in Molecular Neuroscience 10:1–14. https://doi.org/10.3389/fnmol.2017.00286 Sun HJ, Xiang P, Luo J, et al (2016) Mechanisms of arsenic disruption on gonadal, adrenal and thyroid endocrine systems in humans: A review. Environment International 95:61–68. https://doi.org/10.1016/j.envint.2016.07.020 Sung MM, Das SK, Levasseur J, et al (2015) Resveratrol treatment of mice with pressure-overloadinduced heart failure improves diastolic function and cardiac energy metabolism. Circulation: Heart Failure 8:128–137. https://doi.org/10.1161/CIRCHEARTFAILURE.114.001677 Tracy RE (2012) Cardiomyocyte size estimated from noninvasive measurements of left ventricular wall thickness and chamber diameter. Journal of the American Society of Hypertension 6:185–192 Tyler CR, Allan AM (2014) The Effects of Arsenic Exposure on Neurological and Cognitive Dysfunction in Human and Rodent Studies: A Review. Current environmental health reports 1:132–147. https://doi.org/10.1007/s40572-014-0012-1 Unnikrishnan D, Dutcher JP, Garl S, et al (2004) Cardiac monitoring of patients receiving arsenic trioxide therapy. British journal of haematology 124:610–617 Wang R, Zhang J, Wang S, et al (2019) The cardiotoxicity induced by arsenic trioxide is alleviated by salvianolic acid A via maintaining calcium homeostasis and inhibiting endoplasmic reticulum stress. Molecules 24:1–12. https://doi.org/10.3390/molecules24030543 Wang Y, Zhao F, Liao Y, et al (2012) Arsenic exposure and glutamate-induced gliotransmitter release from astrocytes. Neural Regeneration Research 7:2439–2445 Watson WH, Yager JD (2007) Arsenic: Extension of its endocrine disruption potential to interference with estrogen receptor-mediated signaling. Toxicological Sciences 98:1–4. https://doi.org/10.1093/toxsci/kfm111 Wojciechowski P, Juric D, Louis XL, et al (2010) Resveratrol arrests and regresses the development of pressure overload- but not volume overload-induced cardiac hypertrophy in rats. Journal of Nutrition 140:962–968. https://doi.org/10.3945/jn.109.115006 Zhang W, Guo C, Gao R, et al (2013) The protective role of resveratrol against arsenic trioxide-induced cardiotoxicity. Evidence-based Complementary and Alternative Medicine 2013:. https://doi.org/10.1155/2013/407839 Zhang W, Wang L, Fan Q, et al (2011) Arsenic trioxide re-sensitizes ERα-negative breast cancer cells to endocrine therapy by restoring ERα expression in vitro and in vivo. Oncology Reports 26:621–628. https://doi.org/10.3892/or.2011.1352 Supplementary Table Supplementary Table 2 is not available with this version. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 20 May, 2025 Read the published version in Journal of Molecular Histology → Version 1 posted Editorial decision: Revision requested 12 Apr, 2025 Reviews received at journal 07 Apr, 2025 Reviewers agreed at journal 31 Mar, 2025 Reviewers agreed at journal 28 Mar, 2025 Reviewers invited by journal 28 Mar, 2025 Editor assigned by journal 28 Mar, 2025 Submission checks completed at journal 27 Mar, 2025 First submitted to journal 26 Mar, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6316307","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":442132210,"identity":"604937d1-d766-4afa-9df9-0a6c4d61e84c","order_by":0,"name":"Saroj N/A","email":"","orcid":"","institution":"Dr. Rajendra Prasad Government Medical College","correspondingAuthor":false,"prefix":"","firstName":"Saroj","middleName":"","lastName":"N/A","suffix":""},{"id":442132212,"identity":"c681950c-9e73-4c95-838a-ea629bae6080","order_by":1,"name":"Kamakshi Mehta","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAxElEQVRIiWNgGAWjYBACgwMMjA8+8NgwsIG5bERoMWxgYDacIZPGwMbGTKQWY6AyYR6bw0CKWC1m7KfTGGfknM/jk+8/wPCh7DBhLTY8udsefDhzuxjkMMYZ54jRwpC73XBmz+3ENqAWZt42IrSY8b/dJs377xxEy19itBhL5G6T5uE5ANHCSIwWwxlvNxvO4EkG+iXZ4GDPuXTCWgzO524ERqVdnnzzwYcPfpRZE9YCAwkg4gDx6mFaRsEoGAWjYBRgBQCWejknBhvP0QAAAABJRU5ErkJggg==","orcid":"","institution":"University of Pittsburgh School of Medicine, UPMC Vision Institute","correspondingAuthor":true,"prefix":"","firstName":"Kamakshi","middleName":"","lastName":"Mehta","suffix":""},{"id":442132214,"identity":"ab00579d-a5d5-412e-ac9e-d69c3b580990","order_by":2,"name":"Balpreet Kaur","email":"","orcid":"","institution":"University College of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Balpreet","middleName":"","lastName":"Kaur","suffix":""},{"id":442132216,"identity":"1ba22de3-4fa0-4b4d-86dd-aa0438af0f75","order_by":3,"name":"Kamlesh Kumar Pandey","email":"","orcid":"","institution":"All India Institute of Medical Sciences (AIIMS)","correspondingAuthor":false,"prefix":"","firstName":"Kamlesh","middleName":"Kumar","lastName":"Pandey","suffix":""},{"id":442132218,"identity":"8948dddb-8e49-49fc-a3a8-82e18df4b984","order_by":4,"name":"Saroj Kaler","email":"","orcid":"","institution":"All India Institute of Medical Sciences (AIIMS)","correspondingAuthor":false,"prefix":"","firstName":"Saroj","middleName":"","lastName":"Kaler","suffix":""},{"id":442132220,"identity":"de9afdde-7334-4798-b2ae-a8630b9b12fc","order_by":5,"name":"Pushpa Dhar","email":"","orcid":"","institution":"All India Institute of Medical Sciences (AIIMS)","correspondingAuthor":false,"prefix":"","firstName":"Pushpa","middleName":"","lastName":"Dhar","suffix":""}],"badges":[],"createdAt":"2025-03-27 02:53:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6316307/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6316307/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10735-025-10439-x","type":"published","date":"2025-05-20T15:56:53+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":80995123,"identity":"9f901cac-12fe-423f-aa69-aaba55929d9e","added_by":"auto","created_at":"2025-04-21 04:55:10","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":4123905,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative photomicrographs showing areas of interest (AOI) selected for morphological and morphometric studies. (A) Cresyl Violet (CV) stained coronal mouse brain section showing anatomical landmarks for identification of Ventromedial Hypothalamic Nucleus (VMH). (B) Photomicrograph showing the anatomical divisions of Ventromedial Hypothalamic nucleus into VMHDM (dorsomedial), VMHC (central) and VMHVL (ventrolateral). (C) Haematoxylin-eosin (H-E) stained cross-section of mouse heart (1X) showing three cardiac layers and chambers- Right ventricle (RV), Left ventricle (LV), and Interventricular septum (IVS). Scale bar- 20µm, 100µm and 1000µm. Abbreviations: f- Fornix; ME- Median Eminence; mt- mammillothalamic tract.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/58f0b9b763467373b2527432.png"},{"id":80995120,"identity":"2508e819-4dcd-4db1-82a9-19cac90bcba3","added_by":"auto","created_at":"2025-04-21 04:55:10","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":2367532,"visible":true,"origin":"","legend":"\u003cp\u003eSnapshot of screen showing methodology adopted for the measurement of soma size of neurons in VMH nucleus using NIS Element (Version AR3.10) image analysis software.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/7e7984fec385981c59195da5.png"},{"id":80995118,"identity":"7098bfce-6567-4448-b400-c91a118d7340","added_by":"auto","created_at":"2025-04-21 04:55:09","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1087135,"visible":true,"origin":"","legend":"\u003cp\u003eHigh magnification photomicrographs of H\u0026amp;E-stained sections of the Left Ventricular wall from control and experimental groups. (40X, scale bar 50 μm). Cardiomyocyte nuclei (black arrow), Intercalated disc (dashed black arrow); Blood vessel (blue arrow).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/6a04d7c6504c4783d302ac0c.png"},{"id":80995131,"identity":"8cdc3dde-0a79-421c-a863-a6ae7f8d8c5b","added_by":"auto","created_at":"2025-04-21 04:55:10","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1179733,"visible":true,"origin":"","legend":"\u003cp\u003eHigh magnification (100X, 20μm scale bar) photomicrographs of H\u0026amp;E-stained sections of LV wall from control and experimental groups. A-NC; B-ATO alone treated; C- ATO+\u003cem\u003eRes \u003c/em\u003eco-treated. Cardiomyocyte nuclei (black arrow); Intercalated disc (blue arrow); Branching pattern (red arrow); fibroblast (green arrow).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/11adc36e1d7f88753d4a41ea.png"},{"id":80995296,"identity":"9b0c94b0-b94e-4ff8-9252-f34df5f3181e","added_by":"auto","created_at":"2025-04-21 05:03:10","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1318846,"visible":true,"origin":"","legend":"\u003cp\u003eHigh magnification photomicrographs of H \u0026amp; E-stained heart sections showing the Right Ventricular wall from control and experimental groups. (40X, scale bar 50 μm). Cardiomyocyte nuclei (black arrow); Blood vessel (blue arrow); Branching pattern (red arrow) and Fibroblast (green arrow)\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/b948df354cf7b98819f3455e.png"},{"id":80995297,"identity":"46040e09-bb70-464b-853b-effb470bb851","added_by":"auto","created_at":"2025-04-21 05:03:10","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1230040,"visible":true,"origin":"","legend":"\u003cp\u003eHigh magnification (100x, scale bar 20μm) photomicrographs of H\u0026amp;E-stained sections of RV wall from control and experimental groups. A-NC; B-ATO alone treated; C- ATO+\u003cem\u003eRes \u003c/em\u003eco-treated. Cardiomyocyte nuclei (black arrow); Branching pattern (yellow arrow), Intercalated disc (blue arrow); Fibroblast (green arrow)\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/5b24ad131906de07a09702ce.png"},{"id":80995305,"identity":"afc457d6-6cb0-4a09-90b1-496c8d34898d","added_by":"auto","created_at":"2025-04-21 05:03:11","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":2423585,"visible":true,"origin":"","legend":"\u003cp\u003eLow (upper panel) and high (lower panel) magnification photomicrographs of H\u0026amp;E-stained sections of Interventricular septum (IVS) from control and experimental groups. A1, A- NC; B1- VC; C1, C –ATO alone treated; D1- \u003cem\u003eRes \u003c/em\u003ealone treated and E1, E- ATO +\u003cem\u003eRes.\u003c/em\u003eUpper panel (20X, scale bar 200 μm), Lower panel (100x, scale bar 20 μm). Cardiomyocyte nuclei (black arrow); Branching pattern (yellow arrow), Intercalated disc (blue arrow); Fibroblast (green arrow)\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/b89b37b794bb3585b132b097.png"},{"id":80995304,"identity":"0e403faf-3bd1-4e35-9bc5-2bf1ffbafcae","added_by":"auto","created_at":"2025-04-21 05:03:11","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":2346923,"visible":true,"origin":"","legend":"\u003cp\u003eLow (upper panel) and high (lower panel) magnification photomicrographs of Masson’s trichrome stained sections from control and experimental groups. A1, A2-NC; B1, B2-VC; C1, C2-ATO alone; D1, D2-\u003cem\u003eRes\u003c/em\u003e alone; E1, E2-ATO+\u003cem\u003eRes\u003c/em\u003e. Upper panel (20X, scale bar 200μm), Lower panel (40X, scale bar 50 μm). Blood vessel nuclei (black arrow), → increased fibrosis in the vessel wall., Intercalated disc (blue arrow).\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/1c222f398f8bb78fd24cd775.png"},{"id":80995133,"identity":"232c4024-1188-4a13-874b-47b670b40315","added_by":"auto","created_at":"2025-04-21 04:55:10","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":3314180,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative low-power CV-stained coronal sections of Ventromedial Hypothalamic Nucleus (Bregma: -1.70mm) from control (A, B) \u0026amp; experimental groups (C to F) showing dense neuronal core in the vicinity of the third ventricle. Scale bar-20µm.\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/7ebceb60050d8cc2e7424e45.png"},{"id":80995300,"identity":"7aa23416-059d-442c-b3ad-09783fa77005","added_by":"auto","created_at":"2025-04-21 05:03:10","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":2784997,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/16ca06d2f9e8ef3d56f1fe2c.png"},{"id":80995136,"identity":"17b14755-3b54-433e-a205-b4bfded226cc","added_by":"auto","created_at":"2025-04-21 04:55:10","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":488153,"visible":true,"origin":"","legend":"\u003cp\u003eBar diagrams showing the morphometric analysis in VMH nucleus (A) Cell density (cells/mm\u003csup\u003e2\u003c/sup\u003e) (B) Mean soma size (µm\u003csup\u003e2\u003c/sup\u003e). Values are expressed as mean ± SD. Significance levels shown at *p\u0026lt;0.05, ***p\u0026lt;0.001 compared with As\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3 \u003c/sub\u003e2mg/kg bw alone;\u003csup\u003e #\u003c/sup\u003ep\u0026lt;0.05, \u003csup\u003e##\u003c/sup\u003ep\u0026lt;0.01, \u003csup\u003e###\u003c/sup\u003ep\u0026lt;0.001 compared with As\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3 \u003c/sub\u003e4mg/kg bw alone.\u003c/p\u003e","description":"","filename":"11.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/211c201ad6ae30d0a78b1976.png"},{"id":80995132,"identity":"059c3395-8f1d-45da-b295-08fa59354965","added_by":"auto","created_at":"2025-04-21 04:55:10","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":3841679,"visible":true,"origin":"","legend":"\u003cp\u003eHigh-power (40X) photomicrographs of immunostained mouse brain coronal (bregma +0.74 mm) sections showing expression of Estrogen receptor-α in nucleus (↘) and processes (→) of neurons of Preoptic area (Hypothalamus) among control (A, B) and experimental groups (C to F). Scale bar 50 µm.\u003c/p\u003e","description":"","filename":"12.png","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/766acbb02d7b6833a2a61918.png"},{"id":83459947,"identity":"685f446c-9910-4c9b-bc09-5dba0593f113","added_by":"auto","created_at":"2025-05-26 16:03:54","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":37622033,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6316307/v1/628f1d9a-a8ff-4437-b064-a1e9f75f8c48.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Arsenic-Induced Neurocardiac Toxicity and Protective Role of Resveratrol: Histopathological and Molecular Insights","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eExposure to heavy metals (arsenic, lead, cadmium, mercury, and copper) of hydro-geological origin may adversely affect humans (Balali-Mood et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). One of these toxic metalloids is Arsenic with the symbol \u003cem\u003eAs\u003c/em\u003e and an atomic number 33. A significant source of inorganic arsenic (\u003cem\u003eiAs)\u003c/em\u003e exposure is contaminated groundwater with higher \u003cem\u003eAs\u003c/em\u003e concentrations, thereby affecting communities that rely entirely on groundwater for drinking and cooking (Smith et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). According to past surveys, arsenic has been identified as the world\u0026rsquo;s second-leading water-borne cause of mortality (Straif et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2009\u003c/span\u003e)Moreover, several Indian states lying along the GMB (Ganga-Meghna-Brahmputra) belt (West Bengal, Jharkhand, Bihar, Uttar Pradesh, Assam, Manipur, and Chhattisgarh) are reported to be adversely impacted by groundwater contamination with \u003cem\u003eAs\u003c/em\u003e levels that exceed the WHO permissible limit.\u003c/p\u003e \u003cp\u003eDespite its reputation for toxicity, \u003cem\u003eAs\u003c/em\u003e serves paradoxically as a therapeutic agent employed since ancient eras for treating various illnesses (Paul et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Arsenic trioxide (ATO), an inorganic compound of \u003cem\u003eAs\u003c/em\u003e, is currently used as an anti-neoplastic agent in treating acute promyelocytic leukemia (APL). Even though APL patients generally tolerate ATO therapy well, it can also cause toxic effects in some cases. However, its use is limited to a certain extent keeping in consideration the adverse health effects observed in APL patients (Wang et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Various cardiac abnormalities such as prolonged QT interval, arrhythmias, and pericardial effusion have been observed in APL patients following ATO therapy (Ohnishi et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Barbey et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). These cardiac complications from ATO exposure are significant because they may predict a fatal outcome in APL patients.\u003c/p\u003e \u003cp\u003eApart from the heart, other primary targets for \u003cem\u003eiAs\u003c/em\u003e toxicity include the liver, lungs, kidneys, and brain. However, symptoms of \u003cem\u003eiAs\u003c/em\u003e poisoning may take from hours to years to manifest, depending on the dose and length of exposure. With context to the brain, \u003cem\u003eiAs\u003c/em\u003e-induced neurotoxicity leads to neuroinflammation, oxidative stress, and neurodegeneration, particularly in the hippocampus and cortex, resulting in cognitive decline and neuronal loss (Rodr\u0026iacute;guez et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2001\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Luo et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Medda et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Mehta et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2021b\u003c/span\u003e). It disrupts neurotransmission by impairing dopamine, glutamate, and acetylcholine signaling, affecting learning, memory, and motor functions (Wang et al. \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Tyler and Allan \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Sun et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Chandravanshi et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Additionally, \u003cem\u003eAs\u003c/em\u003e increases blood-brain barrier permeability, allowing further toxin entry, and induce mitochondrial dysfunction, leading to neuronal apoptosis. These effects contribute to cognitive and behavioral deficits such as memory loss, attention impairments, anxiety, and depression. Moreover, \u003cem\u003eAs\u003c/em\u003e exposure causes epigenetic alterations, potentially leading to long-term neurodevelopmental consequences and increasing the risk of neurodegenerative diseases like Alzheimer\u0026rsquo;s and Parkinson\u0026rsquo;s (Tyler and Allan \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Medda et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eOur study primarily focuses on \u003cem\u003eiAs\u003c/em\u003e-induced toxicity in the heart and Ventromedial Hypothalamic (VMH) area. Functionally, the VMH is involved in energy homeostasis, feeding behavior, and sympathetic nervous system regulation. VMH is also reported to be targeted in obesity and diabetes, which are major risk factors for cardiovascular disease (CVD). By selecting the VMH, we can investigate how central neural circuits related to metabolic regulation might influence or be influenced by cardiovascular function. Additionally, the VMH is involved in regulating the body\u0026rsquo;s stress response, which has direct implications for both metabolic and cardiovascular health. To date, numerous epidemiological studies have established a strong correlation between chronic arsenic exposure and deteriorated mental health. Nevertheless, there are scanty animal studies that could reveal the association between arsenic exposure and neurocardiac health. Mouse is an ideal species to study the \u003cem\u003ein-vivo\u003c/em\u003e effects of drugs, toxic metals on several organs, including the nervous system.\u003c/p\u003e \u003cp\u003eSeveral mechanisms have been proposed for the cardio- and neurotoxic effects of ATO including oxidative stress, apoptosis, altered DNA methylation, genotoxicity, mitochondrial damage, and calcium dysregulation (Kitchin and Conolly \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Eyvani et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Additionally, \u003cem\u003eAs\u003c/em\u003e also interacts with endogenous cellular antioxidants such as glutathione (GSH) to form a transient compound called glutathione trioxide, thus depleting the cellular GSH levels and resulting in oxidative stress (Ran et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCurrently, clinical medicine has no effective strategies to protect against cardiotoxicity caused by inorganic arsenic (\u003cem\u003eiAs\u003c/em\u003e). Given that oxidative stress is a key mechanism in \u003cem\u003eiAs\u003c/em\u003e-induced toxicity, it is worthwhile to investigate the application of naturally occurring antioxidants in animal models of arsenic toxicity.\u003c/p\u003e \u003cp\u003eResveratrol (3,5,4\u0026prime;-trihydroxy-trans-stilbene) [\u003cem\u003eRes\u003c/em\u003e] is a plant-based polyphenolic stilbene, found abundantly in grapes, all types of berries, soy, and peanuts. Because of its effective antioxidant activity, \u003cem\u003eRes\u003c/em\u003e has been shown to protect against ATO-induced cardiotoxicity as well as neurotoxicity via upregulation of antioxidant enzymes and downregulation of apoptotic levels (Zhang et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Mehta et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2021a\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe present study raises two possible questions: Can Arsenic Trioxide (ATO) induce toxic effects in the heart and ventromedial hypothalamic nucleus (VMH)? Given the known cardiotoxic and neurotoxic effects of ATO in Acute Promyelocytic leukemia (APL) patients and its role in generating oxidative damage, can Resveratrol supplementation alleviate these detrimental effects caused by prolonged exposure to ATO in mice, and if so, how? Hence, we intend to evaluate the potential cardio- and neuroprotective efficacy of Resveratrol, a proven antioxidant in a mouse model of ATO-induced toxicity affecting the heart and hypothalamus.\u003c/p\u003e"},{"header":"2. Methods","content":"\u003cdiv id=\"Sec3\"\u003e\n \u003ch2\u003e2.1 Animals\u003c/h2\u003e\n \u003cp\u003eAdult female Swiss albino mice (weight\u0026thinsp;=\u0026thinsp;25\u0026ndash;30 g) were procured from the Central Animal Facility (CAF) of All India Institute of Medical Sciences (AIIMS) after obtaining ethical clearance from the Institutional Animal Ethics Committee (IAEC). The animals were kept in cages under 12 hr light/12 hr dark cycles and temperature (20\u0026ndash;25˚C) and humidity (50\u0026ndash;60%) controlled rooms of CAF. They were fed routinely with a standard rodent diet (\u003cem\u003echow\u003c/em\u003e) and given free access to drinking water. Animals were acclimatized for 7 days before the start of the treatment regimen.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\"\u003e\n \u003ch2\u003e2.2 Study design\u003c/h2\u003e\n \u003cp\u003eAfter acclimatization, the animals were randomly divided into five groups with six animals per group: Group \u0026Iota; - normal control (no treatment), Group II - vehicle control (5% Gum Acacia), Group III \u0026ndash;Arsenic trioxide, ATO (2mg/kg b.w.) alone, Group IV- \u003cem\u003eRes\u003c/em\u003e (40mg/kg b.w.) alone, and Group V- ATO and \u003cem\u003eRes\u003c/em\u003e (2mg and 40 mg/kg b.w.). Both test substances were administered orally over an experimental period of 45 days.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\"\u003e\n \u003ch2\u003e2.3 Chemicals and reagents\u003c/h2\u003e\n \u003cp\u003eArsenic trioxide (A1010), Resveratrol (R-5010), and Gum acacia (V800016) were purchased from Sigma Chemicals, St. Louis, USA.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\"\u003e\n \u003ch2\u003e2.4 ATO and Resveratrol dose selection\u003c/h2\u003e\n \u003cp\u003eThe dose of ATO used in the present study was 2mg/kg body weight (bw) which is equivalent to the therapeutic dose in humans for the treatment of APL, i.e., 0.15 mg/kg per day for 60 days (Powell et al. \u003cspan\u003e2011\u003c/span\u003e; Lo-Coco et al. \u003cspan\u003e2013\u003c/span\u003e). The conversion of human equivalent dose (HED) to animal equivalent dose (AED) was done by applying a BSA (body surface area) based formula (Shin et al. \u003cspan\u003e2010\u003c/span\u003e). To prepare a 2 mg/kg bw concentration of ATO, 20 mg of ATO was fully dissolved in 500 \u0026micro;l of NaOH, followed by dilution to a final volume of 20 ml with distilled water. The daily ATO dosage was determined based on the body weight of each animal.\u003c/p\u003e\n \u003cp\u003eThe dose for \u003cem\u003eRes\u003c/em\u003e (40 mg/kg bw) was adopted from the previous literature, wherein an oral dose of 40 mg/kg bw has been documented to be cardio- and neuroprotective in animal models (Mukherjee et al. \u003cspan\u003e2010\u003c/span\u003e; Mehta et al. \u003cspan\u003e2021b\u003c/span\u003e, \u003cspan\u003ea\u003c/span\u003e). \u003cem\u003eRes\u003c/em\u003e was freshly prepared in 5% gum acacia and kept on ice until its administration through oral gavage.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\"\u003e\n \u003ch2\u003e2.5 Histology\u003c/h2\u003e\n \u003cp\u003eOn Day 45, the animals were anesthetized with sodium pentobarbitone (60 mg/kg, i.p.) and underwent transcardial perfusion fixation with 4% paraformaldehyde (prepared in 0.1M PBS). The perfusion-fixed heart and brains were later processed for paraplast embedding and staining methods. The boundaries and gross histology of two AOI (Areas of Interest) were identified by different staining methods, as shown in Fig.\u0026nbsp;\u003cspan\u003e1\u003c/span\u003e. Hematoxylin and Eosin (H-E) staining was performed to visualize the cardiac morphology (Fig.\u0026nbsp;\u003cspan\u003e1\u003c/span\u003eC), and cresyl violet staining was done to observe the neuronal morphology in the ventromedial hypothalamic nucleus (VMH) (Fig.\u0026nbsp;\u003cspan\u003e1\u003c/span\u003eA, B). Additionally, we performed Masson\u0026rsquo;s Trichrome staining to determine the status of connective tissue or fibrosis (if any) within the heart\u0026apos;s ventricular walls.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\"\u003e\n \u003ch2\u003e2.6 Morphological observations\u003c/h2\u003e\n \u003cp\u003eMorphological observations were carried out on H-E and Masson\u0026rsquo;s trichrome stained transverse sections (7\u0026micro;m thick) of the mouse heart using a bright field Nikon E-600 microscope fitted with Nikon Digital Camera System (DS-Fil \u0026ndash;U2). We studied various morphological features of VMH neurons and cardiac myocytes, including their branching pattern, shape, and size. The general appearance of fibroblasts and the location of intercalated discs were also observed in the heart sections of control and experimental mice.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\"\u003e\n \u003ch2\u003e2.7 Morphometric analysis in heart\u003c/h2\u003e\n \u003cp\u003eMeasurement of wall thickness and nuclear size was carried out by the attached image analysis system (Nikon Imaging Software, NIS Elements AR 3.1). For determining the thickness of ventricular walls (LV, RV, and IVS), the first section was chosen randomly, and the subsequent section was every sixth section from it, and a total of six sections (6 different sites) were analyzed for further calculations. For each chamber (LV \u0026amp; RV), the diameter of around ten to twelve myocytes was measured at the centre of the nucleus (40X). In the same sections, the area of the nucleus was also determined.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\"\u003e\n \u003ch2\u003e2.8 Morphometric study in Ventromedial Hypothalamic (VMH) Nucleus\u003c/h2\u003e\n \u003cp\u003eThe rostral-caudal limits of VMH were identified from serial brain sections, and the midpoint of the nucleus was located for analysis as previously illustrated (Garris et al. \u003cspan\u003e1982\u003c/span\u003e, \u003cspan\u003e1985\u003c/span\u003e; Garris \u003cspan\u003e1989\u003c/span\u003e). The first section was randomly chosen, followed by every seventh section, so the interval between sections considered for counting was 49 \u0026micro;m. This ensures that the same cell was not counted twice, given that the typical cell diameter in this region is smaller than 49 \u0026micro;m. A total of six sections were evaluated per animal (n\u0026thinsp;=\u0026thinsp;5 animals/group) for the morphometric study.\u003c/p\u003e\u003cbr\u003e\n \u003cp\u003e\u003cstrong\u003e2.8.1 Neuronal density (neurons/unit area)\u003c/strong\u003e: A rectangular reference frame of a defined area (200\u0026times;170 \u0026micro;m\u003csup\u003e2\u003c/sup\u003e) (with solid lines on two adjacent sides and dotted lines on the other two adjacent sides) was placed on the specified region of the digital image taken at 40X (Fig. \u003cspan\u003e2\u003c/span\u003e), and the neuronal cell bodies with nucleoli and lying within the VMH nucleus were considered for counting. According to the Forbidden line rule, the neurons lying along the right and upper lines of the frame were included for counting, while those lying along the left and lower lines of the reference frame were excluded. To standardize the measurements, only neurons exhibiting a well-stained nucleus with a distinct nucleolus were counted. Glia was readily distinguished from neurons by their size, nuclear shape, cytoplasm, location, and characteristic staining (Roy et al. \u003cspan\u003e2005\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e2.8.2 Soma size estimation\u003c/strong\u003e: The Soma size of the VMH neurons was measured on digital images captured at higher magnification (100X) to have a clear and distinct observation of neuronal features such as soma, nucleus, cytoplasm, and nucleoli (Fig. \u003cspan\u003e2\u003c/span\u003e). Cells showing euchromatin material and nucleoli within the nuclei and the nuclei surrounded by cytoplasm were identified as neurons (Benes et al. \u003cspan\u003e2001\u003c/span\u003e; Korbo et al. \u003cspan\u003e2004\u003c/span\u003e; Namavar et al. \u003cspan\u003e2012\u003c/span\u003e). Sections showing the full extent (Bregma \u0026minus;\u0026thinsp;1.06 to -2.06 mm) of VMH nucleus per animal were analyzed for measuring the soma size of neurons in the VMH nucleus. An average of 10\u0026ndash;15 neurons per section was considered for determining soma size estimation. Six sections per animal were considered, and an average of 450 neurons per group (n\u0026thinsp;=\u0026thinsp;5/group) were analyzed to measure soma size.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\"\u003e\n \u003ch2\u003e2.9 Immunohistochemistry\u003c/h2\u003e\n \u003cp\u003eOn day 46, the perfusion-fixed brains (without brainstem and cerebellum) were processed for immunohistochemistry. Briefly, the brains were cryopreserved in 15% and 30% sucrose solutions, embedded in OCT (optimum cutting temperature), and sectioned at 25\u0026micro;m using a cryotome (Leica CM1900). Coronal sections containing the hypothalamus were collected in PBS and treated with 3% H₂O₂ to quench peroxidase activity. Following PBS rinses, sections were blocked with 10% normal goat serum and incubated with rabbit monoclonal anti-ER alpha (estrogen receptor-\u0026alpha;, 1:50, ab32063; Abcam) at 4\u0026deg;C for 48 hours. After incubation with biotinylated IgG peroxidase secondary antibody, immunostaining was visualized using a diaminobenzidine (DAB) reaction. Sections were then washed, dehydrated, cleared in xylene, cover-slipped with DPX, and scanned under a bright-field microscope to capture images.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\"\u003e\n \u003ch2\u003e2.10 Statistical Analysis\u003c/h2\u003e\n \u003cp\u003eThe collected data from the control and experimental animals were analyzed by SPSS 17 (Chicago, IL, USA) statistical software, and graphs were plotted using graph pad prism (Version 9). Body weight, heart weight, and heart heart-to-body weight ratios were analyzed using one-way ANOVA followed by a post hoc LSD test for multiple comparison analysis. The data for the thickness of ventricular walls (LV, RV, IVS), the diameter of myocyte (LV and RV), and the area of the nucleus (LV and RV) were expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD and analyzed by one-way analysis of variance (ANOVA) followed by the Bonferroni test for multiple comparisons among groups. Differences were considered statistically significant at p-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e3.1 General physical characteristics\u003c/h2\u003e \u003cp\u003eDuring the experimental period, the animals were observed for their general well-being, including body weight, state of alertness, hair loss (if any), and physical activities like rearing and grooming. No significant differences were observed regarding these physical features. On day 1 of the experimental period, the body weights of the control and experimental animals were comparable (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). On day 45, an increase in body weight was observed both in the control and the experimental groups, though the increase was not statistically significant (p\u0026thinsp;=\u0026thinsp;0.64). The percentage change in body weight (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) was maximum (12.76\u0026thinsp;\u0026plusmn;\u0026thinsp;4.41%) in group IV (Res alone treated) followed by group III (ATO alone treated) with a value of 11.92\u0026thinsp;\u0026plusmn;\u0026thinsp;3.48%. We herein assume that ATO at this low dose of 2mg/kg b.w. was not sufficient to alter the body weight of mice.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDescriptive statistics of Percentage (%) change in body weight, heart weight (mg), body weight (g), and heart-to-body weight ratio on Day 45 of the experimental period among control and experimental animals\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnimal Groups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBody weight (gm) on day 1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBody weight (gm) on day 45\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e% age change in body weight\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eHeart weight (mg)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHeart weight/Body weight ratio\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNC\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e30.41\u0026thinsp;\u0026plusmn;\u0026thinsp;4.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e33.31\u0026thinsp;\u0026plusmn;\u0026thinsp;2.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e9.39\u0026thinsp;\u0026plusmn;\u0026thinsp;3.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e285.2\u0026thinsp;\u0026plusmn;\u0026thinsp;21.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e8.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.81\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eVC\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e33.36\u0026thinsp;\u0026plusmn;\u0026thinsp;1.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e33.88\u0026thinsp;\u0026plusmn;\u0026thinsp;2.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e1.55\u0026thinsp;\u0026plusmn;\u0026thinsp;2.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e246.28\u0026thinsp;\u0026plusmn;\u0026thinsp;34.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.84\u0026thinsp;\u0026plusmn;\u0026thinsp;2.04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eATO alone\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e29.6\u0026thinsp;\u0026plusmn;\u0026thinsp;4.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e33.13\u0026thinsp;\u0026plusmn;\u0026thinsp;2.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e11.92\u0026thinsp;\u0026plusmn;\u0026thinsp;3.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e310.09\u0026thinsp;\u0026plusmn;\u0026thinsp;19.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eRes alone\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e31. 26\u0026thinsp;\u0026plusmn;\u0026thinsp;1.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e35.25\u0026thinsp;\u0026plusmn;\u0026thinsp;2.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e12.76\u0026thinsp;\u0026plusmn;\u0026thinsp;4.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e241.08\u0026thinsp;\u0026plusmn;\u0026thinsp;16.32\u003csup\u003ec***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.81\u003csup\u003ec***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eATO\u0026thinsp;+\u0026thinsp;Res\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e30.51\u0026thinsp;\u0026plusmn;\u0026thinsp;2.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e33.3\u0026thinsp;\u0026plusmn;\u0026thinsp;3.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e9.14\u0026thinsp;\u0026plusmn;\u0026thinsp;3.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e250.41\u0026thinsp;\u0026plusmn;\u0026thinsp;17.73\u003csup\u003ed***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.99\u003csup\u003ed***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003eData is presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD; n\u0026thinsp;=\u0026thinsp;6/group. Means were compared using one-way ANOVA followed by LSD test for multiple comparisons. \u003csup\u003eb*\u003c/sup\u003e VC group \u003cem\u003evs\u003c/em\u003e ATO alone; \u003csup\u003ec***\u003c/sup\u003eATO alone \u003cem\u003evs Res\u003c/em\u003e alone. \u003csup\u003ed***\u003c/sup\u003eATO alone group \u003cem\u003evs\u003c/em\u003e ATO\u0026thinsp;+\u0026thinsp;\u003cem\u003eRes\u003c/em\u003e co-treated group. \u003csup\u003e*\u003c/sup\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.05; \u003csup\u003e* *\u003c/sup\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.01; \u003csup\u003e***\u003c/sup\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.001. Abbreviations: NC- normal control, VC- vehicle control, ATO \u0026ndash;Arsenic Trioxide, Res- Resveratrol alone, ATO\u0026thinsp;+\u0026thinsp;Res \u0026ndash; ATO \u0026amp; Resveratrol co-treated group.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eNext, we observed that the heart weight was found to be higher in the ATO-alone treated animals (310.09\u0026thinsp;\u0026plusmn;\u0026thinsp;19.13) relative to controls (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e); however, the data was not statistically significant. In Res alone and ATO\u0026thinsp;+\u0026thinsp;Res co-treated animals, the heart weight was significantly lower (241.08\u0026thinsp;\u0026plusmn;\u0026thinsp;16.32; p\u0026thinsp;=\u0026thinsp;0.000) and (250.41\u0026thinsp;\u0026plusmn;\u0026thinsp;17.73; p\u0026thinsp;=\u0026thinsp;0.000), respectively when compared to ATO alone treated animals.\u003c/p\u003e \u003cp\u003eAs shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, the heart to body weight ratio was observed to be significantly higher (9.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48; p\u0026thinsp;=\u0026thinsp;0.03) in ATO-treated animals in comparison to vehicle controls (7.84\u0026thinsp;\u0026plusmn;\u0026thinsp;2.04), suggesting ATO-induced hypertrophy in these animals. However, \u003cem\u003eRes\u003c/em\u003e alone treated animals and ATO\u0026thinsp;+\u0026thinsp;\u003cem\u003eRes\u003c/em\u003e co-treated animals showed significantly lower ratios (6.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.81, p\u0026thinsp;=\u0026thinsp;0.001) (7.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.99, p\u0026thinsp;=\u0026thinsp;0.01) in contrast to ATO alone treated animals, thus indicating the beneficial role of Res against this cardiotoxin.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e3.2 Effects of \u003cem\u003eRes\u003c/em\u003e on Left and right ventricular wall thickness\u003c/h2\u003e \u003cp\u003eThe morphometric study on H\u0026amp;E-stained heart sections showed an apparent decrease in left and right ventricular thickness; however, the difference was found to be non-significant ventricular thickness on treatment with \u003cem\u003eRes\u003c/em\u003e in the ATO-treated compared with ATO alone group (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05; non-significant) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparative analysis of morphometric parameters in mouse heart: Wall Thickness (LV/RV), Interventricular septum (IVS) Thickness, Cardiac Myocyte Nuclear Area and Cardiac Myocyte Diameter in Left Ventricle (LV) and Right Ventricle (RV) across the control and experimental groups.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnimal Groups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eWall thickness (mm)-LV\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eWall\u003c/p\u003e \u003cp\u003ethickness (mm)-RV\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eIVS Thickness (mm)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCardiac myocyte nuclear area in LV (\u0026micro;m\u003csup\u003e2\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCardiac myocyte nuclear area in RV (\u0026micro;m\u003csup\u003e2\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCardiac myocyte diameter in LV (\u0026micro;m)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCardiac myocyte diameter in RV (\u0026micro;m)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNC\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e1.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e35.41\u0026thinsp;\u0026plusmn;\u0026thinsp;10.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e31.74\u0026thinsp;\u0026plusmn;\u0026thinsp;6.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e9.64\u0026thinsp;\u0026plusmn;\u0026thinsp;1.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e8.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eVC\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e1.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e38.24\u0026thinsp;\u0026plusmn;\u0026thinsp;11.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e36.02\u0026thinsp;\u0026plusmn;\u0026thinsp;12.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e10.32\u0026thinsp;\u0026plusmn;\u0026thinsp;1.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.87\u0026thinsp;\u0026plusmn;\u0026thinsp;1.66\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eATO alone\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e1.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e38.36\u0026thinsp;\u0026plusmn;\u0026thinsp;11.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e34.83\u0026thinsp;\u0026plusmn;\u0026thinsp;10.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e10.73\u0026thinsp;\u0026plusmn;\u0026thinsp;1.28 \u003csup\u003ea**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e10.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003ea***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eRes alone\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e1.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e38.77\u0026thinsp;\u0026plusmn;\u0026thinsp;9.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e39.52\u0026thinsp;\u0026plusmn;\u0026thinsp;12.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e9.16\u0026thinsp;\u0026plusmn;\u0026thinsp;1.38 \u003csup\u003ec***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.53\u0026thinsp;\u0026plusmn;\u0026thinsp;1.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eATO\u0026thinsp;+\u0026thinsp;Res\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e1.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003ed***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e31.30\u0026thinsp;\u0026plusmn;\u0026thinsp;6.20\u003csup\u003ed*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e \u003cp\u003e33.23\u0026thinsp;\u0026plusmn;\u0026thinsp;7.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e8.88\u0026thinsp;\u0026plusmn;\u0026thinsp;1.36 \u003csup\u003ed***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e8.90\u0026thinsp;\u0026plusmn;\u0026thinsp;1.27 \u003csup\u003ed***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eValues expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;S.D, n\u0026thinsp;=\u0026thinsp;6/group. Means were compared using one-way ANOVA followed by the Bonferroni test for multiple comparisons. \u003csup\u003ea**\u003c/sup\u003eand \u003csup\u003ea***\u003c/sup\u003e denotes NC \u003cem\u003evs\u003c/em\u003e ATO alone treated group; \u003csup\u003ec***\u003c/sup\u003e denotes ATO alone \u003cem\u003evs\u003c/em\u003e Res alone; \u003csup\u003ed*\u003c/sup\u003eand \u003csup\u003ed***\u003c/sup\u003edenotes ATO alone \u003cem\u003evs\u003c/em\u003e ATO\u0026thinsp;+\u0026thinsp;Res co-treated group (\u003csup\u003e*\u003c/sup\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.05, \u003csup\u003e**\u003c/sup\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.01, \u003csup\u003e***\u003c/sup\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Abbreviations: NC- normal control, VC- vehicle control, ATO \u0026ndash;ATO alone, Res- Resveratrol alone, ATO\u0026thinsp;+\u0026thinsp;Res \u0026ndash; ATO \u0026amp; Resveratrol co-treated group.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Effects of \u003cem\u003eRes\u003c/em\u003e on the Thickness of Interventricular Septum (IVS)\u003c/h2\u003e \u003cp\u003eNo significant difference was observed in the thickness of IVS among control (0.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18) and ATO alone treated (0.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10, p\u0026thinsp;=\u0026thinsp;1.000) animals. A significant decrease in the thickness of IVS was observed in the ATO\u0026thinsp;+\u0026thinsp;\u003cem\u003eRes\u003c/em\u003e co-treated groups (0.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18, p\u0026thinsp;=\u0026thinsp;0.000) compared with the ATO alone (0.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10) group, thereby indicating the beneficial role of \u003cem\u003eRes\u003c/em\u003e on ATO-induced alterations in IVS thickness (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e3.4 Effects of \u003cem\u003eRes\u003c/em\u003e on cardiac myocyte diameter and nuclear area\u003c/h2\u003e \u003cp\u003eThe average values along with descriptive statistics of cardiac myocyte diameter in LV and RV walls are tabulated in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The diameter of cardiac myocytes in LV was found to be significantly increased in ATO-alone treated animals (10.73\u0026thinsp;\u0026plusmn;\u0026thinsp;1.28; p\u0026thinsp;=\u0026thinsp;0.003) when compared to normal control. On the contrary, the animals receiving either \u003cem\u003eRes\u003c/em\u003e alone (9.16\u0026thinsp;\u0026plusmn;\u0026thinsp;1.38; p\u0026thinsp;=\u0026thinsp;0.000) or co-treatment of ATO\u0026thinsp;+\u0026thinsp;\u003cem\u003eRes\u003c/em\u003e showed a significant decrease (8.88\u0026thinsp;\u0026plusmn;\u0026thinsp;1.36; p\u0026thinsp;=\u0026thinsp;0.000) in myocyte diameter compared to ATO alone treated animals. Similarly in the RV region, animals exposed to ATO alone showed a significant increase in myocyte diameter (10.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22, p\u0026thinsp;=\u0026thinsp;0.000) compared to control animals. ATO\u0026thinsp;+\u0026thinsp;\u003cem\u003eRes\u003c/em\u003e co-treated animals revealed a significant decrease (8.90\u0026thinsp;\u0026plusmn;\u0026thinsp;1.27, p\u0026thinsp;=\u0026thinsp;0.001) in RV myocyte diameter compared to alone-treated animals (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eATO alone exposed animals showed an apparent increase in the cardiac myocyte nuclear area in the region of LV (38.24\u0026thinsp;\u0026plusmn;\u0026thinsp;11.94) and RV (34.83\u0026thinsp;\u0026plusmn;\u0026thinsp;10.93) as compared to normal control animals (LV- 35.41\u0026thinsp;\u0026plusmn;\u0026thinsp;10.11; RV- 31.74\u0026thinsp;\u0026plusmn;\u0026thinsp;6.04), respectively, though not statistically significant. ATO\u0026thinsp;+\u0026thinsp;\u003cem\u003eRes\u003c/em\u003e co-treated group showed a significant decrease (31.30\u0026thinsp;\u0026plusmn;\u0026thinsp;6.20, p\u0026thinsp;=\u0026thinsp;0.030) in the nuclear area in the LV region as compared to ATO alone treated animals as shown in Supplementary Table\u0026nbsp;2.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e3.5 General microscopic observations of cardiac tissue\u003c/h2\u003e \u003cp\u003eTo study the morphological features of ventricular myocardium in the control, ATO- alone and ATO\u0026thinsp;+\u0026thinsp;Resveratrol co-treated mice, we examined H \u0026amp; E stained paraplast sections under the bright field microscope. Overall, cardiac muscle fibers were observed to be well arranged across control and experimental groups (Fig.\u0026nbsp;3\u0026ndash;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e7\u003c/span\u003e). Also, the branching patterns of cardiac myocytes along with sites of intercalated discs were evident across all the animal groups (Fig.\u0026nbsp;3\u0026ndash;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e7\u003c/span\u003e). The nuclei of the myocytes as well as fibroblasts presented a normal appearance in control as well as experimental animals (Fig.\u0026nbsp;3\u0026ndash;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e7\u003c/span\u003e). However, in Masson\u0026rsquo;s trichrome stained sections of ATO alone treated animals, substantial fibrosis in the vessel wall was noticed compared to control sections (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e8\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003e3.6 Morphological observations in Ventromedial Nucleus of Hypothalamus (VMH)\u003c/h2\u003e \u003cp\u003eThe ventromedial nucleus of the hypothalamus was identified as a bilateral cell group in the medial basal hypothalamus lying near the third ventricle. The outer limits of the VMH were defined by the encircling stria terminalis fibers, which separated the dense neuronal core of the nucleus from the relatively lesser neuron-concentrated area (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The cells of the ventromedial nucleus appeared to be variable in shape and size. Neuronal profiles in CV-stained sections containing VMH nuclei showed vesicular nuclei and prominent nucleoli under high-power magnification (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e9\u003c/span\u003e, 10). The neuronal population of the VMH nucleus showed variability based on their size and shape. Neurons of various shapes including spherical, round, oval, and triangular, and sizes, such as small, medium, and large, were observed in the VMH nucleus (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e9\u003c/span\u003e, 10). In most of the sections, VMH neurons presented round to oval-shaped cell bodies with round centrally placed nuclei having two or more nucleoli. The neurons were more densely packed in the ventromedial aspect of the third ventricle than in the dorsolateral aspect. Qualitatively, intensely stained small and medium-sized neurons were seen in some of the sections of ATO (2 \u0026amp; 4 mg/kg) alone groups in comparison to the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e9\u003c/span\u003e, 10). Otherwise, no obvious change was observed in the morphological features of VMH neurons among experimental groups receiving ATO (2 \u0026amp; 4 mg/kg) alone in comparison to controls and RES supplemented.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003e3.7 Morphometric observations in Ventromedial Nucleus of Hypothalamus (VMH)\u003c/h2\u003e \u003cp\u003eThe values for neuronal density and soma size of VMH neurons are presented in Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e11\u003c/span\u003e. The neuronal density in VMH nucleus was found to be significantly (p\u0026thinsp;=\u0026thinsp;0.0471, p\u0026thinsp;=\u0026thinsp;0.0303) reduced in ATO alone treated groups (2mg/kg, 6690\u0026thinsp;\u0026plusmn;\u0026thinsp;1410 \u0026amp; 4 mg/kg, 6537\u0026thinsp;\u0026plusmn;\u0026thinsp;827) when compared to normal control (9471\u0026thinsp;\u0026plusmn;\u0026thinsp;1337). The mean neuronal density in groups receiving RES along with ATO (ATO 2\u0026thinsp;+\u0026thinsp;40mg/kg, 9813\u0026thinsp;\u0026plusmn;\u0026thinsp;1400 \u0026amp; ATO 4\u0026thinsp;+\u0026thinsp;40 mg/kg, 9480\u0026thinsp;\u0026plusmn;\u0026thinsp;2001) was significantly (\u003csup\u003ec\u003c/sup\u003ep=0.0174, \u003csup\u003ef\u003c/sup\u003ep=0.0295) greater than ATO (2 and 4 mg/kg) alone treated groups.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe mean soma size of VMH neurons was observed to be significantly (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) decreased in ATO (2 \u0026amp; 4 mg/kg) (246\u0026thinsp;\u0026plusmn;\u0026thinsp;12.21; 222\u0026thinsp;\u0026plusmn;\u0026thinsp;32.89) alone exposed groups in comparison to the normal control group (418\u0026thinsp;\u0026plusmn;\u0026thinsp;48.31). On the contrary, ATO\u0026thinsp;+\u0026thinsp;RES co-treated (ATO 2\u0026thinsp;+\u0026thinsp;40mg/kg\u0026thinsp;=\u0026thinsp;409\u0026thinsp;\u0026plusmn;\u0026thinsp;37.19; ATO 4\u0026thinsp;+\u0026thinsp;40mg/kg\u0026thinsp;=\u0026thinsp;397\u0026thinsp;\u0026plusmn;\u0026thinsp;26.17) groups showed a significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) increase in mean soma size of VMH neurons when compared to the mean of soma size of VMH neurons in ATO 2 and 4 mg alone treated group (409\u0026thinsp;\u0026plusmn;\u0026thinsp;37.19; 397\u0026thinsp;\u0026plusmn;\u0026thinsp;26.17), thereby suggesting RES-induced restoration of soma size of neurons in VMH nucleus.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003e3.8 Effect of ATO\u0026thinsp;+\u0026thinsp;RES on Estrogen Receptor-α Expression\u003c/h2\u003e \u003cp\u003eSince estrogen receptor regulation plays a crucial role in metabolism, neuroendocrine function, and cardiovascular health, we aimed to test the hypothesis that prolonged arsenic exposure disrupts estrogen signaling, potentially affecting both neuroendocrine and cardiac function. Immunohistochemical localization of ERα was observed in the coronal sections (25 \u0026micro;m) of the mouse brain, showing the preoptic area of the hypothalamus (bregma\u0026thinsp;+\u0026thinsp;0.74 mm to -0.58 mm). Specifically, ERα immunoreactivity was observed in the cytoplasm of the neurons as well as in the neuropil among control and experimental animals (Fig.\u0026nbsp;12). Largely, the ERα\u0026thinsp;+\u0026thinsp;ve neurons were observed to be rounded or pyramidal in shape (Fig.\u0026nbsp;12). Qualitatively, ERα expression was less intense in the cytoplasm of the neurons in preoptic areas of the experimental groups receiving ATO (2 and 4 mg/kg bw) alone in comparison to controls and RES-supplemented groups. Hence, prolonged ATO exposure reduces ERα expression in the preoptic area of the hypothalamus, potentially disrupting estrogen signaling and neuroendocrine function, while resveratrol may offer protective effects.\u003c/p\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eRecently, our lab provided evidence of the neuroprotective efficacy of Resveratrol on arsenic-induced cytoarchitectural perturbations, cognitive dysfunction, and disrupted estrogen signaling in the mouse hippocampus (Mehta et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2021b\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003ea\u003c/span\u003e). However, the protective effects of Res on arsenic-induced cardiotoxicity remains poorly understood. Therefore, the present study was designed to determine the beneficial roles of Resveratrol (a potent antioxidant) on the morphological and morphometric features of the heart in adult mice subjected to chronic exposure to Arsenic Trioxide (ATO). We found that Resveratrol supplementation significantly alleviated cardiac tissue damage, reducing cardiac fibrosis and ventricular hypertrophy to some extent. These findings underscore resveratrol's potential as a therapeutic agent against arsenic trioxide-induced cardiotoxicity, highlighting its role in preserving cardiac integrity and function amidst environmental toxic insults. Such insights offer promising avenues for developing preventive strategies and interventions against arsenic trioxide-induced cardiac pathology.\u003c/p\u003e \u003cp\u003eATO is a highly potent anticancer drug being used in the treatment of APL (Emadi and Gore \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). However, cardiotoxicity associated with the use of ATO limits its use to a certain extent, (Saad et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) although ATO has been reported to adversely affect the functions of the Liver, Kidney, and CNS as well (Zhang et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). ATO-induced adverse effects on the heart have been manifested as increased thickness of ventricular walls. To date, the possible mechanisms underlying ATO-induced cardiotoxicity include DNA fragmentation, ROS generation/oxidative stress, cardiac ion channel changes, and apoptosis (Zhang et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePrevious studies examined the effects of \u003cem\u003eRes\u003c/em\u003e on oxidative stress and various signaling pathways in advanced stages of heart failure induced by subcutaneous administration of Isoproterenol (ISO). This study revealed that ISO administration decreased LV function and altered myocardial fibres orientation, while \u003cem\u003eRes\u003c/em\u003e treatment restored LV function and moderated heart failure severity (Chakraborty et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Likewise, another study showed the cardioprotective effects of polyphenol phytoalexin \u003cem\u003eRes\u003c/em\u003e against doxorubicin-induced cardiotoxicity in adult female mice, thus providing in vivo evidence (Osman et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). In the present study, we also observed an increase in LV and RV wall thickness in H\u0026amp;E-stained heart sections of animals exposed to ATO alone when compared to the control group. However, the LV and RV wall thickness in animals co-treated with ATO and \u003cem\u003eRes\u003c/em\u003e was less than in ATO alone treated groups and comparable to wall thickness in controls. These observations do suggest the beneficial role of \u003cem\u003eRes\u003c/em\u003e against ATO-induced adverse effects on cardiac morphology. The observations of our study also revealed a significant increase in cardiac myocyte diameter (LV and RV) in ATO alone exposed animals, thereby suggestive of cardiac hypertrophy compared to normal controls and the animals receiving ATO and \u003cem\u003eRes\u003c/em\u003e simultaneously.\u003c/p\u003e \u003cp\u003ePrior work provided insights into the effects of \u003cem\u003eRes\u003c/em\u003e in a post-infarction heart failure rat model, where isoproterenol was used to induce myocardial infarction and post-infarction remodeling (Riba et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Riba and co-workers also showed that systolic left ventricular function was significantly increased, whereas plasma BNP levels, left ventricular wall thickness and dimensions were decreased after 8 weeks of \u003cem\u003eRes\u003c/em\u003e treatment (15 mg/kg/day) (Riba et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Likewise, attenuation of doxorubicin-induced impairment of cardiac function in aged mice was shown through the restoration of SIRT1 activity (Sin et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Research has extensively documented the impact of \u003cem\u003eRes\u003c/em\u003e on cardiac structure and function in a mouse model of heart failure induced by pressure overload (Sung et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Moreover, the study also observed positive effects on cardiac energy metabolism, suggesting \u003cem\u003eRes\u0026rsquo;s\u003c/em\u003e potential to address both functional and metabolic aspects in the context of heart failure. \u003cem\u003eRes\u003c/em\u003e administration at a dose of 150 mg/kg/day led to improvements in diastolic function, decreased left ventricular diameters and volumes, and mitigated cardiac fibrosis, hypertrophy, and remodeling through its antifibrotic and anti-inflammatory properties (Sung et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). However, systolic function did not change after a 2-week-long \u003cem\u003eRes\u003c/em\u003e supplementation. Similarly, another group found that even the low dose of \u003cem\u003eRes\u003c/em\u003e (2.5 mg/kg/day for 28 days) was able to regress the pressure-overload-induced cardiac hypertrophy and remodeling in Sprague Dawley rats (Wojciechowski et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Previous clinical studies investigated Resveratrol's potential cardioprotective effects in myocardial infarction patients with preserved ejection fraction (baseline: 54.77\u0026thinsp;\u0026plusmn;\u0026thinsp;1.64% in the treated group). They discovered a significant improvement in diastolic function after administering 10 mg/day of Resveratrol for 3 months, yet no significant improvement was noted in systolic function (Magyar et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the current study, the morphometric analysis showed an apparent increase in the width of cardiomyocytes in the lateral wall of the left ventricle of the ATO (2mg/kg) alone treated group compared to other groups, which is suggestive of ATO-induced adverse effects on myocyte size. It has been reported that increased measurement of myocardiocytes is associated with left ventricular hypertrophy (ASHLEY 1945; Tracy \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Several clinical studies have reported that ATO treatment in APL patients is associated with various cardiac complications such as ECG abnormalities, ventricular tachycardia, pericardial effusion, myocardial infarction, and left ventricular (Unnikrishnan et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Ravandi et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Such studies have examined ATO-induced ventricular hypertrophy using the echocardiography (ECG) technique (Bagul et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the present study, morphometric observations carried out on H\u0026amp;E-stained sections (40X) showed increased nucleus size and myocyte size in ATO-alone treated animals. These changes were attenuated to some extent in animals co-treated with ATO and \u003cem\u003eRes\u003c/em\u003e. The results of the current study align with a prior study, showing that supplementing \u003cem\u003eRes\u003c/em\u003e at a dosage of 10 mg/kg/day for 8 weeks led to a reduction in cardiac hypertrophy in rats with diabetes. This reduction was attributed to the antioxidant effects of \u003cem\u003eRes\u003c/em\u003e, which are mediated by SIRT1 (Riba et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). This highlights the fact that the biological activities of \u003cem\u003eRes\u003c/em\u003e may be dependent on its multiple molecular targets including SIRT1.\u003c/p\u003e \u003cp\u003eMoreover, observations (present study) from Masson\u0026rsquo;s Trichrome stained sections showed an increase in interstitial and perivascular fibrosis in the ATO alone treated group compared to the control group. A previous study also showed the accumulation of collagen fibres in transgenic mice contributing to cardiac hypertrophy, in addition, to an increase in cardiomyocyte size (Shi et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Similarly, another group assessed the long-term effects of ATO therapy in Guinea pigs (Chu et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Chu and co-workers observed substantial structural alterations in both Right and Left ventricles of animals treated with ATO based on analysis of Masson\u0026rsquo;s trichrome-stained heart sections compared to controls (Chu et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Nevertheless, future studies are warranted to explore in-depth mechanisms of ATO-induced cardiac damage.\u003c/p\u003e \u003cp\u003eIn this study, we also examined the effects of ATO exposure on morphological features \u0026amp; morphometric parameters of VMH nucleus using Cresyl Violet stained paraplast sections containing region and to note any alteration in these features following co-administration of RES (ATO\u0026thinsp;+\u0026thinsp;RES). Given the potential link between histopathological changes in hypothalamic neurons and disrupted estrogen receptor (ER) signaling, we explored the expression patterns of ER-α within the hypothalamic nuclei. Notably, we observed a qualitative reduction in ER-α expression in the preoptic area of ATO-exposed groups (2 mg/kg and 4 mg/kg bw), suggesting a downregulation of estrogen signaling due to arsenic exposure. However, co-treatment with resveratrol (ATO\u0026thinsp;+\u0026thinsp;RES) appeared to mitigate this effect, partially restoring ER-α expression.\u003c/p\u003e \u003cp\u003eThese findings indicate that ATO exposure disrupts estrogen receptor regulation in the hypothalamus, potentially impacting neuroendocrine function. The partial reversal of ER-α downregulation by resveratrol suggests its neuroprotective role in counteracting arsenic-induced endocrine dysregulation, possibly through its antioxidant and anti-inflammatory mechanisms. Furthermore, these observations support the notion that inorganic arsenic (iAs) may act as an endocrine-disrupting compound (EDC) within specific brain regions. We focused on ER expression due to its highly divergent nature among steroid hormone receptors, its extensive co-regulatory interactions, and its distinct activation and deactivation phases (Stenoien et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Dennis and O\u0026rsquo;Malley \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Chatterjee and Chatterji \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2010a\u003c/span\u003e). Moreover, our previous findings have also demonstrated that ATO disrupts estrogen signaling in the hippocampus, a brain region abundant in estrogen receptors, further reinforcing the potential endocrine-disrupting effects of arsenic (Mehta et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2021b\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTo the best of our knowledge, we could find only one study (Ommati et al., 2020) which could explain the microscopic changes in hypothalamic tissue after \u003cem\u003eiAs\u003c/em\u003e exposure. Recently, Transmission electron microscopy (TEM) findings revealed an increase in the number of double membrane-surrounded autophagosomes (autophagic vacuoles) in HPG tissues of As\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e -treated animals (Ommati et al., 2020). TEM findings further showed that the highest number of autophagic vacuoles was recorded in the mature hypothalamus of F1-male mice exposing to the highest dose of As\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e (20 ppm). Moreover, there are no other reports that have determined the effects of \u003cem\u003eiAs\u003c/em\u003e exposure on the estrogen signaling in brain regions like the hippocampus. However, several in-vitro and in-vivo reports demonstrate the endocrine-disrupting effects of \u003cem\u003eiAs\u003c/em\u003e exposure in other target tissues and cell line models (Watson and Yager \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Akram et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Zhang et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Endocrine disruptors can either mimic or antagonize the effects of hormones in the target tissues. The role of \u003cem\u003eAs\u003c/em\u003e as a potent EDC has been suggested by \u003cem\u003eiAs\u003c/em\u003e-induced alteration of gene regulation by the closely related steroid hormone receptors for glucocorticoids, mineralocorticoids, progesterone, androgen, and estrogen (Bodwell et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2006\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Chatterjee and Chatterji, 2010; Kaltreider et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Sun et al., \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe incidence of ATO-induced cardiotoxicity and neural defects in APL patients is relatively low, but it can be severe when it does occur. Several factors contribute to ATO-induced cardiotoxicity, including aging, pre-existing heart disease, and concurrent use of other cardiotoxic drugs. To mitigate the risk of ATO-induced cardiotoxicity and neural defects, it is important to monitor cardiac and neural functions regularly in APL patients receiving ATO treatment. This can involve performing electrocardiograms (ECGs), echocardiograms, and electroencephalograms (EEG), fMRI tests. In some cases, it may be necessary to adjust the dosage or duration of ATO treatment or to switch to an alternative antioxidative therapy.\u003c/p\u003e \u003cp\u003e Our study presents promising implications for patient care in managing cardiotoxicity associated with Arsenic Trioxide (ATO) treatment for Acute Promyelocytic Leukemia (APL). Investigating the effects of Resveratrol (Res) on mouse hearts exposed chronically to ATO sheds light on potential interventions to mitigate ATO-induced cardiac hypertrophy and fibrosis in APL patients. Implementing Res as a supplementary treatment alongside ATO could potentially enhance oncology patient care by minimizing adverse cardiac effects. Additionally, Res might serve as a dietary supplement in arsenic-endemic areas, potentially mitigating prevailing arsenic-induced cardiotoxicity among inhabitants.\u003c/p\u003e \u003cp\u003eA major limitation of this study is that we did not assess estrogen receptor signaling in the heart, which could provide a more comprehensive understanding of systemic estrogen regulation under ATO exposure. Since estrogen receptors play a crucial role in cardiovascular function, future studies should investigate whether ATO-induced downregulation of ER-α in the hypothalamus also occurs in the heart and whether RES offers similar protective effects. Exploring estrogen receptor-mediated pathways in both the brain and heart could help elucidate the broader endocrine and cardiovascular consequences of arsenic toxicity and the therapeutic potential of RES.\u003c/p\u003e"},{"header":"5. Conclusions","content":"\u003cp\u003eGlobally, millions of people are poisoned by arsenic due to its ecological prevalence and epidemiological importance. Our study revealed the significant cardiac changes in the mouse model of arsenic toxicity (ATO), notably the histological modifications. Meanwhile, resveratrol supplementation following arsenic intoxication in adult mice produced ameliorative effects against cardiac and neuronal morphology. The observed cardio- and neuroprotective effects of Resveratrol could be attributed to its potent antioxidant activity and metal-chelating action against arsenic. It could also be presumed that supplementation of Resveratrol might be an ideal approach for amelioration of ATO-induced cardio- and neurotoxicity, though countless in-depth studies are warranted in this direction to reach any substantial conclusion.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe duly acknowledge the technical help extended by Mr. Kirpal Singh and the financial support provided by the Department of Anatomy at the All India Institute of Medical Sciences, New Delhi, India.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAkram Z, Jalali S, Shami SA, et al (2010) Adverse effects of arsenic exposure on uterine function and structure in female rat. Experimental and Toxicologic Pathology 62:451\u0026ndash;459. https://doi.org/10.1016/j.etp.2009.07.008\u003c/li\u003e\n\u003cli\u003eASHLEY LM (1945) A determination of the diameters of ventricular myocardial fibers in man and other mammals. The American journal of anatomy 77:325\u0026ndash;363. https://doi.org/10.1002/aja.1000770303\u003c/li\u003e\n\u003cli\u003eBagul PK, Deepthi N, Sultana R, Banerjee SK (2015) Resveratrol ameliorates cardiac oxidative stress in diabetes through deacetylation of NFkB-p65 and histone 3. The Journal of nutritional biochemistry 26:1298\u0026ndash;1307\u003c/li\u003e\n\u003cli\u003eBalali-Mood M, Naseri K, Tahergorabi Z, et al (2021) Toxic Mechanisms of Five Heavy Metals: Mercury, Lead, Chromium, Cadmium, and Arsenic. Frontiers in Pharmacology 12:1\u0026ndash;19. https://doi.org/10.3389/fphar.2021.643972\u003c/li\u003e\n\u003cli\u003eBarbey JT, Pezzullo JC, Soignet SL (2003) Effect of arsenic trioxide on QT interval in patients with advanced malignancies. Journal of Clinical Oncology 21:3609\u0026ndash;3615. https://doi.org/10.1200/JCO.2003.10.009\u003c/li\u003e\n\u003cli\u003eBenes FM, Vincent SL, Todtenkopf M (2001) The density of pyramidal and nonpyramidal neurons in anterior cingulate cortex of schizophrenic and bipolar subjects. Biological Psychiatry 50:395\u0026ndash;406. https://doi.org/10.1016/S0006-3223(01)01084-8\u003c/li\u003e\n\u003cli\u003eBodwell JE, Gosse JA, Nomikos AP, Hamilton JW (2006) Arsenic disruption of steroid receptor gene activation: Complex dose - Response effects are shared by several steroid receptors. Chemical Research in Toxicology 19:1619\u0026ndash;1629. https://doi.org/10.1021/tx060122q\u003c/li\u003e\n\u003cli\u003eBodwell JE, Kingsley LA, Hamilton JW (2004) Arsenic at very low concentrations alters glucocorticoid receptor (GR)-mediated gene activation but not GR-mediated gene repression: Complex dose-response effects are closely correlated with levels of activated GR and require a functional GR DNA binding d. Chemical Research in Toxicology 17:1064\u0026ndash;1076. https://doi.org/10.1021/tx0499113\u003c/li\u003e\n\u003cli\u003eChakraborty S, Pujani M, Haque SE (2015) Combinational effect of resveratrol and atorvastatin on isoproterenol-induced cardiac hypertrophy in rats. Journal of pharmacy \u0026amp; bioallied sciences 7:233\u0026ndash;238. https://doi.org/10.4103/0975-7406.160037\u003c/li\u003e\n\u003cli\u003eChandravanshi LP, Gupta R, Shukla RK (2019) Arsenic-Induced Neurotoxicity by Dysfunctioning Cholinergic and Dopaminergic System in Brain of Developing Rats. Biological Trace Element Research 189:118\u0026ndash;133. https://doi.org/10.1007/s12011-018-1452-5\u003c/li\u003e\n\u003cli\u003eChatterjee A, Chatterji U (2010a) Arsenic abrogates the estrogen-signaling pathway in the rat uterus. Reproductive biology and endocrinology : RB\u0026amp;E 8:80. https://doi.org/10.1186/1477-7827-8-80\u003c/li\u003e\n\u003cli\u003eChatterjee A, Chatterji U (2010b) Arsenic abrogates the estrogen-signaling pathway in the rat uterus. Reproductive Biology and Endocrinology 8:1\u0026ndash;11. https://doi.org/10.1186/1477-7827-8-80\u003c/li\u003e\n\u003cli\u003eChu W, Li C, Qu X, et al (2012) Arsenic-induced interstitial myocardial fibrosis reveals a new insight into drug-induced long QT syndrome. Cardiovascular research 96:90\u0026ndash;98\u003c/li\u003e\n\u003cli\u003eDennis AP, O\u0026rsquo;Malley BW (2005) Rush hour at the promoter: How the ubiquitin-proteasome pathway polices the traffic flow of nuclear receptor-dependent transcription. Journal of Steroid Biochemistry and Molecular Biology 93:139\u0026ndash;151. https://doi.org/10.1016/j.jsbmb.2004.12.015\u003c/li\u003e\n\u003cli\u003eEmadi A, Gore SD (2010) Arsenic trioxide - An old drug rediscovered. Blood Reviews. https://doi.org/10.1016/j.blre.2010.04.001\u003c/li\u003e\n\u003cli\u003eEyvani H, Moghaddaskho F, Kabuli M, et al (2016) Arsenic trioxide induces cell cycle arrest and alters DNA methylation patterns of cell cycle regulatory genes in colorectal cancer cells. Life Sciences 167:67\u0026ndash;77. https://doi.org/10.1016/j.lfs.2016.10.020\u003c/li\u003e\n\u003cli\u003eGarris DR (1989) Morphometric analysis of obesity (ob/ob)- and diabetes (db/db)-associated hypothalamic neuronal degeneration in C57BL/KsJ mice. Brain Research 501:162\u0026ndash;170. https://doi.org/10.1016/0006-8993(89)91037-8\u003c/li\u003e\n\u003cli\u003eGarris DR, Diani AR, Smith C, Gerritsen GC (1982) Depopulation of the ventromedial hypothalamic nucleus in the diabetic Chinese hamster. Acta Neuropathologica 56:63\u0026ndash;66. https://doi.org/10.1007/BF00691183\u003c/li\u003e\n\u003cli\u003eGarris DR, West RL, Coleman DL (1985) Morphometric analysis of medial basal hypothalamic neuronal degeneration in diabetes (db/db) mutant C57BL/KsJ mice: Relation to age and hyperglycemia. Developmental Brain Research 20:161\u0026ndash;168. https://doi.org/10.1016/0165-3806(85)90101-4\u003c/li\u003e\n\u003cli\u003eKaltreider RC, Davis AM, Lariviere JP, Hamilton JW (2001) Arsenic alters the function of the glucocorticoid receptor as a transcription factor. Environmental Health Perspectives 109:245\u0026ndash;251. https://doi.org/10.1289/ehp.01109245\u003c/li\u003e\n\u003cli\u003eKitchin KT, Conolly R (2010) Arsenic-induced carcinogenesissoxidative stress as a possible mode of action and future research needs for more biologically based risk assessment. Chemical Research in Toxicology 23:327\u0026ndash;335. https://doi.org/10.1021/tx900343d\u003c/li\u003e\n\u003cli\u003eKorbo L, Amrein I, Lipp HP, et al (2004) No evidence for loss of hippocampal neurons in non-Alzheimer dementia patients. Acta Neurologica Scandinavica 109:132\u0026ndash;139. https://doi.org/10.1034/j.1600-0404.2003.00182.x\u003c/li\u003e\n\u003cli\u003eLo-Coco F, Avvisati G, Vignetti M, et al (2013) Retinoic Acid and Arsenic Trioxide for Acute Promyelocytic Leukemia. New England Journal of Medicine 369:111\u0026ndash;121. https://doi.org/10.1056/nejmoa1300874\u003c/li\u003e\n\u003cli\u003eLuo JH, Qiu ZQ, Zhang L, Shu WQ (2012) Arsenite exposure altered the expression of NMDA receptor and postsynaptic signaling proteins in rat hippocampus. Toxicology Letters 211:39\u0026ndash;44. https://doi.org/10.1016/j.toxlet.2012.02.021\u003c/li\u003e\n\u003cli\u003eMagyar K, Halmosi R, Palfi A, et al (2012) Cardioprotection by resveratrol: A human clinical trial in patients with stable coronary artery disease. Clinical Hemorheology and Microcirculation 50:179\u0026ndash;187. https://doi.org/10.3233/CH-2011-1424\u003c/li\u003e\n\u003cli\u003eMedda N, Patra R, Ghosh TK, Maiti S (2020) Neurotoxic Mechanism of Arsenic: Synergistic Effect of Mitochondrial Instability, Oxidative Stress, and Hormonal-Neurotransmitter Impairment. Biological Trace Element Research 198:8\u0026ndash;15\u003c/li\u003e\n\u003cli\u003eMehta K, Kaur B, Pandey KK, et al (2021a) Resveratrol protects against inorganic arsenic-induced oxidative damage and cytoarchitectural alterations in female mouse hippocampus. Acta Histochemica 123:151792. https://doi.org/10.1016/j.acthis.2021.151792\u003c/li\u003e\n\u003cli\u003eMehta K, Pandey KK, Kaur B, et al (2021b) Resveratrol attenuates arsenic-induced cognitive deficits via modulation of Estrogen-NMDAR-BDNF signalling pathway in female mouse hippocampus. Psychopharmacology 238:2485\u0026ndash;502. https://doi.org/10.1007/s00213-021-05871-2\u003c/li\u003e\n\u003cli\u003eMukherjee S, Dudley JI, Das DK (2010) Dose-dependency of resveratrol in providing health benefits. Dose-Response 8:478\u0026ndash;500. https://doi.org/10.2203/dose-response.09-015.Mukherjee\u003c/li\u003e\n\u003cli\u003eNamavar MR, Raminfard S, Jahromi ZV, Azari H (2012) Effects of high-fat diet on the numerical density and number of neuronal cells and the volume of the mouse hypothalamus: a stereological study. Anatomy \u0026amp; Cell Biology 45:178. https://doi.org/10.5115/acb.2012.45.3.178\u003c/li\u003e\n\u003cli\u003eOhnishi K, Yoshida H, Shigeno K, et al (2000) Prolongation of the QT interval and ventricular tachycardia in patients treated with arsenic trioxide for acute promyelocytic leukemia. Annals of Internal Medicine 133:881\u0026ndash;885. https://doi.org/10.7326/0003-4819-133-11-200012050-00012\u003c/li\u003e\n\u003cli\u003eOsman AMM, Al-Harthi SE, AlArabi OM, et al (2013) Chemosensetizing and cardioprotective effects of resveratrol in doxorubicin- treated animals. Cancer Cell International 13:1. https://doi.org/10.1186/1475-2867-13-52\u003c/li\u003e\n\u003cli\u003ePaul NP, Galv\u0026aacute;n AE, Yoshinaga-Sakurai K, et al (2023) Arsenic in medicine: past, present and future. BioMetals 36:283\u0026ndash;301. https://doi.org/10.1007/s10534-022-00371-y\u003c/li\u003e\n\u003cli\u003ePowell BL, Moser B, Stock W, et al (2011) Arsenic trioxide improves event-free and overall survival for adults with acute promyelocytic leukemia : North American Leukemia Intergroup Study C9710 Arsenic trioxide improves event-free and overall survival for adults with acute promyelocytic leukemia . Cancer 116:3751\u0026ndash;3757. https://doi.org/10.1182/blood-2010-02-269621\u003c/li\u003e\n\u003cli\u003eRan S, Liu J, Li S (2020) A Systematic Review of the Various Effect of Arsenic on Glutathione Synthesis in Vitro and in Vivo. BioMed Research International 2020:. https://doi.org/10.1155/2020/9414196\u003c/li\u003e\n\u003cli\u003eRavandi F, Estey E, Jones D, et al (2009) Effective treatment of acute promyelocytic leukemia with all-trans-retinoic acid, arsenic trioxide, and gemtuzumab ozogamicin. Journal of Clinical Oncology 27:504\u003c/li\u003e\n\u003cli\u003eRiba A, Deres L, Sumegi B, et al (2017) Cardioprotective effect of resveratrol in a postinfarction heart failure model. Oxidative Medicine and Cellular Longevity 2017:. https://doi.org/10.1155/2017/6819281\u003c/li\u003e\n\u003cli\u003eRodr\u0026iacute;guez VM, Carrizales L, Jim\u0026eacute;nez-Capdeville ME, et al (2001) The effects of sodium arsenite exposure on behavioral parameters in the rat. Brain Research Bulletin 55:301\u0026ndash;308. https://doi.org/10.1016/S0361-9230(01)00477-4\u003c/li\u003e\n\u003cli\u003eRodr\u0026iacute;guez VM, Jim\u0026eacute;nez-Capdeville ME, Giordano M (2003) The effects of arsenic exposure on the nervous system. Toxicology Letters 145:1\u0026ndash;18. https://doi.org/10.1016/S0378-4274(03)00262-5\u003c/li\u003e\n\u003cli\u003eRoy TS, Sharma V, Seidler FJ, Slotkin TA (2005) Quantitative morphological assessment reveals neuronal and glial deficits in hippocampus after a brief subtoxic exposure to chlorpyrifos in neonatal rats. Developmental Brain Research 155:71\u0026ndash;80. https://doi.org/10.1016/j.devbrainres.2004.12.004\u003c/li\u003e\n\u003cli\u003eSaad SY, Alkharfy KM, Arafah MM (2010) Cardiotoxic effects of arsenic trioxide/imatinib mesilate combination in rats. Journal of Pharmacy and Pharmacology 58:567\u0026ndash;573. https://doi.org/10.1211/jpp.58.4.0017\u003c/li\u003e\n\u003cli\u003eShi K, Zhao W, Chen Y, et al (2014) Cardiac hypertrophy associated with myeloproliferative neoplasms in JAK2V617F transgenic mice. Journal of Hematology \u0026amp; Oncology 7:1\u0026ndash;8\u003c/li\u003e\n\u003cli\u003eShin J, Seol I, Son C (2010) Interpretation of Animal Dose and Human Equivalent Dose for Drug Development. The Journal of Korean Oriental Medicine 31:1\u0026ndash;7\u003c/li\u003e\n\u003cli\u003eSin TK, Tam BT, Yung BY, et al (2015) Resveratrol protects against doxorubicin-induced cardiotoxicity in aged hearts through the SIRT1-USP7 axis. Journal of Physiology 593:1887\u0026ndash;1899. https://doi.org/10.1113/jphysiol.2014.270101\u003c/li\u003e\n\u003cli\u003eSmith AH, Arroyo AP, Guha Mazumder DN, et al (2000) Arsenic-induced skin lesions among Atacameno people in Northern Chile despite good nutrition and centuries of exposure. Environmental Health Perspectives 108:617\u0026ndash;620. https://doi.org/10.1289/ehp.00108617\u003c/li\u003e\n\u003cli\u003eStenoien DL, Simeoni S, Sharp ZD, Mancini MA (2000) Subnuclear dynamics and transcription factor function. Journal of cellular biochemistry Supplement Suppl 35:99\u0026ndash;106. https://doi.org/10.1002/1097-4644(2000)79:35+\u0026lt;99::aid-jcb1132\u0026gt;3.0.co;2-w\u003c/li\u003e\n\u003cli\u003eStraif K, Benbrahim-Tallaa L, Baan R, et al (2009) A review of human carcinogens--part C: metals, arsenic, dusts, and fibres. The lancet oncology 10:453\u0026ndash;454. https://doi.org/10.1016/s1470-2045(09)70134-2\u003c/li\u003e\n\u003cli\u003eSun H, Yang Y, Shao H, et al (2017) Sodium arsenite-induced learning and memory impairment is associated with endoplasmic reticulum stress-mediated apoptosis in rat hippocampus. Frontiers in Molecular Neuroscience 10:1\u0026ndash;14. https://doi.org/10.3389/fnmol.2017.00286\u003c/li\u003e\n\u003cli\u003eSun HJ, Xiang P, Luo J, et al (2016) Mechanisms of arsenic disruption on gonadal, adrenal and thyroid endocrine systems in humans: A review. Environment International 95:61\u0026ndash;68. https://doi.org/10.1016/j.envint.2016.07.020\u003c/li\u003e\n\u003cli\u003eSung MM, Das SK, Levasseur J, et al (2015) Resveratrol treatment of mice with pressure-overloadinduced heart failure improves diastolic function and cardiac energy metabolism. Circulation: Heart Failure 8:128\u0026ndash;137. https://doi.org/10.1161/CIRCHEARTFAILURE.114.001677\u003c/li\u003e\n\u003cli\u003eTracy RE (2012) Cardiomyocyte size estimated from noninvasive measurements of left ventricular wall thickness and chamber diameter. Journal of the American Society of Hypertension 6:185\u0026ndash;192\u003c/li\u003e\n\u003cli\u003eTyler CR, Allan AM (2014) The Effects of Arsenic Exposure on Neurological and Cognitive Dysfunction in Human and Rodent Studies: A Review. Current environmental health reports 1:132\u0026ndash;147. https://doi.org/10.1007/s40572-014-0012-1\u003c/li\u003e\n\u003cli\u003eUnnikrishnan D, Dutcher JP, Garl S, et al (2004) Cardiac monitoring of patients receiving arsenic trioxide therapy. British journal of haematology 124:610\u0026ndash;617\u003c/li\u003e\n\u003cli\u003eWang R, Zhang J, Wang S, et al (2019) The cardiotoxicity induced by arsenic trioxide is alleviated by salvianolic acid A via maintaining calcium homeostasis and inhibiting endoplasmic reticulum stress. Molecules 24:1\u0026ndash;12. https://doi.org/10.3390/molecules24030543\u003c/li\u003e\n\u003cli\u003eWang Y, Zhao F, Liao Y, et al (2012) Arsenic exposure and glutamate-induced gliotransmitter release from astrocytes. Neural Regeneration Research 7:2439\u0026ndash;2445\u003c/li\u003e\n\u003cli\u003eWatson WH, Yager JD (2007) Arsenic: Extension of its endocrine disruption potential to interference with estrogen receptor-mediated signaling. Toxicological Sciences 98:1\u0026ndash;4. https://doi.org/10.1093/toxsci/kfm111\u003c/li\u003e\n\u003cli\u003eWojciechowski P, Juric D, Louis XL, et al (2010) Resveratrol arrests and regresses the development of pressure overload- but not volume overload-induced cardiac hypertrophy in rats. Journal of Nutrition 140:962\u0026ndash;968. https://doi.org/10.3945/jn.109.115006\u003c/li\u003e\n\u003cli\u003eZhang W, Guo C, Gao R, et al (2013) The protective role of resveratrol against arsenic trioxide-induced cardiotoxicity. Evidence-based Complementary and Alternative Medicine 2013:. https://doi.org/10.1155/2013/407839\u003c/li\u003e\n\u003cli\u003eZhang W, Wang L, Fan Q, et al (2011) Arsenic trioxide re-sensitizes ER\u0026alpha;-negative breast cancer cells to endocrine therapy by restoring ER\u0026alpha; expression in vitro and in vivo. Oncology Reports 26:621\u0026ndash;628. https://doi.org/10.3892/or.2011.1352\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Supplementary Table","content":"\u003cp\u003eSupplementary Table 2 is not available with this version.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-molecular-histology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"hijo","sideBox":"Learn more about [Journal of Molecular Histology](https://www.springer.com/journal/10735)","snPcode":"10735","submissionUrl":"https://submission.springernature.com/new-submission/10735/3","title":"Journal of Molecular Histology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Arsenic trioxide, Cardiotoxicity, Neurotoxicity, Oxidative stress, Resveratrol","lastPublishedDoi":"10.21203/rs.3.rs-6316307/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6316307/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eArsenic toxicity is a global health problem chiefly targeting soft tissues of the body like the brain and heart. The major mechanism underlying arsenic-induced neurotoxicity is oxidative stress. Particularly, the neurons and cardiac myocytes show limitless susceptibility to oxidative stress. Herein, we examined the impact of prolonged arsenic exposure and resveratrol post-treatment on the cardiac and neuronal [Ventromedial hypothalamic nucleus (VMH)] morphology. Adult mice were segregated into control and experimental groups. Controls received distilled water, whereas experimental mice received oral gavage of low (2mg/kg bw) and high (4 mg/kg bw) concentrations of ATO (Arsenic trioxide) for 45 days. Mice were sacrificed on day 45 to obtain perfusion-fixed hearts and brains for histological and morphometric studies. Long-term ATO exposure resulted in a higher heart-to-body weight ratio than controls, suggesting ATO-induced hypertrophy. Microscopic observations revealed a regular arrangement of cardiac muscle fibres, branching patterns of cardiomyocytes, and fibroblasts across all the treatment groups. However, increased cardiac myocyte diameter in ventricles and substantial fibrosis in vessel walls were noticed in ATO-alone exposed hearts relative to controls. Selective vulnerability of hypothalamic neurons following ATO exposure was evident by significant alterations in morphometric parameters (reduced cell density and soma size) in the VMH nucleus of animals receiving ATO (2 and 4 mg/kg) alone. These dramatic histopathological alterations were found to be restored after ATO\u0026thinsp;+\u0026thinsp;\u003cem\u003eRes\u003c/em\u003e co-treatment. We also examined the expression of ER-α in the preoptic area of the hypothalamus and indicated downregulation of ER-α due to prolonged ATO exposure. Our findings highlight Resveratrol as a potent neurocardiac protector against ATO toxicity via estrogen signaling modulation, supporting its therapeutic potential in arsenic poisoning.\u003c/p\u003e","manuscriptTitle":"Arsenic-Induced Neurocardiac Toxicity and Protective Role of Resveratrol: Histopathological and Molecular Insights","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-21 04:55:03","doi":"10.21203/rs.3.rs-6316307/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-04-12T20:34:30+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-04-07T18:16:09+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"145263149892971614366257153856325279964","date":"2025-03-31T06:56:33+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"8535090866267526431024838988622132399","date":"2025-03-28T12:59:20+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-03-28T12:27:33+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-03-28T12:24:50+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-03-28T01:30:29+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Molecular Histology","date":"2025-03-27T02:47:47+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-molecular-histology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"hijo","sideBox":"Learn more about [Journal of Molecular Histology](https://www.springer.com/journal/10735)","snPcode":"10735","submissionUrl":"https://submission.springernature.com/new-submission/10735/3","title":"Journal of Molecular Histology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"32d5d6ef-23dc-4c40-ba3b-035063b56c92","owner":[],"postedDate":"April 21st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-05-26T15:59:08+00:00","versionOfRecord":{"articleIdentity":"rs-6316307","link":"https://doi.org/10.1007/s10735-025-10439-x","journal":{"identity":"journal-of-molecular-histology","isVorOnly":false,"title":"Journal of Molecular Histology"},"publishedOn":"2025-05-20 15:56:53","publishedOnDateReadable":"May 20th, 2025"},"versionCreatedAt":"2025-04-21 04:55:03","video":"","vorDoi":"10.1007/s10735-025-10439-x","vorDoiUrl":"https://doi.org/10.1007/s10735-025-10439-x","workflowStages":[]},"version":"v1","identity":"rs-6316307","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6316307","identity":"rs-6316307","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}