Naringin mitigates Bisphenol A-induced hepatotoxicity in cockerel chicks

Naringin mitigates Bisphenol A-induced hepatotoxicity in cockerel chicks | 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 Naringin mitigates Bisphenol A-induced hepatotoxicity in cockerel chicks Leah Oluwaseyanu Esuola, Oluwaseun Esan, Adamu Shafiu Maikifi, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2129508/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Bisphenol A (BPA) is an environmental pollutant, a monomer used in the polymer industry to produce plastics. BPA has been reported to cause deleterious effects in both humans and animals. Naringin is one of the flavonoids with antioxidant and metal chelating properties. This study was carried out to assess the potential ameliorative effect of naringin on BPA-induced hepatotoxicity in cockerel chicks. Thirty-one-day old cockerel chicks used for this study were randomly divided into 6 groups of five chicks per group as follows: Group 1 (Control), Group 2 (BPA 100 mg/kg), Group 3 (BPA + 100 mg/kg naringin), Groups 4 (BPA + 200 mg/kg naringin), Group 5 (100 mg/kg naringin) and Group 6 (100 mg/kg naringin), respectively. The administration of BPA and naringin was through oral gavage. BPA intoxication precipitated significant (p < 0.05) increased ALT, ALP, AST, TC, TG, LDL, but decreased total protein and HDL-cholesterol when compared with the control. Also, there was a significant increase in hepatic H 2 O 2 generation and MDA content with concomitant decrease in reduce glutathione, glutathione S-transferase, and superoxide dismutase activity in BPA intoxicated chicks. Histology revealed a moderate diffuse sinusoidal congestion, with a severe periportal cellular infiltration in BPA intoxicated chicks. Immunohistochemistry results revealed a higher expression of hepatic caspase 3 and TNF-α in chicks exposed to BPA alone relative to the control and chicks treated with naringin (100 mg/g and 200 mg/kg). Findings from this study showed that naringin administration restored hepatoxicity, improved antioxidant status, and lowered exaggerated values of cholesterol, oxidative stress indices, and ameliorated ultrastructure anarchy. Combining all, the incorporation of naringin into poultry feeds could position the flavonoid as a novel antioxidant and metal chelator with beneficial effects in the reduction of toxicities associated with environmental pollutants such as bisphenol A, thereby boosting production efficiency in livestock production. Bisphenol A naringin hepatotoxicity antioxidant metal binding cockerel chicks Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction Over the years, there has been an increase in the exposure of living beings to some endocrine-disrupting chemicals (EDCs) that are available within the environment. This challenge is of great concern to human and animal health (Kelly et al. 2020 ; Kasonga et al. 2021 ). These EDCs are known to cause interference with the function of the endocrine system, hence, leading to fatal health challenges in any animal or its offspring (EC 1999). Bisphenol-A (BPA) is a typical example of EDC. BPA mimics estrogen which has attracted several research interests among researchers over some decades owing to its unpalatable impacts on human and animal health (Rochester 2013 ). Staples et al. ( 2018 ) reported an increase in the global production of BPA from 5 to 8 million tons (MT) between the years 2010 and 2016. An estimate of 10.2 MT BPA production by the year 2022 has also been made. BPA is one of the organic synthetic compounds that is a key pollutant in the environment (Rahman et al. 2017 ). BPA has structures like oestradiol and diethylstilbestrol; hence, it has the tendency of stimulation of cellular response via osetrogen receptors binding (Rubin 2011 ). As reported by Mohapatra et al. ( 2010 ), the point sources are manufacturing industry effluents, landfill leaching, wastewater treatment plant sludge discharge, dumpsite wastewater, and so on. Some of the areas of the utilization of BPA are pipe materials production, plastic materials, polyesters and polyacrylate production (Tomza-Marcimak et al. 2018). The environmental exposure of humans and animals to BPA is ubiquitous and majorly occurs through oral, respiratory, and dermal routes (Rahman et al. 2017 ). However, dietary ingestion is the main exposure route to living organisms. Birds are exposed BPA since plastic such as overhead water tanks, drinkers and feeding trays are normally utilized in poultry farms for some major activities (Escande et al. 2006 ). Previous studies on animal models reported that BPA-induced oxidative stress caused a reprotoxic effect by affecting reproductive potential and is also hepatotoxic (Oyagbemi et al. 2017 ; Kumar et al. 2019 ). For instance, BPA caused sperm reserves reduction, shorter sperm transit time, and reduction in mitochondrial activity (Matuszczak et al. 2019 ). A study conducted by Abdel-Gwaad et al. ( 2020 ) revealed a significant reduction in the activity of liver catalase following BPA exposure. The study further established that oxidative stress was induced in the liver, and an important alteration in the level of liver enzyme markers in serum was reported. Flavonoids belong to a class of plant secondary metabolites. In plants, they are the origin of bioactive compounds (Ghasemzadeh and Jaafar 2013 ). Flavanoids are a class of polyphenolic secondary metabolites having varying structures in the form of glycones or glycosides in fruits and vegetables (Li et al. 2017 ). They have good health benefits based on their anti-carcinogenic, anti-inflammatory, antioxidant, and anti-mutagenic effects. Naringin is a natural flavonoid. Different studies have established that naringin contains antioxidant, anti-apoptotic, anti-osteoporotic, anti-inflammatory, anti-carcinogenic, and anti-ulcer properties (Wang et al. 2013 ; Chen et al. 2016 ). Naringin also plays a significant function in reducing triglycerides, cholesterol, improvement in immune function, and antioxidant status. Therefore, naringin is a natural antioxidant owing to its ability to scavenge free radicals as well as prevention of lipid peroxidation (Changxing et al. 2018 ). Naringin inhibits radiation-induced reduction of the antioxidant defense system (Chen et al. 2020 ). Some of the studies conducted on the influence of naringin on the organs of some models are highlighted. Goliomytis et al. ( 2019 ) reported that naringin may favorably elongate the shell-life of eggs. Naringin have anti-inflammatory properties as well as the ability to improve yolk colour without any side effects on the quality traits and performance of the eggs. The dietary inclusion of naringin at 100 mg/kg daily lowered plasma fatty acids levels, and improved liver mitochondrial dysfunction and glucose intolerance (Alam et al. 2013 ). Naringin also enhanced the structure and function of both the liver and the heart without having any detrimental effect on the total body weight. Furthermore, naringin was reported to lower the blood sugar level in rat and prevent the development of hyperglycemia (Jung et al. 2014; Parmar et al. 2012 ); and reduce LDL-cholesterol, total cholesterol, and triglycerides levels in chicken (da Silva et al. 2001 ). The consumption of leaked BPA into the feeds and water of poultry animals is detrimental to the health of the animals. Various means of reducing the toxic effect of BPA on different organs through the usage of various flavonoid-based supplements have been devised such as orange-peel powder in a rat model (Abdel-Gwaad et al. 2020 ); naringin in male rats (Elsawy et al. 2021 ); and so on. There are limited studies on the effect of BPA toxicity in birds and how naringin can be used to reverse its deleterious effect. The study was designed to determine the ameliorative effect of naringin and its action of mechanism on BPA toxicity in the liver of cockerel chicks. Materials And Methods Chemicals Naringin, Bisphenol A, Albumin (Sigma Chemical Co., USA), Hydrogen peroxide (H 2 O 2 ), phosphate buffer, sodium hydroxide, copper sulphate, potassium tartarate, Trichloroacetic acid (TCA), Tris-potassium chloride, Ammonium ferrous sulphate, sorbitol, sulphuric acid, carbonate buffer, Adrenaline, Ellman’s reagent (DTNB) reduced glutathione (GST), thiobarbituric acid (TBA), 2-dichloro-4-nitrobensene (CDNB), sodium azide, O-dianiside dihydrochloride were purchased. Biotinylated secondary antibody, Tumour necrosis factor alpha (TNF-α) and Caspase 3 monoclonal antibody were purchased from Elabscience Biotechnology, China. Experimental Animals In this study, thirty one-day-old cockerel chicks purchased from CHI FARMS Limited Ibadan, Nigeria were utilized. The chicks were housed at the experimental rearing facility of Avian diseases unit, Department of Veterinary Medicine, University of Ibadan. Brooding of the chicks was done for 2 weeks by supply of additional heat and then further reared for 4 weeks. The chicks were fed age-appropriate poultry feed from Top Feeds® limited, Ibadan, and clean drinking water ad libtum was given liberally. All the cockerels were kept in open-sided tropical rearing spacious cages with all necessary conditions for their comfort as indicated by Animal Use and Care Research Ethics Committee (ACUREC) Ibadan. The research was done according to the procedures illustrated and approved by the University of Ibadan - ACUREC (UI-ACUREC/048–0521/21). Experimental Design Cockerels with uniform weight were randomly allotted into 6 groups (5 chicks per group). Bisphenol A at 100 mg/kg was administered continuously in drinking water for a period of 6 weeks while the treated groups were administered different doses of Naringin through oral gavage for a period of 2 weeks. Thirty-one-day old cockerel chicks used for this study were randomly divided into 6 groups of five chicks per group as follows: Group 1 (Control), Group 2 (BPA 100 mg/kg), Group 3 (BPA + 100 mg/kg naringin), Groups 4 (BPA + 200 mg/kg naringin), Group 5 (100 mg/kg naringin) and Group 6 (100 mg/kg naringin), respectively. The administration of BPA and naringin was through oral gavage. Sample Collection And Preparation Of Hepatic Post-mitochondrial Fractions Twenty-four hours post administration of the last treatment, chickens were euthanized by quick cervical dislocation. The whole liver was harvested by rapid dissection on ice, rinsed with distilled water, blotted with filter paper, and whole organ weight was determined using an electronic digital scale. Subsequently, about 2 g each of the liver samples were collected inside labeled universal bottles, sliced and blended in 0.1 M aqueous potassium phosphate buffer at pH 7.4. Liver post mitochondrial fractions (PMFs) were obtained using the homogenate obtained after centrifuging at 10,000 g for 10 minutes with a cold centrifuge at -4˚C. The supernatant was collected and stored and used for biochemical assay. Biochemical Assays Total protein concentration in the supernatant was determined using the Biuret method as described by Gornal et al. ( 1949 ). The method described by Wolff ( 1994 ) was used in the determination of hydrogen peroxide generation. The illustrated method of Varshney and Kale ( 1990 ) was employed in the determination of the lipid peroxidation (Malondialdehyde (MDA) concentration). The hepatic reduced glutathione (GSH) concentration determination was determined with the method described by Ellman ( 1959 ). The method of Misra and Fridovich ( 1972 ) with slight modification by Oyagbemi et al. ( 2015 ) was followed in the evaluation of the superoxide dismutase (SOD) activity. The methods described by Buetler et al. ( 1963 ) and Habig et al. ( 1974 ) for the determination of Glutathione peroxidase (GPx) and Glutathione S-transferase (GST) activities, respectively, were adopted. The liver function tests such as total protein, albumin, total bilirubin, conjugated bilirubin, Aspartate transferase (AST), Alanine transferase (ALT), and Alanine phosphatase (ALP) were determined using the commercial colorimetric kits as described by Manterys et al. ( 2016 ) and Adeyemo et al. ( 2018 ). Histopathology Liver tissues were fixed in 10% formalin, embedded in paraffin wax, and sections of 5–6 mm in thickness were made and thereafter stained with Hematoxylin and Eosin (H&E) as previously described by Drury and Walington (1976). Therefore, the sections were examined with light microscopic. Immunohistochemistry In this study, the immunohistochemistry procedure of the liver to determine the hepatic caspase 3 and tumor necrosis factor-alpha (TNF-α) as described by Oyagbemi et al. ( 2019 ) was adopted. Statistical Analysis All values were expressed as mean ± standard deviation (SD) and the test of significance between the experimental groups and the control group was estimated with Student’s t-test. The One-Way Analysis of Variance (ANOVA) with Tukey’s post-hoc test of Graph pad prism 5.0 was also carried out with p-values < 0.05 considered statistically significant. Results Liver function test Figure 1 showed the results obtained for the liver function tests. The values of total protein, albumin, and total bilirubin of chicks intoxicated with BPA (100 mg/kg) was significantly (p < 0.05) lower than that of the control and chicks treated with naringin (Figs. 1a, b, &c). However, there was significant improvement in the values of total protein, albumin, and total bilirubin of chicks treated with naringin (100 & 200 mg/kg) and those that received only naringin (100 & 200 mg/kg). From our results, we observed a significant (p < 0.05) increase in the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase in intoxicated with BPA in comparison to the control. (Fig. 1c, e, & f). Treatment with naringin (100 & 200 mg/kg) significantly reduced the heightened values of AST, ALT, and ALP, respectively, in BPA intoxicated chicks. This is indicative of hepatoprotective action of naringin. Markers Of Oxidative Stress And Antioxidant Defense System In The Liver Hydrogen peroxide (H 2 O 2 ) generation and content of lipid peroxidation product (MDA) in the liver of the BPA intoxicated chicks were significantly (p < 0.05) higher when compared with the control group (Figs. 3a & 3b). Figure 3 also showed that the level of H 2 O 2 and MDA content in the liver of BPA treated chicks with naringin (100 mg/kg & 200 mg/kg) and naringin only (100 mg/kg & 200 mg/kg) in comparison to BPA untreated chicks. The observed significant reduction in the content of MDA was dose-dependent in naringin treated chicks (Fig. 3b), thus, indicating the antioxidative action of naringin. The result showed a significant (p < 0.05) decrease in the level of GSH, GST, and SOD activity in the BPA exposed chicks when compared with the control group (Figs. 4a, 4b, & 4d), while BPA toxicity enhanced the activity of GPx greater than that of the control (Fig. 4c). The noticeable increase in the activity of hepatic GPx might be due to consumption of GSH due to BPA toxicity. However, treated of intoxicated chicks with naringin significantly improved the content of GSH and the activities of GST, SOD, and GPx, respectively, (Figs. 4a, 4b, & 4d). Histopathology Histology revealed a moderate diffuse sinusoidal congestion, with a severe periportal cellular infiltration in BPA intoxicated chicks. However, no visible lesions were observed in chicks in the histopathology of hepatic tissues treated with naringin (Fig. 5). Immunohistochemistry Hepatic caspase 3 and tumor necrosis factor (TNF-α) in the hepatocytes revealed higher expressions in BPA exposed group when compared to the control and groups treated with naringin (Figs. 6 and 7). The reduction in the expressions of TNF-α as well as, Caspase 3 is indicative of amelioration of inflammation, apoptosis and hepatoprotective effects of naringin against bisphenol A toxicity as shown by the reduction in the expression of TNF-α and caspase 3 in the groups treated with naringin and groups administered naringin only. Discussion Different health-related deleterious effects are associated with BPA exposure in living organisms, with BPA showing that endocrine-disrupting chemicals induce oxidative stress (Singh et al. 2016 ; Meli et al. 2020 ). BPA induces oxidative stress by generating reactive oxygen species (ROS) through the generation of free radicals, and thereby causes an imbalance between ROS and antioxidant defenses which then results in oxidative damage (Kobroob et al. 2018 ; Amjad et al. 2020 ). The hepatotoxic effect of BPA was established in this study as demonstrated with a significant (p < 0.05) increase in the activities of ALP, AST, and ALT, and a concomitant decrease in total protein and albumin in BPA-exposed chicks. This is in line with the study carried out by Oguazu et al. ( 2015 ) and Abdel-Gwaad et al. ( 2020 ) as they reported an increase in ALT, AST, and AST which indicates hepatotoxicity action of BPA. Naringin is an example of a flavonoid that has been reported to possess several pharmacological effects such as anti-inflammatory, hypolipidemic, antioxidant, antiviral, antimicrobial, anti- apoptotic, hepatoprotective, and immunomodulatory effects (Changxing et al. 2018 ). Hepatoprotective activities were observed in the naringin-treated groups as there was a significant decrease in ALT, AST, and ALP and a corresponding increase in total protein. This corroborated the earlier study of Imam et al. ( 2016 ) who reported that co-administration of naringin with aluminum ameliorated aluminum-induced liver damage and oxidative stress. Furthermore, the levels of TC, TAG, and LDL-c were significantly (p < 0.05) higher in BPA intoxicated chicks with a concurrent decrease in HDL-c as observed in the present study. This agreed with the findings of Eweda et al. ( 2020 ) who reported that BPA induced dyslipidemic state and increased triglycerides (steatosis) and cholesterol accumulation in the liver tissue of rat. Marmugi et al. ( 2012 ) also reported that the exposure of rat to BPA resulted in hypertriglyceridemia, hypercholesterolemia, and alterations of fatty acids composition in the liver. More so, there was upregulation of genes which are linked with de novo lipogenesis and cholesterol synthesis in the liver. Kumar et al. ( 2019 ) reported that treatment with naringin produced noteworthy developments in altered lipid profiles. The anti-hyperlipidemic effect of naringin was established in this study as naringin reversed the toxicity of BPA by decreasing LDL-c, TC, TG, and increasing HDL-c in chicks treated naringin. The scavenging and neutralization of free radicals such as superoxide radicals, hydrogen peroxides (H 2 O 2 ), and hydroperoxides are done by antioxidants, thereby preventing oxidation reactions. During oxidative stress, free radicals increase in the body (Amjad et al. 2020 ). This study showed an increase in H 2 O 2 contents in the liver of BPA-exposed chicks. This was in concordance with the study of da Silva et al. ( 2018 ) who reported an exaggerated increase H 2 O 2 generation in thyrocytes of female Wistar rats intoxicated with BPA. More so, Kabuto et al. ( 2003 ) observed that BPA caused the overproduction of H 2 O 2 with resultant oxidative stress. In our study, naringin reduced the level of H 2 O 2 in groups treated BPA-treated chicks and naringin-only treated groups. The antioxidant and free radical scavenging power of naringin was earlier reported by Rashmi et al. ( 2018 ) who showed the H 2 O 2 radical scavenging potential of naringin in streptozotocin-induced liver damage. Also, Miles and Calder ( 2021 ) reported that naringin increased liver and kidney expression of anti-inflammatory transcription factors. In this study, the increase in the level of MDA caused lipid peroxidation because of BPA-induced oxidative stress as earlier reported by Kobroob et al. ( 2018 ) and Eweda et al. ( 2020 ). Lipid peroxidation of biological membranes is known to cause loss of membrane fluidity, increases membrane permeability, alters receptor function, and changes in membrane potential. It can be observed in this study that the MDA level was exaggerated in the liver of BPA intoxicated chickens. However, in this study, the group co-treated with naringin caused a significant (p < 0.05) reduction in the MDA level, as previously reported by Rajadurai and Stanely ( 2006 ) & Alam et al. ( 2014 ). Glutathione (GSH) is one of the non-enzymatic antioxidants that is depleted during oxidative stress. In this study, a decreased level of GSH was observed in the liver in the BPA-exposed chicks, which depicts ongoing oxidative stress caused by BPA toxicity. Kobroob et al. ( 2018 ) reported a decrease in GSH in rats exposed to BPA. However, the co-treated with naringin showed improvements in the activities of GSH. This supports the study of Kumar et al. ( 2019 ) who stated that administration of naringin to diabetic rats raised the concentration of GSH. The GPx, SOD, and GST are known to protect tissues from damage caused by oxidative stress by detoxification of several substrates generated from cellular oxidative processes (Sravani et al. 2016 ). The GPx activity in the liver increased in the BPA-exposed chicks in this study, and this might be a result of the adaptive response. We proposed that this could be a result of nuclear translocation of nuclear erythroid-related factor 2 (Nrf2). The first step is the modification of the keap1 segment of Nrf2-Keap1 pathway. Nrf2 is usually low when it is not activated by stress factors. In the presence of ROS such as superoxide, hydroxyl, and peroxyl radicals, cysteine residues (such as Cys 151) are oxidized the sulfhydryl groups on the Keap1 segment. This changes the conformity of Keap1 segment and prevents it from attaching to Nrf2. The Nrf2 released within the cytosol enters the nucleus and heterodimerize with small Maf proteins, and further binds to regulatory gene recognized as antioxidant response elements (ARE). Substances that can initiate activity at the ARE include environmental pollutants, hydrogen peroxide and nitric oxide. The Nrf2-Maf-ARE complex then initiates subsequent specific antioxidant and detoxification genes such as that of GST, GPx, GSH and SOD; thereby, increasing antioxidant activities and inactivating ROS (Baird and Dinkova-Kostova 2011 ; Oyagbemi et al. 2017 ). The antioxidant effect of naringin was established as naringin restored the levels of GSH, GST, GP X , and SOD in the hepatic tissue of BPA-exposed chicks. TNF-α is an example of a pro-inflammatory cytokine, which plays a significant role in the host’s defense against injury. It was stated that BPA toxicity exacerbated the proinflammatory cytokines expression along with interleukins such as IL-1β, IL-6, IL-8, and tumor necrosis factor alpha (TNF-α) as previously reported (Wang et al. 2019 ; Meli et al. 2020 ). This is in agreement with this present study as there was noticeable higher expression of TNF-α in the liver of the BPA-exposed chicks compared to the control and naringin-treated chicks. Naringin has been reported to possess a hepatoprotective effect by regulating inflammatory cytokines and increasing antioxidant enzymes (Caglayan et al. 2018 ). This study establishes that naringin reduces the expression of TNF-α in birds co-administered with naringin and BPA. Caspase-3 is a biomarker of cell death. In this study, there was an increase expression of caspase-3 in the liver of BPA intoxicated chicks. This showed that hepatic apoptosis might have resulted from inflammation and oxidative stress following BPA toxicity (Abdel-Rahman et al. 2018 ; Liu et al. 2022 ). This is in tandem with the work of Elgawish and Abdelrazek ( 2014 ) where increase in expression of caspase-3 in testes of male rats exposed to lead acetate was observed. The anti-apoptotic effect of naringin was exhibited in the BPA group co-treated with naringin and naringin treated groups by cleaving caspase-3 and inhibiting the genes and proteins involved in apoptotic pathways such as P53, P 16 INK 49 (Yuan and Yang 2022 ). Conclusion Based on the results in this present study, it can be inferred and concluded that naringin abated the toxicity of Bisphenol A-induced liver dysfunction. Naringin exhibited its hepatoprotective property by revitalizing cells that were damaged due to BPA-induced oxidative stress. The use of Naringin can serve as a natural and safe feed supplement in poultry nutrition is recommended. Declarations Acknowledgements The authors deeply thank the African Union through the Pan African University Institute of Earth and Life Sciences Institute (PAULESI) for funding this work and Cardio-renal laboratory, University of Ibadan for the facilities used. Ethical approval: The study was conducted following guidelines approved by the Animal Care and Use Research Ethics Committee (ACUREC) of the University of Ibadan (Approval number: UI-ACUREC/048-0521/21). Consent to Participate : Not applicable Consent to Publish: Not applicable Authors Contributions: The authors, Leah Oluwaseyanu Esuola, Oluwaseun Esan, Adamu Shafiu Maikifi , Ademola Adetokunbo Oyagabemi and Temidayo Olutayo Omobowale designed the experiment. Leah Oluwaseyanu Esuola, Oluwaseun Esan, Adamu Shafiu Maikifi performed the immunohistochemistry and biochemical assays. The blood pressure and electrocardiogram were performed by Leah Oluwaseyanu Esuola, Oluwaseun Esan and Temidayo Olutayo Omobowale. Moses Olusola Adetona, Ademola Adetokunbo Oyagabemi, Temidayo Olutayo Omobowale, Oluwafemi Oguntibeju, Momoh Audu Yakubu supervised, proof-read, and approved the submission. Funding: The study was funded by the African Union Competing Interests: The authors declare that they have no competing interest Availability of data and materials: Data will be made available on request References Abdel-Gwaad HMS, AbdEl-Wahab HMF, Mohamed EAK, Sharaf EHAA, Osman AAHM (2020) Modulatory effect of dry orange ( Citrus sinensis) peel powder on bisphenol A- induced hepatic and splenic toxicity in rats. J Basic Appl Zool 81:49 Abdel-Rahman HG, Abdelrazek HMA, Zeidan DW, Mohamed RM, Abdelazim AM (2018) Lycopene: Hepatoprotective and antioxidant effects toward bisphenol A-induced toxicity in female Wistar rats. Oxid Med Cell Longev 2018:5167524 Adeyemo GO, Kabir AO, Tanimowo DA, Ologhobo AD (2018) Performance, blood chemistry, and serum electrolytes of broilers given water from different sources. Nigerian J Anim Sci 20:140–145 Alam AM, Subhan N, Rahman MM, Uddin SJ, Reza HM, Sarker SD (2014) Effect of citrus flavonoids, naringin and naringenin, on metabolic syndrome and their mechanisms of action. Adv Nutr 5:404–417 Alam MA, Kanter K, Brown L (2013) Naringin improves diet induced cardiovasicular dysfunction and obesity in high carbohydrate, high fat diet-fed rats. Nutrients 5:637–650 Amjad S, Rahman MS, Pang MG (2020) Role of antioxidants in alleviating bisphenol A toxicity. Biomolecules 10:1105. doi: 10.3390/biom10081105 Baird L, Dinkova-Kostova AT (2011) The cytoprotective role of the Keap1-Nrf2 pathway. Arch Toxicol 85:241–272 Buetler E, Duron O, Kelly MB (1963) Improved method for determination of blood glutathione reduced. J Lab Clin Med 61:882–888 Caglayan C, Temel Y, Kandemir FH, Kucukler S (2018) Naringin protects against cyclophosphamide-induced hepatotoxicity and nephrotoxicity through modulation of oxidative stress, inflammation, apoptosis, autophagy and DNA damage. Environ Sci Pollut Res 25:20968–20984 Changxing L, Saeed M, Kamboh AA, Alagawany M, Abd El-Hack ME, Arain MA, Babazadeh D, Dhama K, Chenling M (2018) Reconsidering a Citrus Flavonoid Naringin as a Promising Nutritional Supplement and its Beneficial Health Applications in Humans, Animals and Poultry. Int J Pharmacol, 2018:1–10 Chen P, Xiao Z, Wu H, Wang Y, Su W, Li P (2020) The effects of naringin on cigarette smoke-induced dynamic changes in oxidation/antioxidant system in lung of mice. Nat Prod Commun 15:1–9 Chen R, Qi QL, Wang MT, Li QY (2016) Therapeutic potential of naringin: An overview. Pharm Biol 54:3203–3210 da Silva MM, Xavier LLF, Goncalves CFL, Santos-Silva AP, Palva-Melo FD, de Freitas ML, Fortunato RS, Miranda-Alves L, Ferreira ACF (2018) Bisphenol A increases hydrogen peroxide generation by thyrocyte both in vivo and in vitro. Endocrin Connect 7:1196–1207 da Silva RR, de Oliveira TT, Nagem TJ, Pinto AS, Albino LF (2001) Hypo-cholesterolemic effect of naringin and ruttin flavonoid. Arch Latinoam Nutri 51:258–264 Drury RA, Wallington EA (1976) Carlton's Histopathological Techniques, 4th edn. Oxford University Press, London, pp 139–142 Elgawish RAR, Abdelrazek HMA (2014) Effects of lead acetate on testicular function and caspase-3 expression with respect to the protective effect of cinnamon in albino rats. Toxicol Rep 1:795–801 Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77 Elsawy H, Alzahrani AM, Alfwuaires M, Abdel-Moneim AM, Khalil M (2021) Nephroprotective effect of naringin in methotrexate induced renal toxicity in male rats. Biomed Pharmacother 143:112180 Escande A, Pillon A, Servant N, Cravedi JP, Larrea F, Muhn P, Nicolas JC, Cavaillès V, Balaguer P (2006) Evaluation of ligand selectivity using reporter cell lines stably expressing estrogen receptor alpha or beta. Biochem Pharmacol 71:1459–1469 European Commission. Community Strategy for Endocrine Disrupters (1999) A Range of Substances Suspected of Interfering with the Hormone Systems of Humans and Wildlife; Commission of the European Communities: Brussels, Belgium Eweda SM, Newairy AA, Abdou HM, Gaber AS (2020) Bisphenol A-induced oxidative damage in the hepatic and cardiac tissues of rats: The modulatory role of sesame lignans. Exp Th Med 19:33–44 Ghasemzadeh A, Jaafar HZ (2013) Profiling of phenolic compounds and their antioxidant and anticancer activities in pandan ( Pandanus amaryllifolius Roxb.) extracts from different locations of Malaysia. BMC Complement Altern Med 13:341 Goliomytis M, Simitzis P, Papalexi A, Veneti N, Hager-Theolorides AL, Charismadou MA, Deligeogis SG (2019) Influence of citrus flavonoids on laying hen performance, inflammatory immune response, egg quality and yolk oxidative stability. Br Poult Sci 60:272–278 Gornal AG, Bardawill JC, David MM (1949) Determination of serum proteins by means of Biuret reaction. J Biol Chem 177:751–766 Habig WH, Pabst MJ, Jakpoby WB (1974) Glutathione transferase, a first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139 Imam TS, Khalifa HA, Hussein MMA, Ali HA (2016) Aluminum-induced oxidative stress and hepato -renal impairment in male albino rats: Possible. Life Sci J 13:93–104 Jung UJ, Kim SR (2014) Effects of naringin, a flavanone glycoside in grapefruits and citrus fruits, on the nigrostriatal dopaminergic projection in the adult brain. Neural Regen Res 9:1514–1517 Kabuto H, Hasuike S, Minagawa, Shishibori T (2003) Effect of bisphenol A on the metabolisms of active oxygen species in mouse tissues. Environ Res 93:31–35 Kasonga TK, Coetzee MA, Kamika I, Ngole-Jeme VM, Momba MNB (2021) Endocrine-disruptive chemicals as contaminants of emerging concern in wastewater and surface water: A review. J Environ Manag 277:111485 Kelly M, Connolly L, Dean M (2020) Public awareness and risk perceptions of endocrine disrupting chemicals -A qualitative study. Int J Environ Res Public Health 17:7778 Kobroob A, Peerapanyasut W, Chattipakorn N, Wongmekiat O(2018) Damaging effects of Bisphenol A on the kidney and the protection by melatonin: Emerging evidences from in vivo and in vitro studies. Oxidat Med Cell Long 2018; Article ID 3082438, https://doi.org/10.1155/2018/3082438 Kumar SR, Ramli ESM, Abdul-Nasir AA, Ismail NHM, Fahami NAM (2019) Preventive effect of Naringin on metabolic syndrome and its mechanism of action: A systematic review. Evidence-Based Complement Alternat Med 2019:1–11 Li K, Fan H, Yin P, Yang L, Xue Q, Li X, Sun L, Liu Y (2017) Structure-activity relationship of eight high content flavonoids analyzed with a preliminary assign-score method and their contribution to antioxidant ability of flavonoids-rich extract from Scutellana baicalensis shoots. Arab J Chem 11:159–170 Liu R, Cai D, Li X, Liu B, Chen J, Jiang X, Li H, Li Z, Teerds K, Sun J, Bai W, Jin Y (2022) Effects of Bisphenol A on reproductive toxicity and gut microbiota dysbiosis in male rats. Ecotoxicol Environ Safety 239:113623 Manterys A, Francyk-Zarow M, Czyzynska-Cichon I, Drahun A, Kus E, Szymczyk B, Kostogrys RB (2016) Haematological parameters, serum lipid profile, liver function, and fatty acid profile of broiler chickens fed on diets supplemented with pomegranate seed oil and linseed oil. Br Poult Sci 57:771–779 Marmugi A, Ducheix S, Lasserre F, Polizzi A, Paris A, Priymenko N, BertrandMichel J, Pineau T, Guillou H, Martin PG, MselliLakhal L (2012) Low doses of bisphenol A induce gene expression related to lipid synthesis and trigger triglyceride accumulation in adult mouse liver. Hepatology 55:395407 Matuszczak E, Komarowska MD, Debek W, Hermanowicz A(2019) () The impact of bisphenol A on fertility, reproductive system, and development: A review of the literature. Int J Endocrin 2019:4068717 Meli R, Monnolo A, Annunziata C, Pirozzi C, Ferrante MC (2020) Oxidative stress and BPA toxicity: An antioxidant approach for male and female reproductive dysfunction. Antioxidant 9:405 Miles E, Calder PC (2021) Effects of citrus fruit juices and their bioactive components on inflammation and immunity: A narrative review. Front Immunol 12:712608 Misra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 217:3170–3175 Mohapatra DP, Brar SK, Tyagi RD, Surampalli RY (2010) Physicochemical pre-treatment and biotransformation of wastewater and wastewater Sludge – fate of bisphenol A. Chemosphere 78:923–941 Oguazu CE, Ezeonu FE, Ubaoji KI, Anajekwu B (2015) Bisphenol A exerts a transient perturbation of liver function in Wistar albino rats at acute and sub-chronic exposure doses. J Pharmaceut Sci Biosci Res 5:274–278 Oyagbemi AA, Omobowale TO, Asenuga ER, Adejumobi AO, Ajibade TO, Ige TM et al (2019) Sodium fluoride induces hypertension and cardiac complications through generation of reactive oxygen species and activation of nuclear factor kappa beta. Environ Toxicol 32:1089–1101 Oyagbemi AA, Omobowale TO, Akinrinde AS, Saba AB, Ogunpolu BS, Daramola O (2015) Lack of reversal of oxidative damage in renal tissues of lead acetate-treated rats. Environ Toxicol 30:1235–1243 Oyagbemi AA, Omobowale TO, Asenuga ER, Afolabi JM, Adejumobi OA, Adedapo AA, Yakubu MA (2017) Effect of arsenic acid withdrawal on hepatotoxicity and disruption of erythrocyte antioxidant defense system. Toxicol Rep 4:521–529 Parmar HS, Jain P, Chauham DS, Bhinchar DS, Munjal V (2012) DPP-IV inhibitory potential of naringin An in silico, in vitro and in vivo study. Diabetes Res Clin Pract 97:105–111 Rahman S, Kwon WS, Ryu DY, Khatun A, Karmakar PC, Ryu BY, Pang MG (2017) Functional and proteomic alterations of F1 capacitated spermatozoa of adult mice following gestational exposure to Bisphenol A. J Proteome Res 17:524–535 Rajadurai M, Stanely MPP (2006) Preventive effect of naringin on lipid peroxides and antioxidants in isoprote renal-induced cardiotoxicity in Wistar rats: Biochemical and histopathological evidences. Toxicology 228:59–68 Rashmi R, Magesh SB, Ramkumar KM, Suryanarayanan S, SubbaRao MV (2018) Antioxidant potential of naringenin helps to protect liver tissue from streptozotocin-induced damage. Rep Biochem Molecul Biol 7:76–84 Rochester JR (2013) Bisphenol A and human health: A review of the literature. Toxicol 42:132–155 Rubin BS (2011) Bisphenol A: an endocrine disruptor with widespread exposure and multiple effects. J Steroid Biochem Molecular Biol 127:27–34 Singh RP, Shafeeque CM, Sharma SV, Singh R, Kannan M, Sastry KVH, Raghunandanan S, Mohan J, Azeez PA (2016) Effects of bisphenol-A on male reproductive success in adult Kadaknath chicken. Ecotoxicol Environ safety 128:61–66 Sravani J, Padmaja K, Eswara Prasad P, Punya Kumari B (2016) Effect of bisphenol A on antioxidant enzymes and lipid peroxidation in liver of chick embryos. Int J Meat Sci 6:1–5 Staples C, van der Hoeven N, Clark K, Mihaich E, Woelz J, Hentges S (2018) Distributions of concentrations of bisphenol A in North American and European surface waters and sediments determined from 19 years of monitoring data. Chemosphere 201:448–458 Tomza-Marciniak A, St˛epkowska P, Kuba J, Pilarczyk B (2018) Effect of bisphenol A on reproductive processes: A review of in vitro, in vivo and epidemiological studies. J Appl Toxicol 38:51–80 Varshney R, Kale RK (1990) Effect of calmodulin antagonists on radiation induced lipid peroxidation in microsomes. Int J Radiation Biol 58:733–743 Wang D, Ma W, Wang F, Dong J, Wang D, Sun B, Wang B (2015) Stimulation of Wnt/β-catenin signaling to improve bone development by naringin via interacting with AMPK and Akt. Cell Physiol Biochem 36:1563–1576 Wang DM, Yang YJ, Zhang L, Zhang X, Guan FF, Zhang LF (2013) Naringin enhances CaMKII activity and improves long-term memory in a mouse model of Alzheimer’s disease. Int J Mol Sci 14:5576–5586 Wang K, Zhao Z, Ji W (2019) Bisphenol A induces apoptosis oxidative stress and inflammatory response in colon and liver of mice in a mitochondria dependent manner. Bio med Pharmacother 117:109182 Wolff SF (1994) Ferrous ion oxidation in the presence of ferric ion indicator xylenol orange for measurement of hydrogen peroxides. Methods Enzymol 233:182–189 Yuan Z, Yang Z (2022) The effect of naringin on the apoptosis of degenerative nucleus pulposus cells: A study on the function and mechanism. Drug Des Develop Therapy 16:499–508 Cite Share Download PDF Status: Posted Version 1 posted 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-2129508","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":148277404,"identity":"115e85b0-7b8a-43e1-8d40-0704fd4c9180","order_by":0,"name":"Leah Oluwaseyanu Esuola","email":"","orcid":"","institution":"PAU: Pan African University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Leah","middleName":"Oluwaseyanu","lastName":"Esuola","suffix":""},{"id":148277405,"identity":"f2bcac6e-873c-4527-8390-fe32cb9ec386","order_by":1,"name":"Oluwaseun Esan","email":"","orcid":"","institution":"University of Ibadan Faculty of Veterinary Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Oluwaseun","middleName":"","lastName":"Esan","suffix":""},{"id":148277406,"identity":"67d6901f-7a67-43f9-a413-bfeef620056e","order_by":2,"name":"Adamu Shafiu Maikifi","email":"","orcid":"","institution":"Pan African University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Adamu","middleName":"Shafiu","lastName":"Maikifi","suffix":""},{"id":148277407,"identity":"d5b30971-cb0f-4312-b2cf-44d4baf9e8c1","order_by":3,"name":"Temitayo Olabisi Ajibade","email":"","orcid":"","institution":"University of Ibadan Faculty of Veterinary Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Temitayo","middleName":"Olabisi","lastName":"Ajibade","suffix":""},{"id":148277408,"identity":"54de6b0f-d7b6-4590-a76c-9fa3dcc047ec","order_by":4,"name":"Moses Olusola Adetona","email":"","orcid":"","institution":"UCH: University College Hospital Ibadan","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Moses","middleName":"Olusola","lastName":"Adetona","suffix":""},{"id":148277409,"identity":"c3346660-05bc-4147-871b-f0ebb9f16fc7","order_by":5,"name":"Ademola Adetokunbo Oyagbemi","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0002-8996-8610","institution":"University of Ibadan Faculty of Veterinary Medicine","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Ademola","middleName":"Adetokunbo","lastName":"Oyagbemi","suffix":""},{"id":148277410,"identity":"f81984a2-8dc2-4ab0-89ff-e84dc4ddafc7","order_by":6,"name":"Temidayo Olutayo Omobowale","email":"","orcid":"","institution":"University of Ibadan Faculty of Veterinary Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Temidayo","middleName":"Olutayo","lastName":"Omobowale","suffix":""},{"id":148277411,"identity":"1d2fc625-d8f6-4074-89d4-17bb9c563e1a","order_by":7,"name":"Omolade Abodunrin Oladele","email":"","orcid":"","institution":"University of Ibadan Faculty of Veterinary Medicine","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Omolade","middleName":"Abodunrin","lastName":"Oladele","suffix":""},{"id":148277412,"identity":"b770b9bd-9324-42be-943c-3f5f09ee2e2e","order_by":8,"name":"Oluwafemi Omoniyi Oguntibeju","email":"","orcid":"","institution":"Cape Peninsula University of Technology - Bellville Campus","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Oluwafemi","middleName":"Omoniyi","lastName":"Oguntibeju","suffix":""},{"id":148277413,"identity":"53f03ae3-a41e-4fac-9de0-bd6b692a9e4d","order_by":9,"name":"Momoh Audu Yakubu","email":"","orcid":"","institution":"Texas Southern University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Momoh","middleName":"Audu","lastName":"Yakubu","suffix":""}],"badges":[],"createdAt":"2022-10-03 19:18:43","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2129508/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2129508/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":28627623,"identity":"b720556e-3b15-4e20-9ca1-a73e85fccf9b","added_by":"auto","created_at":"2022-11-03 19:02:59","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1992378,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eLiver function tests showing (a) Total protein (b) Albumin (c) Total bilirubin (d) Aspartate aminotransferase (e) Alanine aminotransferase (f) Alkaline phosphatase for various groups\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSuperscript (a) indicates significant difference at p\u0026lt;0.05 when compared with Control group (Group A). Superscript (b) indicates significant difference when compared with Bisphenol A group only (Group B). The results are shown in Mean ± SD (n= 6). Group A: Control, Group B: BPA (100 mg/kg), Group C: BPA and Naringin (100 mg/kg), Group D: BPA and Naringin (200 mg/kg), Group E: Naringin (100 mg/kg), \u0026amp; Group F: Naringin (200 mg/kg).\u003c/p\u003e","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2129508/v1/9f33a7980af61805bd031e4c.jpg"},{"id":28627627,"identity":"30711856-23d0-4614-b4e6-4a8a256f9826","added_by":"auto","created_at":"2022-11-03 19:03:01","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1927564,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eLipid profile showing (a) Cholesterol (b) Triacyglyceride (c) High density lipoprotein (d) Low-density lipoprotein for various groups\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSuperscript (a) indicates significant difference at p\u0026lt;0.05 when compared with Control group (Group A). Superscript (b) indicates significant difference when compared with Bisphenol A group only (Group B). The results are shown in Mean ± SD (n= 6). Group A: Control, Group B: BPA (100 mg/kg), Group C: BPA and Naringin (100 mg/kg), Group D: BPA and Naringin (200 mg/kg), Group E: Naringin (100 mg/kg), \u0026amp; Group F: Naringin (200 mg/kg).\u003c/p\u003e","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2129508/v1/ced1a5c34413ff569140e11f.jpg"},{"id":28627621,"identity":"98d82fef-43e9-4b03-a419-ac2205f18b80","added_by":"auto","created_at":"2022-11-03 19:02:59","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1056094,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMarkers of oxidative stress in liver (a) H2O2(b) MDA\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSuperscript (a) indicates significant difference at p\u0026lt;0.05 when compared with Control group (Group A). Superscript (b) indicates significant difference when compared with Bisphenol A group only (Group B). The results are shown in Mean ± SD (n= 6). Group A: Control, Group B: BPA (100 mg/kg), Group C: BPA and Naringin (100 mg/kg), Group D: BPA and Naringin (200 mg/kg), Group E: Naringin (100 mg/kg), \u0026amp; Group F: Naringin (200 mg/kg).\u003c/p\u003e","description":"","filename":"Figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2129508/v1/604a808fec1e016b0ac2946d.jpg"},{"id":28627622,"identity":"bc5aaf3e-913e-44ec-baf9-3d526593e59e","added_by":"auto","created_at":"2022-11-03 19:02:59","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1720218,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eLiver antioxidant enzyme profile following BPA exposure and naringin application for (a) GSH (b) GPx(c) GST (d) SOD\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSuperscript (a) indicates significant difference at p\u0026lt;0.05 when compared with Control group (Group A). Superscript (b) indicates significant difference when compared with Bisphenol A group only (Group B). The results are shown in Mean ± SD (n= 6). Group A: Control, Group B: BPA (100 mg/kg), Group C: BPA and Naringin (100 mg/kg), Group D: BPA and Naringin (200 mg/kg), Group E: Naringin (100 mg/kg), \u0026amp; Group F: Naringin (200 mg/kg).\u003c/p\u003e","description":"","filename":"Figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2129508/v1/766a2fb4582615eb56fea716.jpg"},{"id":28627624,"identity":"3cd65674-0b31-4a43-9cb3-1cb87f89c73f","added_by":"auto","created_at":"2022-11-03 19:02:59","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":2964641,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe histology of the liver. \u003c/strong\u003eA (Control), B (Bisphenol A; 100 mg/kg), C (Bisphenol A + Naringin 100 mg/kg), D (Bisphenol A + Naringin 200 mg/kg), E (Naringin 100 mg/kg), F (Naringin 2000 mg/kg). Slides stained with Hematoxylin and Eosin. (Magnification x 100)\u003c/p\u003e","description":"","filename":"Figure5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2129508/v1/d7bb2bc5784b60171decfcc4.jpg"},{"id":28627626,"identity":"b3a5e02c-2163-4a5d-b76e-9fdd4c951283","added_by":"auto","created_at":"2022-11-03 19:03:00","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1263459,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe immunohistochemistry of hepatic caspase 3. \u003c/strong\u003eA (Control), B (BisphenolA; 100 mg/kg), C (BisphenolA + Naringin100mg/kg), D (BisphenolA + Naringin200mg/kg), E (Naringin100 mg/kg), F (Naringin2000 mg/kg). Slides stained with high definition Heamtoxylin. (Magnification x 100)\u003c/p\u003e","description":"","filename":"Figure6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2129508/v1/235de16755a4e19ddf97f3d7.jpg"},{"id":28627625,"identity":"7572efb3-48c4-467e-b89c-f3d54feaa734","added_by":"auto","created_at":"2022-11-03 19:02:59","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1464389,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe immunohistochemistry of hepatic Tumour Necrosis Factor Alpha (TNF-α). \u003c/strong\u003eA (Control), B (BisphenolA; 100 mg/kg), C (BisphenolA + Naringin100mg/kg), D (BisphenolA + Naringin200mg/kg), E (Naringin100 mg/kg), F (Naringin2000 mg/kg). Slides stained with high definition Heamtoxylin. (Magnification x 100)\u003c/p\u003e","description":"","filename":"Figure7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2129508/v1/3201b347192fcef7e02d5354.jpg"},{"id":32899210,"identity":"bbe6ea6e-bd68-4e75-9358-7ab5950efcab","added_by":"auto","created_at":"2023-02-14 09:29:28","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1606044,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2129508/v1/d5051155-f6d7-4966-b8ed-2aad38baa196.pdf"}],"financialInterests":"","formattedTitle":"Naringin mitigates Bisphenol A-induced hepatotoxicity in cockerel chicks","fulltext":[{"header":"Introduction","content":"\u003cp\u003eOver the years, there has been an increase in the exposure of living beings to some endocrine-disrupting chemicals (EDCs) that are available within the environment. This challenge is of great concern to human and animal health (Kelly et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Kasonga et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). These EDCs are known to cause interference with the function of the endocrine system, hence, leading to fatal health challenges in any animal or its offspring (EC 1999). Bisphenol-A (BPA) is a typical example of EDC. BPA mimics estrogen which has attracted several research interests among researchers over some decades owing to its unpalatable impacts on human and animal health (Rochester \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Staples et al. (\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) reported an increase in the global production of BPA from 5 to 8\u0026nbsp;million tons (MT) between the years 2010 and 2016. An estimate of 10.2 MT BPA production by the year 2022 has also been made. BPA is one of the organic synthetic compounds that is a key pollutant in the environment (Rahman et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). BPA has structures like oestradiol and diethylstilbestrol; hence, it has the tendency of stimulation of cellular response via osetrogen receptors binding (Rubin \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). As reported by Mohapatra et al. (\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), the point sources are manufacturing industry effluents, landfill leaching, wastewater treatment plant sludge discharge, dumpsite wastewater, and so on. Some of the areas of the utilization of BPA are pipe materials production, plastic materials, polyesters and polyacrylate production (Tomza-Marcimak et al. 2018). The environmental exposure of humans and animals to BPA is ubiquitous and majorly occurs through oral, respiratory, and dermal routes (Rahman et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). However, dietary ingestion is the main exposure route to living organisms. Birds are exposed BPA since plastic such as overhead water tanks, drinkers and feeding trays are normally utilized in poultry farms for some major activities (Escande et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Previous studies on animal models reported that BPA-induced oxidative stress caused a reprotoxic effect by affecting reproductive potential and is also hepatotoxic (Oyagbemi et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Kumar et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). For instance, BPA caused sperm reserves reduction, shorter sperm transit time, and reduction in mitochondrial activity (Matuszczak et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). A study conducted by Abdel-Gwaad et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) revealed a significant reduction in the activity of liver catalase following BPA exposure. The study further established that oxidative stress was induced in the liver, and an important alteration in the level of liver enzyme markers in serum was reported.\u003c/p\u003e \u003cp\u003eFlavonoids belong to a class of plant secondary metabolites. In plants, they are the origin of bioactive compounds (Ghasemzadeh and Jaafar \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Flavanoids are a class of polyphenolic secondary metabolites having varying structures in the form of glycones or glycosides in fruits and vegetables (Li et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). They have good health benefits based on their anti-carcinogenic, anti-inflammatory, antioxidant, and anti-mutagenic effects. Naringin is a natural flavonoid. Different studies have established that naringin contains antioxidant, anti-apoptotic, anti-osteoporotic, anti-inflammatory, anti-carcinogenic, and anti-ulcer properties (Wang et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Chen et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Naringin also plays a significant function in reducing triglycerides, cholesterol, improvement in immune function, and antioxidant status. Therefore, naringin is a natural antioxidant owing to its ability to scavenge free radicals as well as prevention of lipid peroxidation (Changxing et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Naringin inhibits radiation-induced reduction of the antioxidant defense system (Chen et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Some of the studies conducted on the influence of naringin on the organs of some models are highlighted. Goliomytis et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) reported that naringin may favorably elongate the shell-life of eggs. Naringin have anti-inflammatory properties as well as the ability to improve yolk colour without any side effects on the quality traits and performance of the eggs. The dietary inclusion of naringin at 100 mg/kg daily lowered plasma fatty acids levels, and improved liver mitochondrial dysfunction and glucose intolerance (Alam et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Naringin also enhanced the structure and function of both the liver and the heart without having any detrimental effect on the total body weight. Furthermore, naringin was reported to lower the blood sugar level in rat and prevent the development of hyperglycemia (Jung et al. 2014; Parmar et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e); and reduce LDL-cholesterol, total cholesterol, and triglycerides levels in chicken (da Silva et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2001\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe consumption of leaked BPA into the feeds and water of poultry animals is detrimental to the health of the animals. Various means of reducing the toxic effect of BPA on different organs through the usage of various flavonoid-based supplements have been devised such as orange-peel powder in a rat model (Abdel-Gwaad et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2020\u003c/span\u003e); naringin in male rats (Elsawy et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2021\u003c/span\u003e); and so on. There are limited studies on the effect of BPA toxicity in birds and how naringin can be used to reverse its deleterious effect. The study was designed to determine the ameliorative effect of naringin and its action of mechanism on BPA toxicity in the liver of cockerel chicks.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eChemicals\u003c/h2\u003e \u003cp\u003eNaringin, Bisphenol A, Albumin (Sigma Chemical Co., USA), Hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e), phosphate buffer, sodium hydroxide, copper sulphate, potassium tartarate, Trichloroacetic acid (TCA), Tris-potassium chloride, Ammonium ferrous sulphate, sorbitol, sulphuric acid, carbonate buffer, Adrenaline, Ellman\u0026rsquo;s reagent (DTNB) reduced glutathione (GST), thiobarbituric acid (TBA), 2-dichloro-4-nitrobensene (CDNB), sodium azide, O-dianiside dihydrochloride were purchased. Biotinylated secondary antibody, Tumour necrosis factor alpha (TNF-α) and Caspase 3 monoclonal antibody were purchased from Elabscience Biotechnology, China.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eExperimental Animals\u003c/h3\u003e\n\u003cp\u003eIn this study, thirty one-day-old cockerel chicks purchased from CHI FARMS Limited Ibadan, Nigeria were utilized. The chicks were housed at the experimental rearing facility of Avian diseases unit, Department of Veterinary Medicine, University of Ibadan. Brooding of the chicks was done for 2 weeks by supply of additional heat and then further reared for 4 weeks. The chicks were fed age-appropriate poultry feed from Top Feeds\u0026reg; limited, Ibadan, and clean drinking water \u003cem\u003ead libtum\u003c/em\u003e was given liberally. All the cockerels were kept in open-sided tropical rearing spacious cages with all necessary conditions for their comfort as indicated by Animal Use and Care Research Ethics Committee (ACUREC) Ibadan. The research was done according to the procedures illustrated and approved by the University of Ibadan - ACUREC (UI-ACUREC/048\u0026ndash;0521/21).\u003c/p\u003e\n\u003ch3\u003eExperimental Design\u003c/h3\u003e\n\u003cp\u003eCockerels with uniform weight were randomly allotted into 6 groups (5 chicks per group). Bisphenol A at 100 mg/kg was administered continuously in drinking water for a period of 6 weeks while the treated groups were administered different doses of Naringin through oral gavage for a period of 2 weeks. Thirty-one-day old cockerel chicks used for this study were randomly divided into 6 groups of five chicks per group as follows: Group 1 (Control), Group 2 (BPA 100 mg/kg), Group 3 (BPA\u0026thinsp;+\u0026thinsp;100 mg/kg naringin), Groups 4 (BPA\u0026thinsp;+\u0026thinsp;200 mg/kg naringin), Group 5 (100 mg/kg naringin) and Group 6 (100 mg/kg naringin), respectively. The administration of BPA and naringin was through oral gavage.\u003c/p\u003e\n\u003ch3\u003eSample Collection And Preparation Of Hepatic Post-mitochondrial Fractions\u003c/h3\u003e\n\u003cp\u003eTwenty-four hours post administration of the last treatment, chickens were euthanized by quick cervical dislocation. The whole liver was harvested by rapid dissection on ice, rinsed with distilled water, blotted with filter paper, and whole organ weight was determined using an electronic digital scale. Subsequently, about 2 g each of the liver samples were collected inside labeled universal bottles, sliced and blended in 0.1 M aqueous potassium phosphate buffer at pH 7.4. Liver post mitochondrial fractions (PMFs) were obtained using the homogenate obtained after centrifuging at 10,000 \u003cem\u003eg\u003c/em\u003e for 10 minutes with a cold centrifuge at -4˚C. The supernatant was collected and stored and used for biochemical assay.\u003c/p\u003e\n\u003ch3\u003eBiochemical Assays\u003c/h3\u003e\n\u003cp\u003eTotal protein concentration in the supernatant was determined using the Biuret method as described by Gornal et al. (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e1949\u003c/span\u003e). The method described by Wolff (\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e1994\u003c/span\u003e) was used in the determination of hydrogen peroxide generation. The illustrated method of Varshney and Kale (\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e1990\u003c/span\u003e) was employed in the determination of the lipid peroxidation (Malondialdehyde (MDA) concentration). The hepatic reduced glutathione (GSH) concentration determination was determined with the method described by Ellman (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e1959\u003c/span\u003e). The method of Misra and Fridovich (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1972\u003c/span\u003e) with slight modification by Oyagbemi et al. (\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) was followed in the evaluation of the superoxide dismutase (SOD) activity. The methods described by Buetler et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1963\u003c/span\u003e) and Habig et al. (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e1974\u003c/span\u003e) for the determination of Glutathione peroxidase (GPx) and Glutathione S-transferase (GST) activities, respectively, were adopted. The liver function tests such as total protein, albumin, total bilirubin, conjugated bilirubin, Aspartate transferase (AST), Alanine transferase (ALT), and Alanine phosphatase (ALP) were determined using the commercial colorimetric kits as described by Manterys et al. (\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and Adeyemo et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eHistopathology\u003c/h3\u003e\n\u003cp\u003eLiver tissues were fixed in 10% formalin, embedded in paraffin wax, and sections of 5\u0026ndash;6 mm in thickness were made and thereafter stained with Hematoxylin and Eosin (H\u0026amp;E) as previously described by Drury and Walington (1976). Therefore, the sections were examined with light microscopic.\u003c/p\u003e\n\u003ch3\u003eImmunohistochemistry\u003c/h3\u003e\n\u003cp\u003eIn this study, the immunohistochemistry procedure of the liver to determine the hepatic caspase 3 and tumor necrosis factor-alpha (TNF-α) as described by Oyagbemi et al. (\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) was adopted.\u003c/p\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eAll values were expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD) and the test of significance between the experimental groups and the control group was estimated with Student\u0026rsquo;s t-test. The One-Way Analysis of Variance (ANOVA) with Tukey\u0026rsquo;s post-hoc test of Graph pad prism 5.0 was also carried out with p-values\u0026thinsp;\u0026lt;\u0026thinsp;0.05 considered statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eLiver function test\u003c/h2\u003e \u003cp\u003eFigure 1 showed the results obtained for the liver function tests. The values of total protein, albumin, and total bilirubin of chicks intoxicated with BPA (100 mg/kg) was significantly (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) lower than that of the control and chicks treated with naringin (Figs.\u0026nbsp;1a, b, \u0026amp;c). However, there was significant improvement in the values of total protein, albumin, and total bilirubin of chicks treated with naringin (100 \u0026amp; 200 mg/kg) and those that received only naringin (100 \u0026amp; 200 mg/kg). From our results, we observed a significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) increase in the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase in intoxicated with BPA in comparison to the control. (Fig.\u0026nbsp;1c, e, \u0026amp; f). Treatment with naringin (100 \u0026amp; 200 mg/kg) significantly reduced the heightened values of AST, ALT, and ALP, respectively, in BPA intoxicated chicks. This is indicative of hepatoprotective action of naringin.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMarkers Of Oxidative Stress And Antioxidant Defense System In The Liver\u003c/h3\u003e\n\u003cp\u003eHydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) generation and content of lipid peroxidation product (MDA) in the liver of the BPA intoxicated chicks were significantly (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher when compared with the control group (Figs.\u0026nbsp;3a \u0026amp; 3b). Figure\u0026nbsp;3 also showed that the level of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e and MDA content in the liver of BPA treated chicks with naringin (100 mg/kg \u0026amp; 200 mg/kg) and naringin only (100 mg/kg \u0026amp; 200 mg/kg) in comparison to BPA untreated chicks. The observed significant reduction in the content of MDA was dose-dependent in naringin treated chicks (Fig.\u0026nbsp;3b), thus, indicating the antioxidative action of naringin. The result showed a significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) decrease in the level of GSH, GST, and SOD activity in the BPA exposed chicks when compared with the control group (Figs.\u0026nbsp;4a, 4b, \u0026amp; 4d), while BPA toxicity enhanced the activity of GPx greater than that of the control (Fig.\u0026nbsp;4c). The noticeable increase in the activity of hepatic GPx might be due to consumption of GSH due to BPA toxicity. However, treated of intoxicated chicks with naringin significantly improved the content of GSH and the activities of GST, SOD, and GPx, respectively, (Figs.\u0026nbsp;4a, 4b, \u0026amp; 4d).\u003c/p\u003e\n\u003ch3\u003eHistopathology\u003c/h3\u003e\n\u003cp\u003eHistology revealed a moderate diffuse sinusoidal congestion, with a severe periportal cellular infiltration in BPA intoxicated chicks. However, no visible lesions were observed in chicks in the histopathology of hepatic tissues treated with naringin (Fig.\u0026nbsp;5).\u003c/p\u003e\n\u003ch3\u003eImmunohistochemistry\u003c/h3\u003e\n\u003cp\u003eHepatic caspase 3 and tumor necrosis factor (TNF-α) in the hepatocytes revealed higher expressions in BPA exposed group when compared to the control and groups treated with naringin (Figs.\u0026nbsp;6 and 7). The reduction in the expressions of TNF-α as well as, Caspase 3 is indicative of amelioration of inflammation, apoptosis and hepatoprotective effects of naringin against bisphenol A toxicity as shown by the reduction in the expression of TNF-α and caspase 3 in the groups treated with naringin and groups administered naringin only.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eDifferent health-related deleterious effects are associated with BPA exposure in living organisms, with BPA showing that endocrine-disrupting chemicals induce oxidative stress (Singh et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Meli et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). BPA induces oxidative stress by generating reactive oxygen species (ROS) through the generation of free radicals, and thereby causes an imbalance between ROS and antioxidant defenses which then results in oxidative damage (Kobroob et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Amjad et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The hepatotoxic effect of BPA was established in this study as demonstrated with a significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) increase in the activities of ALP, AST, and ALT, and a concomitant decrease in total protein and albumin in BPA-exposed chicks. This is in line with the study carried out by Oguazu et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) and Abdel-Gwaad et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) as they reported an increase in ALT, AST, and AST which indicates hepatotoxicity action of BPA. Naringin is an example of a flavonoid that has been reported to possess several pharmacological effects such as anti-inflammatory, hypolipidemic, antioxidant, antiviral, antimicrobial, anti- apoptotic, hepatoprotective, and immunomodulatory effects (Changxing et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eHepatoprotective activities were observed in the naringin-treated groups as there was a significant decrease in ALT, AST, and ALP and a corresponding increase in total protein. This corroborated the earlier study of Imam et al. (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) who reported that co-administration of naringin with aluminum ameliorated aluminum-induced liver damage and oxidative stress. Furthermore, the levels of TC, TAG, and LDL-c were significantly (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) higher in BPA intoxicated chicks with a concurrent decrease in HDL-c as observed in the present study. This agreed with the findings of Eweda et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) who reported that BPA induced dyslipidemic state and increased triglycerides (steatosis) and cholesterol accumulation in the liver tissue of rat. Marmugi et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) also reported that the exposure of rat to BPA resulted in hypertriglyceridemia, hypercholesterolemia, and alterations of fatty acids composition in the liver. More so, there was upregulation of genes which are linked with \u003cem\u003ede novo\u003c/em\u003e lipogenesis and cholesterol synthesis in the liver. Kumar et al. (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) reported that treatment with naringin produced noteworthy developments in altered lipid profiles. The anti-hyperlipidemic effect of naringin was established in this study as naringin reversed the toxicity of BPA by decreasing LDL-c, TC, TG, and increasing HDL-c in chicks treated naringin.\u003c/p\u003e \u003cp\u003eThe scavenging and neutralization of free radicals such as superoxide radicals, hydrogen peroxides (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e), and hydroperoxides are done by antioxidants, thereby preventing oxidation reactions. During oxidative stress, free radicals increase in the body (Amjad et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). This study showed an increase in H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e contents in the liver of BPA-exposed chicks. This was in concordance with the study of da Silva et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) who reported an exaggerated increase H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e generation in thyrocytes of female Wistar rats intoxicated with BPA. More so, Kabuto et al. (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) observed that BPA caused the overproduction of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e with resultant oxidative stress. In our study, naringin reduced the level of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e in groups treated BPA-treated chicks and naringin-only treated groups. The antioxidant and free radical scavenging power of naringin was earlier reported by Rashmi et al. (\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) who showed the H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e radical scavenging potential of naringin in streptozotocin-induced liver damage. Also, Miles and Calder (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) reported that naringin increased liver and kidney expression of anti-inflammatory transcription factors.\u003c/p\u003e \u003cp\u003eIn this study, the increase in the level of MDA caused lipid peroxidation because of BPA-induced oxidative stress as earlier reported by Kobroob et al. (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) and Eweda et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Lipid peroxidation of biological membranes is known to cause loss of membrane fluidity, increases membrane permeability, alters receptor function, and changes in membrane potential. It can be observed in this study that the MDA level was exaggerated in the liver of BPA intoxicated chickens. However, in this study, the group co-treated with naringin caused a significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) reduction in the MDA level, as previously reported by Rajadurai and Stanely (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) \u0026amp; Alam et al. (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eGlutathione (GSH) is one of the non-enzymatic antioxidants that is depleted during oxidative stress. In this study, a decreased level of GSH was observed in the liver in the BPA-exposed chicks, which depicts ongoing oxidative stress caused by BPA toxicity. Kobroob et al. (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) reported a decrease in GSH in rats exposed to BPA. However, the co-treated with naringin showed improvements in the activities of GSH. This supports the study of Kumar et al. (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) who stated that administration of naringin to diabetic rats raised the concentration of GSH. The GPx, SOD, and GST are known to protect tissues from damage caused by oxidative stress by detoxification of several substrates generated from cellular oxidative processes (Sravani et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The GPx activity in the liver increased in the BPA-exposed chicks in this study, and this might be a result of the adaptive response. We proposed that this could be a result of nuclear translocation of nuclear erythroid-related factor 2 (Nrf2). The first step is the modification of the keap1 segment of Nrf2-Keap1 pathway. Nrf2 is usually low when it is not activated by stress factors. In the presence of ROS such as superoxide, hydroxyl, and peroxyl radicals, cysteine residues (such as Cys 151) are oxidized the sulfhydryl groups on the Keap1 segment. This changes the conformity of Keap1 segment and prevents it from attaching to Nrf2. The Nrf2 released within the cytosol enters the nucleus and heterodimerize with small Maf proteins, and further binds to regulatory gene recognized as antioxidant response elements (ARE). Substances that can initiate activity at the ARE include environmental pollutants, hydrogen peroxide and nitric oxide. The Nrf2-Maf-ARE complex then initiates subsequent specific antioxidant and detoxification genes such as that of GST, GPx, GSH and SOD; thereby, increasing antioxidant activities and inactivating ROS (Baird and Dinkova-Kostova \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Oyagbemi et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The antioxidant effect of naringin was established as naringin restored the levels of GSH, GST, GP\u003csub\u003eX\u003c/sub\u003e, and SOD in the hepatic tissue of BPA-exposed chicks.\u003c/p\u003e \u003cp\u003eTNF-α is an example of a pro-inflammatory cytokine, which plays a significant role in the host\u0026rsquo;s defense against injury. It was stated that BPA toxicity exacerbated the proinflammatory cytokines expression along with interleukins such as IL-1β, IL-6, IL-8, and tumor necrosis factor alpha (TNF-α) as previously reported (Wang et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Meli et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). This is in agreement with this present study as there was noticeable higher expression of TNF-α in the liver of the BPA-exposed chicks compared to the control and naringin-treated chicks. Naringin has been reported to possess a hepatoprotective effect by regulating inflammatory cytokines and increasing antioxidant enzymes (Caglayan et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This study establishes that naringin reduces the expression of TNF-α in birds co-administered with naringin and BPA. Caspase-3 is a biomarker of cell death. In this study, there was an increase expression of caspase-3 in the liver of BPA intoxicated chicks. This showed that hepatic apoptosis might have resulted from inflammation and oxidative stress following BPA toxicity (Abdel-Rahman et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Liu et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). This is in tandem with the work of Elgawish and Abdelrazek (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) where increase in expression of caspase-3 in testes of male rats exposed to lead acetate was observed. The anti-apoptotic effect of naringin was exhibited in the BPA group co-treated with naringin and naringin treated groups by cleaving caspase-3 and inhibiting the genes and proteins involved in apoptotic pathways such as P53, P 16\u003csup\u003eINK 49\u003c/sup\u003e (Yuan and Yang \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eBased on the results in this present study, it can be inferred and concluded that naringin abated the toxicity of Bisphenol A-induced liver dysfunction. Naringin exhibited its hepatoprotective property by revitalizing cells that were damaged due to BPA-induced oxidative stress. The use of Naringin can serve as a natural and safe feed supplement in poultry nutrition is recommended.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors deeply thank the African Union\u0026nbsp;through the\u0026nbsp;Pan African University\u0026nbsp;Institute of Earth and Life Sciences Institute (PAULESI)\u0026nbsp;for funding this work and\u0026nbsp;Cardio-renal laboratory,\u0026nbsp;University of Ibadan for the facilities used.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval:\u0026nbsp;\u003c/strong\u003eThe study was conducted following guidelines approved by the Animal Care and Use Research Ethics Committee (ACUREC) of the University of Ibadan (Approval number: UI-ACUREC/048-0521/21). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Participate\u003c/strong\u003e: Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publish:\u003c/strong\u003e Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors Contributions:\u0026nbsp;\u003c/strong\u003eThe authors, Leah Oluwaseyanu Esuola, Oluwaseun Esan,\u0026nbsp;Adamu Shafiu Maikifi\u003cstrong\u003e\u003csup\u003e,\u0026nbsp;\u003c/sup\u003e\u003c/strong\u003eAdemola Adetokunbo Oyagabemi and\u0026nbsp;Temidayo Olutayo\u003csup\u003e\u0026nbsp;\u003c/sup\u003eOmobowale designed the experiment.\u0026nbsp;Leah Oluwaseyanu Esuola, Oluwaseun Esan,\u0026nbsp;Adamu Shafiu Maikifi\u0026nbsp;performed the immunohistochemistry and biochemical assays. \u0026nbsp;The blood pressure and electrocardiogram were performed by\u0026nbsp;Leah Oluwaseyanu Esuola, Oluwaseun Esan and\u0026nbsp;Temidayo Olutayo\u003csup\u003e\u0026nbsp;\u003c/sup\u003eOmobowale. Moses Olusola Adetona, Ademola Adetokunbo Oyagabemi,\u0026nbsp;Temidayo Olutayo\u003csup\u003e\u0026nbsp;\u003c/sup\u003eOmobowale,\u0026nbsp;Oluwafemi Oguntibeju,\u0026nbsp;Momoh Audu Yakubu supervised, proof-read, and approved the submission.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e The study was funded by the African Union \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests:\u003c/strong\u003e The authors declare that they have no competing interest\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials:\u003c/strong\u003e Data will be made available on request\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbdel-Gwaad HMS, AbdEl-Wahab HMF, Mohamed EAK, Sharaf EHAA, Osman AAHM (2020) Modulatory effect of dry orange (\u003cem\u003eCitrus sinensis)\u003c/em\u003e peel powder on bisphenol A- induced hepatic and splenic toxicity in rats. J Basic Appl Zool 81:49\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbdel-Rahman HG, Abdelrazek HMA, Zeidan DW, Mohamed RM, Abdelazim AM (2018) Lycopene: Hepatoprotective and antioxidant effects toward bisphenol A-induced toxicity in female Wistar rats. Oxid Med Cell Longev 2018:5167524\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAdeyemo GO, Kabir AO, Tanimowo DA, Ologhobo AD (2018) Performance, blood chemistry, and serum electrolytes of broilers given water from different sources. Nigerian J Anim Sci 20:140\u0026ndash;145\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlam AM, Subhan N, Rahman MM, Uddin SJ, Reza HM, Sarker SD (2014) Effect of citrus flavonoids, naringin and naringenin, on metabolic syndrome and their mechanisms of action. Adv Nutr 5:404\u0026ndash;417\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlam MA, Kanter K, Brown L (2013) Naringin improves diet induced cardiovasicular dysfunction and obesity in high carbohydrate, high fat diet-fed rats. Nutrients 5:637\u0026ndash;650\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAmjad S, Rahman MS, Pang MG (2020) Role of antioxidants in alleviating bisphenol A toxicity. Biomolecules 10:1105. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/biom10081105\u003c/span\u003e\u003cspan address=\"10.3390/biom10081105\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBaird L, Dinkova-Kostova AT (2011) The cytoprotective role of the Keap1-Nrf2 pathway. Arch Toxicol 85:241\u0026ndash;272\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBuetler E, Duron O, Kelly MB (1963) Improved method for determination of blood glutathione reduced. J Lab Clin Med 61:882\u0026ndash;888\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCaglayan C, Temel Y, Kandemir FH, Kucukler S (2018) Naringin protects against cyclophosphamide-induced hepatotoxicity and nephrotoxicity through modulation of oxidative stress, inflammation, apoptosis, autophagy and DNA damage. Environ Sci Pollut Res 25:20968\u0026ndash;20984\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChangxing L, Saeed M, Kamboh AA, Alagawany M, Abd El-Hack ME, Arain MA, Babazadeh D, Dhama K, Chenling M (2018) Reconsidering a Citrus Flavonoid Naringin as a Promising Nutritional Supplement and its Beneficial Health Applications in Humans, Animals and Poultry. Int J Pharmacol, 2018:1\u0026ndash;10\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen P, Xiao Z, Wu H, Wang Y, Su W, Li P (2020) The effects of naringin on cigarette smoke-induced dynamic changes in oxidation/antioxidant system in lung of mice. Nat Prod Commun 15:1\u0026ndash;9\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen R, Qi QL, Wang MT, Li QY (2016) Therapeutic potential of naringin: An overview. Pharm Biol 54:3203\u0026ndash;3210\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eda Silva MM, Xavier LLF, Goncalves CFL, Santos-Silva AP, Palva-Melo FD, de Freitas ML, Fortunato RS, Miranda-Alves L, Ferreira ACF (2018) Bisphenol A increases hydrogen peroxide generation by thyrocyte both in vivo and in vitro. Endocrin Connect 7:1196\u0026ndash;1207\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eda Silva RR, de Oliveira TT, Nagem TJ, Pinto AS, Albino LF (2001) Hypo-cholesterolemic effect of naringin and ruttin flavonoid. Arch Latinoam Nutri 51:258\u0026ndash;264\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDrury RA, Wallington EA (1976) Carlton's Histopathological Techniques, 4th edn. Oxford University Press, London, pp 139\u0026ndash;142\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eElgawish RAR, Abdelrazek HMA (2014) Effects of lead acetate on testicular function and caspase-3 expression with respect to the protective effect of cinnamon in albino rats. Toxicol Rep 1:795\u0026ndash;801\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEllman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70\u0026ndash;77\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eElsawy H, Alzahrani AM, Alfwuaires M, Abdel-Moneim AM, Khalil M (2021) Nephroprotective effect of naringin in methotrexate induced renal toxicity in male rats. Biomed Pharmacother 143:112180\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEscande A, Pillon A, Servant N, Cravedi JP, Larrea F, Muhn P, Nicolas JC, Cavaill\u0026egrave;s V, Balaguer P (2006) Evaluation of ligand selectivity using reporter cell lines stably expressing estrogen receptor alpha or beta. Biochem Pharmacol 71:1459\u0026ndash;1469\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEuropean Commission. Community Strategy for Endocrine Disrupters (1999) A Range of Substances Suspected of Interfering with the Hormone Systems of Humans and Wildlife; Commission of the European Communities: Brussels, Belgium\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEweda SM, Newairy AA, Abdou HM, Gaber AS (2020) Bisphenol A-induced oxidative damage in the hepatic and cardiac tissues of rats: The modulatory role of sesame lignans. Exp Th Med 19:33\u0026ndash;44\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhasemzadeh A, Jaafar HZ (2013) Profiling of phenolic compounds and their antioxidant and anticancer activities in pandan (\u003cem\u003ePandanus amaryllifolius Roxb.)\u003c/em\u003e extracts from different locations of Malaysia. BMC Complement Altern Med 13:341\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGoliomytis M, Simitzis P, Papalexi A, Veneti N, Hager-Theolorides AL, Charismadou MA, Deligeogis SG (2019) Influence of citrus flavonoids on laying hen performance, inflammatory immune response, egg quality and yolk oxidative stability. Br Poult Sci 60:272\u0026ndash;278\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGornal AG, Bardawill JC, David MM (1949) Determination of serum proteins by means of Biuret reaction. J Biol Chem 177:751\u0026ndash;766\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHabig WH, Pabst MJ, Jakpoby WB (1974) Glutathione transferase, a first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130\u0026ndash;7139\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eImam TS, Khalifa HA, Hussein MMA, Ali HA (2016) Aluminum-induced oxidative stress and hepato -renal impairment in male albino rats: Possible. Life Sci J 13:93\u0026ndash;104\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJung UJ, Kim SR (2014) Effects of naringin, a flavanone glycoside in grapefruits and citrus fruits, on the nigrostriatal dopaminergic projection in the adult brain. Neural Regen Res 9:1514\u0026ndash;1517\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKabuto H, Hasuike S, Minagawa, Shishibori T (2003) Effect of bisphenol A on the metabolisms of active oxygen species in mouse tissues. Environ Res 93:31\u0026ndash;35\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKasonga TK, Coetzee MA, Kamika I, Ngole-Jeme VM, Momba MNB (2021) Endocrine-disruptive chemicals as contaminants of emerging concern in wastewater and surface water: A review. J Environ Manag 277:111485\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKelly M, Connolly L, Dean M (2020) Public awareness and risk perceptions of endocrine disrupting chemicals -A qualitative study. Int J Environ Res Public Health 17:7778\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKobroob A, Peerapanyasut W, Chattipakorn N, Wongmekiat O(2018) Damaging effects of Bisphenol A on the kidney and the protection by melatonin: Emerging evidences from in vivo and in vitro studies. Oxidat Med Cell Long 2018; Article ID 3082438, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1155/2018/3082438\u003c/span\u003e\u003cspan address=\"10.1155/2018/3082438\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumar SR, Ramli ESM, Abdul-Nasir AA, Ismail NHM, Fahami NAM (2019) Preventive effect of Naringin on metabolic syndrome and its mechanism of action: A systematic review. Evidence-Based Complement Alternat Med 2019:1\u0026ndash;11\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi K, Fan H, Yin P, Yang L, Xue Q, Li X, Sun L, Liu Y (2017) Structure-activity relationship of eight high content flavonoids analyzed with a preliminary assign-score method and their contribution to antioxidant ability of flavonoids-rich extract from Scutellana baicalensis shoots. Arab J Chem 11:159\u0026ndash;170\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu R, Cai D, Li X, Liu B, Chen J, Jiang X, Li H, Li Z, Teerds K, Sun J, Bai W, Jin Y (2022) Effects of Bisphenol A on reproductive toxicity and gut microbiota dysbiosis in male rats. Ecotoxicol Environ Safety 239:113623\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eManterys A, Francyk-Zarow M, Czyzynska-Cichon I, Drahun A, Kus E, Szymczyk B, Kostogrys RB (2016) Haematological parameters, serum lipid profile, liver function, and fatty acid profile of broiler chickens fed on diets supplemented with pomegranate seed oil and linseed oil. Br Poult Sci 57:771\u0026ndash;779\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMarmugi A, Ducheix S, Lasserre F, Polizzi A, Paris A, Priymenko N, BertrandMichel J, Pineau T, Guillou H, Martin PG, MselliLakhal L (2012) Low doses of bisphenol A induce gene expression related to lipid synthesis and trigger triglyceride accumulation in adult mouse liver. Hepatology 55:395407\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMatuszczak E, Komarowska MD, Debek W, Hermanowicz A(2019) \u003cb\u003e()\u003c/b\u003e The impact of bisphenol A on fertility, reproductive system, and development: A review of the literature. Int J Endocrin 2019:4068717\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMeli R, Monnolo A, Annunziata C, Pirozzi C, Ferrante MC (2020) Oxidative stress and BPA toxicity: An antioxidant approach for male and female reproductive dysfunction. Antioxidant 9:405\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMiles E, Calder PC (2021) Effects of citrus fruit juices and their bioactive components on inflammation and immunity: A narrative review. Front Immunol 12:712608\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMisra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 217:3170\u0026ndash;3175\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMohapatra DP, Brar SK, Tyagi RD, Surampalli RY (2010) Physicochemical pre-treatment and biotransformation of wastewater and wastewater Sludge \u0026ndash; fate of bisphenol A. Chemosphere 78:923\u0026ndash;941\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOguazu CE, Ezeonu FE, Ubaoji KI, Anajekwu B (2015) Bisphenol A exerts a transient perturbation of liver function in Wistar albino rats at acute and sub-chronic exposure doses. J Pharmaceut Sci Biosci Res 5:274\u0026ndash;278\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOyagbemi AA, Omobowale TO, Asenuga ER, Adejumobi AO, Ajibade TO, Ige TM et al (2019) Sodium fluoride induces hypertension and cardiac complications through generation of reactive oxygen species and activation of nuclear factor kappa beta. Environ Toxicol 32:1089\u0026ndash;1101\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOyagbemi AA, Omobowale TO, Akinrinde AS, Saba AB, Ogunpolu BS, Daramola O (2015) Lack of reversal of oxidative damage in renal tissues of lead acetate-treated rats. Environ Toxicol 30:1235\u0026ndash;1243\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOyagbemi AA, Omobowale TO, Asenuga ER, Afolabi JM, Adejumobi OA, Adedapo AA, Yakubu MA (2017) Effect of arsenic acid withdrawal on hepatotoxicity and disruption of erythrocyte antioxidant defense system. Toxicol Rep 4:521\u0026ndash;529\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eParmar HS, Jain P, Chauham DS, Bhinchar DS, Munjal V (2012) DPP-IV inhibitory potential of naringin An in silico, in vitro and in vivo study. Diabetes Res Clin Pract 97:105\u0026ndash;111\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRahman S, Kwon WS, Ryu DY, Khatun A, Karmakar PC, Ryu BY, Pang MG (2017) Functional and proteomic alterations of F1 capacitated spermatozoa of adult mice following gestational exposure to Bisphenol A. J Proteome Res 17:524\u0026ndash;535\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRajadurai M, Stanely MPP (2006) Preventive effect of naringin on lipid peroxides and antioxidants in isoprote renal-induced cardiotoxicity in Wistar rats: Biochemical and histopathological evidences. Toxicology 228:59\u0026ndash;68\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRashmi R, Magesh SB, Ramkumar KM, Suryanarayanan S, SubbaRao MV (2018) Antioxidant potential of naringenin helps to protect liver tissue from streptozotocin-induced damage. Rep Biochem Molecul Biol 7:76\u0026ndash;84\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRochester JR (2013) Bisphenol A and human health: A review of the literature. Toxicol 42:132\u0026ndash;155\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRubin BS (2011) Bisphenol A: an endocrine disruptor with widespread exposure and multiple effects. J Steroid Biochem Molecular Biol 127:27\u0026ndash;34\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh RP, Shafeeque CM, Sharma SV, Singh R, Kannan M, Sastry KVH, Raghunandanan S, Mohan J, Azeez PA (2016) Effects of bisphenol-A on male reproductive success in adult Kadaknath chicken. Ecotoxicol Environ safety 128:61\u0026ndash;66\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSravani J, Padmaja K, Eswara Prasad P, Punya Kumari B (2016) Effect of bisphenol A on antioxidant enzymes and lipid peroxidation in liver of chick embryos. Int J Meat Sci 6:1\u0026ndash;5\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStaples C, van der Hoeven N, Clark K, Mihaich E, Woelz J, Hentges S (2018) Distributions of concentrations of bisphenol A in North American and European surface waters and sediments determined from 19 years of monitoring data. Chemosphere 201:448\u0026ndash;458\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTomza-Marciniak A, St˛epkowska P, Kuba J, Pilarczyk B (2018) Effect of bisphenol A on reproductive processes: A review of in vitro, in vivo and epidemiological studies. J Appl Toxicol 38:51\u0026ndash;80\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVarshney R, Kale RK (1990) Effect of calmodulin antagonists on radiation induced lipid peroxidation in microsomes. Int J Radiation Biol 58:733\u0026ndash;743\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang D, Ma W, Wang F, Dong J, Wang D, Sun B, Wang B (2015) Stimulation of Wnt/β-catenin signaling to improve bone development by naringin via interacting with AMPK and Akt. Cell Physiol Biochem 36:1563\u0026ndash;1576\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang DM, Yang YJ, Zhang L, Zhang X, Guan FF, Zhang LF (2013) Naringin enhances CaMKII activity and improves long-term memory in a mouse model of Alzheimer\u0026rsquo;s disease. Int J Mol Sci 14:5576\u0026ndash;5586\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang K, Zhao Z, Ji W (2019) Bisphenol A induces apoptosis oxidative stress and inflammatory response in colon and liver of mice in a mitochondria dependent manner. Bio med Pharmacother 117:109182\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWolff SF (1994) Ferrous ion oxidation in the presence of ferric ion indicator xylenol orange for measurement of hydrogen peroxides. Methods Enzymol 233:182\u0026ndash;189\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYuan Z, Yang Z (2022) The effect of naringin on the apoptosis of degenerative nucleus pulposus cells: A study on the function and mechanism. Drug Des Develop Therapy 16:499\u0026ndash;508\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Bisphenol A, naringin, hepatotoxicity, antioxidant, metal binding, cockerel chicks","lastPublishedDoi":"10.21203/rs.3.rs-2129508/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2129508/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBisphenol A (BPA) is an environmental pollutant, a monomer used in the polymer industry to produce plastics. BPA has been reported to cause deleterious effects in both humans and animals. Naringin is one of the flavonoids with antioxidant and metal chelating properties. This study was carried out to assess the potential ameliorative effect of naringin on BPA-induced hepatotoxicity in cockerel chicks. Thirty-one-day old cockerel chicks used for this study were randomly divided into 6 groups of five chicks per group as follows: Group 1 (Control), Group 2 (BPA 100 mg/kg), Group 3 (BPA\u0026thinsp;+\u0026thinsp;100 mg/kg naringin), Groups 4 (BPA\u0026thinsp;+\u0026thinsp;200 mg/kg naringin), Group 5 (100 mg/kg naringin) and Group 6 (100 mg/kg naringin), respectively. The administration of BPA and naringin was through oral gavage. BPA intoxication precipitated significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) increased ALT, ALP, AST, TC, TG, LDL, but decreased total protein and HDL-cholesterol when compared with the control. Also, there was a significant increase in hepatic H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e generation and MDA content with concomitant decrease in reduce glutathione, glutathione S-transferase, and superoxide dismutase activity in BPA intoxicated chicks. Histology revealed a moderate diffuse sinusoidal congestion, with a severe periportal cellular infiltration in BPA intoxicated chicks. Immunohistochemistry results revealed a higher expression of hepatic caspase 3 and TNF-α in chicks exposed to BPA alone relative to the control and chicks treated with naringin (100 mg/g and 200 mg/kg). Findings from this study showed that naringin administration restored hepatoxicity, improved antioxidant status, and lowered exaggerated values of cholesterol, oxidative stress indices, and ameliorated ultrastructure anarchy. Combining all, the incorporation of naringin into poultry feeds could position the flavonoid as a novel antioxidant and metal chelator with beneficial effects in the reduction of toxicities associated with environmental pollutants such as bisphenol A, thereby boosting production efficiency in livestock production.\u003c/p\u003e","manuscriptTitle":"Naringin mitigates Bisphenol A-induced hepatotoxicity in cockerel chicks","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-11-03 19:02:54","doi":"10.21203/rs.3.rs-2129508/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ac4cadf6-fe4d-4e42-bb3c-a9842ab226d6","owner":[],"postedDate":"November 3rd, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-02-14T09:29:16+00:00","versionOfRecord":[],"versionCreatedAt":"2022-11-03 19:02:54","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2129508","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2129508","identity":"rs-2129508","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}