Efficacy of Bacopa monnieri in Mitigating Lead-Induced Blood Toxicity in Mice Compared to Synthetic Antioxidants

Efficacy of Bacopa monnieri in Mitigating Lead-Induced Blood Toxicity in Mice Compared to Synthetic Antioxidants | 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 Efficacy of Bacopa monnieri in Mitigating Lead-Induced Blood Toxicity in Mice Compared to Synthetic Antioxidants Flora Shah, Venmathi Muthu, Nayan Jain, Sakthivel Muthu, Jamith Basha Abdul Wahid, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7161974/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 06 Nov, 2025 Read the published version in Biological Trace Element Research → Version 1 posted 4 You are reading this latest preprint version Abstract The current study investigates the protective and therapeutic potential of Bacopa monnieri extract and synthetic antioxidants against lead-induced hematological toxicity in mice. HPLC analysis of Bacopa monnieri extract revealed a rich profile of bioactive compounds, including apigenin, luteolin flavonoids, bacosides, and bacopasaponins. In vivo experiments demonstrated significant hematological alterations in lead-exposed groups, with dose-dependent reductions in hemoglobin, red blood cell (RBC) count, platelet count, hematocrit, mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH), alongside increases in white blood cell (WBC) count. Lead exposure resulted in hemoglobin declines of up to 51.82% and RBC reductions of up to 59.32% in high-dose groups, indicating severe hematotoxic effects. Co-administration of Bacopa monnieri extract or synthetic antioxidants mitigated these alterations, with Bacopa monnieri showing superior protection by maintaining hemoglobin, RBC, and platelet levels closer to control values. Curative treatments further restored hematological parameters near baseline, highlighting the efficacy of Bacopa monnieri in reversing lead-induced toxicity. The findings of the study revealed dose-dependent statistically significant (p < 0.001) alterations in hematological parameters of lead intoxicated mice groups as compared to control groups, while co-treatment with synthetic antioxidants or Bacopa monnieri extract conferred protection and reduce these toxic effects. Notably, Bacopa monnieri showed higher efficacy in mitigating lead-induced toxicity to hematological system. These data imply that Bacopa monnieri has better potential to be an effective ameliorative therapeutic agent against lead toxicity as compared to synthetic alternatives. Toxicity Ethnopharmacology bioactive compounds Chromatography Clinical toxicology Chemico-biological interaction Lead Toxicity Bacopa monnieri Hematological parameters Antioxidant Figures Figure 1 1. Introduction The increasing prevalence of environmental toxins, particularly heavy metals like lead, has led to growing concerns regarding their impact on human health. Lead toxicity is considered as a serious environmental hazard of public health concern at global level due to its persistent occupational and industrial exposure [ 1 ]. Strong neurotoxic lead is known to cause a variety of harmful effects, including abnormalities in the hematological system. Exposure to lead upsets the delicate equilibrium between pro-oxidant and antioxidant processes, mostly by producing too many reactive oxygen species [ 2 ]. Oxidative stress brought on by this imbalance oxidizes structural proteins and modifies the permeability of blood cell membranes, ultimately resulting in hematological toxicity. The prevalence of lead poisoning worldwide, especially in areas where it is endemic, highlights the critical need for efficient treatment approaches to lessen the negative consequences of the metal. Bluish-white lustrous lead is extensively preferred metal in the world because of its unique physico-chemical properties of softness, low tensile strength, corrosion resistance, heaviness, malleability, ductility and poor conductibility. Lead is still used vehemently for numerous industrial and domestic activities in the developing countries for the production of explosives, fusible alloys, paint, crystal glass, kitchen utensils, crayons, ceramic glaze, pencil, pesticides, radiation shields, cosmetics, ayurvedic medicines, pipes, ship breaking instruments, building construction material and ammunition [ 3 ]. Major sources of lead exposure involved lead battery factories engaged in manufacturing and recycling, mining activities, smelting, refining, and cottage industries. Environmental contamination associated with non-biodegradable lead metal is a serious danger as its accumulation leads to bio-magnification and pose continuous threat to living organisms via its entry into food chain through myriads of pathways including food, water, soil or air [ 4 ]. Lead poisoning, sometimes referred to as plumbism or saturnism, happens when the body accumulates high concentrations of the heavy element lead [ 5 ]. Lead poisoning has serious, systemic effects on many different organ systems. According to Luo et al. (2007), clinical symptoms of lead poisoning can include anemia, encephalopathy, mental retardation, cognitive deficits, lethargy, loss of appetite, weight loss, dizziness, abdominal pain, constipation, vomiting, irritability, fatigue, anxiety, and the appearance of blue lines on the gums (known as lead lines). In extreme cases, the condition can result in coma and death [ 6 ]. It is believed that in living things, the hematological system is extremely susceptible to lead toxicity [ 7 ]. Blood is an important marker of physiological changes since it is a vital connective tissue that is constantly undergoing metabolic activities. Changes in hematological parameters might therefore function as early and trustworthy markers of the toxicity of lead on different tissues. The hemopoietic system has been demonstrated to be particularly negatively impacted by lead exposure [ 8 ]. As erythrocytes have a significant affinity for lead, they usually contain the majority of the lead metal in the blood. Lead causes elevated levels of aminolevulinic acid (ALA) and excessive formation of reactive oxygen species (ROS) by interfering with the activity of δ-aminolevulinic acid dehydratase (ALAD), which in turn impairs heme synthesis [ 9 ]. Hemoglobin oxidation results from ROS-induced membrane peroxidation, which in turn causes RBC hemolysis and, eventually, anemia. Chelation therapy is the mainstay of current therapeutic techniques to mitigate lead toxicity. However, chelation therapy has severe limits and side effects, even though it is effective in lowering lead levels [ 10 ]. In this regard, there is increasing interest in investigating substitute treatments that may provide a safer and more efficient way to treat lead-induced poisoning. Antioxidant treatment is one of the best alternatives that has shown promise. Synergism of synthetic antioxidants can prove to be beneficial in counteracting the free radical mediated oxidative stress in lead toxicity. Herbal treatments have garnered interest due to their possible therapeutic benefits, in addition to synthetic antioxidants [ 11 ]. Being well-known for its therapeutic properties and long use in traditional medicine, Bacopa monnieri has showed great potential as an antioxidant and neuroprotective agent [ 12 , 13 ]. Bacopa monnieri is a rich source of pharmacologically active phytochemicals, such as luteolin and apigenin, flavonoids, bacosides, and bacopasaponins. It has also demonstrated strong free radical scavenging action and antioxidant properties [ 14 ]. Because of this, it can be a great alternative therapeutic agent for treatment and prevention of oxidative stress and hematological toxicity induced by heavy metal lead. This study was undertaken to thoroughly assess the ameliorative effects of synthetic antioxidants (N-Acetyl Cysteine, Ascorbic acid, Tocopheryl acetate and Thiamine Mixture) as a combinational therapy approach and Bacopa monnieri as herbal antioxidant therapy approach in a model of lead-induced hematological toxicity in male Swiss albino mice. In comparative analysis of effectiveness of lead toxicity treatment with synthetic and herbal antioxidants, the research hopes to shed light on the mechanism via which these ameliorative agents lessen the harmful effects of lead and pinpoint the best therapeutic approaches for treatment of lead poisoning. The results of this study should have a major impact on public health, especially in areas where lead exposure is still a major concern. Administration of Bacopa monnieri could also prove to be new, safe, and very effective treatment for lead intoxication. 2. Materials and Methods 2.1 Experimental Animals Cadila Pharmaceuticals in Dholka, India, supplied healthy male Swiss Albino Mice ( Mus musculus ) weighing between 30–35 gram and aged 4–5 weeks. The animals were cared for in accordance with Animal Maintenance and Registration No. 167/1999/CPCSEA, which was given by the Ministry of Social Justice and Empowerment of the Indian government. 2.2 Animal Care The animals were housed in the animal facility of Department of Zoology, Gujarat University following the criteria established by Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), India. Individually housed in plastic enclosures, they were kept under a controlled 12-hour light-dark cycle at a temperature of 26 ± 2°C, following authorized experimental methods approved by the institutional animal ethics committee. The animals were fed normal commercial laboratory chow (Amrut laboratory animal feed from Pranav Agro Industries, which included 3625 kcal/kg energy, 22.10% crude protein, 4.10% crude oil, 4.45% crude fiber, 9.10% ash, and 0.75% sand silica) and tap water ad libitum . All animals were given a one-week acclimatization period before beginning treatment. The treatment was given orally once a day using a cannula attached to a hypodermic syringe, before their feeding routine. 2.3 Chemicals Analytical grade reagents with 99% purity were used in this study. The required chemicals, such as Tocopheryl acetate (Vitamin E), Ascorbic acid (Vitamin C), N-acetyl Cysteine (NAC) and Thiamine (Vitamin B 1 ) were obtained from reliable standard companies like Hi-Media, Merck Laboratory Pvt. Ltd., India and Sigma-Aldrich St. Louis, MO, USA. 2.4 Preparation of Reagents Lead acetate trihydrate, Ascorbic acid, N-Acetyl Cysteine, Thiamine, etc. reagents utilized for present in vivo study were prepared by dissolving in double distilled water in order to obtain the solutions of required concentrations. Tocopheryl acetate was given directly to the subjects in this study as Evion-emulsion syrup, which was obtained from Merck Laboratory. 2.5 Plant Collection and Preparation of Plant Extract In December, whole plant of Bacopa monnieri was harvested from the botanical garden of Gujarat University, Ahmedabad, India. The plant was identified and authenticated by the University's botany department, and a voucher specimen was preserved in the herbarium. Following collection, the plant was washed with double-distilled water, cut into small pieces, and air-dried in a shaded, dust-free area for one week at room temperature. After drying, the plant material was ground into a coarse powder. Soxhlet extraction was then conducted with 10 grams of this powder using 100 ml of 90% ethanol at 80°C for 15 hours. The crude extract was concentrated, air-dried, and stored at -10°C in a dark container for future analysis. 2.7 Characterization of Bacopa monnieri Extract by HPLC Analysis The primary bioactive components of the Bacopa monnieri ethanolic extract were identified and characterized using High Performance Liquid Chromatography (HPLC) with a Shimadzu Model LC 2010CHT and LC Solution software. A C-18 Phenomenex column (150 mm x 4.6 mm, 5 µm) was included in the HPLC system. 500 mg of the ethanolic extract was dissolved in 50 ml of HPLC-grade methanol in a 100 ml volumetric flask to prepare the sample. The extract was subjected to sonication for 10–15 minutes, cooled and made up to 100 ml with methanol. A 0.45 µm membrane filter was then used to filter the mixture. The plant extract was injected into the HPLC column in a 20 µl aliquot, and eluted using a mobile phase that contained 0.25% orthophosphoric acid in water and acetonitrile at a flow rate of 1.5 ml/min. The column oven was maintained at 25°C. The separation was carried out using gradient chromatographic technique for 45 minutes of run time. The separated components were detected using PDA detector at the wavelength of 205nm. The compounds in the chromatogram have been identified by comparing their retention time with the standard curve [ 15 ]. 2.8 Experimental Design Experimental design comprised of a total of 100 healthy adult Swiss strain male albino mice equally divided into ten experimental groups. Group-I to VIII were orally treated for 4-weeks. Group I animals were maintained without any treatment and represented untreated control group. They were given free access to feed and water ad libitum . Group II animals were orally administered mixture of synthetic antioxidants (N-Acetyl Cysteine-5.5mM/kg/day, Ascorbic acid-200mg/kg/day, Tocopheryl acetate-160mg/kg/day and Thiamine-30mg/kg/day) which served as synthetic antidote control group. Group III animals were orally treated with ethanolic extract of Bacopa monnieri (10mg/kg/day) which served as herbal antidote control group. Group IV, V and VI orally received low dose (160mg/kg/day), mid dose (266mg/kg/day) and high dose (320mg/kg/day) of lead acetate, respectively. Group VII animals were orally administered high dose (320mg/kg/day) of lead acetate along with synthetic antioxidants mixture in prescribed dosage for synergistic study. Group VIII animals were orally treated with high dose (320mg/kg/day) of lead acetate along with Bacopa monnieri ethanolic extract (10mg/kg/day) for synergistic study. Group IX animals were administered high dose (320mg/kg/day) of lead acetate along with prescribed dosage of synthetic antioxidants mixture for 4-weeks and supplementation of synthetic antioxidants was continued for next 2-weeks in order to analyze curative effect of synthetic antidote. Group X animals were treated with high dose (320mg/kg/day) of lead acetate along with Bacopa monnieri extract (10mg/kg/day) for 4-weeks and supplementation of plant extract was continued for next 2-weeks in order to analyze the curative potential of herbal antidote. With the use of pilot studies and the body of available literature, the experimental dosages for lead acetate and antidotes were chosen based on LD 50 values and laboratory standardization [ 16 – 18 ]. Table 1 provides a full description of the in vivo study's experimental design. Table 1 Experimental Design for In Vivo Study Type of Study Groups Experimental Group No. Experimental Groups No. of Animals Treatment Duration Day of Autopsy Control Study Group - I Untreated Control [UC] 10 4 - Weeks 29th Groups Group - II Synthetic Antidote Control [A-I] (NAC-5.5mM/kg/day + Ascorbic acid-200mg/kg/day + Tocopheryl acetate-160mg/kg/day + Thiamine-30mg/kg/day) 10 4 - Weeks 29th Group - III Herbal Antidote Control [A-II] ( Bacopa monnieri ethanolic Extract) (10mg/kg/day) 10 4 - Weeks 29th Toxin Exposed Group - IV Lead Acetate Low Dose [LD] (160mg/kg/day) 10 4 - Weeks 29th Groups Group - V Lead Acetate Mid Dose [MD] (266mg/kg/day) 10 4 - Weeks 29th Group - VI Lead Acetate High Dose [HD] (320mg/kg/day) 10 4 - Weeks 29th Synergistic Study Group - VII Lead Acetate High Dose + Synthetic Antidote [HD + A-I] 10 4 - Weeks 29th Groups Group -VIII Lead Acetate High Dose + Herbal Antidote [HD + A-II] 10 4 - Weeks 29th Curative Study Group - IX Lead Acetate High Dose and Synthetic Antidote (4-Weeks) + Only Synthetic Antidote (2-Weeks) [HD + A-I] © 10 6 - Weeks 43rd Groups Group - X Lead Acetate High Dose and Herbal Antidote (4-Weeks) + Only Herbal Antidote (2-Weeks) [HD + A-II] © 10 6 - Weeks 43rd 2.9 Sample Collection After an experimental period of four weeks for synergistic investigation and six weeks for curative study animals were fasted overnight and next day sacrificed by subjecting them to an excessive dosage of diethyl ether. For the purpose of estimating hematological parameters, blood samples from each experimental group were obtained by cardiac puncture and preserved in EDTA (Ethylene Diamine Tetra Acetic Acid) vials. Collected blood samples were stored at -20°C for further analysis. 2.10 Experimental Parameters In this study, hematological parameters of Hemoglobin (Hb), Total RBC, Total WBC, Platelets, Hematocrit (HCT), Mean Corpuscular Volume (MCV) and Mean Corpuscular Hemoglobin (MCH) were analyzed from blood samples of all experimental animals using automated hematology analyzer [Model: CELL-DYN 3700 SYSTEM] working on the principle of measurement of blood parameters by the Electrical Impedance method and equipped with an automated sample loader module. 2.11 Statistical Analysis The in vivo study data were statistically analysed using GraphPad Prism software, version 5.03. The results were represented as mean ± standard error of mean (SEM) for each parameter (n = 10). Comparison among different groups was made by One-way Analysis of Variance (ANOVA) followed by Tukey’s post hoc test. At the significance level of p < 0.05, a statistically significant difference was considered. 3. Results 3.1 Characterization of the Ethanolic Extract of Bacopa monnieri HPLC analysis of an ethanolic extract of Bacopa monnieri confirmed the presence of considerable number of bioactive phytochemicals in the forms of apigenin and luteolin flavonoids, as well as derivatives of jujubogenin in the forms of bacosides and bacopasaponins (Fig. 1 ). 3.2 Hematological Parameters Results of present in vivo study revealed statistically significant (p < 0.001) alteration in hematological parameters of lead administered groups of animals as compared to control groups. Results emphasized statistically significant decline in Hemoglobin level (Table-2), Total RBC count, Platelet count (Table-3), Hematocrit value, Mean corpuscular volume and Means corpuscular hemoglobin level (Table-4) as well as increment in Total WBC count (Table-3) in lead acetate exposed Groups IV, V and VI of mice as compared to control groups. The toxic effect observed was dose-dependent in manner (Table-5). Table 2 presents the effect of lead and antioxidants on blood hemoglobin levels in control and treated mice in vivo . The untreated control group (Group I) had an average hemoglobin level of 13.70 ± 0.060 g/dL. Mice administered with a synthetic antioxidant mixture (Group II) showed a hemoglobin level of 13.40 ± 0.075 g/dL, while those treated with Bacopa monnieri extract (Group III) had a level of 13.50 ± 0.080 g/dL. In contrast, mice exposed to low, mid, and high doses of lead acetate (Groups IV, V, and VI) exhibited significantly lower hemoglobin levels, with values of 11.40 ± 0.063 g/dL, 8.29 ± 0.009 g/dL, and 6.60 ± 0.058 g/dL, respectively (p < 0.001). Co-administration of high-dose lead with synthetic antioxidants (Group VII) or Bacopa monnieri extract (Group VIII) protected against reduction and maintained hemoglobin levels to 11.90 ± 0.092 g/dL and 12.50 ± 0.071 g/dL, respectively. Additionally, curative study groups with high-dose lead-exposure and co-treatment with synthetic antioxidants (Group IX) or Bacopa monnieri extract (Group X) for 6-weeks duration resulted in restoration of hemoglobin levels to 13.10 ± 0.042 g/dL and 13.30 ± 0.080 g/dL, respectively. Table 2 Effect of Lead and Antioxidants on Blood Hemoglobin Level of Control and Treated Mice In Vivo Group No. Experimental Groups Experimental Parameter Hemoglobin (Hb) g/dL Group-I Untreated Control (UC) 13.70 ± 0.060 Group-II Synthetic Antioxidants Mixture (A-I) 13.40 ± 0.075 Group-III Bacopa monnieri Extract (A-II) 13.50 ± 0.080 Group-IV Lead Acetate Low Dose (LD) 11.40 ± 0.063 a* Group-V Lead Acetate Mid Dose (MD) 8.29 ± 0.009 a* Group-VI Lead Acetate High Dose (HD) 6.60 ± 0.058 a* Group-VII HD + A-I 11.90 ± 0.092 b* Group-VIII HD + A-II 12.50 ± 0.071 b* Group-IX HD + A-I © 13.10 ± 0.042 b* Group-X HD + A-II © 13.30 ± 0.080 b* p – Values: * p < 0.001 Values are expressed as mean ± SEM; (n = 10) a as compared to control group b as compared to toxin (high dose lead acetate) exposed group (Group VI) No significant difference was noted between untreated control and antidotes control groups Table 3 illustrates the impact of lead exposure and antioxidant treatments on various hematological parameters in mice. In the untreated control group (Group I), baseline values were Total RBC at 7.67 ± 0.010 M/µl, Total WBC at 0.82 ± 0.002 K/µl, and Platelets at 1093.0 ± 1.578 K/µl. The synthetic antioxidants mixture (Group II) and Bacopa monnieri extract (Group III) had similar values to the control group, indicating no significant changes. However, lead acetate exposure significantly altered these parameters. Mice exposed to low dose lead acetate (Group IV) exhibited reduced Total RBC (5.03 ± 0.002 M/µl), increased Total WBC (1.54 ± 0.009 K/µl), and decreased Platelets (962.00 ± 0.955 K/µl). Mid dose (Group V) and high dose lead acetate (Group VI) resulted in even more pronounced reductions in Total RBC, with values of 4.84 ± 0.004 M/µl and 3.12 ± 0.002 M/µl, respectively, and Platelets dropping to 610.00 ± 0.558 K/µl and 466.50 ± 0.004 K/µl. Total WBC counts increased to 2.01 ± 0.002 K/µl and 2.36 ± 0.001 K/µl in these groups. Co-administration of antioxidants improved these hematological parameters, with high dose lead plus synthetic antioxidants (Group VII) showing Total RBC at 7.05 ± 0.001 M/µl, Total WBC at 0.98 ± 0.001 K/µl, and Platelets at 966.00 ± 1.000 K/µl, and high dose lead with Bacopa monnieri (Group VIII) showing even better results with Total RBC at 7.12 ± 0.001 M/µl, Total WBC at 0.97 ± 0.001 K/µl, and Platelets at 980.90 ± 1.149 K/µl. Continued treatment with antioxidants for curative study (Groups IX and X) showed further improvements, with Total RBC returning to 7.63 ± 0.001 M/µl and 7.64 ± 0.001 M/µl, Total WBC decreasing to 0.92 ± 0.002 K/µl and 0.91 ± 0.001 K/µl, and Platelets increasing to 1058.0 ± 1.174 K/µl and 1075.0 ± 0.919 K/µl. Table 3 Effect of Lead and Antioxidants on Hematological Parameters of Control and Treated Mice In Vivo Group No. Experimental Groups Experimental Parameters Total RBC M/µl Total WBC K/ µl Platelets K/µl Group-I Untreated Control (UC) 7.67 ± 0.010 0.82 ± 0.002 1093.0 ± 1.578 Group-II Synthetic Antioxidants Mixture (A-I) 7.64 ± 0.010 0.83 ± 0.001 1086.0 ± 0.856 Group-III Bacopa monnieri Extract (A-II) 7.66 ± 0.008 0.82 ± 0.001 1089.0 ± 0.895 Group-IV Lead Acetate Low Dose (LD) 5.03 ± 0.002 a* 1.54 ± 0.009 a* 962.00 ± 0.955 a* Group-V Lead Acetate Mid Dose (MD) 4.84 ± 0.004 a* 2.01 ± 0.002 a* 610.00 ± 0.558 a* Group-VI Lead Acetate High Dose (HD) 3.12 ± 0.002 a* 2.36 ± 0.001 a* 466.50 ± 0.004 a* Group-VII HD + A-I 7.05 ± 0.001 b* 0.98 ± 0.001 b* 966.00 ± 1.000 b* Group-VIII HD + A-II 7.12 ± 0.001 b* 0.97 ± 0.001 b* 980.90 ± 1.149 b* Group-IX HD + A-I © 7.63 ± 0.001 b* 0.92 ± 0.002 b* 1058.0 ± 1.174 b* Group-X HD + A-II © 7.64 ± 0.001 b* 0.91 ± 0.001 b* 1075.0 ± 0.919 b* p – Values: * p < 0.001 Values are expressed as mean ± SEM; (n = 10) a as compared to control group b as compared to toxin (high dose lead acetate) exposed group (Group VI) No significant difference was noted between untreated control and antidotes control groups Table 4 presents the impact of lead exposure and antioxidant treatments on various hematological parameters in mice. In the untreated control group (Group I), the hematocrit (HCT) was 35.80 ± 0.004%, mean corpuscular volume (MCV) was 50.60 ± 0.086 fL, and mean corpuscular hemoglobin (MCH) was 17.40 ± 0.041 pg. The synthetic antioxidants mixture (Group II) and Bacopa monnieri extract (Group III) showed similar values to the control group. Lead acetate exposure led to significant reductions in these parameters: low dose (Group IV) resulted in HCT at 24.00 ± 0.387%, MCV at 47.00 ± 0.258 fL, and MCH at 16.90 ± 0.027 pg; mid dose (Group V) showed HCT at 21.60 ± 0.074%, MCV at 45.50 ± 0.073 fL, and MCH at 16.70 ± 0.056 pg; and high dose (Group VI) resulted in HCT at 18.10 ± 0.107%, MCV at 44.00 ± 0.152 fL, and MCH at 15.50 ± 0.073 pg. Co-administration of high dose lead with antioxidants demonstrated improved parameters: high dose lead with synthetic antioxidants (Group VII) had HCT at 34.50 ± 0.081%, MCV at 49.00 ± 0.106 fL, and MCH at 16.20 ± 0.056 pg; high dose lead and Bacopa monnieri (Group VIII) showed HCT at 34.80 ± 0.104%, MCV at 49.10 ± 0.087 fL, and MCH at 16.50 ± 0.073 pg. Continued treatment with antioxidants for curative study of 6-weeks (Groups IX and X) further improved values, with HCT at 35.10 ± 0.032% and 35.30 ± 0.101%, MCV at 50.00 ± 0.191 fL and 50.00 ± 0.134 fL, and MCH at 16.90 ± 0.095 pg and 17.10 ± 0.101 pg, respectively. Table 4 Effect of Lead and Antioxidants on Hematological Parameters of Control and Treated Mice In Vivo Group No. Experimental Groups Experimental Parameters HCT (%) MCV (fL) MCH (pg) Group-I Untreated Control (UC) 35.80 ± 0.004 50.60 ± 0.086 17.40 ± 0.041 Group-II Synthetic Antioxidants Mixture (A-I) 35.90 ± 0.001 50.10 ± 0.111 17.00 ± 0.094 Group-III Bacopa monnieri Extract (A-II) 35.90 ± 0.002 50.40 ± 0.067 17.30 ± 0.047 Group-IV Lead Acetate Low Dose (LD) 24.00 ± 0.387 a* 47.00 ± 0.258 a* 16.90 ± 0.027 a* Group-V Lead Acetate Mid Dose (MD) 21.60 ± 0.074 a* 45.50 ± 0.073 a* 16.70 ± 0.056 a* Group-VI Lead Acetate High Dose (HD) 18.10 ± 0.107 a* 44.00 ± 0.152 a* 15.50 ± 0.073 a* Group-VII HD + A-I 34.50 ± 0.081 b* 49.00 ± 0.106 b* 16.20 ± 0.056 b* Group-VIII HD + A-II 34.80 ± 0.104 b* 49.10 ± 0.087 b* 16.50 ± 0.073 b* Group-IX HD + A-I © 35.10 ± 0.032 b* 50.00 ± 0.191 b* 16.90 ± 0.095 b* Group-X HD + A-II © 35.30 ± 0.101 b* 50.00 ± 0.134 b* 17.10 ± 0.101 b* p – Values: * p < 0.001 Values are expressed as mean ± SEM; (n = 10) a as compared to control group b as compared to toxin (high dose lead acetate) exposed group (Group VI) No significant difference was noted between untreated control and antidotes control groups Table − 5 Gross Effect of Lead and Antioxidants on Mice Blood In Vivo. (% of difference with respect to their control groups) Sr. No. Biochemical Parameters Group IV (LD) Group V (MD) Group VI (HD) Group VII (HD + A-I) Group VIII (HD + A-II) Group IX (HD + A-I) © Group X (HD + A-II) © 1. Hemoglobin 16.79 39.49 51.82 13.14 8.76 4.38 2.92 2. Total RBC 34.42 36.90 59.32 8.08 7.17 0.52 0.39 3. Total WBC 87.80* 145.12* 187.80* 19.51* 18.29* 12.20* 10.98* 4. Platelets 11.99 44.19 57.32 11.62 10.26 18.13 1.65 5. Hematocrit 32.96 39.66 49.44 3.63 2.79 1.96 1.40 6. Mean Corpuscular Volume 7.11 10.08 13.04 3.16 2.96 1.19 1.19 7. Mean Corpuscular Hemoglobin 2.87 4.02 10.92 6.90 5.17 2.87 1.72 All values are expressed in % of decrease or *increase Table 5 summarizes the gross effects of lead exposure and antioxidant treatments on various blood parameters in mice, expressed as the percentage difference compared to control groups. Lead acetate administration resulted in significant alterations: low dose (Group IV) led to a decrease in hemoglobin by 16.79%, decrease in total RBC by 34.42%, increase in total WBC by 87.80%, decrease in platelets by 11.99%, decrease in hematocrit by 32.96%, decrease in mean corpuscular volume (MCV) by 7.11%, and decrease in mean corpuscular hemoglobin (MCH) by 2.87% as compared to control group of animals. The mid dose (Group V) showed a decrease in hemoglobin by 39.49%, decrease in total RBC by 36.90%, increase in total WBC by 145.12%, decrease in platelets by 44.19%, decrease in hematocrit by 39.66%, decrease in MCV by 10.08%, and decrease in MCH by 4.02% as compared to control group. The high dose (Group VI) resulted in a decrease in hemoglobin by 51.82%, decrease in total RBC by 59.32%, increase in total WBC by 187.80%, decrease in platelets by 57.32%, decrease in hematocrit by 49.44%, decrease in MCV by 13.04%, and decrease in MCH by 10.92% as compared to control group of mice. In contrast, co-administration of high dose lead with synthetic antioxidants (Group VII) and high dose lead with Bacopa monnieri (Group VIII) mitigated these effects and protected against reductions in hemoglobin, total RBC, and other parameters being less pronounced. For instance, Group VII (HD + A-I) showed a decrease in hemoglobin by 13.14%, decrease in total RBC by 8.08%, increase in total WBC by 19.51%, decrease in platelets by 11.62%, decrease in hematocrit by 3.63%, decrease in MCV by 3.16%, and decrease in MCH by 6.90%. Group VIII (HD + A-II) displayed a decrease in hemoglobin by 8.76%, decrease in total RBC by 7.17%, increase in total WBC by 18.29%, decrease in platelets by 10.26%, decrease in hematocrit by 2.79%, decrease in MCV by 2.96%, and decrease in MCH by 5.17% as compared to control group. Curative treatment with antioxidants (Groups IX and X) showed even smaller deviations, with Group IX (HD + A-I ©) and Group X (HD + A-II ©) exhibiting minimal differences compared to controls, indicating significant ameliorative effects. Table 6 presents the gross effects of lead exposure and antioxidant treatments on mice blood parameters, expressed as percentage differences with respect to high dose lead acetate exposed Group VI. In Group VI mice, which were exposed to high dose lead acetate, significant changes were observed in various hematological parameters. In Group VI animals, the hemoglobin level was reduced by 51.82%, while total RBC decreased by 59.32%, and total WBC increased by 187.80% as compared to control group. Platelets decreased by 57.32%, hematocrit dropped by 49.44%, mean corpuscular volume (MCV) fell by 13.04%, and mean corpuscular hemoglobin (MCH) was reduced by 10.92% as compared to control group. In contrast, the addition of antioxidants showed considerable protection against alteration in hematological parameters. For instance, Group VII (HD + A-I) demonstrated an increase in hemoglobin by 80.30%, an increase in total RBC by 125.96%, and a decrease in total WBC by 58.47% compared to the high dose lead group. Platelets and hematocrit also improved by 107.07% and 90.61%, respectively in Group VII animals as compared to Group VI. MCV and MCH showed decrease by 11.36% and 4.52%, respectively in Group VII animals as compared to Group VI. Group VIII (HD + A-II) displayed an even more pronounced effect, with an increase in hemoglobin by 89.39%, an increase in total RBC by 128.21%, and a decrease in total WBC by 58.90% as compared to Group VI. Platelets and hematocrit increased by 110.27% and 92.27%, respectively, while MCV and MCH decreased by 11.59% and 6.45%, respectively in Group VIII animals as compared to Group VI. Groups IX and X, which received continuous antioxidant treatment after withdrawal of lead acetate showed near-normalization of these parameters to control group. Group IX (HD + A-I ©) achieved an increase in hemoglobin by 98.48%, an increase in total RBC by 144.55%, and a decrease in total WBC by 61.02% as compared to Group-VI. Platelets and hematocrit increased by 126.80% and 93.92%, respectively, with MCV and MCH decrease by 13.64% and 9.03%, respectively in Group IX animals as compared to Group VI. Similarly, Group X (HD + A-II ©) exhibited an increase in hemoglobin by 101.52%, an increase in total RBC by 144.87%, and a decrease in total WBC by 61.44% in Group X animals as compared to Group VI. Platelets and hematocrit rose by 130.44% and 95.03%, respectively, while MCV and MCH showed decrease by 13.64% and 10.32%, respectively in Group X animals as compared to Group VI. These results highlight the efficacy of antioxidants in mitigating the adverse effects of high dose lead exposure on blood parameters. Table − 6 Gross Effect of Lead and Antioxidants on Mice Blood In Vivo. (% of difference with respect to high dose lead acetate exposed groups) Sr. No. Biochemical Parameters Group VI (HD) (Relative to Control) Group VII (HD + A-I) Group VIII (HD + A-II) Group IX (HD + A-I) © Group X (HD + A-II) © 1. Hemoglobin 51.82 80.30* 89.39* 98.48* 101.52* 2. Total RBC 59.32 125.96* 128.21* 144.55* 144.87* 3. Total WBC 187.80* 58.47 58.90 61.02 61.44 4. Platelets 57.32 107.07* 110.27* 126.80* 130.44* 5. Hematocrit 49.44 90.61* 92.27* 93.92* 95.03* 6. Mean Corpuscular Volume 13.04 11.36* 11.59* 13.64* 13.64* 7. Mean Corpuscular Hemoglobin 10.92 4.52* 6.45* 9.03* 10.32* All values are expressed in % of decrease or *increase Results of present study has emphasized significant role of co-administration of synthetic antioxidant mixture or Bacopa monnieri extract in high dose lead-exposed mice groups of synergistic study. Group VII and Group VIII revealed statistically significant protective effect against alteration in hematological parameters as depicted by increase in hemoglobin (80.30% and 89.39% respectively), Total RBC (125.96% and 128.21% respectively), Platelets (107.07% and 110.27% respectively), Hematocrit (90.61% and 92.27% respectively), MCV (11.36% and 11.59% respectively), MCH (4.52% and 6.45% respectively) and reduction in Total WBC (58.47% and 58.90% respectively) as compared to high dose lead exposed mice Group VI (Table-6). Comparing the mice in the curative study groups IX and X to the mice in the high dose lead administered group VI, the curative study groups animals showed statistically significant amelioration against reductions in hemoglobin, total RBC, platelets, hematocrit, MCV, and MCH as well as elevation in total WBC. Groups VII and VIII, which received high doses of lead with synthetic and herbal antidotes respectively, did not exhibit the same improvements as compared to Groups IX and X in the curative study. Curative study Groups IX and X depicted significant increase in Hemoglobin content (98.48% and 101.52% respectively), Total RBC (144.55% and 144.87% respectively), Platelet count (126.80% and 130.44% respectively), Hematocrit value (93.92% and 95.03% respectively), MCV (13.64% and 13.64% respectively), MCH (9.03% and 10.32% respectively) and decrease in Total WBC (61.02% and 61.44% respectively) as compared to their respective high dose lead exposed Group VI (Table-6). These findings demonstrated that synthetic antioxidants mixture and Bacopa monnieri extract administration for 2-weeks aftermath withdrawal of lead acetate in respective group IX and Group X of animals resulted into normalization of all hematological parameters nearest to control group with more pronounced and remarkable curative effect of herbal antidote as compared to synthetic antioxidants mixture against lead induced hematological toxicity. The findings of the study demonstrated that the co-administration of a high dose of lead acetate with either a synthetic antioxidant mixture or Bacopa monnieri extract produced notable protective effects against alteration in the hematological damage caused by lead. In particular, the body's capacity to combat oxidative stress was improved when lead acetate and synthetic antioxidants were combined, leading to improvements in important blood parameters like hemoglobin and red blood cell counts due to synergistic effect. In a similar vein, significant protection against lead toxicity was obtained by combining lead acetate with Bacopa monnieri extract, which is well known for its strong antioxidant qualities. The abundant range of flavonoids and bioactive substances included in the plant extract facilitated the neutralization of reactive oxygen species leading to the restoration of normal hematological function. These results show that both therapeutic approaches greatly reduce the negative consequences of exposure to lead, indicating that they may be useful as therapeutic agents to prevent or ameliorate lead-induced oxidative damage and enhance blood health in general. 4. Discussion The findings of current in vivo study showed that, as compared to control groups, mice administered with lead acetate had significant alterations in hematological parameters of hemoglobin, total RBC count, total WBC count, platelets, hematocrit, mean corpuscular volume, and mean corpuscular hemoglobin. Exposure to lead has a substantial effect on biosynthesis of heme, peroxidase, cytochrome and catalase [ 19 ]. Lead obstructs a number of enzymatic stages in heme biosynthesis pathway; most notably delta-aminolevulinic acid dehydratase (ALAD), which is extremely vulnerable to the harmful effects of lead. Due to the fact that 70% of blood lead binds to ALAD, lead interferes with the enzyme's ability to function by directly attaching to the -SH groups that are necessary for the catalytic activity of the enzyme [ 20 ]. ALAD catalyzes formation of porphobilinogen from delta-aminolevulinic acid (ALA) and ferrochelatase, which incorporates iron into protoporphyrin. Failure to condense two molecules of delta-aminolevulinic acid (ALA) for formation of porphobilinogen by ALAD and insertion of iron into protoporphyrin by ferrochelatase results in depressed heme formation [ 21 ]. Heme synthesis impairment prevents the improvement in red blood cell population. Lead is also known to readily inhibit porphobilinogen synthase in erythrocytes responsible for regulating H b synthesis in vivo and in vitro [ 22 ]. The decline in Hb aftermath lead administration was also noted in other scientific studies in accordance of our results [ 23 , 24 ]. Because of their strong affinity for lead, red blood cells usually contain most of the circulating lead in the blood stream [ 25 ]. Due to a number of key factors, including the inhibition of heme and hemoglobin biosynthesis, induction of hemoglobin auto-oxidation, alteration of erythrocyte morphology by direct interaction of lead metal with RBC membranes, induction of lipid peroxidation [ 26 ] and limited ability of RBCs to repair their damaged components, a severe oxidative stress is generated in lead-exposed RBCs, which ultimately results in their decreased survival (Caylak et al. 2008). Lead exposure may account for increasing fragility of erythrocytes [ 27 ] and inducing hemolysis as well as origin of defective cells that can be eliminated by spleen [ 28 ]. Lead's inhibitory effect on the essential erythrocyte enzyme glucose-6-phosphate dehydrogenase and the erythrocytes' shortened life span [ 29 ] could be the cause of the decline in hemoglobin and total red blood cell count. Results of lead toxicity mediated reduction in RBC observed in our study corroborates with the findings of other scientific researchers [ 30 , 31 ]. Reduced heme synthesis, which results in anemia, is one of the most significant impacts of lead exposure on the hematological system. Decrease in red blood cell survival, increase in the rate of RBC destruction, decrease in the rate of RBC synthesis, decrease in the production of hemoglobin in the bone marrow, or combination of all these mechanisms may be responsible for the development of anemia in lead-induced toxicity [ 32 ]. Given that the red blood cell count, hemoglobin concentration, and packed cell volume all affect the validity of these indexes, lead acetate's toxic effects on hemoglobin concentration and red blood cell count may have contributed to the changes in mean corpuscular volume and mean corpuscular hemoglobin concentration observed in this study. Following exposure to lead, the resulted alteration in Platelet count, hematocrit, MCV, and MCH were consistent with findings of prior studies [ 33 ]. Lead-induced tissue inflammation could be the cause of the elevated Total WBC levels. Increase in Total WBC count obtained in our study also corroborates with the findings of other researchers [ 34 , 35 ]. Lead induced alterations in hematological parameters were ameliorated by co-supplementation of mixture of synthetic antioxidants comprising of Ascorbic acid, Tocopheryl acetate, N-acetyl cysteine and Thiamine (Group-VII and IX) or ethanolic extract of Bacopa monnieri (Group-VIII and X). Vitamin-C has been reported as a good antioxidant to overcome lead induced hematotoxicity (Sharma and Panwar, 2013). Animals are more vulnerable to the hemolytic effects of lead poisoning when they are deficient in Vitamin-E. Our results corroborate with the reports of other researchers which highlight that, in lead-treated animals, Vitamin-C and Vitamin-E significantly increased packed cell volume, hemoglobin concentration, red blood cell count, and neutrophil percentage while significantly decreasing white blood cell count and lymphocyte percentage. Kamruzmman (2006) and Haque (2005) also reported that lead content in blood, liver and kidney was significantly reduced following administration of Vitamin-C and Vitamin-E. N-Acetyl Cysteine exhibited antioxidant capacity against lead toxicity via promoting maintenance of intracellular reduced glutathione levels and scavenging of free radicals [ 33 , 36 ]. Thiamine has been shown to have antioxidant potential because of its ability to scavenge free radicals and ability to form complex with lead metal resulting into its excretion. It has been demonstrated that Bacopa monnieri , a multi-purpose traditional herb with a wide range of medicinal uses, exhibits antioxidant effects by chelating metal ions, breaking oxidative chain reactions, scavenging reactive oxygen species such as peroxides, superoxides, and hydroxyl radicals, and enhancing the activities of antioxidative defense enzymes [ 37 ]. It also functions as a potent blood purifier. Phytochemical screening of Bacopa monnieri extract has revealed presence of variety of primary bioactive constituents. These included flavonoids such as glucoronyl-7-apigenin and glucortonyl-7-luteolin, alkaloids, D-mannitol, potassium salts, and steroids, as well as saponins such as hersaponin, jujubogenin , and pseudojujubogenin [ 32 ]. Numerous studies revealed that the presence of distinctive active constituent, a dammarane type tri-terpenoid saponin known as "Bacoside-A" as chemical derivative of jujubogenin in Bacopa monnieri was primarily responsible for the pharmacological actions of plant, including its antioxidant activity. HPLC analysis data also supported the presence of these phytochemicals in Bacopa monnieri extract in our study. Ameliorative potential of the synthetic antioxidant’s mixture and Bacopa monnieri exhibited protective role against lead induced hematotoxicity in synergistic study groups. A significant amelioration observed in all studied hematological parameters of curative study groups receiving 4-weeks exposure of lead acetate and 6-weeks antidote treatment suggested that lead toxicity mediated oxidative stress in vivo could be reversible by exogenous supplementation of pharmacological manipulations rich in antioxidant potential. The antioxidant qualities of both synthetic and herbal antidotes may be responsible for lowering of lipid peroxidation, preserving the activity of delta-aminolevulinic acid dehydratase (ALAD), stabilizing the plasma membrane of red blood cells, and lowering hemolysis in these cells, which in turn preserve blood hemoglobin, total WBC count, platelets, MCV, and MHC nearest to the control group. Therefore, by reducing oxidative stress and ensuring the preservation of antioxidant equilibrium in Swiss albino mice cells, synthetic antioxidants and Bacopa monnieri functioned as mitigating agents against lead-induced hematological toxicity. This work clearly shows that lead exposure severely exaggerated the hematological system in Swiss albino mice by inducing oxidative stress, which changes blood cell membrane permeability and oxidizes structural proteins. Lead co-administration with synthetic antioxidants or Bacopa monnieri extract provided significant protection against this toxicity, with the latter proving to be more effective. This therapeutic efficacy is most likely owing to the wide range of active phytochemicals present in Bacopa monnieri , which function as powerful free radical scavengers. The findings indicate that lead-induced hematological damage can be both transient and reversible through use of Bacopa monnieri and synthetic antioxidants, emphasizing their ameliorative potential as safer antidotes against lead toxicity. This study provides important information for establishing novel treatment strategies in occupational toxicology and pharmacology to combat lead poisoning. Declarations Acknowledgement Dr. FS is thankful to the Department of Life-Science, University School of Sciences, Gujarat University, Gujarat, India for providing lab and research facility. The authors extend their appreciation to the Deanship of Scientific Research at Northern Border University, Arar, KSA for funding this research work through the project number "NBU-FFR-2024-1329-16". The authors extend their appreciation to the Deanship of Scientific Research at King Khalid University for funding this work through the General Research Project under the grant number (RGP 2/252/45). Competing Interests: Authors have declared that there are no conflicts of interests. Ethical approval: N/A Informed consent: N/A Author Contributions: Conceptualization, Writing- Original Draft and Supervision: FS and NJ.; Writing - Review & Editing: KN, JBAW, MS, Data curation, Validation: SM.; Resources, Project Administration: VM. Data Availability Statement: N/A Funding statement: Deanship of Scientific Research at Northern Border University, Arar, KSA (NBU-FFR-2024-1329-16). Deanship of Scientific Research at King Khalid University (RGP 2/252/45). 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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-7161974","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":488634425,"identity":"7e01082e-411d-4d8a-8b60-c2e6bde64e26","order_by":0,"name":"Flora Shah","email":"","orcid":"","institution":"Gujarat University","correspondingAuthor":false,"prefix":"","firstName":"Flora","middleName":"","lastName":"Shah","suffix":""},{"id":488634426,"identity":"91333fe8-dcd3-45da-9f79-439dce03c3dd","order_by":1,"name":"Venmathi Muthu","email":"","orcid":"","institution":"Saveetha Institute of Medical and Technical Sciences (SIMATS)","correspondingAuthor":false,"prefix":"","firstName":"Venmathi","middleName":"","lastName":"Muthu","suffix":""},{"id":488634427,"identity":"a848e59f-de68-42ba-935a-04df37e033f3","order_by":2,"name":"Nayan Jain","email":"","orcid":"","institution":"Gujarat University","correspondingAuthor":false,"prefix":"","firstName":"Nayan","middleName":"","lastName":"Jain","suffix":""},{"id":488634428,"identity":"956ab134-af03-4205-b92b-9e2a96e01dda","order_by":3,"name":"Sakthivel Muthu","email":"","orcid":"","institution":"Saveetha Medical College and Hospital, Saveetha Institute of Medical and Technical Sciences (SIMATS)","correspondingAuthor":false,"prefix":"","firstName":"Sakthivel","middleName":"","lastName":"Muthu","suffix":""},{"id":488634429,"identity":"9e29074c-b194-42db-9055-33d7441c750a","order_by":4,"name":"Jamith Basha Abdul Wahid","email":"","orcid":"","institution":"Northern Border University","correspondingAuthor":false,"prefix":"","firstName":"Jamith","middleName":"Basha Abdul","lastName":"Wahid","suffix":""},{"id":488634430,"identity":"73cb12b5-bf23-4fee-9eb3-76bca7994713","order_by":5,"name":"Mumtaj Shah","email":"","orcid":"","institution":"King khalid University","correspondingAuthor":false,"prefix":"","firstName":"Mumtaj","middleName":"","lastName":"Shah","suffix":""},{"id":488634431,"identity":"7db6029f-0961-4d4b-ab5a-8d08b9dd6a40","order_by":6,"name":"Karuppiah Nagaraj","email":"data:image/png;base64,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","orcid":"","institution":"Saveetha Medical College and Hospital, Saveetha Institute of Medical and Technical Sciences (SIMATS)","correspondingAuthor":true,"prefix":"","firstName":"Karuppiah","middleName":"","lastName":"Nagaraj","suffix":""}],"badges":[],"createdAt":"2025-07-19 05:23:20","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7161974/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7161974/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12011-025-04849-x","type":"published","date":"2025-11-06T15:57:30+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":88044773,"identity":"2c5e63ae-ee24-432a-b8a9-64e63055f50d","added_by":"auto","created_at":"2025-07-31 18:03:26","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":55020,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eHPLC Analysis of Ethanolic Extract of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBacopa monnieri\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7161974/v1/59d9a8b9fbcdb132d8e01e0b.png"},{"id":95564126,"identity":"3ff5b10b-f75f-4c50-9753-ba2d41c8f803","added_by":"auto","created_at":"2025-11-10 16:08:08","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1241573,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7161974/v1/f3765e32-574f-440b-b1a8-fa12dd44b8f7.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Efficacy of Bacopa monnieri in Mitigating Lead-Induced Blood Toxicity in Mice Compared to Synthetic Antioxidants","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe increasing prevalence of environmental toxins, particularly heavy metals like lead, has led to growing concerns regarding their impact on human health. Lead toxicity is considered as a serious environmental hazard of public health concern at global level due to its persistent occupational and industrial exposure [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Strong neurotoxic lead is known to cause a variety of harmful effects, including abnormalities in the hematological system. Exposure to lead upsets the delicate equilibrium between pro-oxidant and antioxidant processes, mostly by producing too many reactive oxygen species [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Oxidative stress brought on by this imbalance oxidizes structural proteins and modifies the permeability of blood cell membranes, ultimately resulting in hematological toxicity. The prevalence of lead poisoning worldwide, especially in areas where it is endemic, highlights the critical need for efficient treatment approaches to lessen the negative consequences of the metal.\u003c/p\u003e\u003cp\u003eBluish-white lustrous lead is extensively preferred metal in the world because of its unique physico-chemical properties of softness, low tensile strength, corrosion resistance, heaviness, malleability, ductility and poor conductibility. Lead is still used vehemently for numerous industrial and domestic activities in the developing countries for the production of explosives, fusible alloys, paint, crystal glass, kitchen utensils, crayons, ceramic glaze, pencil, pesticides, radiation shields, cosmetics, ayurvedic medicines, pipes, ship breaking instruments, building construction material and ammunition [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Major sources of lead exposure involved lead battery factories engaged in manufacturing and recycling, mining activities, smelting, refining, and cottage industries. Environmental contamination associated with non-biodegradable lead metal is a serious danger as its accumulation leads to bio-magnification and pose continuous threat to living organisms via its entry into food chain through myriads of pathways including food, water, soil or air [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eLead poisoning, sometimes referred to as plumbism or saturnism, happens when the body accumulates high concentrations of the heavy element lead [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Lead poisoning has serious, systemic effects on many different organ systems. According to Luo \u003cem\u003eet al.\u003c/em\u003e (2007), clinical symptoms of lead poisoning can include anemia, encephalopathy, mental retardation, cognitive deficits, lethargy, loss of appetite, weight loss, dizziness, abdominal pain, constipation, vomiting, irritability, fatigue, anxiety, and the appearance of blue lines on the gums (known as lead lines). In extreme cases, the condition can result in coma and death [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIt is believed that in living things, the hematological system is extremely susceptible to lead toxicity [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Blood is an important marker of physiological changes since it is a vital connective tissue that is constantly undergoing metabolic activities. Changes in hematological parameters might therefore function as early and trustworthy markers of the toxicity of lead on different tissues. The hemopoietic system has been demonstrated to be particularly negatively impacted by lead exposure [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. As erythrocytes have a significant affinity for lead, they usually contain the majority of the lead metal in the blood. Lead causes elevated levels of aminolevulinic acid (ALA) and excessive formation of reactive oxygen species (ROS) by interfering with the activity of δ-aminolevulinic acid dehydratase (ALAD), which in turn impairs heme synthesis [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Hemoglobin oxidation results from ROS-induced membrane peroxidation, which in turn causes RBC hemolysis and, eventually, anemia.\u003c/p\u003e\u003cp\u003eChelation therapy is the mainstay of current therapeutic techniques to mitigate lead toxicity. However, chelation therapy has severe limits and side effects, even though it is effective in lowering lead levels [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. In this regard, there is increasing interest in investigating substitute treatments that may provide a safer and more efficient way to treat lead-induced poisoning. Antioxidant treatment is one of the best alternatives that has shown promise. Synergism of synthetic antioxidants can prove to be beneficial in counteracting the free radical mediated oxidative stress in lead toxicity. Herbal treatments have garnered interest due to their possible therapeutic benefits, in addition to synthetic antioxidants [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Being well-known for its therapeutic properties and long use in traditional medicine, \u003cem\u003eBacopa monnieri\u003c/em\u003e has showed great potential as an antioxidant and neuroprotective agent [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. \u003cem\u003eBacopa monnieri\u003c/em\u003e is a rich source of pharmacologically active phytochemicals, such as luteolin and apigenin, flavonoids, bacosides, and bacopasaponins. It has also demonstrated strong free radical scavenging action and antioxidant properties [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Because of this, it can be a great alternative therapeutic agent for treatment and prevention of oxidative stress and hematological toxicity induced by heavy metal lead.\u003c/p\u003e\u003cp\u003eThis study was undertaken to thoroughly assess the ameliorative effects of synthetic antioxidants (N-Acetyl Cysteine, Ascorbic acid, Tocopheryl acetate and Thiamine Mixture) as a combinational therapy approach and \u003cem\u003eBacopa monnieri\u003c/em\u003e as herbal antioxidant therapy approach in a model of lead-induced hematological toxicity in male Swiss albino mice. In comparative analysis of effectiveness of lead toxicity treatment with synthetic and herbal antioxidants, the research hopes to shed light on the mechanism \u003cem\u003evia\u003c/em\u003e which these ameliorative agents lessen the harmful effects of lead and pinpoint the best therapeutic approaches for treatment of lead poisoning. The results of this study should have a major impact on public health, especially in areas where lead exposure is still a major concern. Administration of \u003cem\u003eBacopa monnieri\u003c/em\u003e could also prove to be new, safe, and very effective treatment for lead intoxication.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1 Experimental Animals\u003c/h2\u003e\u003cp\u003eCadila Pharmaceuticals in Dholka, India, supplied healthy male Swiss Albino Mice (\u003cem\u003eMus musculus\u003c/em\u003e) weighing between 30\u0026ndash;35 gram and aged 4\u0026ndash;5 weeks. The animals were cared for in accordance with Animal Maintenance and Registration No. 167/1999/CPCSEA, which was given by the Ministry of Social Justice and Empowerment of the Indian government.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2 Animal Care\u003c/h2\u003e\u003cp\u003e The animals were housed in the animal facility of Department of Zoology, Gujarat University following the criteria established by Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), India. Individually housed in plastic enclosures, they were kept under a controlled 12-hour light-dark cycle at a temperature of 26\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C, following authorized experimental methods approved by the institutional animal ethics committee. The animals were fed normal commercial laboratory chow (Amrut laboratory animal feed from Pranav Agro Industries, which included 3625 kcal/kg energy, 22.10% crude protein, 4.10% crude oil, 4.45% crude fiber, 9.10% ash, and 0.75% sand silica) and tap water \u003cem\u003ead libitum\u003c/em\u003e. All animals were given a one-week acclimatization period before beginning treatment. The treatment was given orally once a day using a cannula attached to a hypodermic syringe, before their feeding routine.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3 Chemicals\u003c/h2\u003e\u003cp\u003eAnalytical grade reagents with 99% purity were used in this study. The required chemicals, such as Tocopheryl acetate (Vitamin E), Ascorbic acid (Vitamin C), N-acetyl Cysteine (NAC) and Thiamine (Vitamin B\u003csub\u003e1\u003c/sub\u003e) were obtained from reliable standard companies like Hi-Media, Merck Laboratory Pvt. Ltd., India and Sigma-Aldrich St. Louis, MO, USA.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.4 Preparation of Reagents\u003c/h2\u003e\u003cp\u003eLead acetate trihydrate, Ascorbic acid, N-Acetyl Cysteine, Thiamine, etc. reagents utilized for present \u003cem\u003ein vivo\u003c/em\u003e study were prepared by dissolving in double distilled water in order to obtain the solutions of required concentrations. Tocopheryl acetate was given directly to the subjects in this study as Evion-emulsion syrup, which was obtained from Merck Laboratory.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.5 Plant Collection and Preparation of Plant Extract\u003c/h2\u003e\u003cp\u003eIn December, whole plant of \u003cem\u003eBacopa monnieri\u003c/em\u003e was harvested from the botanical garden of Gujarat University, Ahmedabad, India. The plant was identified and authenticated by the University's botany department, and a voucher specimen was preserved in the herbarium. Following collection, the plant was washed with double-distilled water, cut into small pieces, and air-dried in a shaded, dust-free area for one week at room temperature. After drying, the plant material was ground into a coarse powder. Soxhlet extraction was then conducted with 10 grams of this powder using 100 ml of 90% ethanol at 80\u0026deg;C for 15 hours. The crude extract was concentrated, air-dried, and stored at -10\u0026deg;C in a dark container for future analysis.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003e2.7 Characterization of Bacopa monnieri Extract by HPLC Analysis\u003c/h2\u003e\u003cp\u003eThe primary bioactive components of the \u003cem\u003eBacopa monnieri\u003c/em\u003e ethanolic extract were identified and characterized using High Performance Liquid Chromatography (HPLC) with a Shimadzu Model LC 2010CHT and LC Solution software. A C-18 Phenomenex column (150 mm x 4.6 mm, 5 \u0026micro;m) was included in the HPLC system. 500 mg of the ethanolic extract was dissolved in 50 ml of HPLC-grade methanol in a 100 ml volumetric flask to prepare the sample. The extract was subjected to sonication for 10\u0026ndash;15 minutes, cooled and made up to 100 ml with methanol. A 0.45 \u0026micro;m membrane filter was then used to filter the mixture. The plant extract was injected into the HPLC column in a 20 \u0026micro;l aliquot, and eluted using a mobile phase that contained 0.25% orthophosphoric acid in water and acetonitrile at a flow rate of 1.5 ml/min. The column oven was maintained at 25\u0026deg;C. The separation was carried out using gradient chromatographic technique for 45 minutes of run time. The separated components were detected using PDA detector at the wavelength of 205nm. The compounds in the chromatogram have been identified by comparing their retention time with the standard curve [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003e2.8 Experimental Design\u003c/h2\u003e\u003cp\u003eExperimental design comprised of a total of 100 healthy adult Swiss strain male albino mice equally divided into ten experimental groups. Group-I to VIII were orally treated for 4-weeks. Group I animals were maintained without any treatment and represented untreated control group. They were given free access to feed and water \u003cem\u003ead libitum\u003c/em\u003e. Group II animals were orally administered mixture of synthetic antioxidants (N-Acetyl Cysteine-5.5mM/kg/day, Ascorbic acid-200mg/kg/day, Tocopheryl acetate-160mg/kg/day and Thiamine-30mg/kg/day) which served as synthetic antidote control group. Group III animals were orally treated with ethanolic extract of \u003cem\u003eBacopa monnieri\u003c/em\u003e (10mg/kg/day) which served as herbal antidote control group. Group IV, V and VI orally received low dose (160mg/kg/day), mid dose (266mg/kg/day) and high dose (320mg/kg/day) of lead acetate, respectively. Group VII animals were orally administered high dose (320mg/kg/day) of lead acetate along with synthetic antioxidants mixture in prescribed dosage for synergistic study. Group VIII animals were orally treated with high dose (320mg/kg/day) of lead acetate along with \u003cem\u003eBacopa monnieri\u003c/em\u003e ethanolic extract (10mg/kg/day) for synergistic study. Group IX animals were administered high dose (320mg/kg/day) of lead acetate along with prescribed dosage of synthetic antioxidants mixture for 4-weeks and supplementation of synthetic antioxidants was continued for next 2-weeks in order to analyze curative effect of synthetic antidote. Group X animals were treated with high dose (320mg/kg/day) of lead acetate along with \u003cem\u003eBacopa monnieri\u003c/em\u003e extract (10mg/kg/day) for 4-weeks and supplementation of plant extract was continued for next 2-weeks in order to analyze the curative potential of herbal antidote. With the use of pilot studies and the body of available literature, the experimental dosages for lead acetate and antidotes were chosen based on LD\u003csub\u003e50\u003c/sub\u003e values and laboratory standardization [\u003cspan additionalcitationids=\"CR17\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e provides a full description of the \u003cem\u003ein vivo\u003c/em\u003e study's experimental design.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eExperimental Design for \u003cem\u003eIn Vivo\u003c/em\u003e Study\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eType of\u003c/p\u003e\u003cp\u003eStudy Groups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eExperimental Group No.\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eExperimental Groups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eNo.\u003c/p\u003e\u003cp\u003eof\u003c/p\u003e\u003cp\u003eAnimals\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eTreatment Duration\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eDay\u003c/p\u003e\u003cp\u003eof\u003c/p\u003e\u003cp\u003eAutopsy\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eControl\u003c/p\u003e\u003cp\u003eStudy\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup - I\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eUntreated Control [UC]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29th\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroups\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup - II\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSynthetic Antidote Control [A-I]\u003c/p\u003e\u003cp\u003e(NAC-5.5mM/kg/day\u0026thinsp;+\u0026thinsp;Ascorbic acid-200mg/kg/day\u0026thinsp;+\u0026thinsp;Tocopheryl acetate-160mg/kg/day\u0026thinsp;+\u0026thinsp;Thiamine-30mg/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29th\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup - III\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eHerbal Antidote Control [A-II]\u003c/p\u003e\u003cp\u003e(\u003cem\u003eBacopa monnieri\u003c/em\u003e ethanolic Extract) (10mg/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29th\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eToxin\u003c/p\u003e\u003cp\u003eExposed\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup - IV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eLead Acetate Low Dose [LD]\u003c/p\u003e\u003cp\u003e(160mg/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29th\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroups\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup - V\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eLead Acetate Mid Dose [MD]\u003c/p\u003e\u003cp\u003e(266mg/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29th\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup - VI\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eLead Acetate High Dose [HD]\u003c/p\u003e\u003cp\u003e(320mg/kg/day)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29th\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSynergistic\u003c/p\u003e\u003cp\u003eStudy\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup - VII\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eLead Acetate High Dose\u0026thinsp;+\u0026thinsp;Synthetic Antidote [HD\u0026thinsp;+\u0026thinsp;A-I]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29th\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroups\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup -VIII\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eLead Acetate High Dose\u0026thinsp;+\u0026thinsp;Herbal Antidote [HD\u0026thinsp;+\u0026thinsp;A-II]\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29th\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCurative\u003c/p\u003e\u003cp\u003eStudy\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup - IX\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eLead Acetate High Dose and Synthetic Antidote (4-Weeks) +\u003c/p\u003e\u003cp\u003eOnly Synthetic Antidote (2-Weeks) [HD\u0026thinsp;+\u0026thinsp;A-I] \u0026copy;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e43rd\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroups\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGroup - X\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eLead Acetate High Dose and Herbal Antidote (4-Weeks) +\u003c/p\u003e\u003cp\u003eOnly Herbal Antidote (2-Weeks) [HD\u0026thinsp;+\u0026thinsp;A-II] \u0026copy;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6 - Weeks\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e43rd\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\u003ch2\u003e2.9 Sample Collection\u003c/h2\u003e\u003cp\u003eAfter an experimental period of four weeks for synergistic investigation and six weeks for curative study animals were fasted overnight and next day sacrificed by subjecting them to an excessive dosage of diethyl ether. For the purpose of estimating hematological parameters, blood samples from each experimental group were obtained by cardiac puncture and preserved in EDTA (Ethylene Diamine Tetra Acetic Acid) vials. Collected blood samples were stored at -20\u0026deg;C for further analysis.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003e2.10 Experimental Parameters\u003c/h2\u003e\u003cp\u003eIn this study, hematological parameters of Hemoglobin (Hb), Total RBC, Total WBC, Platelets, Hematocrit (HCT), Mean Corpuscular Volume (MCV) and Mean Corpuscular Hemoglobin (MCH) were analyzed from blood samples of all experimental animals using automated hematology analyzer [Model: CELL-DYN 3700 SYSTEM] working on the principle of measurement of blood parameters by the Electrical Impedance method and equipped with an automated sample loader module.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003e2.11 Statistical Analysis\u003c/h2\u003e\u003cp\u003eThe \u003cem\u003ein vivo\u003c/em\u003e study data were statistically analysed using GraphPad Prism software, version 5.03. The results were represented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of mean (SEM) for each parameter (n\u0026thinsp;=\u0026thinsp;10). Comparison among different groups was made by One-way Analysis of Variance (ANOVA) followed by Tukey\u0026rsquo;s \u003cem\u003epost hoc\u003c/em\u003e test. At the significance level of p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, a statistically significant difference was considered.\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003e3.1 Characterization of the Ethanolic Extract of Bacopa monnieri\u003c/h2\u003e\u003cp\u003eHPLC analysis of an ethanolic extract of \u003cem\u003eBacopa monnieri\u003c/em\u003e confirmed the presence of considerable number of bioactive phytochemicals in the forms of apigenin and luteolin flavonoids, as well as derivatives of \u003cem\u003ejujubogenin\u003c/em\u003e in the forms of bacosides and bacopasaponins (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003e3.2 Hematological Parameters\u003c/h2\u003e\u003cp\u003eResults of present \u003cem\u003ein vivo\u003c/em\u003e study revealed statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) alteration in hematological parameters of lead administered groups of animals as compared to control groups. Results emphasized statistically significant decline in Hemoglobin level (Table-2), Total RBC count, Platelet count (Table-3), Hematocrit value, Mean corpuscular volume and Means corpuscular hemoglobin level (Table-4) as well as increment in Total WBC count (Table-3) in lead acetate exposed Groups IV, V and VI of mice as compared to control groups. The toxic effect observed was dose-dependent in manner (Table-5).\u003c/p\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e presents the effect of lead and antioxidants on blood hemoglobin levels in control and treated mice \u003cem\u003ein vivo\u003c/em\u003e. The untreated control group (Group I) had an average hemoglobin level of 13.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.060 g/dL. Mice administered with a synthetic antioxidant mixture (Group II) showed a hemoglobin level of 13.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.075 g/dL, while those treated with \u003cem\u003eBacopa monnieri\u003c/em\u003e extract (Group III) had a level of 13.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.080 g/dL. In contrast, mice exposed to low, mid, and high doses of lead acetate (Groups IV, V, and VI) exhibited significantly lower hemoglobin levels, with values of 11.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.063 g/dL, 8.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.009 g/dL, and 6.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.058 g/dL, respectively (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Co-administration of high-dose lead with synthetic antioxidants (Group VII) or \u003cem\u003eBacopa monnieri\u003c/em\u003e extract (Group VIII) protected against reduction and maintained hemoglobin levels to 11.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.092 g/dL and 12.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.071 g/dL, respectively. Additionally, curative study groups with high-dose lead-exposure and co-treatment with synthetic antioxidants (Group IX) or \u003cem\u003eBacopa monnieri\u003c/em\u003e extract (Group X) for 6-weeks duration resulted in restoration of hemoglobin levels to 13.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.042 g/dL and 13.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.080 g/dL, respectively.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eEffect of Lead and Antioxidants on Blood Hemoglobin Level of Control and Treated Mice \u003cem\u003eIn Vivo\u003c/em\u003e\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eGroup No.\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eExperimental Groups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eExperimental Parameter\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eHemoglobin (Hb)\u003c/p\u003e\u003cp\u003eg/dL\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-I\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eUntreated Control (UC)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.70 \u0026plusmn; 0.060\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-II\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSynthetic Antioxidants Mixture (A-I)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.40 \u0026plusmn; 0.075\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-III\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cem\u003eBacopa monnieri\u003c/em\u003e Extract (A-II)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.50 \u0026plusmn; 0.080\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-IV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLead Acetate Low Dose (LD)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.40 \u0026plusmn; 0.063\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-V\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLead Acetate Mid Dose (MD)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e8.29 \u0026plusmn; 0.009\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-VI\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLead Acetate High Dose (HD)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.60 \u0026plusmn; 0.058\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-VII\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHD\u0026thinsp;+\u0026thinsp;A-I\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e11.90 \u0026plusmn; 0.092\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-VIII\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHD\u0026thinsp;+\u0026thinsp;A-II\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12.50 \u0026plusmn; 0.071\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-IX\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHD\u0026thinsp;+\u0026thinsp;A-I \u0026copy;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.10 \u0026plusmn; 0.042\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroup-X\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHD\u0026thinsp;+\u0026thinsp;A-II \u0026copy;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13.30 \u0026plusmn; 0.080\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"3\"\u003ep \u0026ndash; Values: \u003csup\u003e*\u003c/sup\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"3\"\u003eValues are expressed as mean \u0026plusmn; SEM; (n\u0026thinsp;=\u0026thinsp;10)\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"3\"\u003e\u003csup\u003ea\u003c/sup\u003e as compared to control group\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd colspan=\"3\"\u003e\u003csup\u003eb\u003c/sup\u003e as compared to toxin (high dose lead acetate) exposed group (Group VI)\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eNo significant difference was noted between untreated control and antidotes control groups\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eTable 3 illustrates the impact of lead exposure and antioxidant treatments on various hematological parameters in mice. In the untreated control group (Group I), baseline values were Total RBC at 7.67 ± 0.010 M/µl, Total WBC at 0.82 ± 0.002 K/µl, and Platelets at 1093.0 ± 1.578 K/µl. The synthetic antioxidants mixture (Group II) and \u003cem\u003eBacopa monnieri\u003c/em\u003e extract (Group III) had similar values to the control group, indicating no significant changes. However, lead acetate exposure significantly altered these parameters. Mice exposed to low dose lead acetate (Group IV) exhibited reduced Total RBC (5.03 ± 0.002 M/µl), increased Total WBC (1.54 ± 0.009 K/µl), and decreased Platelets (962.00 ± 0.955 K/µl). Mid dose (Group V) and high dose lead acetate (Group VI) resulted in even more pronounced reductions in Total RBC, with values of 4.84 ± 0.004 M/µl and 3.12 ± 0.002 M/µl, respectively, and Platelets dropping to 610.00 ± 0.558 K/µl and 466.50 ± 0.004 K/µl. Total WBC counts increased to 2.01 ± 0.002 K/µl and 2.36 ± 0.001 K/µl in these groups. Co-administration of antioxidants improved these hematological parameters, with high dose lead plus synthetic antioxidants (Group VII) showing Total RBC at 7.05 ± 0.001 M/µl, Total WBC at 0.98 ± 0.001 K/µl, and Platelets at 966.00 ± 1.000 K/µl, and high dose lead with \u003cem\u003eBacopa monnieri\u003c/em\u003e (Group VIII) showing even better results with Total RBC at 7.12 ± 0.001 M/µl, Total WBC at 0.97 ± 0.001 K/µl, and Platelets at 980.90 ± 1.149 K/µl. Continued treatment with antioxidants for curative study (Groups IX and X) showed further improvements, with Total RBC returning to 7.63 ± 0.001 M/µl and 7.64 ± 0.001 M/µl, Total WBC decreasing to 0.92 ± 0.002 K/µl and 0.91 ± 0.001 K/µl, and Platelets increasing to 1058.0 ± 1.174 K/µl and 1075.0 ± 0.919 K/µl.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 3\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eEffect of Lead and Antioxidants on Hematological Parameters of Control and Treated Mice \u003cem\u003eIn Vivo\u003c/em\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eGroup No.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eExperimental Groups\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eExperimental Parameters\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTotal RBC\u003c/p\u003e\n \u003cp\u003eM/µl\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTotal WBC\u003c/p\u003e\n \u003cp\u003eK/ µl\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePlatelets\u003c/p\u003e\n \u003cp\u003eK/µl\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-I\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUntreated Control (UC)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.67 ± 0.010\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.82 ± 0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1093.0 ± 1.578\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-II\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSynthetic Antioxidants Mixture (A-I)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.64 ± 0.010\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.83 ± 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1086.0 ± 0.856\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-III\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eBacopa monnieri\u003c/em\u003e Extract (A-II)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.66 ± 0.008\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.82 ± 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1089.0 ± 0.895\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-IV\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLead Acetate Low Dose (LD)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.03 ± 0.002\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.54 ± 0.009\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e962.00 ± 0.955\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-V\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLead Acetate Mid Dose (MD)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.84 ± 0.004\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.01 ± 0.002\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e610.00 ± 0.558\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-VI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLead Acetate High Dose (HD)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.12 ± 0.002\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.36 ± 0.001\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e466.50 ± 0.004\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-VII\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHD + A-I\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.05 ± 0.001\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.98 ± 0.001\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e966.00 ± 1.000\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-VIII\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHD + A-II\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.12 ± 0.001\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.97 ± 0.001\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e980.90 ± 1.149\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-IX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHD + A-I ©\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.63 ± 0.001\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.92 ± 0.002\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1058.0 ± 1.174\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-X\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHD + A-II ©\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.64 ± 0.001\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.91 ± 0.001\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1075.0 ± 0.919\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003ep – Values: \u003csup\u003e*\u003c/sup\u003ep \u0026lt; 0.001\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eValues are expressed as mean ± SEM; (n = 10)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e\u003csup\u003ea\u003c/sup\u003e as compared to control group\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e\u003csup\u003eb\u003c/sup\u003e as compared to toxin (high dose lead acetate) exposed group (Group VI)\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv\u003e\n \u003cp\u003eNo significant difference was noted between untreated control and antidotes control groups\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003eTable 4 presents the impact of lead exposure and antioxidant treatments on various hematological parameters in mice. In the untreated control group (Group I), the hematocrit (HCT) was 35.80 ± 0.004%, mean corpuscular volume (MCV) was 50.60 ± 0.086 fL, and mean corpuscular hemoglobin (MCH) was 17.40 ± 0.041 pg. The synthetic antioxidants mixture (Group II) and \u003cem\u003eBacopa monnieri\u003c/em\u003e extract (Group III) showed similar values to the control group. Lead acetate exposure led to significant reductions in these parameters: low dose (Group IV) resulted in HCT at 24.00 ± 0.387%, MCV at 47.00 ± 0.258 fL, and MCH at 16.90 ± 0.027 pg; mid dose (Group V) showed HCT at 21.60 ± 0.074%, MCV at 45.50 ± 0.073 fL, and MCH at 16.70 ± 0.056 pg; and high dose (Group VI) resulted in HCT at 18.10 ± 0.107%, MCV at 44.00 ± 0.152 fL, and MCH at 15.50 ± 0.073 pg. Co-administration of high dose lead with antioxidants demonstrated improved parameters: high dose lead with synthetic antioxidants (Group VII) had HCT at 34.50 ± 0.081%, MCV at 49.00 ± 0.106 fL, and MCH at 16.20 ± 0.056 pg; high dose lead and \u003cem\u003eBacopa monnieri\u003c/em\u003e (Group VIII) showed HCT at 34.80 ± 0.104%, MCV at 49.10 ± 0.087 fL, and MCH at 16.50 ± 0.073 pg. Continued treatment with antioxidants for curative study of 6-weeks (Groups IX and X) further improved values, with HCT at 35.10 ± 0.032% and 35.30 ± 0.101%, MCV at 50.00 ± 0.191 fL and 50.00 ± 0.134 fL, and MCH at 16.90 ± 0.095 pg and 17.10 ± 0.101 pg, respectively.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 4\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eEffect of Lead and Antioxidants on Hematological Parameters of Control and Treated Mice \u003cem\u003eIn Vivo\u003c/em\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eGroup No.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eExperimental Groups\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eExperimental Parameters\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHCT\u003c/p\u003e\n \u003cp\u003e(%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMCV\u003c/p\u003e\n \u003cp\u003e(fL)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMCH\u003c/p\u003e\n \u003cp\u003e(pg)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-I\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUntreated Control (UC)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e35.80 ± 0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50.60 ± 0.086\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.40 ± 0.041\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-II\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSynthetic Antioxidants Mixture (A-I)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e35.90 ± 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50.10 ± 0.111\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.00 ± 0.094\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-III\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eBacopa monnieri\u003c/em\u003e Extract (A-II)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e35.90 ± 0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50.40 ± 0.067\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.30 ± 0.047\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-IV\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLead Acetate Low Dose (LD)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e24.00 ± 0.387\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e47.00 ± 0.258\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.90 ± 0.027\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-V\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLead Acetate Mid Dose (MD)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.60 ± 0.074\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e45.50 ± 0.073\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.70 ± 0.056\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-VI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLead Acetate High Dose (HD)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18.10 ± 0.107\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44.00 ± 0.152\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.50 ± 0.073\u003csup\u003ea*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-VII\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHD + A-I\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e34.50 ± 0.081\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e49.00 ± 0.106\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.20 ± 0.056\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-VIII\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHD + A-II\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e34.80 ± 0.104\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e49.10 ± 0.087\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.50 ± 0.073\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-IX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHD + A-I ©\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e35.10 ± 0.032\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50.00 ± 0.191\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.90 ± 0.095\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup-X\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHD + A-II ©\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e35.30 ± 0.101\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50.00 ± 0.134\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.10 ± 0.101\u003csup\u003eb*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003ep – Values: \u003csup\u003e*\u003c/sup\u003ep \u0026lt; 0.001\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eValues are expressed as mean ± SEM; (n = 10)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e\u003csup\u003ea\u003c/sup\u003e as compared to control group\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e\u003csup\u003eb\u003c/sup\u003e as compared to toxin (high dose lead acetate) exposed group (Group VI)\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eNo significant difference was noted between untreated control and antidotes control groups\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable id=\"Tab5\" border=\"1\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"9\"\u003e\n \u003cp\u003eTable − 5 Gross Effect of Lead and Antioxidants on Mice Blood \u003cem\u003eIn Vivo.\u003c/em\u003e (% of difference with respect to their control groups)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSr.\u003c/p\u003e\n \u003cp\u003eNo.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBiochemical\u003c/p\u003e\n \u003cp\u003eParameters\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup IV\u003c/p\u003e\n \u003cp\u003e(LD)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup V\u003c/p\u003e\n \u003cp\u003e(MD)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup VI\u003c/p\u003e\n \u003cp\u003e(HD)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup VII\u003c/p\u003e\n \u003cp\u003e(HD + A-I)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup VIII\u003c/p\u003e\n \u003cp\u003e(HD + A-II)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup IX\u003c/p\u003e\n \u003cp\u003e(HD + A-I) ©\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup X\u003c/p\u003e\n \u003cp\u003e(HD + A-II) ©\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHemoglobin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e39.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e51.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.92\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal RBC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e34.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e36.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e59.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal WBC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e87.80*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e145.12*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e187.80*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19.51*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18.29*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.20*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.98*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePlatelets\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e57.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.65\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHematocrit\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e32.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e39.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e49.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean Corpuscular Volume\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.19\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean Corpuscular Hemoglobin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"9\"\u003e\n \u003cp\u003eAll values are expressed in % of decrease or *increase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eTable 5 summarizes the gross effects of lead exposure and antioxidant treatments on various blood parameters in mice, expressed as the percentage difference compared to control groups. Lead acetate administration resulted in significant alterations: low dose (Group IV) led to a decrease in hemoglobin by 16.79%, decrease in total RBC by 34.42%, increase in total WBC by 87.80%, decrease in platelets by 11.99%, decrease in hematocrit by 32.96%, decrease in mean corpuscular volume (MCV) by 7.11%, and decrease in mean corpuscular hemoglobin (MCH) by 2.87% as compared to control group of animals. The mid dose (Group V) showed a decrease in hemoglobin by 39.49%, decrease in total RBC by 36.90%, increase in total WBC by 145.12%, decrease in platelets by 44.19%, decrease in hematocrit by 39.66%, decrease in MCV by 10.08%, and decrease in MCH by 4.02% as compared to control group. The high dose (Group VI) resulted in a decrease in hemoglobin by 51.82%, decrease in total RBC by 59.32%, increase in total WBC by 187.80%, decrease in platelets by 57.32%, decrease in hematocrit by 49.44%, decrease in MCV by 13.04%, and decrease in MCH by 10.92% as compared to control group of mice. In contrast, co-administration of high dose lead with synthetic antioxidants (Group VII) and high dose lead with \u003cem\u003eBacopa monnieri\u0026nbsp;\u003c/em\u003e(Group VIII) mitigated these effects and protected against reductions in hemoglobin, total RBC, and other parameters being less pronounced. For instance, Group VII (HD + A-I) showed a decrease in hemoglobin by 13.14%, decrease in total RBC by 8.08%, increase in total WBC by 19.51%, decrease in platelets by 11.62%, decrease in hematocrit by 3.63%, decrease in MCV by 3.16%, and decrease in MCH by 6.90%. Group VIII (HD + A-II) displayed a decrease in hemoglobin by 8.76%, decrease in total RBC by 7.17%, increase in total WBC by 18.29%, decrease in platelets by 10.26%, decrease in hematocrit by 2.79%, decrease in MCV by 2.96%, and decrease in MCH by 5.17% as compared to control group. Curative treatment with antioxidants (Groups IX and X) showed even smaller deviations, with Group IX (HD + A-I ©) and Group X (HD + A-II ©) exhibiting minimal differences compared to controls, indicating significant ameliorative effects.\u0026nbsp;\u003c/p\u003e\n\u003cdiv align=\"left\"\u003eTable 6 presents the gross effects of lead exposure and antioxidant treatments on mice blood parameters, expressed as percentage differences with respect to high dose lead acetate exposed Group VI. In Group VI mice, which were exposed to high dose lead acetate, significant changes were observed in various hematological parameters. In Group VI animals, the hemoglobin level was reduced by 51.82%, while total RBC decreased by 59.32%, and total WBC increased by 187.80% as compared to control group. Platelets decreased by 57.32%, hematocrit dropped by 49.44%, mean corpuscular volume (MCV) fell by 13.04%, and mean corpuscular hemoglobin (MCH) was reduced by 10.92% as compared to control group. In contrast, the addition of antioxidants showed considerable protection against alteration in hematological parameters. For instance, Group VII (HD + A-I) demonstrated an increase in hemoglobin by 80.30%, an increase in total RBC by 125.96%, and a decrease in total WBC by 58.47% compared to the high dose lead group. Platelets and hematocrit also improved by 107.07% and 90.61%, respectively in Group VII animals as compared to Group VI. MCV and MCH showed decrease by 11.36% and 4.52%, respectively in Group VII animals as compared to Group VI. Group VIII (HD + A-II) displayed an even more pronounced effect, with an increase in hemoglobin by 89.39%, an increase in total RBC by 128.21%, and a decrease in total WBC by 58.90% as compared to Group VI. Platelets and hematocrit increased by 110.27% and 92.27%, respectively, while MCV and MCH decreased by 11.59% and 6.45%, respectively in Group VIII animals as compared to Group VI. Groups IX and X, which received continuous antioxidant treatment after withdrawal of lead acetate showed near-normalization of these parameters to control group. Group IX (HD + A-I ©) achieved an increase in hemoglobin by 98.48%, an increase in total RBC by 144.55%, and a decrease in total WBC by 61.02% as compared to Group-VI. Platelets and hematocrit increased by 126.80% and 93.92%, respectively, with MCV and MCH decrease by 13.64% and 9.03%, respectively in Group IX animals as compared to Group VI. Similarly, Group X (HD + A-II ©) exhibited an increase in hemoglobin by 101.52%, an increase in total RBC by 144.87%, and a decrease in total WBC by 61.44% in Group X animals as compared to Group VI. Platelets and hematocrit rose by 130.44% and 95.03%, respectively, while MCV and MCH showed decrease by 13.64% and 10.32%, respectively in Group X animals as compared to Group VI. These results highlight the efficacy of antioxidants in mitigating the adverse effects of high dose lead exposure on blood parameters.\u0026nbsp;\u003c/div\u003e\n\u003cdiv align=\"left\"\u003e\u003cbr\u003e\u003c/div\u003e\n\u003ctable id=\"Tab6\" border=\"1\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003eTable − 6 Gross Effect of Lead and Antioxidants on Mice Blood \u003cem\u003eIn Vivo.\u003c/em\u003e (% of difference with respect to high dose lead acetate exposed groups)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSr.\u003c/p\u003e\n \u003cp\u003eNo.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBiochemical\u003c/p\u003e\n \u003cp\u003eParameters\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup VI (HD)\u003c/p\u003e\n \u003cp\u003e(Relative to Control)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup VII\u003c/p\u003e\n \u003cp\u003e(HD + A-I)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup VIII\u003c/p\u003e\n \u003cp\u003e(HD + A-II)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup IX\u003c/p\u003e\n \u003cp\u003e(HD + A-I) ©\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGroup X\u003c/p\u003e\n \u003cp\u003e(HD + A-II) ©\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHemoglobin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e51.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e80.30*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e89.39*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e98.48*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e101.52*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal RBC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e59.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e125.96*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e128.21*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e144.55*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e144.87*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal WBC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e187.80*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e58.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e58.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e61.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e61.44\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePlatelets\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e57.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e107.07*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e110.27*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e126.80*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e130.44*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHematocrit\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e49.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e90.61*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e92.27*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e93.92*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e95.03*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean Corpuscular Volume\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.36*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.59*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.64*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.64*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMean Corpuscular Hemoglobin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.52*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.45*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9.03*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.32*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003eAll values are expressed in % of decrease or *increase\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eResults of present study has emphasized significant role of co-administration of synthetic antioxidant mixture or \u003cem\u003eBacopa monnieri\u003c/em\u003e extract in high dose lead-exposed mice groups of synergistic study. Group VII and Group VIII revealed statistically significant protective effect against alteration in hematological parameters as depicted by increase in hemoglobin (80.30% and 89.39% respectively), Total RBC (125.96% and 128.21% respectively), Platelets (107.07% and 110.27% respectively), Hematocrit (90.61% and 92.27% respectively), MCV (11.36% and 11.59% respectively), MCH (4.52% and 6.45% respectively) and reduction in Total WBC (58.47% and 58.90% respectively) as compared to high dose lead exposed mice Group VI (Table-6).\u003c/p\u003e\n\u003cp\u003eComparing the mice in the curative study groups IX and X to the mice in the high dose lead administered group VI, the curative study groups animals showed statistically significant amelioration against reductions in hemoglobin, total RBC, platelets, hematocrit, MCV, and MCH as well as elevation in total WBC. Groups VII and VIII, which received high doses of lead with synthetic and herbal antidotes respectively, did not exhibit the same improvements as compared to Groups IX and X in the curative study. Curative study Groups IX and X depicted significant increase in Hemoglobin content (98.48% and 101.52% respectively), Total RBC (144.55% and 144.87% respectively), Platelet count (126.80% and 130.44% respectively), Hematocrit value (93.92% and 95.03% respectively), MCV (13.64% and 13.64% respectively), MCH (9.03% and 10.32% respectively) and decrease in Total WBC (61.02% and 61.44% respectively) as compared to their respective high dose lead exposed Group VI (Table-6). These findings demonstrated that synthetic antioxidants mixture and \u003cem\u003eBacopa monnieri\u003c/em\u003e extract administration for 2-weeks aftermath withdrawal of lead acetate in respective group IX and Group X of animals resulted into normalization of all hematological parameters nearest to control group with more pronounced and remarkable curative effect of herbal antidote as compared to synthetic antioxidants mixture against lead induced hematological toxicity.\u003c/p\u003e\n\u003cp\u003eThe findings of the study demonstrated that the co-administration of a high dose of lead acetate with either a synthetic antioxidant mixture or \u003cem\u003eBacopa monnieri\u003c/em\u003e extract produced notable protective effects against alteration in the hematological damage caused by lead. In particular, the body's capacity to combat oxidative stress was improved when lead acetate and synthetic antioxidants were combined, leading to improvements in important blood parameters like hemoglobin and red blood cell counts due to synergistic effect. In a similar vein, significant protection against lead toxicity was obtained by combining lead acetate with \u003cem\u003eBacopa monnieri\u003c/em\u003e extract, which is well known for its strong antioxidant qualities. The abundant range of flavonoids and bioactive substances included in the plant extract facilitated the neutralization of reactive oxygen species leading to the restoration of normal hematological function. These results show that both therapeutic approaches greatly reduce the negative consequences of exposure to lead, indicating that they may be useful as therapeutic agents to prevent or ameliorate lead-induced oxidative damage and enhance blood health in general.\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThe findings of current \u003cem\u003ein vivo\u003c/em\u003e study showed that, as compared to control groups, mice administered with lead acetate had significant alterations in hematological parameters of hemoglobin, total RBC count, total WBC count, platelets, hematocrit, mean corpuscular volume, and mean corpuscular hemoglobin. Exposure to lead has a substantial effect on biosynthesis of heme, peroxidase, cytochrome and catalase [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Lead obstructs a number of enzymatic stages in heme biosynthesis pathway; most notably delta-aminolevulinic acid dehydratase (ALAD), which is extremely vulnerable to the harmful effects of lead. Due to the fact that 70% of blood lead binds to ALAD, lead interferes with the enzyme's ability to function by directly attaching to the -SH groups that are necessary for the catalytic activity of the enzyme [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. ALAD catalyzes formation of porphobilinogen from delta-aminolevulinic acid (ALA) and ferrochelatase, which incorporates iron into protoporphyrin. Failure to condense two molecules of delta-aminolevulinic acid (ALA) for formation of porphobilinogen by ALAD and insertion of iron into protoporphyrin by ferrochelatase results in depressed heme formation [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Heme synthesis impairment prevents the improvement in red blood cell population. Lead is also known to readily inhibit porphobilinogen synthase in erythrocytes responsible for regulating H\u003csub\u003eb\u003c/sub\u003e synthesis \u003cem\u003ein vivo\u003c/em\u003e and \u003cem\u003ein vitro\u003c/em\u003e [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. The decline in Hb aftermath lead administration was also noted in other scientific studies in accordance of our results [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Because of their strong affinity for lead, red blood cells usually contain most of the circulating lead in the blood stream [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Due to a number of key factors, including the inhibition of heme and hemoglobin biosynthesis, induction of hemoglobin auto-oxidation, alteration of erythrocyte morphology by direct interaction of lead metal with RBC membranes, induction of lipid peroxidation [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e] and limited ability of RBCs to repair their damaged components, a severe oxidative stress is generated in lead-exposed RBCs, which ultimately results in their decreased survival (Caylak \u003cem\u003eet al.\u003c/em\u003e 2008).\u003c/p\u003e\u003cp\u003eLead exposure may account for increasing fragility of erythrocytes [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e] and inducing hemolysis as well as origin of defective cells that can be eliminated by spleen [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Lead's inhibitory effect on the essential erythrocyte enzyme glucose-6-phosphate dehydrogenase and the erythrocytes' shortened life span [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e] could be the cause of the decline in hemoglobin and total red blood cell count. Results of lead toxicity mediated reduction in RBC observed in our study corroborates with the findings of other scientific researchers [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Reduced heme synthesis, which results in anemia, is one of the most significant impacts of lead exposure on the hematological system. Decrease in red blood cell survival, increase in the rate of RBC destruction, decrease in the rate of RBC synthesis, decrease in the production of hemoglobin in the bone marrow, or combination of all these mechanisms may be responsible for the development of anemia in lead-induced toxicity [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eGiven that the red blood cell count, hemoglobin concentration, and packed cell volume all affect the validity of these indexes, lead acetate's toxic effects on hemoglobin concentration and red blood cell count may have contributed to the changes in mean corpuscular volume and mean corpuscular hemoglobin concentration observed in this study. Following exposure to lead, the resulted alteration in Platelet count, hematocrit, MCV, and MCH were consistent with findings of prior studies [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Lead-induced tissue inflammation could be the cause of the elevated Total WBC levels. Increase in Total WBC count obtained in our study also corroborates with the findings of other researchers [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eLead induced alterations in hematological parameters were ameliorated by co-supplementation of mixture of synthetic antioxidants comprising of Ascorbic acid, Tocopheryl acetate, N-acetyl cysteine and Thiamine (Group-VII and IX) or ethanolic extract of \u003cem\u003eBacopa monnieri\u003c/em\u003e (Group-VIII and X). Vitamin-C has been reported as a good antioxidant to overcome lead induced hematotoxicity (Sharma and Panwar, 2013). Animals are more vulnerable to the hemolytic effects of lead poisoning when they are deficient in Vitamin-E. Our results corroborate with the reports of other researchers which highlight that, in lead-treated animals, Vitamin-C and Vitamin-E significantly increased packed cell volume, hemoglobin concentration, red blood cell count, and neutrophil percentage while significantly decreasing white blood cell count and lymphocyte percentage. Kamruzmman (2006) and Haque (2005) also reported that lead content in blood, liver and kidney was significantly reduced following administration of Vitamin-C and Vitamin-E. N-Acetyl Cysteine exhibited antioxidant capacity against lead toxicity via promoting maintenance of intracellular reduced glutathione levels and scavenging of free radicals [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Thiamine has been shown to have antioxidant potential because of its ability to scavenge free radicals and ability to form complex with lead metal resulting into its excretion.\u003c/p\u003e\u003cp\u003eIt has been demonstrated that \u003cem\u003eBacopa monnieri\u003c/em\u003e, a multi-purpose traditional herb with a wide range of medicinal uses, exhibits antioxidant effects by chelating metal ions, breaking oxidative chain reactions, scavenging reactive oxygen species such as peroxides, superoxides, and hydroxyl radicals, and enhancing the activities of antioxidative defense enzymes [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. It also functions as a potent blood purifier. Phytochemical screening of \u003cem\u003eBacopa monnieri\u003c/em\u003e extract has revealed presence of variety of primary bioactive constituents. These included flavonoids such as glucoronyl-7-apigenin and glucortonyl-7-luteolin, alkaloids, D-mannitol, potassium salts, and steroids, as well as saponins such as \u003cem\u003ehersaponin, jujubogenin\u003c/em\u003e, and \u003cem\u003epseudojujubogenin\u003c/em\u003e [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Numerous studies revealed that the presence of distinctive active constituent, a dammarane type tri-terpenoid saponin known as \"Bacoside-A\" as chemical derivative of \u003cem\u003ejujubogenin\u003c/em\u003e in \u003cem\u003eBacopa monnieri\u003c/em\u003e was primarily responsible for the pharmacological actions of plant, including its antioxidant activity. HPLC analysis data also supported the presence of these phytochemicals in \u003cem\u003eBacopa monnieri\u003c/em\u003e extract in our study.\u003c/p\u003e\u003cp\u003eAmeliorative potential of the synthetic antioxidant\u0026rsquo;s mixture and \u003cem\u003eBacopa monnieri\u003c/em\u003e exhibited protective role against lead induced hematotoxicity in synergistic study groups. A significant amelioration observed in all studied hematological parameters of curative study groups receiving 4-weeks exposure of lead acetate and 6-weeks antidote treatment suggested that lead toxicity mediated oxidative stress \u003cem\u003ein vivo\u003c/em\u003e could be reversible by exogenous supplementation of pharmacological manipulations rich in antioxidant potential. The antioxidant qualities of both synthetic and herbal antidotes may be responsible for lowering of lipid peroxidation, preserving the activity of delta-aminolevulinic acid dehydratase (ALAD), stabilizing the plasma membrane of red blood cells, and lowering hemolysis in these cells, which in turn preserve blood hemoglobin, total WBC count, platelets, MCV, and MHC nearest to the control group. Therefore, by reducing oxidative stress and ensuring the preservation of antioxidant equilibrium in Swiss albino mice cells, synthetic antioxidants and \u003cem\u003eBacopa monnieri\u003c/em\u003e functioned as mitigating agents against lead-induced hematological toxicity. This work clearly shows that lead exposure severely exaggerated the hematological system in Swiss albino mice by inducing oxidative stress, which changes blood cell membrane permeability and oxidizes structural proteins. Lead co-administration with synthetic antioxidants or \u003cem\u003eBacopa monnieri\u003c/em\u003e extract provided significant protection against this toxicity, with the latter proving to be more effective. This therapeutic efficacy is most likely owing to the wide range of active phytochemicals present in \u003cem\u003eBacopa monnieri\u003c/em\u003e, which function as powerful free radical scavengers. The findings indicate that lead-induced hematological damage can be both transient and reversible through use of \u003cem\u003eBacopa monnieri\u003c/em\u003e and synthetic antioxidants, emphasizing their ameliorative potential as safer antidotes against lead toxicity. This study provides important information for establishing novel treatment strategies in occupational toxicology and pharmacology to combat lead poisoning.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDr. FS is thankful to the Department of Life-Science, University School of Sciences, Gujarat University, Gujarat, India for providing lab and research facility. The authors extend their appreciation to the Deanship of Scientific Research at Northern Border University, Arar, KSA for funding this research work through the project number \u0026quot;NBU-FFR-2024-1329-16\u0026quot;. The authors extend their appreciation to the Deanship of Scientific Research at King Khalid University for funding this work through the General Research Project under the grant number (RGP 2/252/45).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003c/strong\u003e\u003cstrong\u003eCompeting Interests:\u0026nbsp;\u003c/strong\u003eAuthors have declared that there are no conflicts of interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval:\u003c/strong\u003e N/A\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed consent:\u003c/strong\u003e N/A\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions:\u003c/strong\u003e Conceptualization, Writing- Original Draft and Supervision: FS and NJ.; Writing - Review \u0026amp; Editing: KN, JBAW, MS, Data curation, Validation: SM.; Resources, Project Administration: VM.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement:\u003c/strong\u003e N/A\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding statement:\u003c/strong\u003e Deanship of Scientific Research at Northern Border University, Arar, KSA (NBU-FFR-2024-1329-16). Deanship of Scientific Research at King Khalid University (RGP 2/252/45).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAugustyniak, A., Bartosz, G., Cipak, A., Duburs, G., Hor\u0026aacute;kova, L.U., Luczaj, W., Majekova, M., Odysseos, A.D., Rackova, L., Skrzydlewska, E. and Stefek, M. 2010. Natural and synthetic antioxidants: an updated overview. \u003cem\u003eFree Radical Research\u003c/em\u003e, 44(10), pp.1216-1262.\u003c/li\u003e\n\u003cli\u003eATSDR (Agency for Toxic Substances and Disease Registry). 2001. The nature and extent of lead poisoning in children in the United States: A report to Congress. \u003cem\u003eAtlanta: US Department of Health and Human Services; DHHS Report\u003c/em\u003e, no. 99-2966. \u003c/li\u003e\n\u003cli\u003eBhattacharya, S.K., Kumar, A., and Ghosal, S., 2000. Effect of\u003cem\u003e Bacopa monniera\u003c/em\u003e on animal models of Alzheimer disease and perturbed central cholinergic markers of congnition in rats. 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Impacts of ascorbic acid and thiamine supplementation at different concentrations on lead toxicity in testis. \u003cem\u003eClin Chim Acta\u003c/em\u003e, 370: 82-88.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"biological-trace-element-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bter","sideBox":"Learn more about [Biological Trace Element Research](https://www.springer.com/journal/12011)","snPcode":"12011","submissionUrl":"https://submission.nature.com/new-submission/12011/3","title":"Biological Trace Element Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Toxicity, Ethnopharmacology, bioactive compounds, Chromatography, Clinical toxicology, Chemico-biological interaction, Lead Toxicity, Bacopa monnieri, Hematological parameters, Antioxidant","lastPublishedDoi":"10.21203/rs.3.rs-7161974/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7161974/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe current study investigates the protective and therapeutic potential of Bacopa monnieri extract and synthetic antioxidants against lead-induced hematological toxicity in mice. HPLC analysis of Bacopa monnieri extract revealed a rich profile of bioactive compounds, including apigenin, luteolin flavonoids, bacosides, and bacopasaponins. In vivo experiments demonstrated significant hematological alterations in lead-exposed groups, with dose-dependent reductions in hemoglobin, red blood cell (RBC) count, platelet count, hematocrit, mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH), alongside increases in white blood cell (WBC) count. Lead exposure resulted in hemoglobin declines of up to 51.82% and RBC reductions of up to 59.32% in high-dose groups, indicating severe hematotoxic effects. Co-administration of Bacopa monnieri extract or synthetic antioxidants mitigated these alterations, with Bacopa monnieri showing superior protection by maintaining hemoglobin, RBC, and platelet levels closer to control values. Curative treatments further restored hematological parameters near baseline, highlighting the efficacy of Bacopa monnieri in reversing lead-induced toxicity. The findings of the study revealed dose-dependent statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) alterations in hematological parameters of lead intoxicated mice groups as compared to control groups, while co-treatment with synthetic antioxidants or Bacopa monnieri extract conferred protection and reduce these toxic effects. Notably, Bacopa monnieri showed higher efficacy in mitigating lead-induced toxicity to hematological system. These data imply that Bacopa monnieri has better potential to be an effective ameliorative therapeutic agent against lead toxicity as compared to synthetic alternatives.\u003c/p\u003e","manuscriptTitle":"Efficacy of Bacopa monnieri in Mitigating Lead-Induced Blood Toxicity in Mice Compared to Synthetic Antioxidants","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-31 18:03:21","doi":"10.21203/rs.3.rs-7161974/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-07-21T14:15:35+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-07-21T12:35:28+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-07-21T03:16:26+00:00","index":"","fulltext":""},{"type":"submitted","content":"Biological Trace Element Research","date":"2025-07-19T05:19:54+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"biological-trace-element-research","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bter","sideBox":"Learn more about [Biological Trace Element Research](https://www.springer.com/journal/12011)","snPcode":"12011","submissionUrl":"https://submission.nature.com/new-submission/12011/3","title":"Biological Trace Element Research","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"6c5c18d5-6029-4cf4-8140-65007a07d516","owner":[],"postedDate":"July 31st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-11-10T16:04:22+00:00","versionOfRecord":{"articleIdentity":"rs-7161974","link":"https://doi.org/10.1007/s12011-025-04849-x","journal":{"identity":"biological-trace-element-research","isVorOnly":false,"title":"Biological Trace Element Research"},"publishedOn":"2025-11-06 15:57:30","publishedOnDateReadable":"November 6th, 2025"},"versionCreatedAt":"2025-07-31 18:03:21","video":"","vorDoi":"10.1007/s12011-025-04849-x","vorDoiUrl":"https://doi.org/10.1007/s12011-025-04849-x","workflowStages":[]},"version":"v1","identity":"rs-7161974","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7161974","identity":"rs-7161974","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}