EFFECT OF VITAMIN C AND THYMOQUINONE ON EXPERIMENTALLY BISPHENOL A INDUCED HEPATO - RENAL TOXICITY IN ADULT MALE ALBINO RATS

EFFECT OF  VITAMIN C AND THYMOQUINONE ON EXPERIMENTALLY BISPHENOL A  INDUCED HEPATO - RENAL TOXICITY IN ADULT MALE ALBINO RATS

By

Ahmad Gad Alla¹ and Ahmad M Gad Alla²

Department of Physiology¹ and  Forensic Medicine², Faculty of Medicine,

Al- Azhar University

ABSTRACT

Background: Bisphenol-A (BPA) is the building block of polycarbonate plastics, a hard plastic used to make numerous consumer products, including most baby  bottles and water bottles. Objective: Assessing the adverse effect of oral administration of   BPA on liver and kidney of adult male albino rats and evaluation of  the  role  of  vitamin C and thymoquinone (TQ) in alleviating the possible detrimental effects of  BPA. Material and Methods: Fifty six adult  male  albino  rats of  local strain were housed in 14 suitable metal cages(20 ×32× 20 cm for every 4 rats).They were divided into seven equal groups : The 1st group was served as a control group, the 2nd group was treated with bisphenol-A, the 3

 group was normal and treated with vitamin C alone ,the 4

group was  normal and treated with thymoquinone  alone, the 5

group  was  administered by bisphenol-A and treated with vitamin C, the  6

group was administered by  bisphenol-A and treated with  thymoquinone and the 7

 group was administered by bisphenol-A and treated with  both vitamin C and thymoquinone. All animals were treated for four weeks. Results: Administration of  bisphenol-A (50mg/Kg body weight orally) to rats resulted in an increase in alanine transaminase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), urea , creatine, and hepatic and renal MDA level, and decreased reduced glutathione (GSH) and superoxide dismutase (SOD) contents of  the liver and  kidney as compared to control group. In contrast, the administration of vitamin C (500 mg/ kg body weight) and/ or thymoquinone(10 mg /kg body weight) to bisphenol-A treated rats attenuated the toxicity of  bisphenol. This alleviation was more pronounced in the   bisphenol-A group treated with both vitamin C and  thymoquinone. Conclusion: BPA has detrimental effect on liver and kidney of  adult male albino rats. It supported the possibility that the synergistic effect of thymoquinone and vitamin C played a notable role in protecting the liver and kidney against BPA-induced toxicity and oxidative stress in male rats.

Key words: Bisphenol-A, vitamin C, thymoquinone.

INTRODUCTION

    BPA is an additive that has been used for more than 40 years to harden plastics, keep bacteria from growing in foods and prevent cans from rusting (Maćczak et al., 2017). BPA is the building block for polycarbonate, a hard strong plastic used to make compact disks, eye glass lenses, drinking glasses, water bottles, epoxy resins and baby bottles (Crain et al., 2007). Human exposure to BPA occurs through multiple routes. However, oral exposure is considered the major route of  exposure, while air, dust, and water are other possible sources of exposure (Di Donato et al., 2017).

    BPA may be absorbed in the gastrointestinal tract after ingesting products packed in plastic containers. Like intestinal phenols, BPA is conjugated by glucuronic acid in intestine and liver, and excreted in urine within 24 as BPA- glucuronide . BPA is only biologically active in its unconjugated form (Wang et al., 2017).

    Reactive oxygen species (ROS) are cytotoxic agents causing oxidative damage by attacking cell membrane and DNA (Ameziane et al., 2017). Antioxidants are scavengers that prevent cell and tissue damage that could lead to cellular damage and disease (Cooper  et al., 2011).Antioxidant agents have been used to prevent tissue damage in various clinical settings and experimental models, and could help in preventing lipid peroxidation and hydrogen peroxide levels, resulting  in  reduced  kidney and hepatic injury (Gong et al., 2013).

   Vitamin C is a potent natural antioxidant capable of reducing oxidative stress, scavenging free radicals and reestablishing anti oxidative systems (Masaki, 2010). Even in small amounts, vitamin C can protect indispensable molecules in the body such as proteins, lipids, carbohydrates, and nucleic acids (DNA and RNA) from damage by free radicals and reactive oxygen species (ROS) that are generated during normal metabolism (Sarita et al., 2011). Thymoquinone (TQ) was shown to inhibit tissue inflammation and oxidative stress (Muhammad et al., 2017).The protecting effects of  vitamin C and  thymoquinone  in the  kidney and liver are  reported  to be due to their antioxidant activity (Ayman et al., 2014).

    The present study aimed to evaluate the effect of vitamin C and thymoquinone  on  BPA-induced  hepato– renal toxicity in adult male albino rats.

MATERIAL AND METHODS

Chemicals: Bisphenol-A, vitamin C and  thymoquinone were purchased from Sigma –Aldrich Chemical CO.(USA). All other chemicals were of analytical grade and were obtained from commercial kits purchased from (Biodiagnostics Co. (Dokky, Giza, Egypt). Half gram of  Bisphenol-A powder  were dissolved in 50 cm sterile water, and 1 cm of solution contained 10 mg of  Bisphenol-A. Half gram of vitamin C powder was dissolved in 5 cm sterile water, and 1 cm of solution contained 100 mg of vitamin C. Half gram of thymoquinone powder was dissolved in 50 cm sterile water and 1 cm of solution contained 10 mg of  thymoquinone.

Animals: Fifty six adult male albino rats of  local strain weighing 180 to 200 grams. They were chosen as an animal model for this study. They were brought from animal house, Faculty of Medicine, Assiute University, Assiute, Egypt. They were housed in metal cages (20 ×32× 20 cm for every 4 rats) at room temperature, with the natural light/dark cycle in animal laboratory of Pharmacology Department, Al-Azhar University of Medicine (Assiute). They were maintained on dry chow pellets and water ad libitum throughout the experimental period.  All the experiments were performed during the same time of  day, between 9 a.m. and 12 p.m. to avoid variations due to diurnal rhythms (Shimizu et al., 2015).They were kept for two weeks under this condition to adapt the laboratory conditions before the start of the experiment.

Methods: After 2 weeks of acclimatiza-tion, male albino rats were randomly divided into 7 equal groups as follows:

Group І was served as a control group.

Group ІІ was administered freshly prepared BPA 50 mg /Kg body weight (5cm for each 100 g body weight of rat) in water once daily for four weeks orally via orogastric gavage (Da Chen et al., 2016).

Group ІІІ was administered vitamin C at  a dose of  500 mg /kg body weight (1cm for each 50 g body weight of rat) once daily for four weeks orally via oro gastric gavage (Adikwu and Deo, 2013).

Group ІV was administered  thymo-quinone (TQ) 10 mg /kg body weight (1cm for each 100 g from weight of rat) once daily for four weeks orally via oro gastric gavage (Aycan et al .,2015 ).

Group V received the same previous doses of BPA and vitamin C at the same time for four weeks orally via oro gastric gavage.

Group VІ received the same previous doses of BPA and TQ at the same time for four weeks orally via oro gastric gavage.

Group VІІ received the same previous doses of  BPA, vitamin C and TQ at the same time for four weeks orally via oro gastric gavage.  

Collection of blood samples for laboratory assessment: After 24 hours of receiving the last dose, blood samples were obtained from the retroorbital venous plexus of the eye. Blood was collected into non heparinzed tubes. Serum was separated from blood by centrifugation at 5000 rpm for 10 minutes and was stored at -20 C until used for estimation of alanine transaminase (ALT), aspartate  aminotransferase (AST), alkaline phosphatase (ALP), and urea and creatinine levels.

Tissue sampling and preparation of liver and kidney homogenate: Immediately after blood samples were collected, under ether anesthesia, the abdomens of the rats were immediately opened after reaching the stage of surgical anesthesia as evident by loss of withdrawal reflex. Then the liver and kidney of  different groups were excised, weighed,  perfused with normal saline to remove blood ,blotted  between filter papers and used for the preparation of tissue homogenate .About 0.5 gm of each  organ was homogenized in 4.5 ml of phosphate –buffered saline  (pH 7).The crude tissue homogenate was then centrifuged at 8,000 rpm for 30 minutes. The clear supernatant   from  liver and kidney homogenate  were stored at 4◦C and were used for the assay of reduced glutathione (GSH) (Beutler et al., 1963), superoxide dismutase (SOD) (Nishikimi et al., 1972), and malondialdehyde (MDA)(Ohkawa et al., 1978) by colorimetric method.

Statistical analysis was done using the computer program (SPSS). The quantitative data were presented in the form of mean ± standard error (S.E). Statistical analysis of  the difference between groups was performed by using One –way analysis of variance (ANOVA) followed by Tukey-Kramer test for differences between means. A value of P<0.05 was used as the limit for statistical significance.

RESULTS

The liver functions (ALT, AST and ALP) in BPA treated group(ІІ) resulted in significant elevation compared to control group (І).Also, liver functions (ALT, AST and ALP) in BPA treated group(ІІ) resulted in significant elevation as compared to group treated with either vitamin C(ІІІ) or thymoquinone (ІV) each  alone. Meanwhile, administration of  either vitamin C(ІІІ) or thymoquinone (ІV) each alone revealed non-significant changes in liver functions (ALT, AST and ALP) as compared to a normal control group (І). Also, administration of thymoquinone alone revealed non-significant changes in liver functions (ALT, AST and ALP) as compared to group treated with vitamin C (ІІІ) alone.The  BPA group treated with either vitamin C (V)or thymoquinone (VІ) each alone afforded significant decrease in liver functions (ALT, AST and ALP) as compared to bisphenol-A group (ІІ). Administration of a mixture of antioxidant elements (vitamin C + thymoquinone) into bisphenol  A  group(VІІ) achieved a significant reduction in liver functions (ALT, AST and ALP) as compared to the  Bisphenol-A group (ІІ), it was obvious that bisphenol -A group treated with a mixture of antioxidant element(vitamin C + thymoquinone) (VІІ) induced more decrement in liver functions levels than Bisphenol-A group  treated  with either vitamin C (V) or thymoquinone (VІ) (Table 1).

Table (1): Changes in ALT, AST and ALP in response to Bisphenol-A, vitamin C and  thymoquinone  administration in different groups (Mean± SE).

- Significant result as compared to group V, VІ ±

- Significant result as compared to group ІІ #

- Significant result as compared to group І *

     Urea and creatine in BPA-treated group (ІІ) showed significant elevation compared to control group (І). Also, urea and creatine in BPA treated group group (ІІ) resulted in significant elevation as compared to group treated with either vitamin C group (ІІІ) or thymoquinone (ІV) each alone. Meanwhile, administra-tion of either vitamin C (ІІІ) or  thymoquinone (ІV)  each alone revealed non-significant changes in kidney functions (urea and creatinine)  as compared to control group (І). Also, administration of thymoquinone (ІV) alone revealed non-significant changes in kidney functions (urea and  creatinine) as compared to group treated with vitamin C(ІІІ) alone. The BPA group treated with either vitamin C (V) or thymoquinone(VІ)  each alone afforded significant decrease in kidney functions (urea and  creatine) as  compared  to the BPA group(ІІ). It was obvious that  BPA group treated with a mixture of antioxidant elements (vitamin C + thymoquinone) (VІІ)   induced more decrement in kidney functions levels than BPA group treated with either vitamin C(V) or thymoquinone(VІ) each alone. Administration of a mixture of antioxidant elements (vitamin C + thymoquinone) in BPA group (VІІ) succeeded in restoring kidney functions (urea and creatine) to normal values as compared to the untreated BPA group (ІІ) (Table 2).

Table (2): Changes in urea and creatine in response to Bisphenol-A, vitamin C and thymoquinone administration  in different groups (Mean± SE).

- Significant result as compared to group V, VІ ±

- Significant result as compared to group ІІ #

- Significant result as compared to group І *

     The level of liver GSH   and  SOD  in the BPA group (ІІ) significantly decreased as compared to control group(І). Also, level of  liver  GSH and SOD in BPA treated group(ІІ) resulted in significant decrease as compared to group treated with either vitamin C (ІІІ)  or  thymo-quinone (ІV)   each alone. Meanwhile, administration of either vitamin C (ІІІ)  or  thymoquinone (ІV)  each alone revealed  non-significant changes in  liver GSH  and   SOD  level as compared to control group (І). Also, administration of thymoquinone (ІV) alone induced non-significant changes in liver GSH and SOD level as compared to group treated with vitamin C (ІІІ) alone. The treatment of  BPA  group with either  vitamin C (V) or thymoquinone (VІ) each antioxidant alone elicited a significant increase in liver GSH  and SOD level as compared to BPA group (ІІ). It was obvious that BPA group treated with a mixture of  both  vitamin C  and  thymoquinone (VІІ)  induced more increment in liver GSH and SOD level than BPA  group  treated with either vitamin C (V) or  thymoquinone (VІ) each alone. The treatment of  BPA group with both vitamin C and thymoquinone (VІІ) significantly increased level of  GSH and SOD in liver in comparison with the untreated  BPA group(ІІ) (Table 3).

     The level of  liver MDA in the BPA-treated rats (ІІ) was significantly higher than that of control  group (І).  Also, level of   liver MDA in BPA treated group (ІІ) resulted in significant elevation as compared to group treated with either vitamin C (ІІІ) or thymoquinone (ІV) each alone. Meanwhile, administration of either vitamin C (ІІІ)  or  thymoquinone (ІV)  each alone induced non-significant changes in liver MDA level as compared to the  normal control group (І). Also, administration of thymoquinone (ІV) alone revealed non-significant changes in liver MDA as compared to group treated with vitamin C (ІІІ) alone. The treatment of  BPA  group with either vitamin C (V) or thymoquinone (VІ)  each antioxidant alone elicited a significant decrease in liver MDA level as compared to BPA group (ІІ). It was obvious that BPA group treated with both vitamin C and thymoquinone (VІІ) induced more decrement in liver MDA level than BPA group treated with either vitamin C (V)  or thymoquinone (VІ) each antioxidant alone. The treatment of  BPA group with both vitamin C and  thymoquinone(VІІ) significantly inhibited the increase of  liver MDA in comparison with BPA group (ІІ).

Table (3): Changes in liver GSH, SOD and MDA activities in response to Bisphenol-A, vitamin C and thymoquinone administration in different groups (Mean± SE).

- Significant result as compared to group V, VІ ±

- Significant result as compared to group ІІ #

- Significant result as compared to group І *

    The levels of kidney GSH and SOD in the BPA group (ІІ) were significantly decreased as compared to control group (І). Also, levels of kidney GSH and SOD in BPA treated group (ІІ) resulted in significant decrease as compared to group treated with either vitamin C (ІІІ)  or  thymoquinone (ІV) each alone. Mean-while, administration of either vitamin C (ІІІ) or thymoquinone (ІV) each alone revealed non-significant changes in kidney GSH and SOD levels as compared to  the  normal control group (І). Also, administration of  thymoquinone (ІV) alone induced non-significant changes in kidney GSH and SOD as compared to group treated with vitamin C(ІІІ)  alone. The treatment of  BPA  group with either vitamin C (V) or thymoquinone (VІ)each antioxidant alone elicited a significant increase in kidney GSH and SOD levels as compared to BPA group(ІІ) .It was obvious that  BPA group treated with both vitamin C + thymoquinone(VІІ) induced more increment in GSH and SOD levels than BPA group treated with either vitamin C (V) or thymoquinone (VІ) each  antioxidant alone. The treatment of   BPA group with both vitamin C and thymoquinone (VІІ) significantly increased level of  GSH and SOD in kidney in comparison with the BPA group (ІІ).

    The levels of kidney MDA in BPA-treated   rats (ІІ) was significantly higher than that of control group (І). Also, levels of  kidney MDA in BPA treated group(ІІ) resulted in significant elevation as compared to group treated with either vitamin C (ІІІ)  or  thymoquinone (ІV)   each alone. Meanwhile, administration of either vitamin C (ІІІ) or  thymoquinone (ІV)  each alone induced  non-significant changes in kidney MDA level as compared to  the normal control group (І). Also, administration of thymoquinone (ІV) alone revealed non-significant changes in kidney MDA   as compared to group   treated with vitamin C (ІІІ) alone. The treatment of   BPA  group with either vitamin C (V) or thymoquinone (VІ) each alone elicited a significant decrease in kidney MDA level as compared  to BPA group (ІІ).  It was obvious that BPA group treated with both vitamin C+ thymo-quinone (VІІ) induced more decrement in kidney MDA   level than BPA group treated with either vitamin C (V) or thymoquinone (VІ) each antioxidant alone. The treatment of   BPA group with a mixture of   antioxidant elements (vitamin C+ thymoquinone) (VІІ) significantly inhibited  the increase of  kidney MDA in comparison with the  BPA group (ІІ).

Table (4): Changes in kidney GSH, SOD and MDA activities in response to Bisphenol-A, vitamin C and thymoquinone administration in different groups (Mean ± SE).

- Significant result as compared to group V, VІ ±

- Significant result as compared to group ІІ #

- Significant result as compared to group І *

DISCUSSION

    This study set out to assess the impact of vitamin C and thymoquinone on bisphenol A - induced hepato- renal toxicity in adult male albino rats. The current study demonstrated that BPA caused elevation of kidney and liver enzymes (damage markers).  Also, this study has revealed disturbances in oxidative/ anti oxidative status in rat liver and kidney with concomitant impairment in its proper functioning as a result of  BPA administration. Supplementation with vitamin C and thymoquinone protected liver and kidney against BPA induced hepato–renal toxicity and improved its antioxidant status. In the present study, it has been observed that administration of a mixture of antioxidant elements (vitamin C + thymoquinone)  in  bisphenol  A  group succeeded in restoring liver and kidney functions to normal values.

    Liver function tests are routinely used as diagnostic markers for hepatotoxicity.  Aminotransfersases (ALT and AST) and alkaline phosphatase (ALP) are the major markers in monitoring the functional status of   liver.  The ALT activity is an important index to measure the degree of cell membrane damage while AST is an indicator of mitochondrial damage. The activity of ALP is often employed to assess the integrity of   plasma membrane (Shi et al., 2015).

    BPA administration significantly increased the serum indices of   liver function as indicated by elevation in the activity of ALT, AST and ALP as compared to the control group. The high levels of  ALT, AST and  ALP are attributed to damage in liver.

     Moon et al., (2012) mentioned that   BPA can induce hepatic damage and mitochondrial dysfunction by increasing oxidative stress in the liver.  Similarly, elevated levels of serum indices as a result of liver damage in rats due to BPA toxicity have been previously reported by Korkmaz et al. (2010). Ronn et al. (2013) also reported an increase in AST activity in male rats treated with BPA 200 mg/kg/day and  increased  ALP ,γ-glutamyl transpeptidase activity in male rats treated with 600 mg/kg/day .

  The mechanisms involved in  Bisphenol-induced hepato- renal damage were investigated  and our results showed  that  BPA induced oxidative stress, evident by decreased GSH  level and  SOD activity with increased lipid peroxidation  product, MDA, which was in consistent  with previous studies of  Zeinab et al.(2012 ) and Ozra et al.(2015). These results are consistent with the previous findings released by some research group who had found an association between  Bisphenol A toxicity and the increased oxidative stress in rats (El-Megharbel et al .,2015). In the current study , it was noted that  the  elevation  in  liver enzymes was decreased  significantly after treatment of  Bisphenol rats with either vitamin C or  thymo-quinone  each alone or their combination as compared to Bisphenol  group. Similar results were obtained  by Wesam (2014) who reported  that  treatment  with thymoquinone at a dose of  10 mg/kg  BW significantly suppressed the bisphenol-induced elevation in these hepatic biomarkers.

    The efficacy of thymoquinone in protecting   hepatic enzyme leakage may be related to its ability to preserve the structural and functional integrity of the liver against the adverse effects of   BPA as well as repair of  hepatic tissue damage caused  by BPA. These results come in accordance with studies which indicated   the hepatoprotective effect of  thymo-quinone in models of liver injury (Hamid et al.(2014). This result was in agreement with Korkmaz  et al.( 2010) who reported that  vitamin C co-administration along with  BPA protect against liver damage in  male rats. Milad  et al. (2015) also reported that the administration of  vitamin C or vitamin C combined with chitosan restored  MDA levels, which reveals that vitamin C preserves the integrity of  cellular membrane  and  the normal physiological functions of  hepatocyte.

    Hepatoprotective property of   vitamin C is attributed to its antioxidant property. Vitamin C which is a major water-soluble antioxidant is believed to decrease lipid peroxidation either directly or indirectly by regenerating vitamin E. Vitamin C is an important free radical scavenger in extracellular fluids, trapping radicals and   protecting biomembranes  from peroxide damage. (Adikwu and Deo, 2013).

     Furthermore, BPA administration elevated   levels of serum renal injury markers such as urea and creatine as compared to control group.  These results are in agreement with Hassan et al. (2012) and Ahmed et al. (2015) who reported that  Bisphenol has a nephrotoxic effect and result in affecting the glomerular filtration. Also, these results were in agreement with the results of   Sangai et al., (2012) and Yldiz et al., (2013) who reported that the bisphenol- treated rats showed severe degenerative changes in the renal corpuscles and the renal tubules. Jyothi et al. (2009) observed significantly increased values in the concentration of serum creatinine when exposed  to  bisphenol  in rats. The most likely explanation  for these finding  is by ELadak et al.(2015)  who reported that  Bisphenol A has  a nephrotoxic effect due to accumulation of  BPA toxic metabolites and inability of  the kidney to eliminate them and reported that  bisphenol  A induces mitochondrial dysfunction and rough endoplasmic damage which in turn is  important for protein pathway.

    In the present study ,it was noted that the elevation in kidney function markers was decreased  significantly after treatment of  Bisphenol rats with either vitamin C or  thymoquinone  each alone or their combination as compared  to Bisphenol  group. The result of  the present work was in agreement with Mohamed  and Emad, (2015) where they noticed the protective effects of  Nigella sativa oil (NSO) and/or ascorbic acid (AA), against oxytetracycline( OTC)-induced  nephrotoxicity in rabbits. The activities of  the enzymes involved in glutathione pathways were also disrupted in the bisphenol  treated group (tab 3, 4),  indicating the involvement of oxidative stress in hepato-renal damage.

    Our results showed   that administration of Bisphenol caused a significant increase in hepatic lipid peroxidation level (MDA), depletion of  GSH contents and decreased  SOD activity (Table 3) as compared to control group. These findings come in agreement with the results obtained by Raluca et al. (2014)  who found that  Bisphenol had  increased  the hepatic oxidative stress and mitochondrial dysfunction leading to structural changes of  rat liver and alteration of  hepatic reduced glutathione (GSH) contents, glutathione peroxidase (GPx) and  glutathione reductase (GR) activities. The result of   the present work was in agreement with Hassan et  al.,(2012)  who  noticed that  Bisphenol induced  hepatotoxicity  through oxidative stress.

    This study demonstrated that adminis-tration of  Bisphenol caused a significant increase in  renal lipid peroxidation level (MDA), depletion of GSH  contents and decreased SOD activity (Table 4) as compared to that of rats of control group. This result was in disagreement with (Mourad and Khadrawy (2012) who reported   that there were non-significant changes in oxidative stress parameters in the kidney and liver due to BPA treatment. All these effects are involved in Bisphenol -induced hepato-renal oxidative damage and toxicity, as a result of excessive generation of free radicals, which have been reported to affect various biological molecules, including lipids and induce lipid peroxidation.

    Oxidative stress, mitochondrial damage and intracellular glutathione depletion are the most important factors contributing to the prediction of hepato-renal toxicity (Ahmad and Basma2016). Reactive oxygen species (ROS) are instantly produced in the body due to exposure to a wide range of exogenous chemicals, drugs and xenobiotics.

    In the current study, treatment with vitamin C or thymoquinone each alone or their combination played a role in ameliorating bisphenol-induced hepato-renal toxicity. Their free radical scaveng-ing abilities seem to mediate such a protective effect, indicated by the reduction of   MDA as well as the elevation of GSH and SOD levels in hepatic and renal tissue. But, this alleviation is more pronounced in the bisphenol group treated with both of the antioxidants. Also the improvement was more pronounced when both vitamin C and thymoquinone were used together in bisphenol group.

     Thus, the synergistic effect of vitamin C and thymoquinone is most powerful in reducing the hepato-renal toxicity induced by BPA and improving the liver and kidney antioxidant status. This result  was in agreement with Saleem et al., (2012) who noticed nephro-protective effect of vitamin C and Nigella sativa oil on gentamicin associated nephrotoxicity in rabbits.

     The protective effect of vitamin C and thymoquinone against BPA-induced oxidative stress in our rat model could be either direct by inhibiting lipid peroxidation and scavenging free radicals, or indirect through the enhancement of SOD and GSH activities; the enzymatic free radical scavengers in the cells. Therefore, vitamin C and thymiquinone could be used in combination to prevent and treat hepatic and renal diseases, especially those induced by oxidative damage.

CONCLUSION

     Based on the data of the current study, it may be concluded that BPA has detrimental effect on liver and kidney of male rats. Also it supports the possibility that the synergistic effect of thymo-quinone and vitamin C played a notable role in protecting the liver and kidney against BPA-induced toxicity and oxidative stress in male rats. The strong ant oxidative activity could partially be the mechanism underlying the protective effects of thymoquinone and vitamin C. The versatility and potency of thymo-quinone and vitamin C make it potential candidates for therapeutic and preventive drugs for hepato-ranal toxicity resulting from toxicants, including BPA. To mimic the study's result, drink tap water or rely on BPA-free stainless steel water bottles. To be safe, avoid all canned foods and replace with non-canned variations.

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