The Alleviative Roles Of Organic Selenium Sources And Gum Arabic On CCL₄-induced Hepatonephrotoxicity And Apoptosis In Wistar Rats

Liver and kidney are the most important organs in term of their contribution in metabolism and detoxification of various materials.The diseases of liver and kidney are one of the critical health problems found in human and animals.In the small pets,animals such as cats and dogs the diseases of liver and kidney seem to be serious compared to large animals.Exposure to toxic agents through different route of administration will lead to severe damage of organs and cells of the body.Carbon tetrachloride（CCL₄）is toxic chemical agent that cause damage to the different organs of the body especially liver and kidney.selenium-enriched yeast（an organic selenium source,SY）,Gum Arabic（GA）and selenizing astragalus polysaccharides（another organic selenium source）have various degree of antioxidant and anti-apoptosis effects,therefore the synergistic effect of them will be stronger compared to each material alone.Accordingly,our present research was to verify the effect of GA+SY or selenizing astragalus polysaccharides（SAP）into CCL₄-induced liver or kidney injuries and apoptosis in rats,which will be presented in the following experiments:Experiment 1:The potential roles of Gum Arabic and SY combination on CCL₄-induced nephrotoxicity and apoptosis in Wistar ratsIn this study,the protective effects of Gum Arabic and selenium-enriched yeast（GA+SY）on renal oxidative stress and apoptosis in the kidney of wistar rats,which was induced by CCL₄,were determined.The results showed that GA+SY alleviated significantly CCL₄-induced renal damage and apoptosis,specified by the histopathological examination and ultrastructural remark.Moreover,treatment with GA+SY also inhibited significantly CCL₄-induced remarkable enhancement of oxidative stress,such as the increase of lipid peroxidation level and the depletion of intracellular reduced glutathione GSH,SOD and TO AC level in kidney.Additionally the levels of creatinine and urea were significantly（P<0.05）elevated in rats administered CCL₄ only.However,GA+SY significantly decreased the levels of urea,creatinine and increased the total protein which was affected by CCL₄ administration.Moreover,to explore the underlying mechanisms of GA+SY action,we found that activities of caspase-3 and caspase-9 were notably inhibited by treatment of GA+SY.Furthermore,gum Arabic and selenium yeast significantly corrected the changes of antiapoptotic genes（bcl2 and bcl-xl）,proapoptotic（bax）and apoptotic（caspase-3 and caspase-9）genes expressions that was down and up-regulated by CCL₄ treatment consecutively.In conclusion,Gum Arabic and selenium yeast combination significantly reduced nephrotoxicity and apoptosis induced by CCL₄ administration,through improvement of antioxidant status,reduction of inflammation,and antiapoptosis effects.Experiment 2:Roles of gum Arabic and SY in reduction of CCL₄-induced hepatic function damage,oxidative stress and apoptosis in the Wistar ratsIncrease in serum liver enzymes,oxidative stress markers,and apoptosis are characteristics features for acute hepatic injury,which is a universal problem,that results from exposure to oxidative stress agents and toxins such as carbon tetrachloride.In this study,40 male Wistar,rats of 12 weeks of age were randomly divided into five groups（each group have 8 rat）.Control group,CCL₄ group,（CCL₄+GA）group,（CCL₄+SY）group and（CCL₄+GA+SY）group.The experiment continued for 21 days after adaptation.Blood and liver samples were collected at the end of the experiment for biochemical analysis,antioxidants capacity,genes,proteins expression and histopathology.Data analysis showed that GA+SY significantly（P<0.05）attenuated histopathological abnormalities,decreased the elevation of ALT（72±2.88 U/L）,AST（124.37±19.18 U/L）,ALP（167.33±3.18）and MDA（4.23 ± 0.2 nmol/mg）levels in serum,and increased the inhibited of GSH-PX（226.47± 10.49 U/mg）,SOD（38.33±0.88 U/mg）,and GSH levels（15.13a±1.01 nmol/g）in serum of rats caused by CCL₄ treatment.Moreover,GA+SY inhibited elevation of caspase-3 expressions caused by CCL₄ treatment by using immunohistochemistry and western blotting.Furthermore,administration of GA+SY significantly（P<0.05）corrected the changes of apoptosis related genes expressions（bcl2,bcl-xl,bax and caspase-3,9）which was caused by CCL₄ treatment.In Conclusion:our findings indicate that GA+SY exert protective effect against CCL₄-induced hepatic injury by reduction of inflammation,suppressing apoptosis,and enhancing of antioxidation status.Experiment 3:Roles of selenizing astragalus polysaccharides in alleviation of CCL₄-induced kidney injuries and apoptosisAcute kidney injury is a universal problem,which results from exposure to oxidative stress agents and toxins such as carbon tetrachloride.Kidney injuries are characterized by an increase in serum creatinine,urea,oxidative stress markers,and apoptosisIn the present study,12 weeks of age forty male Wistar rats were divided randomly into five groups（8 in each group）.After adaptation,the experiment continued for 21 days.At the end of the experiment,blood and kidney samples were collected for chemical analysis,antioxidants capability,genes,and proteins expressions.Kidney cell apoptosis was evaluated by immunohistochemistry of caspase-3.Analysis of data showed that sAPS3 were significantly（P<0.05）attenuated histopathological abnormalities,decreased serum urea,creatinine and MDA levels and increased antioxidant capacity:GSH-Px,SOD,and GSH levels in rats treated by CCL₄.Moreover,sAPS3 significantly（P<0.05）regulated the antiapoptotic（bcl2 and bcl-xl）,proapoptotic（bax）and apoptotic（caspase-3,caspase-9）genes expression.In summary,sAPS3 significantly ameliorated the effects of CCL₄ treatment through,regulation of genes expression,improvement of antioxidant status and reduction the levels of urea and creatinine.