Protective Effects Of Aspongopus Chinensis On Testicular Damage Induced By Manganese In Rats

Objective: To investigate the protective effect of Aspongopus chinensis extract on male rat with reproductive system damage.Methods: Fifty Male Sprague-Dawley rats(180–200g)were randomly divided into control group,Mn group,low,medium and high dose of intervention group with Aspongopus chinensis(which are Asp50+Mn,Asp100+Mn,Asp200+Mn),ten rats in each group.Firstly,isovolumetric Manganese dichloride dehydrate(Mn Cl2·4H2O)of 30mg/kg and normal saline(NS)were separately injected into enterocoelia 1/d,7d/w.Meanwhile,the quite dosage of extracting solution from Aspongopus chinensis(50mg/kg,100mg/kg,200mg/kg)and distilled water were given to the rats by the means of gavage 1/d,7d/w.Observing and detecting male rats reproductive parameters including sperm concentration,viscera coefficient of testis and epididymis.Detecting with the testis and serum of male SD rats: 1.The staining of hematoxylin-eosin(HE)was employed to observe the structure of testis.2.The immunohistochemical(IHC)and western blot were used to evaluate the expression of apoptosis-related genes including Bcl-2,Bax,P-p53,Cyt C,C-Cas3.3.WST-1 method was employed to detect the total superoxide dismutase(T-SOD)activity of rat testis and serum.4.Enzymatic reaction method was employed to detect the glutathione peroxidase(GSH-PX)activity of rat testis and serum.5.Chemical colorimetry was used to detect the total antioxidant capacity(T-AOC)activity of rat testis and serum.6.Glucosinolates barbituric acid(TBA)method was used to detect MDAlevel of rat testis and serum.Results: 1.The effects of Aspongopus chinensis extract on sperm concentration,viscera coefficient of testis and epididymis with damaged by Mn was that the sperm concentration of Mn group and viscera coefficient of testis and epididymis were lower than that of Aspongopus chinensis treatment group and control group(P<0.05).2.The effects of Aspongopus chinensis extract on the testicular tissue morphology with damaged by Mn were that testicular tissue morphology had significantly improvement,raw seminiferoustubule business-like gradually smaller,basal membrane gradually complete,layer of seminiferous epithelia cell was increased,the arranged of all levels of sperm cells was relatively orderly,cell boundary became clear after the treatment of Aspongopus chinensis extract,and the morphological structure of high dose of intervention group with Aspongopus chinensis was same with the control group.3.The effects of Apongopus chinensis extract on antioxidase system of testis and serumwith damaged by Mn were that the T-SOD,T-AOC,GSH-PX activity of testis and serum of Mn group were lower than that of Aspongopus chinensis intervention group(P<0.05),while the contains of MDA of Mn group were higher than that of control group and the Aspongopus chinensis intervention group(P<0.05).4.The effects of Aspongopus chinensis extract on the expression of Bcl-2with damaged by Mn were that the immunohistochemical positive staining and the protein expression quantity of Bcl-2 from the control group and Aspongopus chinensis intervention group were higher than that of Mn group,and they were gradually higher as the Aspongopus chinensis extract dosage increased(P<0.05).5.The effects of Aspongopus chinensis extract on the expression of testicular apoptosis-related protein of BAX,P-P53,CYT C,C-CAS 3 with damaged by Mn were that the immunohistochemical positive staining and the relative expression quantity of BAX,P-P53,CYT C,C-CAS 3 in control group and Aspongopus chinensis intervention group were lower than that of Mn group(P<0.05),and they were gradually lower as the Aspongopus chinensis extract dosage increased(P<0.05),interestingly,the relative expression of Caspase-3 was no significantly difference(there is no date suggest).Conclusion: 1.The Aspongopus chinensis extract has significant improvement to male rat with gential system damage of the sperm concentration,viscera coefficient of testis and epididymis.2.The possible mechanisms of the Aspongopus chinensis extract intervene the male rat with reproductive system damage were:(1)reducing the oxidative stress state by increasing the antioxidant enzyme activities of rat testis and serum,and stopping lipid peroxidation of reproductive system to inhibit the apoptosis of spermatogenic cells.(2)restraining the expression of BAX to increase the expression of BCL-2 by down-regulating the P-P53 to weaken the apoptosis trend;Meanwhile,up-regulating the expression ofBCL-2and inhibiting the Cyt C up-regulation to stop the delivery of mitochondrial apoptotic signals,while the low expression of downstream signal Cleaved Caspase-3 can inhibit the apoptosis of testis cells.