The Effect Of Acrylonitrile Exposure On Male Sperm Quality And Experimental Study On Genistein Antagonize Testicular Leydig Toxicity Induced By Acrylonitrile

Partâ… Effect of acrylonitrile exposure on semen quality among occupational workersAim To investigate the effects of acrylonitrile exposure on the semen quality of occupational workers.Materials and methods Sixty-seven male workers who were exposure to acrylonitrile and 27 male healthy volunteers in the same area were selected as the exposure group and control group,respectively.The men provided information relating to lifestyle,occupational details.After the semen was collected according to the standard method,the workers' semen qualities were analyzed by the WLJY-9000 CASA(computer-assisted semen analysis) system.Serum concentrations of thyrotropic stimulating hormone(TSH),follicle stimulating hormone(FSH),luteotrophic hormone(LH),testosterone(T) and estradiol(E₂) were analyzed by electro-chemiluminescence immunoassay.Sperm DNA integrity was assessed using the single-cell gel electrophoresis(SCGE) assays.Results Significant differences were observed in the sperm concentration,the concentration of motile and progressive sperm,straight line velocity(VSL),average path velocity(VAP),linearity(LIN) and wobble coefficient(WOB).Through the analysis of multiple regression,we obtained age and exposure time(seniority) showed a significant relationship with the sperm concentration,the concentration of progressive sperm,and average path velocity (VAP).In exposure group,the levels of E₂ increased significantly and T decreased, although there was no significant decrease in the T levels between two groups. Statistically significant increasing was found in sperm DNA damage in exposure group compared to control group for the tail moment,otherwise tail length and Olive tail moment have no significant increasing.Conclusions Occupational exposure to acrylonitrile could affect the semen quality and sexual hormone levels of the workers, especially,the sperm movement parameters.This appears to be the first study showing the value of CASA in observation of reproductive toxicity of acrylonitrile exposure in the male workers.The application of CASA provides a plausible and objective method to predict the fertility of male AN-exposed workers. Partâ…¡Experimental study of testicular toxicity induced by acrylonitrileAim To investigate the acrylonitrile induced testicular toxicity and oxidative stress and associated Fas/Fas ligand(FasL) expression.Material and methods Sprague-Dawley rats were administer drinking water by gavage formulated to contain 0,10,15,20 mg per kg body weight per day for ten weeks,respectively.Rats were closely monitored clinically with body weight every week.After ten weeks,all rats were necropsied and then genital organ weights and histopathology were examined. Ultrastructural studies of testes were observed under transmission electron microscope. The epididymal sperm motile parameters were analyzed by computer-assisted semen analysis system.Oxidative stress was assessed in testes by determining glutathione (GSH) level,glutathione peroxidase(GSH-Px) activity,superoxide dismutase(SOD) activity and lipid peroxidation as indicated by malondialdehyde(MDA) production. Serum testosterone level was assessed by enzyme linked immunosorbent assay (ELISA).Fas and FasL expression in rat testis were examined by immunohistochemistry.Expression levels of Fas,FasL and 3β-hydroxysteroid dehydrogenase(3β-HSD) were analyzed by RT-PCR and Western blotting.Results Our data showed significant changes in the weight of the body,however,no obvious differences in the weight of testis and epididymis were found after AN treatment. Significant reducing was observed in the parameters of the percentage of motile sperm,the percentage of progressive motile sperm,VAP,VSL and LIN in a dose dependent manner.Histopathologic evaluations demonstrated that massive vacuolization appeared in the seminiferous epithelium and Leydig cells apoptosis were found in interstitial tissue of AN treated rat testis which consistent with the decreasing serum testosterone level in AN treated rat.Immunohistochemistry experiments in AN treated rat testis demonstrated the increasing expression of FasL in Sertoli and Leydig cells whereas the increasing expression of Fas was confined to the Leydig cells.AN reated testis revealed a significant decrease in testicular GSH content as well as a significant enhancement of MDA accumulation in a dose-dependent manner.We found a significant increase in expression of Fas and FasL mRNA and protein in AN treated rat testis.Moreover,there was a significant decrease in the levels of 3beta-HSD mRNA and protein of AN treated testis.Conclusions Our results suggest that AN exposure had side effect on sperm mobility of rats.Oxidative stress may play a role in the testicular toxicity induced by AN,and the Fas/FasL system may be involved in the oxidative damage and the apoptosis of Leydig cells.Partâ…¢Genistein prevents acrylonitrile-induced toxicity in primary cultured rat Leydig cellsAim To determine the effect of AN on the viability of primary-cultured rat Leydig cells as well as the oxidative damage generated by AN by measuring MDA and GSH levels in primary cultured Leydig cells.We also analyzed whether the AN toxicity could be avoided by using antioxidant genistein.Material and methods Leydig cells were separated by enzymatic digestion combined with discontinuous Percoll density gradient centrifugation.Cells were exposed in vitro to different concentrations of AN for different time intervals.Cell toxicity was measured by CCK8 assay.Oxidative stress was assessed by determining glutathione(GSH) level and malondialdehyde (MDA) production.Expression levels of Fas and FasL were assessed by RT-PCR and Western blotting.Apoptosis rate was analyzed by Annexin V-FITC/PI flow cytometry (FCM) and the expression of NF-κB p65 was measured by Western blot.Results Our data show that AN inhibited the growth in a time and dose dependent manner.The expression of Fas and FasL was significantly elevated in a time dependent manner. AN enhanced oxidative stress,as indicated by malondialdehyde(MDA) accumulation, and depleted reduced glutathione(GSH) level.Leydig cell exposed to AN treated with genistein afford some protection,as indicated by reducing MDA level and elevating level of GSH.Furthermore,co-treatment with genistein and AN cause a decrease in apoptosis rate of Leydig cells.Meanwhile,the expression of NF-κB p65 increased significantly.Conclusions Our study indicated the apoptotic effect of acrylonitrile on Leydig cells might be through oxidative stress and Fas/FasL system.Genistein could prevent acrylonitrile-induced oxidative stress and apoptosis of Leydig cells,which may be involved NF-κB signal transduction pathways.