Study On Toxic Effects Of Manganese On Sertoli Cells In Vitro

objective: To study effects of Mn (manganese chloride) on the expression of MIS,ABP and FSHR in Sertoli cells of Sprague Dawley (SD) rats in vitro.Method: Testicular Sertoli cells from 20-day-old male SD rats were prepared (by digeston with coUagenase and trypsin) and cultured.Electron microscope was used to identify the Sertoli cells. After 6 days' cell culture.Sertoli cells were supplemented with different concentrations of Mn (0,1,10,50,100, 200μmol/L) for 24hours. 100μmol/L Mn concentration was used for time course (0,6h,24h,48h) study.The expression of ABP,FSHR and MIS mRNA was measeured by Real-time quantitative PCR. FSHR protein in the cultured supernatant was determined with ELISA after Sertoli cells were exposed to different concentrations of Mn for 24h.Results:With the increase of the Mn concentration(1,10μmol/L),the expression of ABPmRNA in Sertoli cells increased (p<0.01),but decreased as further increasing of higher Mn concentrations (50,100,200μmol/L) in a dose-dependent manner (p<0.01).Compared with 10μmol/L Mn dose,the expression of ABP mRNA decreased at the higher doses (50,100,200μmol/L) (p<0.01).The same dose-effect relationships tendency of FSHR were observed both at the mRNA level in cells and protein in the supernatant of Mn treated Sertoli cells.No significant alterations were observed in the expression of MIS mRNA(p>0.05) after Mn treatment.Conclusion:50μmol/L and higher concentrations of Mn could inhibit the expression of ABP and FSHR,and this function could work as one of the reproductive toxicity mechanisms of Mn on Sertoli cells.