Effects Of 4-BDE On Apoptosis Of Sertoli Cells And Changes Of Fas/Fasl Signaling Pathway

Objective:To study the effect of 4-BDE on testicular Sertoli cell apoptosis,and to explore the mechanism of Fas/Fasl signaling pathway in the regulation of testicular Sertoli cell apoptosis induced by 4-BDE,so as to provide theoretical and experimental basis for male reproductive toxicity induced by 4-BDE.Methods:1.Select different concentrations of 4-BDE to infect rat testicular Sertoli cells for24 h,and determine the cell survival rate by CCK-8 method.Select the appropriate concentration of 4-BDE(low,medium and high concentration groups),and set the blank group and DMSO solvent group;2.The apoptosis of Sertoli cells was detected by flow cytometry after Annexin V-FITC/PI double staining;3.The expression levels of Fas,FasL,FADD,Caspase-3 and Caspase-8 mRNA in Sertoli cells were detected by q-PCR;4.WB was used to detect the expression of Fas,FasL,FADD,Procaspase-3 and Procaspase-8 proteins in Sertoli cells of testis.Results:1.4-BDE induced apoptosis of Sertoli cells in a dose-dependent manner,that is,the total apoptosis rate increased with the increase of 4-BDE concentration.The apoptosis rate in 160 μg/ml and 320 μg/ml group was significantly higher than that in0 μg/ml group(p < 0.05),and the apoptosis rate in 80 μg/ml group was slightly higher than that in 0 μg/ml group,but the difference was not significant(P > 0.05).2.4-BDE induced the increase of transcription level of apoptosis-related mRNA in Sertoli cells.The mRNA transcription of Fas、Fasl、FADD、Caspase-3 and Caspase-8 increased with the increase of the dose of 4-BDE.Compared with 0 μg/ml group,the mRNA transcripts of Fasl,FADD,Caspase-3 and Caspase-8 in 80 μg/ml and 160 μg/ml groups were significantly increased(p < 0.01);In 320 μg/ml group,the transcription of Fas、Fasl、FADD、Caspase-3 and Caspase-8 mRNA increased significantly(p < 0.01).3.4-BDE induced the expression of apoptosis-related proteins and the activation of zymogen in Sertoli cells With the increase of the dose of 4-BDE,the expression of Fas,Fasl and FADD protein in Sertoli cells treated with 4-BDE increased significantly,while the expression of Procaspase-3 and Procaspase-8 protein decreased significantly.Specifically: compared with the 0 μg/ml group,the expression of Fas and FADD protein was significantly increased in 80 μg/ml group(p < 0.01).The expression of Fasl protein was increased,but there was no significant difference(P >0.05).The expression of Procaspase-8 and Procaspase-3 protein was decreased in 80μg/ml group,the expression levels of as,Fasl and FADD protein were significantly increased(p < 0.01),while the expression levels of Procaspase-3 protein were significantly decreased(p < 0.05),the expression levels of Fas,Fasl and FADD in320 μg/ml group were significantly higher than those in 320 μg/ml group(p < 0.01),and the expression levels of Procaspase-3 and Procaspase-8 were significantly lower than those in 320 μg/ml group(p < 0.01),the difference was significant(p < 0.01).Conclusion:1.4-BDE can induce apoptosis of Sertoli cells in a dose-dependent manner.2.4-BDE may regulate caspase through Fas/Fasl,and finally participate in the process of testicular Sertoli cell apoptosis.