Effects Of Fluoride On Apoptosis Lymphocyte Mediated By FasL In Sertoli Cells And Lymphocyte Co-cultured System

[Objective] To study the effects of fluoride on immune privilege of sertoli cells, and to explore the mechanism of fluoride toxicity to immune privilege, the expression of FasL associated with immune privilege in Sertoli cells as well as Fas in lymphocytes were detected, thus providing a theoretical basis for organ transplantation.[Method] In the present study, sertoli cells treated with vary dose of fluoride was determined for their function. To examine the toxicology effects of HE staining and Giemsa staining assay was performed; mRNA expression of FasL in sertoli cells exposed to fluoride as well as Fas、caspase3 and caspase8 in lymphocytes after co-cultured with sertoli cells were determined by quantitative real-time RT-PCR respectively; The apoptosis rate of lymphocytes was detected by TUNEL assay; Meanwhile, the expression of FasL and Fas were detected by western blot.[Result] (1)The identification of sertoli cells:IF of sertoli cells with GATA4 showed that:sertoli cells can stably expressed GATA4, and the number of sertoli cells reached 95%. Thus cell model can be established by sertoli cells.(2)The result of MTT assay showed that:Compared to the control group, relative survival of lymphocytes treated with 10-6、10-5、10-4mol/L NaF concentration was decreased, and decreased significantly with 10-3mol/L concentration.(3) Relative survival of co-cultured lymphocytes with sertoli cells by MTT:Compared to the control group, relative survival of lymphocytes co-cultured with sertoli cells treated with 10-5、10-4mol/L NaF concentration was decreased, and decreased significantly with 10-3mol/L concentration.(4) Apoptosis rate of co-cultured lymphocytes with sertoli cells byTUNEL:Compared to the control group, apoptosis rate of lymphocytes co-cultured with sertoli cells treated with 10-5、10-4and 10-3mol/L NaF concentration was decreased significantly.(5)The result of western blot showed that:Compared to the control group, the expression of FasL and Fas in the experimental group were decreased; Compared to the control group, expression of FasL was prominently dereased in sertoli cells treated with the lowest concentration of NaF; meanwhile, the expression of Fas was decreased significantly in lymphocytes with 10-4 and 10-3mol/L NaF.(6)The result of Real-time-PCR showed that:Compared to the control group, mRNA expression of FasL、 Fas、caspase3 and caspase8 in the experimental group were decreased; Compared to the control group, expression of FasL was prominently dereased in sertoli cells treated with 10-4 and 10-5mol/L NaF; meanwhile, mRNA expression of Fas、caspase3 and caspase8 were decreased significantly in lymphocytes with the highest concentration of NaF.[Conclusion] Fluoride reduced the expression of FasL gene and protein in Sertoli cells, affecting the function of Sertoli cells killing lymphocytes, and reducing immune privilege function of sertoli cells.