氟对体外培养的小鼠睾丸间质细胞增殖和细胞凋亡的影响

目的 观察不同浓度氟化钠对睾丸间质细胞增殖和细胞凋亡的影响,为氟中毒的机制研究提供依据.方法 取体外培养的睾丸间质细胞,胰酶消化后制成单细胞悬液,常规培养,待细胞融合率达80％,且未出现细胞分化时,将细胞分4组,加入不同浓度的氟化钠染毒(0,5,10,20 mg/L)睾丸间质细胞,分别干预0,24,48,72,96,1...

Effect of fluoride at different concentrations on proliferation and apoptosis of mouse Leydig cells in vitro

Objective To explore the effect of fluoride at different concentrations on proliferation and apoptosis of mouse Leydig cells in vitro.Methods Mouse Leydig cells were collected by trypsinization to form monoplast suspension,centrifuged to float again,and then cultured in vitro.The cell numbers were counted,the cell suspension density adjusted,and the cells were inoculated into 96-shadow mask according to 1×106 per milliliter.All cells were cultured routinely.When the cell fusion ratio reached 80% and the cells did not start to differentiate,the cells were divided into 4 groups,cultured in DMEM-F12 medium containing 0,5,10,20 mg/ L fluoride,respectively.After each group was treated for 0,24,48,72,96,and 120 h,MTT test was used to detect the cell proliferation in each group.Flow cytometry（FCM） with annexin V-PI double staining was applied to detect the apoptosis in Leydig cells of each group after treatment for 48 h.Results 1） After the cells were treated for 24 h and 48 h,fluoride（20 mg/L） significantly inhibited proliferation of the Leydig cells,but 5 mg/L and10 mg/L had a less effect on the Leydig cells.After the cells were treated for 72 h,fluoride（10 mg/L） significantly inhibited proliferation of the Leydig cells.After the cells were treated for 96 h and 120 h,fluoride（5 mg/L,10 mg/L,20 mg/L） significantly inhibited Leydig cell proliferation.2） When the cells were treated by fluoride for 48 h,apoptosis was induced in the Leydig cells of all groups.The Leydig cells showed a highest late apoptosis rate at 10 mg/L fluoride,while the highest early apoptosis rate in Leydig cells was after 20 mg/L fluoride treatment.Conclusion Fluoride can effiectively inhibit proliferation and promote apoptosis of Leydig cells in mice.