摘要
目的：探讨环境污染物三丁基氯化锡（TBT）和氯化三苯锡（TPT）对大鼠睾丸Leydig细胞的影响。方法：①用0～80nmol／L浓度的TBT和TPT处理大鼠睾丸Leydig（LC-540）细胞24～96h，用四唑蓝（MTT）法确定细胞的存活率；②用DNA片段法确定是否存在细胞凋亡；③观察细胞内Ca^2＋螯合剂氨基苯乙烷四乙酸（BAPTA）和蛋白激酶A（PKA）抑制剂H-89、蛋白激酶C（PKC）抑制剂GF109203X、酪氨酸蛋白激酶（TPK）抑制剂Genistein是否可以阻断TBT所致的细胞凋亡；④检测TBT处理的原代大鼠睾丸Leydig细胞分泌睾酮的变化。结果：@TBT和TPT影响Leydig细胞存活率的强度基本相同，在20～80nmol／L浓度时细胞存活率呈剂量和时间依赖性降低；②DNA片段法研究证实TBT和TPT可引起细胞凋亡；③BAPTA可阻断20nmol／L的TBT所致的细胞凋亡，而PKA、PKC和TPK抑制剂对细胞存活率没有影响；@TBT可减少Leydig细胞分泌睾酮，并使其对人绒毛膜促性腺激素（hCG）刺激的反应性降低。结论：环境污染物TBT和TPT能直接导致睾丸Leydig细胞凋亡，并抑制睾酮分泌，这种致凋亡作用可能与细胞内Ca^2＋浓度增加有关。
Objective： To investigate the effects of tributyltin chloride （TBT） and triphenyltin chloride （TPT） on rat testicular Leydig cells. Methods ： The rat Leydig cells （ LC-540 ） were incubated with 0 to 80 nmol/L TBT and TPT for 24 - 96 h, and then the cell viability was determined by MTT. DNA fragmentation ladder formation of cell apoptosis was examined by. agarose electrophoresis. Effects of chelator of intracellular Ca^2＋ （BAPTA） and the inhibitors of PKA, PKC and TPK on cell apoptosis induced by TBT were observed. Effects of TBT on testosterone production in primary cultured rat Leydig cells treated with or without hCG were detected. Resuits ： TBT and TPT suppressed Leydig cell survival in a time-and dose-dependent manner. The suppressive effects of TBT and TPT on the cell survival was caused by apoptosis which was determined by DNA ladder formation. The apoptotic effect of TBT was possibly mediated by the rise in intracellular Ca^2＋ because it could be blocked by BAPTA, the chelator of intracellular Ca^2＋ ; PKA, PKC and TPK inhibitors did not prevent the apoptotic effects induced by TBT. TBT markedly suppressed testosterone production of primary cultured rat Leydig cells with or without hCG stimulation. Conclusion ： TBT and TPT induced apoptosis in rat testicular Leydig cells possibly through increasing intracellular Ca^2＋. TBT reduced the testosterone production of rat Leydig cells.
出处
《中华男科学杂志》
CAS
CSCD
2006年第6期516-519,共4页
National Journal of Andrology
