氯化三丁基锡对大鼠睾丸支持细胞的影响
Effects of Tributyltin Chloride on Rat Sertoli Cell Function
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摘要:
目的 探讨氯化三丁基锡对雄性SD大鼠睾丸支持细胞活力和增殖的影响。 方法 无菌制备20~23天龄SD大鼠的睾丸支持细胞,用HE染色、FAS-L免疫组化染色进行细胞鉴定。设空白对照组、溶剂对照组和20& #215;10-9、40& #215;10-9、80& #215;10-9、120& #215;10-9 mol/L氯化三丁基锡染毒组,用四唑盐比色法(MTT)、免疫细胞化学法观察细胞增殖及增殖细胞核抗原(PCNA)的表达情况,流式细胞仪分析细胞周期分布。 结果 各染毒组睾丸支持细胞的存活率明显低于对照组(P<0.05),G0/G1期细胞构成比增加,细胞增殖指数下降,80& #215;10-9、120& #215;10-9 mol/L染毒组PCNA表达量下降。 结论 氯化三丁基锡可通过抑制睾丸支持细胞增殖,影响细胞周期分布和PCNA表达而表现出对雄性大鼠的生殖毒性。 Abstract:Objective To detect the toxic effects of tributyltin chloride (TBT) on cultured rat Sertoli cells and its injury mechanism. Methods Sertoli cells were isolated from the testes of 20-23 day-old male SD rats and identified with hematoxylineosin staining (HE) and fas ligand immunohistochemical staining (FAS-L). To obtain more Sertoli cells, cultures were hypotonically treated with Tris-HCl. One blank control group, one solvent control group and four tributyltin chloride groups (20& #215;10-9, 40& #215;10-9, 80& #215;10-9, and 120& #215;10-9 mol/L, respectively) were set. Cell viability were determined by methyl thiazolyl tetrazolium (MTT), expression of proliferating cell nuclear antigen (PCNA) were determined by streptavidin-perosidase (SP), and the distribution of the cell cycles were analyzed by flow cytometry (FCM). Results TBT can decrease the survival rate of Sertoli cells and interfere with the distribution of cell cycles. The result of immunocytochemistry showed that the expression of PCNA was significantly inhibited in the 80& #215;10-9 mol/L and 120& #215;10-9 mol/L dose groups. Conclusion TBT can inhibit the proliferation and affect the cell distribution and PCNA expression of Sertoli cells.
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