Qian Jie, Bian Qian, Cui Lunbiao, Chen Jianfeng, Song Ling, Wang Xinru
Key Laboratory of Reproductive Medicine of Jiangsu Province, Nanjing Medical University, Nanjing 210029, China.
Toxicol Lett. 2006 Oct 10;166(2):178-86. doi: 10.1016/j.toxlet.2006.06.646. Epub 2006 Jul 3.
In this study, the effects of 4-tert-octylphenol (OP) were examined on the viability of rat cultured Sertoli cells using the MTT assay and OP-induced apoptosis was detected by transmission electron microscope (TEM), flow cytometric analysis and Hoechst staining. In addition, RT-PCR was used to analyze the levels of Bcl-2 and Bax mRNA. Bcl-2, Bax and caspase-3 protein expressions were determined by Western blot analysis. Sertoli cells were treated with OP from 30 to 60microM for 6-24h. Decreased viability of Sertoli cells and increased apoptosis occurred in a concentration- and a time-dependent manner. The expression of Bcl-2 was down-regulated, while the expression of Bax up-regulated. OP also down-regulated the expression of 32kDa procaspase-3, which was cleaved to generate active subunit (17kDa). These results suggest that OP may induce Sertoli cell apoptosis by regulation of Bcl-2/Bax and caspase-3 activation.
在本研究中，使用MTT法检测了4-叔辛基苯酚（OP）对大鼠培养的支持细胞活力的影响，并通过透射电子显微镜（TEM）、流式细胞术分析和Hoechst染色检测了OP诱导的细胞凋亡。此外，采用RT-PCR分析Bcl-2和Bax mRNA的水平。通过蛋白质印迹分析确定Bcl-2、Bax和半胱天冬酶-3蛋白的表达。支持细胞用30至60微摩尔的OP处理6至24小时。支持细胞活力降低和细胞凋亡增加呈浓度和时间依赖性。Bcl-2的表达下调，而Bax的表达上调。OP还下调了32 kDa前体半胱天冬酶-3的表达，该酶被切割产生活性亚基（17 kDa）。这些结果表明，OP可能通过调节Bcl-2/Bax和半胱天冬酶-3的激活来诱导支持细胞凋亡。
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