玉米赤霉烯酮对大鼠睾丸支持细胞Fas/FasL凋亡信号通路的影响

Effects of zearalenone on apoptosis via the Fas/FasL pathway in sertoli cells

为探讨玉米赤霉烯酮（zearalenon,ZEA）诱导睾丸支持细胞（sertoli cell,SC）凋亡的影响,试验以分离大鼠原代SC为材料,不同浓度ZEA处理SC 24 h,采用CCK-8法观察支持细胞活力的变化,流式细胞术检测细胞凋亡率的影响,q RT-PCR检测Fas、Fas L转录水平的变化,Western-blot法检测Fas、Fas L等凋亡相关蛋白表达的情况。结果显示,与对照组相比,随着ZEA浓度的升高,睾丸支持细胞的活力受到明显的抑制（P〈0.05或P〈0.01）;流式细胞术检测结果显示,与对照组相比,10、20 mol/L ZEA染毒组细胞凋亡率呈极显著升高（P〈0.01）;q RT-PCR结果分析显示,10、20 mol/L染毒组细胞Fas和Fas L转录水平呈显著或极显著升高（P〈0.05或P〈0.01）;凋亡相关蛋白检测结果显示,与对照组相比,5 mol/L以上ZEA染毒组Fas、Fas L、FADD、Cleaved-caspase8、Cleaved-caspase3蛋白表达均出现显著或极显著性升高（P〈0.05或P〈0.01）。结果表明,ZEA可以通过Fas/Fas L途径诱导睾丸支持细胞发生凋亡。

In order to investigate the apoptosis of sertoli cell（SC）induced by zearalenon（ZEA）,the primary SC was used as the experiment material,and exposed to different concentration of ZEA（0,5,10, 20μmol/L）for 24 h. The CCK-8 method was used to estimate cells viability;the flow cytometry was used to detect an apoptotic rate. The level of gene transcription ofF as and FasL was detected by qRT-PCR and the expression of apoptosis-related proteins was detected by Western-blot. The results showed that,compared with the control group,the viability of the SC in the exposed group was inhibited （P〈0.05 or P〈0.01） with the increase of ZEA concentration. The flow cytometry showed that the apoptotic rate was increased significantly（P〈0. 01）in ZEA group. The qRT-PCR results showed that the level of gene transcription of Fas and FasL was increased significantly with 10,20〈mol/L of ZEA（P〈0.05 or P〈0.01）. Westernblot analysis showed that the expression of Fas,FasL,FADD,Cleaved-caspase8 and Cleaved-caspase3 was increased in cells treaed with 5μmol/L or more ZEA（P〈0.05 or P〈0.01）. All these results clarified that ZEA would induce the apoptosis of sertoli cell via the Fas/FasL pathway.