活性氧在p,p'-DDE诱导大鼠睾丸支持细胞凋亡中的作用
The Role of Reactive Oxygen Species in p, p'-DDE-induced Apoptosis of Sertoli Cells in Rats
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摘要:
目的 研究活性氧在2,2-双-(对氯苯基)-1,1-二氯乙稀(p,p'-DDE)诱导大鼠睾丸支持细胞凋亡中的作用。 方法 从大鼠睾丸组织中分离支持细胞进行离体原代培养3d,用0、10、30、50μmol/L的p,p'-DDE及加有300μmol/L的抗氧化剂N-乙酰-L-半胱氨酸(NAC)的p,p'-DDE (50μmol/L)继续培养24h,应用流式细胞仪测定细胞内活性氧(ROS)水平、线粒体膜势能(△ψm)、凋亡发生率。 结果 50μmol/L的p,p'-DDE可以诱导支持细胞ROS显著升高,10、30、50μmol/L p,p'-DDE处理组的线粒体膜势能下降,30、50μmol/L的p,p'-DDE可以诱导支持细胞凋亡,与对照组相比,差异有统计学意义(P<0.05);NAC有效抑制了ROS升高,削弱线粒体膜势能下降,减少了凋亡发生率,与50μmol/L的p,p'-DDE处理组相比,差异有显著意义(P<0.05)。 结论 p,p'-DDE可以诱导大鼠睾丸支持细胞凋亡,ROS产生可能是其重要原因之一。 Abstract:Objective To investigate the mechanism of p,p'-DDE-induced apoptosis of Sertoli cells and the role of reactive oxygen species. Methods Sertoli cells from testes of rats were treated with 0, 10, 30, and 50 μmol/L p,p'-DDE, or with 50 μmol/L p,p'-DDE pretreated by 300 μmolFL NAC (N-acetyl-L-cysteine). Then the fluoresence intensity of reactive oxygen species(ROS), mitochondria membrane potential(△ψm)and apoptosis percentage was detected by a FACS Calibur flow cytometer. Results 50 μmol/L p,p'-DDE increased the ROS level. The mitochondria membrane potential was decreased after all p,p'-DDE's treatments. The percentage of apoptosis was increased after treatment with 30 or 50 μmol/L p,p'-DDE. Compared with the control group, the difference was statistically significant(P<0.05). The antioxidant NAC inhibited the increased ROS generation, decreased the mitochondria membrane potential and apoptosis percentage (P<0.05). Conclusion p,p'-DDE could induce apoptosis in Sertoli cells. The generation of ROS may be an important reason in the apoptosis.
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