新型大气污染物MMT诱导人SK-N-SH细胞凋亡的分子机制

The Mechanism of MMT-induced Apoptosis in SK-N-SH Cells

目的研究甲基环戊二烯羰基锰(MMT)诱导人SK-N-SH细胞凋亡的分子机制。方法MMT诱导SK-N-SH细胞后,MTT法检测细胞存活率,分光光度法检测活性氧(ROS)生成;化学发光法检测Caspase-3活性;Western blot法检测p38 MAPK磷酸化程度。结果在MMT诱导SK-N-SH细胞过程中,细胞ROS生成量为正常组的5.2倍(P<0.01);抗氧化剂N-乙酰半胱氨酸(NAC)及二硫苏糖醇(DTT)可有效抑制细胞ROS生成(P<0.01),提高细胞存活率(P<0.01);在这一过程中,Capase-3活性明显增强,为对照组的3.01倍(P<0.01);同时p38 MAPK磷酸化程度加强;p38 MAPK特异性抑制剂SB203580可提高细胞存活率(P<0.05),抑制Caspase-3的活性(P<0.01)。结论MMT诱导的SK-N-SH细胞凋亡与ROS升高,激活Caspase-3有关,这一过程有p38 MAPK相关信号转导途径参与。

Objective To explore the molecular mechanism of MMT-induced apoptosis in SK-N-SH cells. Method The cells viability was determined by MTT Assay. The spectrophotometer was used to check the production of ROS. The change on the phosphorylation of p38-MAPK was determined by Western blot. And the activity of Caspase-3 was also measured. Results In the apoptotic process, the ROS production ascended to 5.2 times of the control group（ P 〈 0.01 ）. Antioxidants, such as NAC and DTT, inhibited the production of ROS（ P 〈 0.01 ）and improved the viability of cells（ P 〈 0.01 ）. At the same time, the activity of Caspase-3 grew to 3.01 times of the control（ P 〈 0.01 ）. The phosphorylation of p38-MAPK could also be found in this process and could be prevented by the special inhibitor SB203580. SB203580 could also improve the viability of cells（ P 〈 0.05） and inhibit the activity of Caspase-3（ P 〈 0.01）. Conclusion During MMT-induced apoptosis in SK-N-SH cells, the production of ROS increases. And the activity of Capase-3 also ascends. The p38 MAPK-related signal pathway is involved in this process.