二甲基甲酰胺致心肌细胞凋亡线粒体通路的体外实验研究

目的 探讨二甲基甲酰胺（dimethylformamide，DMF）致H9c2心肌细胞凋亡线粒体通路相关蛋白的影响。方法 采用蛋白免疫印迹法（western blot，WB）检测不同剂量DMF（0 mM、50 mM、100 mM、200 mM）染毒24 h后胞浆内活化的Caspase-3（cleaved caspase-3）、Bax、Bcl-2以及细胞色素C（Cyt-c）水平的改变。结果 不同剂量DMF（0 mM，50 mM，100 mM，200 mM）染毒24 h后胞浆Cleaved caspase-3、Bax和Cyt-c水平升高，Bcl-2水平降低，组间差异均有统计学意义（Cleaved caspase-3：F=23.33，P<0.001；Bax：F=60.31，P<0.001；Bcl-2：F=577.8，P<0.001；Cyt-c：F=104.0，P<0.001）。Cleaved caspase-3和Cyt-c水平在100 mM、200 mM染毒组较对照组升高，差异均有统计学意义（均有P<0.05）。Bax在50 mM、100 mM染毒组较对照组升高，差异均有统计学意义（均有P<0.05），200 mM染毒组与对照组相比差异无统计学意义（P=1.000）。Bcl-2水平在50 mM、100 mM、200 mM染毒组较对照组均降低，差异均有统计学意义（均有P<0.05）。Bax/Bcl-2比值增加，100 mM，200 mM组较对照组差异均有统计学意义（均有P<0.05）。结论 DMF染毒后可上调H9c2心肌细胞Cleaved caspase-3、Cyt-c以及Bax/Bcl-2水平，下调Bcl-2水平。线粒体通路参与了DMF诱导的H9c2心肌细胞凋亡的过程。

In vitro study of mitochondrial pathway involved cardiomyocytes apoptosis induced by N,N-dimethylformamide

Objective To investigate Dimethylformamide (DMF) on apoptosis-related proteins about mitochondrial pathway in cardiomyocytes. Methods Western Blotting was used to detect the the levels of Cleaved caspase-3, Bax, Bcl-2 and Cytochrome C(Cyt-c) in cytoplasm after different doses of DMF (0 mmol/L, 50 mmol/L, 100 mmol/L, 200 mmol/L) exposure for 24 h. Results After different doses of DMF (0 mM, 50 mM, 100 mM, 200 mM) exposure for 24h, the levels of Cleaved caspase-3, Bax and Cyt-c increased,while the level of Bax decreased. The differences between groups were statistically different (Cleaved caspase-3: F=23.33, P<0.001; Bax:F=60.31, P<0.001; Bcl-2: F=577.8, P<0.001; Cyt-c: F=104.0, P<0.001). Cleaved caspase-3 and Cyt-c increased after 100 mM and 200 mM exposure compared with control group, the differences were significant (all P<0.05). The levels of Bax increased after 50 mM and 100 mM DMF exposure compared with control group (all P<0.05), while the difference between 200 mM DMF exposure and control group were not statistically significant(P=1.000). The level of Bcl-2 exposed under 50 mM, 100 mM and 200 mM DMF decreased compared with the control group, the differences were statistically significant (all P<0.05). The ratio of Bax/Bcl-2 in 100 mM and 200 mM exposure group increased compared with control group, the differences were significant(all P<0.05). Conclusions DMF up-regulated the level of Cleaved caspase-3, Cyt-c and Bax/Bcl-2, meanwhile down-regulated the level of Bcl-2.Mitochondrial pathway was involved in H9c2 cardiomyocytes apoptosis induced by DMF.