In vitro study of mitochondrial pathway involved cardiomyocytes apoptosis induced by N, N-dimethylformamide
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摘要: 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。Abstract: 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.
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Key words:
- Dimethylformamide /
- Myocytes, cardiac /
- Apoptosis /
- In vitro
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王玉荣. 国内外DMF的生产与消费 [J]. 化工科技市场, 2004,27(12):32-37. 成振江,范竹玉. 二甲基甲酰胺中毒研究进展 [J]. 安全、健康和环境, 2011,11(12):44-46,49. 何月莹,吕艳,王德伟,等. 1990-2007年国内二甲基甲酰胺职业中毒案例分析 [J]. 工业卫生与职业病, 2009,35(3):184-189. 李冬梅,高丽蕙,朱兰英,等. 急性二甲基甲酰胺中毒性心肌损害1例报告 [J]. 职业与健康, 2007,23(10):804-805. 向梅. 二甲基甲酰胺对心肌细胞毒性及其氧化应激机制研究 [D]. 合肥:安徽医科大学, 2013. 詹凤侠,杨永坚,方四新,等. 653例二甲基甲酰胺暴露工人职业健康监护资料分析 [J]. 中国现代医学杂志, 2009,19(7):1081-1084. 向梅,杨永坚,王取南,等. 二甲基甲酰胺对心肌细胞的毒性作用 [J]. 中华疾病控制杂志, 2013,17(7):581-584. Hirsch T, Marzo I, Kroemer G. Role of the mitochondrial permeability transition pore in apoptosis:reactive oxygen species: membrane aspects [J]. Bioscience Reports, 1997,17(1):67-76. 郑天胜,李翔. 线粒体凋亡通路的研究进展 [J]. 医学综述, 2013,19(18):3282-3285. Joseph EK, Levine JD. Caspase signalling in neuropathic and inflammatory pain in the rat [J]. Eur J Neurosci, 2004,20(11):2896-2902 吴勃岩,王雪,董静,等. 齐墩果酸对S180荷瘤小鼠细胞色素C和Caspase-3表达的影响 [J]. 中医药信息, 2014,31(4):14-16. 路艳艳. 二甲基甲酰胺致人肝细胞凋亡及对Bcl-2、Bax和Caspase-3表达影响的研究 [D]. 杭州:浙江大学, 2007. Renault TT, Teijido O, Antonsson B, et al. Regulation of bax mitochondrial localization by Bcl-2 and Bcl-x (L): keep your friends close but your enemies closer [J]. Int J Biochem Cell Biol, 2013,45(1):64-67. 许茸茸,李应东. Bcl-2家族与线粒体凋亡通路相互作用研究进展 [J]. 中国老年学杂志, 2013,33(12):2977-2979. Tsai CJ, Liu S, Hung CL, et al. BAX-Induced Apoptosis Can Be Initiated through a Conformational Selection Mechanism [J]. Structure, 2015,23(1):139-148.
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