二甲基甲酰胺通过活性氧介导线粒体通路诱导H9c2心肌细胞凋亡研究

发布时间：2018-04-18 21:08

本文选题：二甲基甲酰胺 + 肌细胞　；参考：《中华疾病控制杂志》2017年04期

【摘要】：目的初步探讨二甲基甲酰胺(dimethylformamide,DMF)是否通过线粒体通路致H9c2心肌细胞凋亡及可能机制。方法 H9c2心肌细胞体外培养,分组给予不同处理:正常对照组,N-乙酰半胱氨酸(N-acetyl-L-cysteine,NAC)(4 mmol/L)组,DMF(200 mmol/L)组,NAC(4 mmol/L)+DMF(200 mmol/L)组,24 h后使用免疫荧光细胞化学法(immune-fluorescence cytochemistry,IFC)检测细胞内细胞色素C(cytochrome C,Cyt-c)定位,蛋白免疫印迹法检测胞浆Cyt-c、Bcl-2、Cleaved caspse-3水平。结果 IFC观察发现,正常对照及NAC组Cyt-c呈点状或斑块状分布于胞浆,DMF作用24 h导致Cyt-c弥散分布于胞浆,而NAC可抑制DMF的这种作用;蛋白免疫印迹结果表明,Cyt-c蛋白水平组间差异具有统计学意义(F=26.14,P0.001),与对照组相比,DMF组Cyt-c水平升高,NAC+DMF组较DMF组Cyt-c水平降低,差异均有统计学意义(均有P0.001);Bcl-2蛋白水平组间差异具有统计学意义(F=28.92,P0.001),与对照组相比,DMF组Bcl-2蛋白水平降低,与DMF组相比,NAC+DMF组Bcl-2蛋白水平升高,差异均有统计学意义(均有P0.001);Cleaved caspase-3蛋白水平组间差异具有统计学意义(F=28.98,P0.001)),与对照组相比,DMF组Cleaved caspase-3水平上升,NAC+DMF组较DMF组Cleaved caspase-3水平降低,差异均具有统计学意义(均有P0.001)。结论 DMF可以通过线粒体通路诱导H9c2心肌细胞凋亡,氧化应激可能参与该通路的发生。
[Abstract]:Aim to investigate whether dimethylformamide (DMF) can induce apoptosis of H9c2 cardiomyocytes through mitochondrial pathway and its possible mechanism.Methods H9c2 cardiomyocytes were cultured in vitro and divided into two groups: the normal control group was treated with N-acetyl-L-cysteine N-acetyl-L-cysteine (NACX / L) 4 mmol / L) group (n = 4 mmol / L). After 24 h, the cytochrome C(cytochrome (C(cytochrome) was detected by immuno-fluorescence cytochemistry.The level of cytoplasmic Cyt-c-2 Bcl-2Cleaved caspse-3 was detected by Western blot.Results IFC observation showed that the distribution of Cyt-c in the cytoplasm of normal control group and NAC group was punctate or plaques after 24 h treatment, which resulted in the distribution of Cyt-c in the cytoplasm, whereas NAC could inhibit the effect of DMF.The results of Western blot showed that there was significant difference in the level of Cyt-c protein between the two groups. Compared with the control group, the level of Cyt-c in the Cyt-c group increased and the Cyt-c level in the NAC DMF group was lower than that in the DMF group.There were significant differences between the two groups in the level of Bcl-2 protein (P 0.001). Compared with the control group, the level of Bcl-2 protein in the DMF group was lower than that in the control group, and the level of Bcl-2 protein in the DMF group was higher than that in the DMF group.The difference was statistically significant (P 0.001). The level of Cleaved caspase-3 in Cleaved DMF group was significantly lower than that in DMF group (P 0.001), and the difference was statistically significant (P 0.001). The difference was statistically significant (all P 0.001), and the difference was statistically significant (all P 0.001) compared with the control group, the level of Cleaved caspase-3 in the DMF group was significantly lower than that in the DMF group (P < 0.05), and there was no significant difference between the two groups (P > 0.05).Conclusion DMF can induce apoptosis of H9c2 cardiomyocytes through mitochondrial pathway, and oxidative stress may be involved in this pathway.
【作者单位】：安徽医科大学公共卫生学院劳动卫生与环境卫生学系;安徽国际旅行卫生保健中心;安徽医科大学第一附属医院感染管理科;
【分类号】：R114

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