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,P<0.001),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P<0.001);Bcl-2蛋白水平组间差异具有统计学意义(F=28.92,P<0.001),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P<0.001);Cleaved caspase-3蛋白水平组间差异具有统计学意义(F=28.98,P<0.001)),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P<0.001)。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。
Objective To explore whether N,N-dimethylformamide （DMF） can induce apoptosis in H9c2 cardiomyocytes through mitochondrial pathway and its potential mechanism preliminarily.Methods H9c2 Cardiomyocytes were cultured in vitro and divided into groups with different treatments：control group,NAC group （4 mmol/L）,DMF group （200 mmol/L）,NAC （4 mmol/L） ＋ DMF （200 mmol/L） group.After treating with 24 h,the location of Cyt-c was detected by immunofluorescence cytochemistry （IFC） and the levels of Cyt-c,Bcl-2,Cleaved caspase-3 were detected by western blotting.Results IFC showed that the control and NAC group of Cyt-c were distributed in spotted or patchy shape in the cytoplasm,and the DMF group of Cyt-c was distributed in the cytoplasm diffusely,while NAC could inhibit the effect induced by DMF.Western blotting showed that the levels of Cyt-c different groups were significantly different （F =26.14,P 〈0.001）.Comparing with the control group,as for the level of Cyt-c,the DMF group was significantly increased and the NAC ＋ DMF group was significantly decreased than DMF group （all P 〈0.001）;as for the level of Bcl-2 among different groups were significantly different （F =28.92,P 〈0.001）.Compare with control group,the DMF group was significantly decreased and the NAC ＋ DMF group was significantly increased than DMF group （all P 〈 0.001）;as for the level of Cleaved caspase-3,there were significantly different （F =28.98,P 〈 0.001） among different groups.Compare with control group,the DMF group was significantly increased and the NAC ＋ DMF group was significantly decreased than DMF group （all P 〈0.001）.Conclusions DMF can induce apoptosis in H9c2 Cardiomyocytes through mitochondrial pathway.Oxidative stress may be involved in the pathogenesis.
作者
马晓丽
杨永坚
王娟
荣青秀
王璐璐
孙文静
向梅
MA Xiao-li YANG Yong-jian WANG Juan RONG Qing- xiu WANG Lu-lu SUN Wen-jing XIANG Mei(Department of Occupational and Environmental Health, School of Public Health, Anhui Medical University, Hefei 230032, China Anhui International Travel Healthcare Center, Hefei 230001, China Department of lnfection Management, the First Affiliated Hospital of Anhui Medical University, Hefei 230022, China)
出处
《中华疾病控制杂志》
CAS
CSCD
北大核心
2017年第4期404-408,共5页
Chinese Journal of Disease Control & Prevention
