Study on apoptosis mechanism of H9c2 cardiomyocytes induced by N, N-dimethylformamide
10.3760/cma.j.issn.1001-9391.2017.02.006
- VernacularTitle: 二甲基甲酰胺诱导H9c2心肌细胞凋亡机制研究
- Author:
Qingxiu RONG
1
;
Yongjian YANG
;
Wanwan ZHANG
;
Juan WANG
;
Xiaoli MA
;
Mei XIANG
;
Lulu WANG
;
Wenjing SUN
Author Information
1. Anhui Medical University, Department of Occupational and Environmental Health, Hefei 230032, China
- Publication Type:Journal Article
- Keywords:
Dimethylformamide;
H9c2 cardiomyocytes;
Apoptosis;
NF-kappa B;
Mitochondria
- From:
Chinese Journal of Industrial Hygiene and Occupational Diseases
2017;35(2):106-110
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the change levels of nuclear factor-kappa B (NF-κB) p65 protein in cytoplasm and nuclear, phosphorylation of inhibitor of kappa B (p-IκB) protein and cytochrome C (Cyt-c) , cleaved cysteinyl aspartate specific proteinase-3 (Cleaved caspase-3) , B-cell lymphoma/leukemia-2 (Bcl-2) in cytoplasm in the process of N, N-dimethylformamide (DMF) -induced apoptosis in H9c2 cardiomyocytes, and explore the tentative mechanism of apoptosis.
Methods:H9c2 cardiomyocytes were exposed to 200 mmol/L DMF. Western blotting was used to detect the protein expression levels of p65 in cytoplasm and nuclear, p-IκB after exposure for 0, 2, 4, 6, 8, 12 h, and the protein expression levels of Cyt-c, Cleaved caspase-3, Bcl-2 in cytoplasm after exposure for 0, 2, 4, 8, 12, 24 h. Immunofluorescencecytochemistry (IFC) was used to observe the location of Cyt-c after 200 mmol/L DMF exposure for different times.
Results:The levels of p65 in cytoplasm and nuclear and p-IκB among groups were statistically significant (F were 7.79, 33.11, 90.25, respectively, all P<0.01) . Compared with the control group, the levels of p65 in cytoplasm of 2, 4, 6 h group were significantly decreased (all P<0.01) ; the levels of p65 in nuclear of 2, 4, 6, 8 h were significantly increased (all P<0.01) ; the levels of p-IκB of 2, 4, 6 h group were significantly increased (all P<0.01) . The levels of Cyt-c, Cleaved caspase-3 and Bcl-2 among groups were statistically significant (F were 51.42, 503.68, 73.37, respectively, all P<0.01) . Compared with the control group, the levels of Cyt-c of 8, 12 h group were significantly increased (both P<0.01) ; the levels of Cleaved caspase-3 of 2, 4, 8, 12, 24 h were significantly increased (all P<0.01) ; the levels of Bcl-2 of 2, 4, 8, 12, 24 h group were significantly decreased (all P<0.01) . IFC showed that Cyt-c was released from the mitochondria to the cytoplasm gradually as the extension of the exposure time.
Conclusion:NF-κB signaling pathway and mitochondrial pathway are involved in the mechanism of DMF-induced apoptosis in H9c2 cardiomyocytes.
