title: [Study on apoptosis mechanism of H9c2 cardiomyocytes induced by N, N-dimethylformamide].
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 (Fwere 7.79, 33.11, 90.25, respectively, allP<0.01) . Compared with the control group, the levels of p65 in cytoplasm of 2, 4, 6 h group were significantly decreased (allP<0.01) ; the levels of p65 in nuclear of 2, 4, 6, 8 h were significantly increased (allP<0.01) ; the levels of p-IκB of 2, 4, 6 h group were significantly increased (allP<0.01) . The levels of Cyt-c, Cleaved caspase-3 and Bcl-2 among groups were statistically significant (Fwere 51.42, 503.68, 73.37, respectively, allP<0.01) . Compared with the control group, the levels of Cyt-c of 8, 12 h group were significantly increased (bothP<0.01) ; the levels of Cleaved caspase-3 of 2, 4, 8, 12, 24 h were significantly increased (allP<0.01) ; the levels of Bcl-2 of 2, 4, 8, 12, 24 h group were significantly decreased (allP<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.
authors.affiliation: Anhui Medical University, Department of Occupational and Environmental Health, Hefei 230032, China.
authors.forename: Q X
authors.initials: QX
authors.lastname: Rong
authors.forename: Y J
authors.initials: YJ
authors.lastname: Yang
authors.forename: W W
authors.initials: WW
authors.lastname: Zhang
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.forename: X L
authors.initials: XL
authors.lastname: Ma
authors.forename: M
authors.initials: M
authors.lastname: Xiang
authors.forename: L L
authors.initials: LL
authors.lastname: Wang
authors.forename: W J
authors.initials: WJ
authors.lastname: Sun
mesh_terms: D017209:Apoptosis; D015153:Blotting, Western; D004126:Dimethylformamide / Q000494:pharmacology*; D020672:I-kappa B Proteins / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*
chemical_list: D020672:I-kappa B Proteins; D016328:NF-kappa B; D004126:Dimethylformamide
keywords: Apoptosis; Dimethylformamide; H9c2 cardiomyocytes; Mitochondria; NF-kappa B