1. Protective effect of cholecystokinin octapeptide on angiotensin II‐induced apoptosis in H9c2 cardiomyoblast cells
- Author
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Can Wang, Xiaoying Dong, Mingyang Hong, Limu Wei, Huan Yu, Lu Fu, Lin Chen, and Jingqi Zhang
- Subjects
0301 basic medicine ,Cell Survival ,Tetrazolium Salts ,Apoptosis ,Devazepide ,digestive system ,Biochemistry ,Sincalide ,Cell Line ,Phosphatidylinositol 3-Kinases ,03 medical and health sciences ,0302 clinical medicine ,Animals ,Myocytes, Cardiac ,Receptor ,Molecular Biology ,Protein kinase B ,Cholecystokinin ,Chemistry ,Angiotensin II ,Gene Expression Profiling ,Myocardium ,digestive, oral, and skin physiology ,Cell Biology ,Molecular biology ,Rats ,Thiazoles ,030104 developmental biology ,Gene Expression Regulation ,030220 oncology & carcinogenesis ,Cholecystokinin B receptor ,Benzimidazoles ,Signal transduction ,Peptides ,hormones, hormone substitutes, and hormone antagonists ,Signal Transduction - Abstract
Cholecystokinin (CCK) and its receptors are expressed in mammalian cardiomyocytes and are involved in cardiovascular system regulation; however, the exact effect and underlying mechanism of CCK in cardiomyocyte apoptosis remain to be elucidated. We examined whether sulfated CCK octapeptide (CCK-8) protects H9c2 cardiomyoblast cells against angiotensin II (Ang II)-induced apoptosis. The H9c2 cardiomyoblasts were subjected to Ang II with or without CCK-8 and the viability and apoptotic rate were detected using a Cell Counting Kit-8 assay, Hoechst 33342 staining, terminal deoxyribonucleotide transferase-mediated nick-end labeling assays, and flow cytometry. In addition, specific antiapoptotic mechanisms of CCK-8 were investigated using specific CCK1 (Devazepide) or CCK2 (L365260) receptor antagonists, or the PI3K inhibitor LY294002. The expression of CCK, CCK1 receptor, CCK2 receptor, Akt, p-Akt, Bad, p-Bad, Bax, Bcl-2, and caspase-3 were detected by Western blot analysis and real-time polymerase chain reaction. We found that CCK and its receptor messenger RNA (mRNA) and protein are expressed in H9c2 cardiomyoblasts. Ang II-induced increased levels of CCK mRNA and protein expression and decreased levels of CCK1 receptor protein and mRNA. Pretreatment of CCK-8 attenuated Ang II-induced cell toxicity and apoptosis. In addition, pretreatment of H9c2 cells with CCK-8 markedly induced expression of p-Akt, p-bad, and Bcl-2 and decreased the expression levels of Bax and caspase-3. The protective effects of CCK-8 were partly abolished by Devazepide or LY294002. Our results suggest that CCK-8 protects H9c2 cardiomyoblasts from Ang II-induced apoptosis partly via activation of the CCK1 receptor and the phosphatidyqinositol-3 kinase/protein kinase B (PI3K/Akt) signaling pathway.
- Published
- 2019