Hirsutine mitigates high-fat-induced apoptosis in H9C2 cells by activating mitophagy
SHENG Sunpeng1,CHEN Shijia2, DONG Xicheng3, DING Lu4, MA Jianshe4, JIN Peifeng1.
1.Department of Cardiac Surgery,the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China; 2.The First School of Medicine, School of Information and Engineering, Wenzhou Medical University, Wenzhou 325035, China;3.School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325035, China; 4.School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, China
SHENG Sunpeng,CHEN Shijia,DONG Xicheng, et al. Hirsutine mitigates high-fat-induced apoptosis in H9C2 cells by activating mitophagy[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2024, 54(10): 791-797,805.
Abstract：Objective: To explore the effect of Hirsutine on mitophagy and apoptosis in H9C2 cells induced by high-fat. Methods: With untreated H9C2 cells served as the control group (Control), H9C2 cells stimulated by palmitic acid were used as the high-fat group (PA), H9C2 cells treated with Hirsutine as the treatment group (PA+HS) and H9C2 cells treated with the Rho kinase inhibitor Y-27632 as the inhibitor group (PA+HS+Y-27632).CCK8 assay was used to evaluate cell viability; TUNEL staining was used to detect cell apoptosis; TMRE staining was used to detect changes in mitochondrial membrane potential (mPTP); Western blot was used to detect the expression of mitophagy proteins LC3I/II, Pink 1 and Parkin as well as apoptosis-related proteins Cleavedcaspase 3, caspase 3, Bax, and Bcl-2. Results: Compared with the control group, the viability of cells treated with 200, 400, and 800 μmol/L PA decreased (P<0.05); the TMRE fluorescence intensity was significantly weaker,while the expression of LC3-I/II, Pink , and Parkin was significantly lower (P<0.05); the TUNEL-positive cells increased significantly, and the expression of Cleaved-caspase 3/caspase 3 and Bax/Bcl-2 increased significantly (P<0.05) in the PA group. Compared with the PA group, the TMRE fluorescence intensity was significantly stronger; the expression of LC3-I/II, Pink , and Parkin was significantly higher (P<0.05); the TUNEL-positive cells decreased significantly, and the expression of Cleaved-caspase 3/caspase 3 and Bax/Bcl-2 decreased significantly (P<0.05) in the PA+HS group. Compared with the PA+HS group, the TMRE fluorescence intensity was weaker; the expression of LC3-I/II, Pink , and Parkin was significantly lower (P<0.05); the TUNELpositive cells increased significantly, and the expression of Cleaved-caspase 3/caspase 3 and Bax/Bcl-2 increased significantly (P<0.05) in the PA+HS+Y-27632 group. Conclusion: Hirsutine alleviates apoptosis of H9C2 cells induced by high-fat through the activation of mitophagy.
