Protective effect of up-regulation of Yes-associated protein on cardiomyocyte injury induced by hypoxia reoxygenation
Gao Jie
Wei Zhenheng
Yang Chunhua
Wang Ke
Abstract：Objective To study the effect of Yes-associated protein (YAP) on cardiomyocyte injury induced by hypoxia/reoxygenation (H/R). Methods The cardiomyocytes were infected with over-expressed YAP recombinant lentivirus and treated with H/R. The expression of YAP was detected by using real-time fluorescence quantitative polymerase chain reaction (RT-PCR) and Western blotting assay. The proliferation changes of cardiomyocytes were detected by using CCK8 method, and leakage rate of lactic dehydrogenase (LDH) was detected by using dinitrophenylhydrazine (DNPH) method. The apoptosis changes of cardiomyocytes were detected by using flow cytometry (FCM), and levels of activated Caspase-3 protein and Caspase-9 protein were detected by using Western blotting assay. The level of reactive oxygen species (ROS) was detected by using DCFH-DA method, activity of superoxide dismutase (SOD) was detected by using xanthine oxidation (XTO) method, mitochondrial membrane potential (MMP) was detected by using JC-1 method, and levels of Cytochrome C in cytoplasma and mitochondrion were detected by using Western blotting assay. Results The infection of over-expressed YAP recombinant lentivirus increased YAP expression in cardiomyocytes under H/R condition. The proliferation activity decreased, leakage rate of LDH increased, apoptosis rate increased, levels of activated Caspase-3 protein and Caspase-9 protein increased, and ROS level increased in cardiomyocytes after H/R treatment. At the same time SOD activity decreased, MMP was reduced, and level of Cytochrome C protein increased in cytoplasma and decreased in mitochondrion. After up-regulating YAP, the proliferation activity was improved, leakage rate of LDH was decreased, apoptosis was reduced and expressions of Caspase-3 and Caspase-9 were decreased in cardiomyocytes under H/R condition. And SOD activity was increased, ROS level was decreased, MMP was promoted, and level of Cytochrome C protein was decreased in cytoplasma and increased in mitochondrion. Conclusion The up-regulation of YAP can relieve H/R cardiomyocyte injury and reduce apoptosis, and the mechanism may be related to the improvement of antioxidant enzyme activity, decrease of ROS level and inhibition of mitochondrial apoptosis pathway.
Keywords：Hypoxia/reoxygenationYes-associated proteinCardiomyocytesApoptosis
Publication Date：2019-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 825-829 )