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Experimental study on the effect of N-acetylcysteine on H9C2 cardiomyocyte injury induced by high-fat and high-glucose combined with hypoxia/reoxygenation |
LI Yuan XIA Zhongyuan |
Department of Anesthesiology, Renmin Hospital of Wuhan University, Hubei Province, Wuhan 430060, China |
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Abstract Objective To investigate the effect of antioxidant N-acetylcysteine (NAC) on H9C2 cardiomyocyte injury induced by high-fat and high-glucose combined with hypoxia/reoxygenation and its effect on the expression of clock gene BMAL1. Methods H9C2 cells were divided into control group (N group), high-fat and high-glucose combined with hypoxia/reoxygenation group (HFHG+H/R group), high-fat and high-glucose combined with hypoxia/reoxygenation +NAC group (HFHG+H/R+NAC group). Cell viability was detected by CCK-8 method, cell apoptosis was detected by the flow cytometry, reactive oxygen species (ROS) level was observed by DCFH-DA staining, mitochondrial membrane potential (MMP) was detected by JC-1 staining, and the expression level of BMAL1 was determined by Western blot. Results Compared with N group, the cell viability of HFHG+H/R group was significantly lower (P < 0.01), the apoptosis rate and ROS level were significantly increased (P < 0.01), while the expression level of BMAL1 was significantly reduced (P < 0.01). After NAC treatment, the changes of indicators above in the HFHG+H/R+NAC group were significantly reduced, the differences were highly statistically significant (P < 0.01). MMP in the HFHG+H/R group was significantly lower than that in the N group. After NAC treatment, MMP of HFHG+H/R+NAC group was significantly increased. Conclusion NAC may reduce the oxidative stress level of cardiomyocytes and promote the expression level of clock gene BMAL1, thereby reducing cardiomyocyte damage induced by high-fat and high-glucose combined with hypoxia/reoxygenation.
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