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- X L Zhou, Y H Fang, Y Zhao, B Zou, Q R Xu, H Xu, C X Rao, and J C Liu.
- Department of Cardiovascular Surgery, the First Affiliated Hospital of Nanchang University, Nanchang 330006, China.
- Zhonghua Yi Xue Za Zhi. 2016 May 31; 96 (20): 1591-6.
ObjectiveTo investigate the protective effects of Notch1 on myocardial ischemia reperfusion injury and explore underlying molecular mechanism.MethodsH9c2 cells were exposed to hypoxia/reoxygenation (H/R), ischemic preconditioning (IPC) and ischemic postconditioning (IPost) treatment following infection with lentiviral vector-Notch1 intracellular domain (Lv-N1ICD) or Lv-N1ICD-shRNA, and divided into control group, H/R group, H/R+ N1ICD group, IPC group, IPC+ N1ICD-shRNA group, IPost group, IPost+ N1ICD-shRNA group, respectively. Apoptosis was detected by Annexin V/propidium iodide (PI), and reactive oxygen species (ROS) was detected by 2', 7'dichlorofluorescin-diacetate (DCFH-DA). The expression of p-glycogen synthase kinase-3β (p-GSK-3β)/GSK-3β were detected by Western blot analysis.ResultsThe apoptosis rate of cardiomyocytes in control group, H/R group, H/R+ N1ICD group, IPC group, IPC+ N1ICD-shRNA group, IPost group, IPost+ N1ICD-shRNA group was (5.34±1.70)%, (47.03±4.10)%, (33.89±12.25)%, (35.85±3.17)%, (51.12±8.22)%, (37.35±3.82)%, (47.90±3.08)%, respectively, showing significant differences in all group (F=18.47, P<0.05). ROS in control group, H/R group, H/R+ N1ICD group, IPC group, IPC+ N1ICD-shRNA group, IPost group, IPost+ N1ICD-shRNA group was 624.66±79.52, 1 221.87±63.66, 913.12±115.82, 935.85±201.62, 1 204.03±113.82, 967.15±106.11, 1 296.59±222.38, respectively, showing significant differences in all group (F=8.77, P<0.05). The ratio of p-GSK-3β to GSK-3β in control group, H/R group, H/R+ N1ICD group, IPC group, IPC+ N1ICD-shRNA group, IPost group, IPost+ N1ICD-shRNA group was 0.58±0.12, 0.62±0.20, 1.24±0.09, 1.16±0.12, 0.72±0.15, 1.16±0.12, 0.75±0.12, respectively, showing significant differences in all group (F=11.21, P<0.05).ConclusionAs an endogenous cardioprotective factor, Notch1 reduces myocardial ischemia reperfusion injury by promoting GSK-3β phosphorylation, inhibiting cardiomyocyte apoptosis and reducing ROS formation.
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