Change of subunit of NADPH oxidation enzyme complex nox-1 protein in cardiocyte hypoxia-reoxygenation injury and the role of cardiotrophin-1 |
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| Authors: | WAN Lei LI Ju-xiang HONG Kui DING Hao XIA Zi-rong SU Hai YAN Su-juan WU Yan-qing WU Qing-hua CHENG Xiao-shu |
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| Institution: | Department of Cardiology, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, China. E-mail: ljx912@126.com |
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| Abstract: | AIM: To observe the change of subunit of NADPH oxidation enzyme complex nox-1 protein in cardiocyte hypoxia-reoxygenation injury and the role of cardiotrophin-1.METHODS: Cardiomyocytes from the hearts of 1-3 d old neonatal rats were prepared by a modified method. Five groups were included in the study: control; hypoxia/reoxygenation; hypoxia/reoxygenation+CT-1; CT-1+hypoxia/reoxygenation+LY294002 (PIK3/Akt inhibitor); CT-1+hypoxia/reoxygenation+PD98059 (ERK inhibitor); CT-1+hypoxia/reoxygenation+DMSO. The concentration of CT-1 was 10 μg/L. The survival rate of myocytes was evaluated by MTS method. Apoptosis, mitochondrial permeability transition pore (Δψm) and reactive oxygen species (ROS) were detected by flow cytometry. Nox-1 protein was determined by Western blotting.RESULTS: Apoptosis of cardiomyocytes and the level of ROS (19.7%±1.4% vs 2.1%±0.5%, 14.07%±1.25% vs 3.54%±0.86%, P<0.05) increased markedly after hypoxia/reoxygenation, but cardiomyocyte survival rate and the level of Δψm (40.55%±4.25% vs 86.28%±7.15%, P<0.01) decreased significantly. The expression of nox-1 protein was upregulated markedly. With CT-1 intervention, cardiomyocyte survival rate increased markedly, apoptosis, both ROS and expression of nox-1 protein reduced significantly. The level of Δψm increased obviously. The effect of CT-1 was inhibited by LY294002. No significant effect was observed on cells survival in DMSO group, which confirmed that LY294002 was specifically involved in blocking the protective effect of CT-1.CONCLUSION: The expression of subunit of NADPH oxidation enzyme complex nox-1 protein is upregulated markedly in cardiocyte hypoxia-reoxygenation injury. CT-1 protects cardiac cells against hypoxia-reoxygenation injury by downregulating the expression of nox-1 protein to decrease the level of ROS. |
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