Propofol Through Upregulating Caveolin-3 Attenuates Post-Hypoxic Mitochondrial Damage and Cell Death in H9C2 Cardiomyocytes During Hyperglycemia
Propofol Through Upregulating Caveolin-3 Attenuates Post-Hypoxic Mitochondrial Damage and Cell Death in H9C2 Cardiomyocytes During Hyperglycemia
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异丙酚通过上调 Caveolin-3 减轻高血糖期间 H9C2 心肌细胞缺氧后线粒体损伤和细胞死亡
DOI：
10.1159/000484680
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发表时间：
2017-01-01
影响因子：
--
通讯作者：
Xia, Zhengyuan
中科院分区：
文献类型：
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作者：
Deng, Fan;Wang, Shuang;Xia, Zhengyuan
背景/目的：糖尿病受试者的心脏易受心肌缺血再灌注（I/R）损伤。丙泊酚因其抗氧化特性而被证明可保护心肌I/R损伤，但其潜在机制仍不完全清楚。因此，本研究旨在确定丙泊酚是否可以通过在高血糖下上调Caveolin（Cav）-3来减轻线粒体功能障碍/损伤，从而减轻心肌I/R损伤。研究方法：采用酶联免疫吸附法（ELISA）检测H9 C2心肌细胞缺氧/复氧（H/R）后细胞活力、乳酸脱氢酶（LDH）、线粒体活力、肌酸激酶MB（CK-MB）、心肌肌钙蛋白I（cTnI）和细胞内三磷酸腺苷（ATP）含量的变化。使用2，7-二氯二氢荧光素二乙酸酯（DCF-DA）荧光染色评估细胞内的氧化应激水平，并通过检测线粒体膜电位和凋亡半胱氨酸蛋白酶3和9的活化来评估细胞依赖性凋亡。结果如下：细胞暴露于HG没有或与H/R都显着增加细胞损伤，细胞凋亡和增强的氧化应激，与线粒体功能障碍和Cav-3蛋白表达下降。所有这些变化在HG下H/R后进一步加剧。丙泊酚浓度为12.5 - 50 µM（但非100 µM）时，可显著减轻与Cav-3表达增加和促生存蛋白Akt和STAT 3活化相关的H/R损伤，在50 µM丙泊酚时观察到最佳保护作用（P25）。用β-甲基-环糊精破坏Cav-3可消除丙泊酚（P25）的有益作用。结论：丙泊酚通过在高血糖期间上调Cav-3减轻线粒体损伤并改善线粒体生物合成来抵消心肌细胞H/R损伤。
Background/Aims: Hearts from diabetic subjects are susceptible to myocardial ischemia reperfusion (I/R) injury. Propofol has been shown to protect against myocardial I/R injury due to its antioxidant properties while the underlying mechanism remained incompletely understood. Thus, this study aimed to determine whether or not propofol could attenuate myocardial I/R injury by attenuating mitochondrial dysfunction/damage through upregulating Caveolin (Cav)-3 under hyperglycemia. Methods: Cultured rat cardiomyocyte H9C2 cells were subjected to hypoxia/reoxygenation (H/R) in the absence or presence of propofol under high glucose (HG), and cell viability, lactate dehydrogenase (LDH) and mitochondrial viability as well as creatine kinase-MB (CK-MB), cardiac troponin I (cTnI) and intracellular adenosine triphosphate (ATP) content were measured with colorimetric Enzyme-Linked Immunosorbent Assays. Intracellular levels of oxidative stress was assessed using 2,7-dichlorodihydrofluorescein diacetate (DCF-DA) fluorescent staining and mitochondrial-dependent apoptosis was assessed by detecting mitochondrial membrane potential and the activation of apoptotic caspases 3 and 9. Results: Exposure of cells to HG without or with H/R both significantly increased cell injury, cell apoptosis and enhanced oxidative stress that were associated with mitochondrial dysfunction and decreased Cav-3 protein expression. All these changes were further exacerbated following H/R under HG. Administration of propofol at concentrations from 12.5 to 50 µM but not 100 µM significantly attenuated H/R injury that was associated with increased Cav-3 expression and activation of the prosurvival proteins Akt and STAT3 with the optimal protective effects seen at 50 µM of propofol (P25). The beneficial effects of propofol(P25) were abrogated by Cav-3 disruption with β-methyl-cyclodextrin. Conclusion: Propofol counteracts cardiomyocyte H/R injury by attenuating mitochondrial damage and improving mitochondrial biogenesis through upregulating Cav-3 during hyperglycemia.