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p38MAPK信号通路介导右美托咪定对缺氧/复氧心肌细胞损伤的保护作用 预览

Experimental study on the protective effect of dexmedetomidine on hypoxia/reoxygenation myocardial injury induced by p38MAPK signaling pathway
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摘要 目的探讨右美托咪定(Dex)是否通过p38丝裂原活化蛋白激酶(MAPK)信号通路发挥对缺氧/复氧(H/R)心肌细胞损伤的保护作用。方法培养大鼠心肌细胞H9c2,随机分为空白对照组、H/R组、Dex预处理(Dex+H/R)组、抑制剂SB203580(H/R+SB)组和Dex预处理+SB203580(Dex+H/R+SB)组。采用噻唑蓝(MTT)法检测各组细胞活力,流式细胞仪检测细胞凋亡率,试剂盒检测细胞培养液中乳酸脱氢酶(LDH)和肌酸激酶(CK)含量,分光光度计检测超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-Px)含量,Western印迹检测细胞中磷酸化(p)-p38MAPK、细胞色素(Cyt)C和活化的含半胱氨酸的天冬氨酸蛋白水解酶(酶切Caspase)-3蛋白水平。结果与H/R组比,Dex预处理可提高H9c2细胞活力,降低细胞凋亡率,下调LDH和CK的漏出量,减少MDA含量,上调SOD和GSH-Px活性,抑制p-p38MAPK、CytC和酶切Caspase-3蛋白表达,差异有统计学意义(P<0.05)。p38MAPK信号通路抑制剂SB203580可明显增强Dex对H/R心肌细胞损伤的保护作用。结论Dex可能通过p38MAPK信号通路抑制细胞中的氧化应激,下调CytC和酶切Caspase-3蛋白的表达,抑制H9c2细胞凋亡对H/R心肌细胞损伤的保护作用。 ObjectiveTo investigate whether dexmedetomidine (Dex) protects against hypoxia/reoxygenation myocardial injury through p38 mitogen-activated protein kinase (p38MAPK) signaling pathway. MethodsRat cardiomyocytes H9c2 were cultured and H9c2 cells were randomly divided into 5 groups: blank control (control group), hypoxia/reoxygenation (H/R group) and dexmedetomidine pretreatment (Dex+H/R group), inhibitor SB203580 (H/R+SB group) and dexmedetomidine pretreatment + SB203580 (Dex+H/R+SB group). The viability of each group was detected by MTT assay. The apoptosis rate was detected by flow cytometry. Content of lactate dehydrogenase (LDH) and creatine kinase (CK) in cell culture medium, the superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxide (GSH-Px) content were detected. Western blot analysis was used to detect phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK), cytochrome C (CytC) and activated cysteine-containing Caspase-3 protein levels. ResultsCompared with the H/R group, Dex pretreatment could increase the viability of H9c2 cells, decrease the apoptosis rate, down-regulate the leakage of LDH and CK, reduce the MDA content, up-regulate the activity of SOD and GSH-Px, and inhibit the p-p38MAPK, CytC and Cleaved Caspase-3. The expression of Caspase-3 protein was statistically significant ( P <0.05). The p38MAPK signaling pathway inhibitor SB203580 significantly enhanced the protective effect of Dex on hypoxia/reoxygenation myocardial injury. ConclusionsDexmedetomidine might inhibit oxidative stress in cells, down-regulate the expression of CytC and Cleaved Caspase-3 protein, and inhibit the protective effect of H9c2 cell apoptosis on hypoxia/reoxygenation cardiomyocyte injury through p38MAPK signaling pathway.
作者 赵彦芬 赵峰 刘鹏森 ZHAO Yan-Fen;ZHAO Feng;LIU Peng-Sen(Department of Anesthesiology, the Scond Affiliated Hospital, Henan Province Hospital, Henan Traditional Chinese Medicine University, Zheng Zhou 450002, Henan, China)
出处 《中国老年学杂志》 CAS 北大核心 2019年第14期3528-3532,共5页 Chinese Journal of Gerontology
关键词 右美托咪定 p38丝裂原活化蛋白激酶信号通路 缺氧/复氧 心肌细胞 氧化应激 Dexmedetomidine p38MAPK signaling pathway Hypoxia/reoxygenation Cardiomyocytes Oxidative stress
作者简介 第一作者:赵彦芬(1984-),女,主治医师,主要从事器官保护研究。
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