PDF(Pubmed)
Abstract：
Myocardial injury is an indicator of poor prognosis in sepsis, whereas propofol has been reported to protect the myocardium. Therefore, the present study investigated the effect of propofol on myocardial injury in sepsis and its mechanism. An in vitro model of myocardial cell injury was established in myocardial H9C2 cells using lipopolysaccharide (LPS). The Cell Counting Kit 8 (CCK8) assay was used to investigate the effect of propofol pretreatment on the viability of normal and LPS-challenged H9C2 cells, whereas the lactate dehydrogenase (LDH) detection kit was used to measure the levels of LDH. The expression levels of LC3 were analyzed using an immunofluorescence assay. Western blotting was performed to analyze the expression levels of autophagy-related proteins. Following treatment with the autophagy inhibitor 3-methyladenine, CCK8 assay, TUNEL assay, western blotting, 2,7-dichlorohydrofluorescein diacetate assay and ELISA were performed to investigate whether propofol exerted its effects on cell viability, apoptosis, oxidative stress and inflammation via autophagy. Moreover, to further explore the regulatory mechanism of propofol in myocardial injury, sirtuin 1 (SIRT1) was knocked down via transfection with small interfering RNA, and SIRT1 protein was inhibited via the addition of the SIRT1 inhibitor EX527. The present study demonstrated that propofol activated autophagy in LPS-induced cardiomyocytes, and reversed the effects of LPS on viability, apoptosis, oxidative stress and the inflammatory response. Moreover, SIRT1 knockdown and inhibition decreased the activation of autophagy and the protective effect of propofol on LPS-induced cardiomyocytes. In conclusion, propofol reduced LPS-induced cardiomyocyte injury by activating SIRT1-mediated autophagy.
摘要：
心肌损伤是脓毒症患者预后不良的指标,而异丙酚有保护心肌的报道。因此,本研究探讨异丙酚对脓毒症心肌损伤的影响及其机制。利用脂多糖(LPS)在心肌H9C2细胞中建立了心肌细胞损伤的体外模型。细胞计数试剂盒8（CCK8）用于研究异丙酚预处理对正常和LPS攻击的H9C2细胞活力的影响。而乳酸脱氢酶（LDH）检测试剂盒用于测量LDH的水平。使用免疫荧光测定法分析LC3的表达水平。蛋白质印迹法分析自噬相关蛋白的表达水平。用自噬抑制剂3-甲基腺嘌呤治疗后，CCK8测定，TUNEL检测,西方印迹,采用2,7-二氯氢荧光素二乙酸酯法和ELISA法研究异丙酚对细胞活力的影响,凋亡，氧化应激和炎症通过自噬。此外，进一步探讨异丙酚在心肌损伤中的调控机制,沉默蛋白1(SIRT1)通过小干扰RNA转染敲低，和SIRT1蛋白通过添加SIRT1抑制剂EX527被抑制。本研究表明丙泊酚激活LPS诱导的心肌细胞自噬,并逆转了LPS对生存力的影响，凋亡，氧化应激和炎症反应。此外，SIRT1敲除和抑制能降低自噬的激活,并能降低异丙酚对LPS诱导的心肌细胞的保护作用。总之,异丙酚通过激活SIRT1介导的自噬减轻LPS诱导的心肌细胞损伤。
