Department of Anesthesiology, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai 264000, Shandong, P.R. China.
J Microbiol Biotechnol. 2021 Aug 28;31(8):1069-1078. doi: 10.4014/jmb.2103.03040.
Sevoflurane postconditioning (SPostC) has been proved effective in cardioprotection against myocardial ischemia/reperfusion injury. It was also reported that heat shock protein 70 (HSP70) could be induced by sevoflurane, which played a crucial role in hypoxic/reoxygenation (HR) injury of cardiomyocytes. However, the mechanism by which sevoflurane protects cardiomyocytes via HSP70 is still not understood. Here, we aimed to investigate the related mechanisms of SPostC inducing HSP70 expression to reduce the HR injury of cardiomyocytes. After the HR cardiomyocytes model was established, the cells transfected with siRNA for HSP70 (siHSP70) or not were treated with sevoflurane during reoxygenation. The lactate dehydrogenase (LDH) level was detected by colorimetry while cell viability and apoptosis were detected by MTT and flow cytometry. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blotting were used to detect HSP70, apoptosis-, cell cycle-associated factors, iNOS, and Cox-2 expressions. Enzyme-linked immuno sorbent assay (ELISA) was used to measure malondialdehyde (MDA) and superoxide dismutase (SOD). SPostC decreased apoptosis, cell injury, oxidative stress and inflammation and increased viability of HR-induced cardiomyocytes. In addition, SPostC downregulated Bax and cleaved caspase-3 levels, while SPostC upregulated Bcl-2, CDK-4, Cyclin D1, and HSP70 levels. SiHSP70 had the opposite effect that SPostC had on HR-induced cardiomyocytes. Moreover, siHSP70 further reversed the effect of SPostC on apoptosis, cell injury, oxidative stress, inflammation, viability and the expressions of HSP70, apoptosis-, and cell cycle-associated factors in HR-induced cardiomyocytes. In conclusion, this study demonstrates that SPostC can reduce the HR injury of cardiomyocytes by inducing HSP70 expression.
七氟醚后处理（SPostC）已被证明可有效对抗心肌缺血/再灌注损伤。有报道称，七氟醚可以诱导热休克蛋白 70（HSP70），这在心肌细胞缺氧/复氧（HR）损伤中发挥着关键作用。然而，七氟醚通过 HSP70 保护心肌细胞的机制仍不清楚。在这里，我们旨在研究 SPostC 诱导 HSP70 表达减少心肌细胞 HR 损伤的相关机制。在建立 HR 心肌细胞模型后，用 HSP70 的 siRNA（siHSP70）转染或未转染的细胞在再氧合时用七氟醚处理。通过比色法检测乳酸脱氢酶（LDH）水平，通过 MTT 和流式细胞术检测细胞活力和凋亡。逆转录-定量聚合酶链反应（RT-qPCR）和 Western blot 用于检测 HSP70、凋亡、细胞周期相关因子、iNOS 和 Cox-2 的表达。酶联免疫吸附试验（ELISA）用于测量丙二醛（MDA）和超氧化物歧化酶（SOD）。SPostC 降低了 HR 诱导的心肌细胞的凋亡、细胞损伤、氧化应激和炎症，并增加了其活力。此外，SPostC 下调 Bax 和 cleaved caspase-3 水平，而上调 Bcl-2、CDK-4、Cyclin D1 和 HSP70 水平。siHSP70 对 HR 诱导的心肌细胞产生了与 SPostC 相反的作用。此外，siHSP70 进一步逆转了 SPostC 对 HR 诱导的心肌细胞凋亡、细胞损伤、氧化应激、炎症、活力和 HSP70、凋亡和细胞周期相关因子表达的影响。总之，本研究表明，SPostC 通过诱导 HSP70 表达来减轻心肌细胞的 HR 损伤。
