Previous studies failed to elucidate the detailed mechanisms of anesthetic preconditioning as a protective approach against ischemic/reperfusion (I/R) injury in cells. This study mainly centered on discovering the mechanisms of Sevoflurane (Sev) in preventing cardiomyocytes against I/R injury. Human cardiomyocyte AC16 cell line was used to simulate I/R injury based on a hypoxia/reperfusion (H/R) model. After Sev treatment, cell viability and apoptosis were detected by MTT assay and flow cytometry,respectively. Lactate dehydrogenase (LDH) content was measured using an LDH detection kit. Relative mRNA and protein expressions of LINC01133, miR-30a-5p and apoptosis-related proteins were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot as needed. Target gene of miR-30a-5p and their potential binding sites were predicted using Starbase and confirmed by dual-luciferase reporter assay. Cell behaviors were assessed again after miR-30a-5p and LINC01133 transfection. Sev could improve cell viability, reduce LDH leakage, and downregulate the expressions of apoptosis-related proteins (Bax, cleaved caspase-3 and cleaved caspase-9) and LINC01133 as well as upregulate miR-30a-5p and Bcl-2 expressions in H/R cells. MiR-30a-5p was the target of LINC01133, and upregulating miR-30a-5p enhanced the effects of Sev in H/R cells, with a suppression on H/R-induced activation of the p53 signaling pathway. However, upregulating LINC01133 reversed the enhancing effects of miR-30a-5p on Sev pretreatment in H/R cells. Sev could protect cardiomyocytes against H/R injury through the miR-30a-5p/LINC01133 axis, which may provide a possible therapeutic method for curing cardiovascular I/R injury.

中文翻译：

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七氟醚通过LINC01133 / miR-30a-5p轴保护心肌细胞免受缺氧/再灌注损伤。

先前的研究未能阐明麻醉剂预处理的详细机制，将其作为针对细胞中缺血/再灌注（I / R）损伤的保护性方法。这项研究主要集中于发现七氟醚（Sev）预防心肌细胞抗I / R损伤的机制。基于缺氧/再灌注（H / R）模型，将人类心肌细胞AC16细胞系用于模拟I / R损伤。Sev处理后，分别通过MTT法和流式细胞术检测细胞活力和凋亡。使用LDH检测试剂盒测量乳酸脱氢酶（LDH）含量。使用定量实时聚合酶链反应（qRT-PCR）和Western印迹根据需要检测LINC01133，miR-30a-5p和凋亡相关蛋白的相对mRNA和蛋白表达。使用Starbase预测miR-30a-5p的靶基因及其潜在的结合位点，并通过双荧光素酶报告基因测定法进行确认。miR-30a-5p和LINC01133转染后再次评估细胞行为。Sev可以改善细胞活力，减少LDH泄漏，并下调凋亡相关蛋白（Bax，caspase-3裂解和caspase-9裂解）和LINC01133的表达，并上调miR-30a-5p和Bcl-2的表达/ R单元格。MiR-30a-5p是LINC01133的靶标，而上调miR-30a-5p可增强Sev在H / R细胞中的作用，并抑制H / R诱导的p53信号通路的激活。但是，上调LINC01133逆转了miR-30a-5p对H / R细胞中Sev预处理的增强作用。Sev可通过miR-30a-5p / LINC01133轴保护心肌细胞免受H / R损伤，