Department of Anesthesiology, The First Hospital of Lanzhou University, No. 1, Donggang West Road, Lanzhou, 730000, China.
Anesthesia and Operation Centre, the First Medical Center, Chinese PLA General Hospital, Beijing, 100853, China.
Cardiovasc Toxicol. 2021 Oct;21(10):800-807. doi: 10.1007/s12012-021-09670-1. Epub 2021 Jun 28.
The protective effect and mechanism of isoflurane on myocardial injury was investigated by constructing in vitro hypoxia/reoxygenation (HR) cell model. HR cell models were established in vitro and treated with isoflurane (ISO). qRT-PCR was used to detect the relative expression of miR-18a-5p. CCK-8 kit and flow cytometry were performed to evaluate cell proliferation and apoptosis. The myocardial injury related markers, such as Cκ-MB, cTnI and LDH were detected by ELISA. Luciferase reporter gene assay was used to verify the interaction between miR-18a-5p and target genes. The expression of miR-18a-5p was significantly increased in hypoxic cardiomyocytes compared with control group (P < 0.001). Meanwhile, cardiomyocytes in the HR group showed inhibition of proliferation, a significant increase in cell apoptosis and in myocardial injury indicators, such as Cκ-MB, cTnI and LDH (P < 0.001). However, 1% ISO treatment alleviated myocardial cell injury induced by HR. Transfection of miR-18a-5p under ISO reduced the protective effect of 1% ISO against myocardial cell damage. Luciferase report gene assay confirmed that CCND2 might be the target gene of miR-18a-5p. In the in vitro cell model of myocardium, ISO alleviated cardiomyocyte injury caused by hypoxia/reoxygenation by down-regulating the expression of miR-18a-5p.
本研究通过构建体外缺氧/复氧（HR）细胞模型来探讨异氟醚对心肌损伤的保护作用及其机制。体外建立 HR 细胞模型并给予异氟醚（ISO）处理。采用 qRT-PCR 检测 miR-18a-5p 的相对表达水平。使用 CCK-8 试剂盒和流式细胞术评估细胞增殖和凋亡。通过 ELISA 检测心肌损伤相关标志物，如 Cκ-MB、cTnI 和 LDH。采用荧光素酶报告基因实验验证 miR-18a-5p 与靶基因的相互作用。与对照组相比，缺氧心肌细胞中 miR-18a-5p 的表达明显增加（P<0.001）。同时，HR 组的心肌细胞增殖受到抑制，细胞凋亡显著增加，心肌损伤标志物如 Cκ-MB、cTnI 和 LDH 也显著增加（P<0.001）。然而，1%ISO 处理可减轻 HR 诱导的心肌细胞损伤。在 ISO 下转染 miR-18a-5p 可降低 1%ISO 对心肌细胞损伤的保护作用。荧光素酶报告基因实验证实 CCND2 可能是 miR-18a-5p 的靶基因。在体外心肌细胞模型中，ISO 通过下调 miR-18a-5p 的表达来减轻缺氧/复氧引起的心肌细胞损伤。
