Department of Anesthesiology, Beijing Stomatological Hospital Affiliated to Capital Medical University, Beijing, 100050, China.
Department of Anesthesiology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Jinan, 250014, China.
Cardiovasc Toxicol. 2024 Jul;24(7):637-645. doi: 10.1007/s12012-024-09869-y. Epub 2024 May 8.
To investigate the role of microRNA-195-3p (miR-195-3p) in hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury. AC16 human cardiomyocyte cells were cultured and pretreated with different concentrations of isoflurane (ISO) (1%, 2%, and 3%), followed by 6 h each of hypoxia and reoxygenation to construct H/R cell models. The optimum ISO concentration was assessed based on the cell viability. miR-195-3p expression was regulated by in vitro cell transfection. Cell viability was determined by MTT assay, and apoptosis was evaluated by flow cytometry. The levels of myocardial injury and inflammation were determined by enzyme-linked immunosorbent assay. Compared with the control group, the cell viability of the H/R group had significantly decreased and that of ISO pretreatment had increased in a dose-dependent manner. Therefore, we selected a 2% ISO concentration for pretreatment. MiR-195-3p expression had significantly increased in the H/R group and decreased after 2% ISO pretreatment. Additionally, the number of apoptotic cells and the levels of lactate dehydrogenase, creatine kinase-myoglobin binding, cardiac troponin I, interleukin (IL)-1β, IL-6, and tumor necrosis factor-α had increased significantly. ISO preconditioning inhibited H/R-induced AC16 cell damage, whereas miR-195-3p overexpression reversed the protective effects of ISO on cardiomyocytes. The expression of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) was reduced in the H/R-induced AC16 cells, and PTEN is a downstream target gene of miR-195-3p. Preconditioning with 2% ISO plays a protective role in H/R-induced AC16 cell damage by inhibiting miR-195-3p expression.
探讨微小 RNA-195-3p（miR-195-3p）在缺氧/复氧（H/R）诱导的心肌细胞损伤中的作用。
培养 AC16 人心肌细胞，用不同浓度的异氟醚（ISO）（1%、2%和 3%）预处理，然后进行 6 h 缺氧和复氧，构建 H/R 细胞模型。根据细胞活力评估最佳 ISO 浓度。通过体外细胞转染调节 miR-195-3p 的表达。通过 MTT 测定法测定细胞活力，通过流式细胞术评估细胞凋亡。通过酶联免疫吸附试验测定心肌损伤和炎症水平。
与对照组相比，H/R 组细胞活力明显下降，ISO 预处理呈剂量依赖性增加。因此，我们选择 2% ISO 浓度进行预处理。H/R 组 miR-195-3p 表达明显增加，2% ISO 预处理后表达降低。此外，凋亡细胞数和乳酸脱氢酶、肌红蛋白结合肌酸激酶、心肌肌钙蛋白 I、白细胞介素（IL）-1β、IL-6 和肿瘤坏死因子-α水平明显升高。ISO 预处理抑制 H/R 诱导的 AC16 细胞损伤，而过表达 miR-195-3p 则逆转 ISO 对心肌细胞的保护作用。在 H/R 诱导的 AC16 细胞中，磷酸酶和张力蛋白同源物缺失的染色体 10（PTEN）表达减少，PTEN 是 miR-195-3p 的下游靶基因。用 2% ISO 预处理通过抑制 miR-195-3p 的表达对 H/R 诱导的 AC16 细胞损伤起保护作用。
