Department of Cardiology, Panyu Central Hospital, Guangzhou, 511400, Guangdong, PR China.
Department of Respiratory, The First Affiliated Hospital of Jinan University, Guangzhou, 511632, Guangdong, PR China.
J Biomed Nanotechnol. 2022 Jan 1;18(1):202-210. doi: 10.1166/jbn.2022.3231.
To explore the effect of microRNA-455-5p (miR-455-5p) and Cytokine Signaling-3 (SOCS3) expression, a model of the cell damage induced during myocardial infarction was established using H₂O₂. The cell counting Kit-8 (CCK-8) and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assays were used to detect the cell viability and the expression of miR-455-5p and SOCS3 in cells cultured with different concentrations of H₂O₂. After the selection of the optimum culture concentration, a dual-luciferase reporter gene assay was used to detect the binding between and miR-455-5p and its potential target SOCS3. SOCS3 siRNA was transfected into cardiomyocytes using chitosan nanoparticles as a gene carrier, which led to the knockdown of SOCS3 expression, and the cells were transfected with miR-455-5p mimics and inhibitors. The expression of cardiac protective proteins was detected by western blotting, cell viability was detected by CCK8, and cell apoptosis was detected by flow cytometry. The aim of this study was to investigate the effect of miR-455-5p and SOCS3 expression on the activity and apoptosis of damaged cardiomyocytes, and to identify any protective effect on cardiomyocytes. Finally, after the simultaneous overexpression of SOCS3 and miR-455-5p, and the expression of cardiac protective proteins, cell activity, and apoptosis rate were detected. The results showed that the expression of miR-455-5p decreased in a concentration-dependent manner and that the expression of SOCS3 increased in a concentration-dependent manner when the cells were cultured in different concentrations of H₂O₂. The knockdown of SOCS3 expression promoted an increase in cell activity, an increase in cardiac protective proteins, and a decrease in apoptosis. The upregulation of miR-455-5p significantly inhibited the expression of SOCS3, increased cell activity, inhibited apoptosis, and exerted protective effects in myocardial cells. The overexpression of SOCS3 reversed the inhibition of SOCS3 by miR-455-5p and reduced the protective effect of miR-455-5p on myocardial cells. Therefore, this study showed that the upregulation of miR-455-5p significantly inhibited the expression of SOCS3 and resulted in the increased protection of cells damaged by H₂O₂, which was used as a model of myocardial infarction. These results indicate the potential of miR-455-5p in myocardial protection, suggesting that miRNA may be a resource for myocardial therapy.
为了探讨微小 RNA-455-5p（miR-455-5p）和细胞因子信号转导 3（SOCS3）表达的影响，我们使用 H₂O₂建立了心肌梗死细胞损伤模型。细胞计数试剂盒-8（CCK-8）和实时定量逆转录聚合酶链反应（qRT-PCR）检测不同浓度 H₂O₂培养的细胞活力以及 miR-455-5p 和 SOCS3 的表达。选择最佳培养浓度后，使用双荧光素酶报告基因检测 miR-455-5p 与其潜在靶标 SOCS3 之间的结合。使用壳聚糖纳米粒作为基因载体将 SOCS3 siRNA 转染到心肌细胞中，导致 SOCS3 表达下调，然后转染 miR-455-5p 模拟物和抑制剂。通过 Western blot 检测心脏保护蛋白的表达，通过 CCK8 检测细胞活力，通过流式细胞术检测细胞凋亡。本研究旨在探讨 miR-455-5p 和 SOCS3 表达对损伤心肌细胞活性和凋亡的影响，以及对心肌细胞的任何保护作用。最后，同时过表达 SOCS3 和 miR-455-5p 后，检测心脏保护蛋白的表达、细胞活性和凋亡率。结果表明，细胞在不同浓度 H₂O₂中培养时，miR-455-5p 的表达呈浓度依赖性降低，SOCS3 的表达呈浓度依赖性增加。SOCS3 表达下调促进细胞活力增加、心脏保护蛋白增加和凋亡减少。miR-455-5p 的上调显著抑制 SOCS3 的表达，增加细胞活力，抑制凋亡，并对心肌细胞发挥保护作用。SOCS3 的过表达逆转了 miR-455-5p 对 SOCS3 的抑制作用，并降低了 miR-455-5p 对心肌细胞的保护作用。因此，本研究表明，miR-455-5p 的上调显著抑制 SOCS3 的表达，并导致 H₂O₂诱导的细胞损伤得到增强保护，该模型用于心肌梗死。这些结果表明 miR-455-5p 在心肌保护中的潜力，提示 miRNA 可能是心肌治疗的资源。
