Department of Neurosurgery, The Second Hospital of Jilin University, Changchun, Jilin, China.
Department of Cardiology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
J Cell Physiol. 2020 Jan;235(1):176-184. doi: 10.1002/jcp.28956. Epub 2019 Jun 18.
Myocardial ischemia/reperfusion injury (MIRI) is a clinically familiar disease, which possesses a great negative impact on human health. But, the effective treatment is still absent. MicroRNAs (miRNAs) have been testified to play a momentous role in MIRI. The purpose of the study aimed to probe the functions of miR-132 in oxygen and glucose deprivation (OGD)-evoked injury in H9c2 cells. miR-132 expression in H9c2 cells accompanied by OGD disposition was evaluated via real-time quantitative polymerase chain reaction. After miR-132 mimic and inhibitor transfections, the impacts of miR-132 on OGD-affected H9c2 cell viability, apoptosis, cell cycle, and the interrelated factors were appraised by exploiting cell counting kit-8, flow cytometry, and western blot analysis. FOXO3A expression was estimated in above-transfected cells, meanwhile, the correlation between miR-132 and FOXO3A was probed by dual-luciferase report assay. Ultimately, above mentioned cell processes were reassessed in H9c2 cells after preprocessing OGD administration and transfection with si-FOXO3A and si-NC plasmids. We got that OGD disposition obviously enhanced miR-132 expression in H9c2 cells. Overexpressed miR-132 evidently reversed OGD-evoked cell viability repression and apoptosis induction in H9c2 cells. In addition, overexpressed miR-132 mitigated OGD-evoked G0/G1 cell arrest by mediating p21, p27, and cyclin D1 expression. Repression of FOXO3A was observed in miR-132 mimic-transfected cells, which was also predicated as a direct gene of miR-132. We discovered that silenced FOXO3A alleviated OGD-evoked cell injury in H9c2 cells via facilitating cell viability, hindering apoptosis and restraining cell arrest at G0/G1 phase. In conclusion, these investigations corroborated that miR-132 exhibited the protective impacts on H9c2 cells against OGD-evoked injury via targeting FOXO3A.
心肌缺血/再灌注损伤（MIRI）是一种临床上常见的疾病，对人类健康有很大的负面影响。但是，有效的治疗方法仍然缺乏。microRNAs（miRNAs）已被证明在 MIRI 中发挥重要作用。本研究旨在探讨 miR-132 在氧葡萄糖剥夺（OGD）诱导的 H9c2 细胞损伤中的作用。通过实时定量聚合酶链反应评估 H9c2 细胞中 miR-132 在 OGD 处理下的表达。转染 miR-132 模拟物和抑制剂后，通过细胞计数试剂盒-8、流式细胞术和 Western blot 分析评估 miR-132 对 OGD 影响的 H9c2 细胞活力、凋亡、细胞周期及相关因素的影响。在上述转染细胞中评估 FOXO3A 的表达，同时通过双荧光素酶报告实验探讨 miR-132 与 FOXO3A 的相关性。最后，在预处理 OGD 给药和转染 si-FOXO3A 和 si-NC 质粒后，重新评估上述细胞过程在 H9c2 细胞中的作用。结果显示，OGD 处理明显增强了 H9c2 细胞中 miR-132 的表达。过表达 miR-132 可明显逆转 OGD 诱导的 H9c2 细胞活力抑制和凋亡诱导。此外，过表达 miR-132 通过调节 p21、p27 和 cyclin D1 的表达减轻 OGD 诱导的 G0/G1 细胞阻滞。在 miR-132 模拟物转染细胞中观察到 FOXO3A 的抑制，这也被预测为 miR-132 的直接靶基因。研究发现，沉默 FOXO3A 通过促进细胞活力、抑制凋亡和阻止 G0/G1 期细胞阻滞，减轻 H9c2 细胞中 OGD 诱导的细胞损伤。综上所述，这些研究结果证实，miR-132 通过靶向 FOXO3A 对 H9c2 细胞对抗 OGD 诱导损伤具有保护作用。
