Department of Cardiology, Binzhou People's Hospital, Binzhou, Shandong 256610, P.R. China.
Department of Anesthesiology, Binzhou People's Hospital, Binzhou, Shandong 256610, P.R. China.
Mol Med Rep. 2017 Oct;16(4):5363-5369. doi: 10.3892/mmr.2017.7243. Epub 2017 Aug 14.
Krüppel-like factors (KLFs) regulate a wide variety of cellular functions and modulate pathological processes. In the present study, a post‑translational mechanism of microRNAs (miRs) was investigated in H2O2-induced human cardiomyocyte (HCM) injury. In H2O2‑cultured HCM cells, reactive oxygen species and apoptotic cells were measured via flow cytometry. miR‑506/‑124 mimics and inhibitors were transfected to induce gain or loss of miR‑506/‑124 function. Cell proliferation was analyzed by an MTT assay. The targeted genes were predicted by a bioinformatics algorithm and confirmed by a dual luciferase reporter assay. The mRNA and protein expression levels were measured by reverse transcription‑polymerse chain reaction analysis and western blotting, respectively. The results indicated that H2O2 induced significant apoptosis and increased the concentration of reactive oxygen species (ROS) in HCMs. H2O2 markedly upregulated the expression levels of KLF4 and KLF5, and downregulated the expression levels of miR‑506 and miR‑124 in the HCMs. In addition, bioinformatics analysis showed the potential miR‑506 and miR‑124 binding sites within the 3'‑untranslated region of KLF4 and KLF5 in the HCMs. The overexpression of miR‑506 and miR‑124 inhibited the H2O2‑induced upregulation of KLF4 and KLF5 in the HCMs. The overexpression of miR‑506 and miR‑214 reversed the H2O2‑induced apoptosis and increase of ROS in the HCMs. In conclusion, the overexpression of miR‑506 and miR‑214 were confirmed to have a protective effect against H2O2‑induced HCM injury by suppressing the expression of KLF4 and KLF5.
Krüppel 样因子 (KLFs) 调节广泛的细胞功能并调节病理过程。在本研究中，研究了氧化氢 (H2O2) 诱导的人心肌细胞 (HCM) 损伤中的 miRNA (miRs) 的翻译后机制。在 H2O2 培养的 HCM 细胞中，通过流式细胞术测量活性氧和凋亡细胞。转染 miR-506/-124 模拟物和抑制剂以诱导 miR-506/-124 功能的获得或丧失。通过 MTT 分析测定细胞增殖。通过生物信息学算法预测靶基因，并通过双荧光素酶报告基因测定证实。通过逆转录-聚合酶链反应分析和 Western blot 分别测定 mRNA 和蛋白质表达水平。结果表明，H2O2 诱导 HCM 显著凋亡并增加活性氧 (ROS) 浓度。H2O2 显著上调 KLF4 和 KLF5 的表达水平，并下调 HCM 中 miR-506 和 miR-124 的表达水平。此外，生物信息学分析显示了 HCM 中 KLF4 和 KLF5 的 3'-UTR 中潜在的 miR-506 和 miR-124 结合位点。miR-506 和 miR-124 的过表达抑制了 H2O2 诱导的 HCM 中 KLF4 和 KLF5 的上调。miR-506 和 miR-214 的过表达逆转了 H2O2 诱导的 HCM 中细胞凋亡和 ROS 的增加。总之，miR-506 和 miR-214 的过表达通过抑制 KLF4 和 KLF5 的表达被证实对 H2O2 诱导的 HCM 损伤具有保护作用。
