长链非编码RNA FTX通过调控miR-150/KLF13轴减轻过氧化氢诱导的心肌细胞损伤。
Long noncoding RNA FTX ameliorates hydrogen peroxide-induced cardiomyocyte injury by regulating the miR-150/KLF13 axis.
作者信息
Zhang Yamin, Fan Xiaoying, Yang Hua
机构信息
Department of Cardiology, The First Affiliated Hospital of Airforce Military Medical University, No. 127, Changle West Road, Xi'an, Shaanxi, 710032, China.
出版信息
Open Life Sci. 2020 Dec 31;15(1):1000-1012. doi: 10.1515/biol-2020-0100. eCollection 2020.
BACKGROUND
Myocardial reperfusion is an effective therapy for acute myocardial infarction (AMI). However, ischemia/reperfusion (I/R) injury following myocardial reperfusion is a significant limitation for AMI treatment. Five prime to Xist (FTX) was recognized as a biomarker of multiple diseases, including heart disease. However, the molecular mechanism of FTX in I/R injury is unclear.
METHODS
Cell viability was evaluated by using cell counting kit-8 (CCK-8) assay. Apoptosis was analyzed by using a caspase-3 activity detection kit and flow cytometry. The expression of FTX, microRNA (miR)-150, and Kruppel-like factor 13 (KLF13) was measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The interaction of miR-150 and FTX or KLF13 was confirmed by a dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Protein expression of KLF13 was examined by Western blot. The role of FTX was detected in I/R-injured heart tissues .
RESULTS
Hydrogen peroxide (HO) induced cardiomyocyte injury by decreasing cell viability and expediting cell apoptosis. However, FTX alleviated cardiomyocyte injury by promoting cell proliferation and restricting cell apoptosis of H9C2 cells that were treated with HO. In addition, we discovered that FTX directly interacted with miR-150, while KLF13 was a target of miR-150. Rescue experiments showed that miR-150 neutralized the FTX-mediated promotion of cell progression and restriction of cell apoptosis in H9C2 cells treated with HO. KLF13 knockdown restored the effect of miR-150 on increased proliferation and decrease in apoptosis in HO-treated cardiomyocytes. Furthermore, FTX enhanced the expression of KLF13 protein through interaction with miR-150. Upregulation of FTX repressed apoptosis in I/R-injured heart tissues .
CONCLUSION
FTX relieves HO-induced cardiomyocyte injury by increasing KLF13 expression via depletion of miR-150, thus providing a novel therapeutic target for the alleviation of I/R injury.
背景
心肌再灌注是急性心肌梗死（AMI）的有效治疗方法。然而，心肌再灌注后的缺血/再灌注（I/R）损伤是AMI治疗的一个重大限制。5'至Xist（FTX）被认为是包括心脏病在内的多种疾病的生物标志物。然而，FTX在I/R损伤中的分子机制尚不清楚。
方法
使用细胞计数试剂盒-8（CCK-8）检测法评估细胞活力。使用caspase-3活性检测试剂盒和流式细胞术分析细胞凋亡。通过定量逆转录-聚合酶链反应（qRT-PCR）检测FTX、微小RNA（miR）-150和Kruppel样因子13（KLF13）的表达。通过双荧光素酶报告基因和RNA免疫沉淀（RIP）检测法确认miR-150与FTX或KLF13的相互作用。通过蛋白质印迹法检测KLF13的蛋白表达。在I/R损伤的心脏组织中检测FTX的作用。
结果
过氧化氢（HO）通过降低细胞活力和加速细胞凋亡诱导心肌细胞损伤。然而，FTX通过促进细胞增殖和限制HO处理的H9C2细胞的细胞凋亡减轻心肌细胞损伤。此外，我们发现FTX直接与miR-150相互作用，而KLF13是miR-150的靶标。挽救实验表明，miR-150中和了FTX介导的HO处理的H9C2细胞中细胞增殖的促进作用和细胞凋亡的限制作用。KLF13敲低恢复了miR-150对HO处理的心肌细胞增殖增加和凋亡减少的作用。此外，FTX通过与miR-150相互作用增强KLF13蛋白的表达。FTX上调抑制了I/R损伤心脏组织中的细胞凋亡。
结论
FTX通过消耗miR-150增加KLF13表达来减轻HO诱导的心肌细胞损伤，从而为减轻I/R损伤提供了一个新的治疗靶点。
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