Luteolin Inhibits Ischemia/Reperfusion-Induced Myocardial Injury in Rats via Downregulation of microRNA-208b-3p.
Luteolin Inhibits Ischemia/Reperfusion-Induced Myocardial Injury in Rats via Downregulation of microRNA-208b-3p.
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DOI：
10.1371/journal.pone.0144877
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Li D
中科院分区：
文献类型：
--
作者：
Bian C;Xu T;Zhu H;Pan D;Liu Y;Luo Y;Wu P;Li D
木犀草素（LUT）是一种从多种膳食中提取的黄酮类化合物，据报道具有多种疾病的保护作用。近年来的研究表明，LUT在缺血/再灌注（I/R）过程中具有心脏保护作用。然而，LUT是否通过特异性microRNAs（miRs）发挥抗心肌I/R损伤的作用，目前尚未见报道。本研究的目的是确定哪些miR和靶基因LUT发挥这种功能。利用基因芯片检测不同miR在灌流大鼠心脏中的表达。使用TargetScan、MiRDB和米兰达预测靶基因。采用缺氧/复氧模拟I/R。用Ets 1（禽成红细胞增多症病毒E26（vets）癌基因同源物1）的miR-208 b-3 p模拟物、抑制剂和小干扰RNA转染细胞。通过实时定量聚合酶链反应定量miR-208 b-3 p和Ets 1 mRNA。Annexin V-异硫氰酸荧光素/碘化丙啶染色及流式细胞仪检测凋亡细胞百分率。Western blot检测caspase-3、Bcl-2、Bax和Ets 1蛋白表达水平。使用荧光素酶报告基因测定来验证miR-208 b-3 p与Ets 1的3 '-非翻译区之间的组合。与I/R组相比，LUT预处理降低了心肌组织中miR-208 b-3 p的表达。LUT可降低缺血再灌注引起的miR-208 b-3 p表达和细胞凋亡。而miR-208 b-3 p的过表达进一步加重了I/R引起的变化，并阻断了LUT的所有作用。miR-208 b-3 p表达的敲低也减弱了细胞凋亡，而Ets 1的敲低促进了细胞凋亡。荧光素酶报告基因检测结果显示，miR-208 b-3 p能够抑制Ets 1的表达。LUT预处理通过降低miR-208 b-3 p和增加Ets 1表达水平来表达心肌I/R损伤后的抗凋亡作用。
Luteolin (LUT), a kind of flavonoid which is extracted from a variety of diets, has been reported to convey protective effects of various diseases. Recent researches have suggested that LUT can carry out cardioprotective effects during ischemia/reperfusion (I/R). However, there have no reports on whether LUT can exert protective effects against myocardial I/R injury through the actions of specific microRNAs (miRs). The purpose of this study was to determine which miRs and target genes LUT exerted such function through. Expression of various miRs in perfused rat hearts was detected using a gene chip. Target genes were predicted with TargetScan, MiRDB and MiRanda. Anoxia/reoxygenation was used to simulate I/R. Cells were transfected by miR-208b-3p mimic, inhibitor and small interfering RNA of Ets1 (avian erythroblastosis virus E26 (v ets) oncogene homolog 1). MiR-208b-3p and Ets1 mRNA were quantified by real-time quantitative polymerase chain reaction. The percentage of apoptotic cells was detected by annexin V-fluorescein isothiocyanate/propidium iodide dyeing and flow cytometry. The protein expression levels of cleaved caspase-3, Bcl-2, Bax, and Ets1 were examined by western blot analysis. A luciferase reporter assay was used to verify the combination between miR-208b-3p and the 3’-untranslated region of Ets1. LUT pretreatment reduced miR-208b-3p expression in myocardial tissue, as compared to the I/R group. And LUT decreased miR-208b-3p expression and apoptosis caused by I/R. However, overexpression of miR-208b-3p further aggravated the changes caused by I/R and blocked all the effects of LUT. Knockdown of miR-208b-3p expression also attenuated apoptosis, while knockdown of Ets1 promoted apoptosis. Further, the luciferase reporter assay showed that miR-208b-3p could inhibit Ets1 expression. LUT pretreatment conveys anti-apoptotic effects after myocardial I/R injury by decreasing miR-208b-3p and increasing Ets1 expression levels.