LncRNA MEG3靶向miR-21介导TLR4/NF-κB通路减轻小鼠心肌缺血再灌注损伤

doi:10.3969/j.issn.1003-9198.2026.03.006

摘要/Abstract

摘要： 目的观察长链非编码核糖核酸人母系表达基因3(LncRNA MEG3)靶向微小核糖核酸-21(miR-21)介导Toll样受体4(TLR4)/核因子-κB(NF-κB)通路对小鼠心肌缺血再灌注损伤(MIRI)的影响。方法将54只BAB/c小鼠采用随机数表法分为LncRNA MEG3上调组、LncRNA MEG3下调组、miR-21上调组、miR-21下调组、双上调组(LncRNA MEG3上调+miR-21上调)、双下调组(LncRNA MEG3下调+miR-21下调)、空载组、模型组和假手术组,每组6只。除假手术组外均建立MIRI模型。建模24 h后,各组小鼠处死取腹主动脉血,采用ELISA法检测心肌损伤标志物[肌钙蛋白I(cTnI)、肌酸激酶同工酶(CK-MB)、脑钠肽(BNP)和肌红蛋白(Mb)]和炎症因子(TNF-α、IL-6、IL-8)水平。通过苏木素-伊红(HE)染色观察心肌组织形态学改变。检测心肌组织TLR4、NF-κB p65基因与TLR4蛋白、细胞质及细胞核NF-κB p65蛋白表达。采用双荧光素酶基因报告实验与RNA pull-down实验验证LncRNA MEG3靶向调控miR-21。结果各组心肌损伤标志物、炎症因子水平比较:LncRNA MEG3下调组＞miR-21上调组＞双下调组＞模型组/空载组＞双上调组＞miR-21下调组＞LncRNA MEG3上调组＞假手术组(P＜0.05)。心肌组织TLR4基因与蛋白、NF-κB p65基因与细胞核NF-κB p65蛋白表达比较:LncRNA MEG3下调组＞miR-21上调组＞双下调组＞模型组＞双上调组＞miR-21下调组＞LncRNA MEG3上调组＞假手术组/空载组(P＜0.05),各组细胞质NF-κB p65蛋白表达趋势正相反(P＜0.05)。假手术组心肌组织形态学无改变,模型组/空载组、双下调组、miR-21上调组与LncRNA MEG3下调组心肌组织形态学改变逐渐严重,双上调组、miR-21下调组、LncRNA MEG3上调组心肌组织形态学改变逐渐减轻。双荧光素酶基因报告实验与RNA pull-down实验显示,LncRNA MEG3可负反馈靶向调控miR-21。结论上调LncRNA MEG3表达可抑制miR-21,下调TLR4/NF-κB通路,减轻小鼠MIRI及炎症反应,还可拮抗因miR-21上调引起的心肌损伤。

关键词: 长链非编码核糖核酸人母系表达基因3, 微小核糖核酸-21, Toll样受体4, 核因子-κB, 心肌缺血再灌注损伤, 炎症反应

Abstract: Objective To observe the effect of long non-coding ribonucleic acid human maternal expressed gene 3 (LncRNA MEG3) targeting microribonuclease-21 (miR-21) mediating Toll like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) pathway on myocardial ischemia-reperfusion injury (MIRI) in mice. Methods A total of 54 BAB/c mice were randomly divided into LncRNA MEG3 upregulation group, LncRNA MEG3 downregulation group, miR-21 upregulation group, miR-21 downregulation group, both upregulation group, both downregulation group, empty load group, model group, and sham surgery group, with 6 mice in each group. Except for the sham group, MIRI models were established for all mice. After 24 hours of modeling, mice in each group were euthanized. Abdominal aortic blood was collected and enzyme-linked immunosorbent assay was used to detect the levels of myocardial injury markers [cardiac troponin I (cTnI), creatine kinase isoenzyme (CK-MB), brain natriuretic peptide (BNP), and myoglobin (Mb)] and inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-8]. Morphological changes in myocardial tissue were observed using hematoxylin eosin (HE) staining. The expressions of TLR4 and NF-κB p65 genes and the expressions of TLR4, NF-κB p65 protein in cytoplasm and nucleus in myocardial tissues were detected. The dual luciferase gene reporter experiment and RNA pull-down experiment were used to validate the targeted regulation of miR-21 by LncRNA MEG3. Results Myocardial injury markers, inflammatory factor levels among different groups showed LncRNA MEG3 downregulation group>miR-21 upregulation group>both downregulation group>model group/empty vector group>both upregulation group>miR-21 downregulation group>LncRNA MEG3 upregulation group>sham group (P<0.05). TLR4 gene and protein in myocardial tissues, NF-κB p65 gene and nuclear NF-κB p65 protein expression showed LncRNA MEG3 downregulation group>miR-21 upregulation group>both downregulation group>model group>both upregulation group>miR-21 downregulation group>LncRNA MEG3 upregulation group>sham surgery group/empty load group (P<0.05), and the trend of cytoplasmic NF-κB p65 protein expression was positively opposite (P<0.05). There were no changes in the morphology of myocardial tissue in the sham surgery group, and the changes in the model group/empty load group, both downregulation group, miR-21 upregulation group, and LncRNA MEG3 downregulation group gradually became increasingly severe, while the changes in the both upregulation group, miR-21 downregulation group, and LncRNA MEG3 upregulation group gradually decreased. The dual luciferase gene reporter experiment and RNA pull-down experiment revealed that LncRNA MEG3 could negatively feedback and target miR-21 regulation. Conclusions Upregulation of LncRNA MEG3 expression can inhibit miR-21, downregulate TLR4/NF-κB pathway, alleviate MIRI and inflammatory response in mice, and also antagonize myocardial injury caused by miR-21 upregulation.

Key words: long non-coding ribonucleic acid human maternal expression gene 3, microribonuclease-21, Toll like receptor 4, nuclear factor-κB, myocardial ischemia-reperfusion injury, inflammatory reaction

中图分类号:

R 363.14

邢智, 沙吉旦·阿不都热衣木, 帕丽达·阿布来提, 王瑜, 吕茂琳, 高颖. LncRNA MEG3靶向miR-21介导TLR4/NF-κB通路减轻小鼠心肌缺血再灌注损伤[J]. 实用老年医学, 2026, 40(3): 247-253.

XING Zhi, Shajidan Abudureyimu, Palida Abulaiti, WANG Yu, LYU Maolin, GAO Ying. LncRNA MEG3 targets miR-21 to mediate TLR4/NF-κB pathway and alleviate myocardial ischemia-reperfusion injury in mice[J]. Practical Geriatrics, 2026, 40(3): 247-253.

参考文献

[1] SALARI N, MORDDARVANJOGHI F, ABDOLMALEKI A, et al. The global prevalence of myocardial infarction: a systematic review and meta-analysis[J]. BMC Cardiovasc Disord, 2023, 23(1): 206.
[2] 国家心血管病中心, 中国心血管健康与疾病报告编写组, 胡盛寿. 中国心血管健康与疾病报告2023概要[J]. 中国循环杂志, 2024, 39(7): 625-660.
[3] XIANG Q, YI X, ZHU X H, et al. Regulated cell death in myocardial ischemia-reperfusion injury[J]. Trends Endocrinol Metab, 2024, 35(3): 219-234.
[4] ALGOET M, JANSSENS S, HIMMELREICH U, et al. Myocardial ischemia-reperfusion injury and the influence of inflammation[J]. Trends Cardiovasc Med, 2023, 33(6): 357-366.
[5] 余敏, 崔跃, 黎鹏飞, 等. 上调lncRNA MEG3对大鼠心肌细胞系缺氧/复氧损伤的影响[J]. 中西医结合心脑血管病杂志, 2024, 22(4): 660-664.
[6] 张胜, 贾恩志, 王连生. 老年急性心肌梗死病人冠状动脉的蛋白质翻译后修饰定量分析[J]. 实用老年医学, 2023, 37(4): 373-377.
[7] 朱智显, 张小琴, 蔡虎志, 等. 天牛止眩胶囊对自发性高血压大鼠心肌lncRNA MEG3/miR-21调控PTEN诱导左心室肥厚的影响[J]. 湖南中医药大学学报, 2023, 43(3): 437-442.
[8] SHEN S, ZHANG M, WANG X, et al. Single-cell RNA sequencing reveals S100a9(hi) macrophages promote the transition from acute inflammation to fibrotic remodeling after myocardial ischemia-reperfusion[J]. Theranostics, 2024, 14(3): 1241-1259.
[9] CAI W, LIU L, SHI X, et al. Alox15/15-HpETE aggravates myocardial ischemia-reperfusion injury by promoting cardiomyocyte ferroptosis[J]. Circulation, 2023, 147(19): 1444-1460.
[10] ZENG J J, SHI H Q, REN F F, et al. Notoginsenoside R1 protects against myocardial ischemia/reperfusion injury in mice via suppressing TAK1-JNK/p38 signaling[J]. Acta Pharmacol Sin, 2023, 44(7): 1366-1379.
[11] 章颖, 谢玲, 叶佳琦, 等. 层粘连蛋白与老年急性心肌梗死病人主要不良心血管事件的相关性[J]. 实用老年医学, 2024, 38(2): 149-151.
[12] 王昕瑜, 张婷婷, 邢邯英, 等. miR-21在心肌缺血损伤相关信号通路中的作用[J]. 心血管病学进展, 2024, 45(11): 998-1002.
[13] ZHANG Q, LIU F, WANG X, et al. Circulating miR-21 and lncRNA MEG3 as novel prognostic biomarkers in acute myocardial infarction: a multi-center cohort study[J]. Circulation, 2024, 149(12): e500-e510.
[14] XU B W, RAO Y, WANG L, et al. LncRNA MEG3 inhibits renal fibrinoid necrosis of diabetic nephropathy via the MEG3/miR-21/ORAI1 axis[J]. Mol Biol Rep, 2023, 50(4): 3283-3295.
[15] MORADI A, ASLANI M R, MIRSHEKARI JAHANGIRI H, et al. Protective effects of 4-methylumbelliferone on myocardial ischemia/reperfusion injury in rats through inhibition of oxidative stress and downregulation of TLR4/NF-κB/NLRP3 signaling pathway[J]. Naunyn Schmiedebergs Arch Pharmacol, 2024, 397(7): 5015-5027.
[16] MI S, HUANG F, JIAO M, et al. Inhibition of MEG3 ameliorates cardiomyocyte apoptosis and autophagy by regulating the expression of miRNA-129-5p in a mouse model of heart failure[J]. Redox Rep, 2023, 28(1): 2224607.