LncRNA MIAT靶向调节miR-128-3p对心房颤动大鼠心室重构和心肌纤维化的影响

doi:10.11958/20220015

天津医药 ›› 2022, Vol. 50 ›› Issue (9): 932-937.doi: 10.11958/20220015

LncRNA MIAT靶向调节miR-128-3p对心房颤动大鼠心室重构和心肌纤维化的影响

邢佳侬¹(), 梁卓², 邢爱君³, 刘俊兰¹, 彭宏超¹, 张天桦¹, 张春来¹

1 唐山中心医院心内科（邮编063008）
2 北京安贞医院心律失常中心
3 开滦总医院心内科

收稿日期:2022-01-04 修回日期:2022-03-08 出版日期:2022-09-15 发布日期:2022-09-05
作者简介:邢佳侬（1973）,女,主任医师,主要从事心律失常方面研究。E-mail： xjn19730827@163.com
基金资助:
河北省卫生健康委科研基金项目(20211249)

Effects of lncRNA MIAT on ventricular remodeling and myocardial fibrosis in rats with atrial fibrillation through targeting regulation of miR-128-3p

XING Jianong¹(), LIANG Zhuo², XING Aijun³, LIU Junlan¹, PENG Hongchao¹, ZHANG Tianhua¹, ZHANG Chunlai¹

1 Department of Cardiology, Tangshan Central Hospital, Tangshan 063008, China
2 Department of Arrhythmia Center, Beijing Anzhen Hospital
3 Department of Cardiology, Kailuan General Hospital

Received:2022-01-04 Revised:2022-03-08 Published:2022-09-15 Online:2022-09-05

邢佳侬, 梁卓, 邢爱君, 刘俊兰, 彭宏超, 张天桦, 张春来. LncRNA MIAT靶向调节miR-128-3p对心房颤动大鼠心室重构和心肌纤维化的影响[J]. 天津医药, 2022, 50(9): 932-937.

XING Jianong, LIANG Zhuo, XING Aijun, LIU Junlan, PENG Hongchao, ZHANG Tianhua, ZHANG Chunlai. Effects of lncRNA MIAT on ventricular remodeling and myocardial fibrosis in rats with atrial fibrillation through targeting regulation of miR-128-3p[J]. Tianjin Medical Journal, 2022, 50(9): 932-937.

摘要/Abstract

摘要：

目的探讨保守的长链非编码RNA心肌梗死相关转录本（LncRNA MIAT）靶向调节miR-128-3p对心房颤动（AF）大鼠心室重构和心肌纤维化的影响。方法通过舌下静脉注射氯化钙-乙酰胆碱混合液,诱导构建AF大鼠模型,以随机数字表法将其平均分为模型组、LncRNA MIAT siRNA质粒组（MIAT组）、miR-128-3p siRNA质粒组（miR-128-3p组）、LncRNA MIAT siRNA质粒+miR-128-3p siRNA质粒组（MIAT+miR-128-3p组）、空载质粒组,每组12只。另12只大鼠舌静脉注射等剂量生理盐水,作为对照组。分组干预处理后,检测大鼠心房肌电生理水平,比较各组有效不应期（ERP）和90%动作电位时程（APD90）;计算各组大鼠左心室质量指数;天狼星红染色检测大鼠心肌组织纤维化程度,比较各组心肌胶原容积分数（CVF）;使用试剂盒测量各组大鼠血清白细胞介素（IL）-18、IL-6、转化生长因子-β1（TGF-β1）水平;实时荧光定量PCR检测各组大鼠心肌组织miR-128-3p表达;双荧光素酶报告基因试验检测LncRNA MIAT对miR-128-3p的靶向调节作用。结果与对照组比较,模型组大鼠ERP、APD90、心肌组织miR-128-3p表达降低（P<0.05）,左心室质量指数、CVF、血清IL-18、IL-6及TGF-β1水平升高（P<0.05）。分别与模型组、MIAT+miR-128-3p组比较,MIAT组大鼠ERP、APD90、心肌组织miR-128-3p表达均升高,而左心室质量指数、CVF、血清IL-18、IL-6及TGF-β1水平均降低（P<0.05）;miR-128-3p组大鼠ERP、APD90、心肌组织miR-128-3p表达均降低,而左心室质量指数、CVF、血清IL-18、IL-6及TGF-β1水平均升高（P<0.05）;空载质粒组大鼠各指标水平差异均无统计学意义（P>0.05）。LncRNA MIAT可靶向下调miR-128-3p的表达。结论 LncRNA MIAT可通过靶向下调miR-128-3p的表达而参与AF发病,下调LncRNA-MIAT可通过促进miR-128-3p表达来抑制炎症,减少房颤大鼠的心肌纤维化,改善其心房肌电生理水平,延缓大鼠心室重构过程。

关键词: 心房颤动, 心室重构, 心内膜心肌纤维化症, 基因治疗, 疾病模型,动物, 长链非编码RNA心肌梗死相关转录本, miR-128-3p

Abstract:

Objective To investigate the effect of the conserved long non-coding RNA myocardial infarction associate transcript (lncRNA MIAT) targeting and regulating miR-128-3p on ventricular remodeling and myocardial fibrosis in atrial fibrillation (AF) in rats. Methods The calcium chloride-acetylcholine mixture was injected into the sublingual vein to induce AF rat models. The rat models were divided into five groups by random number table method: the model group, the lncRNA MIAT siRNA plasmid group (MIAT group), the miR-128-3p siRNA plasmid group (miR-128-3p group), the lncRNA MIAT siRNA plasmid + miR-128-3p siRNA plasmid group (MIAT+miR-128-3p group) and the empty plasmid group, with 12 rats in each group. Another 12 rats were given with the same dose of normal saline through sublingual vein and were used as the control group. After intervention, the atrial myocardial electrophysiological level of rats was detected, and the effective refractory period (ERP) and 90% action potential duration (APD90) were compared between groups. The left ventricular mass index was also measured in five groups of rats. Sirius red staining was used to detect the degree of myocardial tissue fibrosis in rats. The myocardial collagen volume fraction (CVF) was compared between groups. Serum levels of interleukin (IL)-18, IL-6 and transforming growth factor-β1 (TGF-β1) were detected by kit. Real-time quantitative PCR (qRT-PCR) was used to detect the expression of miR-128-3p in myocardial tissues in each group. Dual luciferase reporter gene assay was used to detect the targeted regulation effect of lncRNA MIAT on miR-128-3p. Results Compared with the control group, the ERP, APD90, and myocardial tissue miR-128-3p expression were significantly reduced in the model group (P<0.05), and the left ventricular mass index, CVF, serum IL-18, IL-6 and TGF-β1 levels were significantly increased (P<0.05). Compared with the model group and the lncRNA MIAT siRNA plasmid+miR-128-3p siRNA plasmid group, the ERP, APD90, and myocardial tissue miR-128-3p expression were significantly increased in the lncRNA MIAT siRNA plasmid group (P<0.05), and the left ventricular mass index, CVF, serum IL-18, IL-6 and TGF-β1 levels were significantly reduced (P<0.05). The ERP, APD90 and myocardial tissue miR-128-3p expression were significantly decreased in the miR-128-3p siRNA plasmid group (P<0.05), and the left ventricular mass index, CVF, serum IL-18, IL-6 and TGF-β1 levels were significantly increased (P<0.05). There were no significant differences in the indicators between the empty plasmid groups (P>0.05). LncRNA MIAT could down-regulate the expression of miR-128-3p by targeting. Conclusion LncRNA MIAT can participate in the pathogenesis of AF by targeting down-regulation of miR-128-3p expression. The down-regulation of lncRNA MIAT can inhibit inflammation by promoting miR-128-3p expression, reduce myocardial fibrosis in AF rats, improve the atrial myocardial electrophysiological level and delay the process of ventricular remodeling in rats.

Key words: atrial fibrillation, ventricular remodeling, endomyocardial fibrosis, genetic therapy, disease models,animal, long non-coding RNA myocardial infarction-related transcript, miR-128-3p

中图分类号:

R541.7

图/表 6

表1 qPCR引物序列

Tab.1 qPCR primer sequence

引物名称	引物序列（5'→3'）	产物大小（bp）
miR-128-3p	上游：AGGCTCACAGTGAACCGGTC 下游：CGCAGGGTCCGAGGTATTC	145
U6	上游：CGCTTCGGCAGCACATATAC 下游：AAATATGGAACGCTTCACGA	132

表2 各组大鼠心房肌电生理水平指标、左心室质量指数、心肌CVF比较（n=12,$\bar{x}±s$）

Tab.2 Comparison of atrial electrophysiological indexes, left ventricular mass index and myocardial CVF between the six groups of rats

组别	ERP （ms）	APD90 （ms）	左心室质量指数（mg/g）	心肌CVF （%）
对照组	46.01±3.02	65.13±4.81	12.20±2.01	0.52±0.17
模型组	20.15±2.58^a	48.94±3.97^a	21.05±3.48^a	20.53±2.04^a
MIAT组	43.57±3.64^b	62.87±4.56^b	13.97±2.46^b	3.74±1.23^b
miR-128-3p组	12.63±0.82^b	37.58±2.01^b	29.03±2.85^b	31.67±3.52^b
MIAT+miR- 128-3p组	22.08±2.16^cd	51.89±2.68^cd	20.01±2.62^cd	18.91±2.19^cd
空载质粒组	19.78±2.27	49.72±4.05	21.67±2.34	21.06±2.23
F	351.599^＊＊	84.145^＊＊	61.401^＊＊	356.630^＊＊

图1 天狼星红染色检测大鼠心肌纤维化程度（×200）

Fig.1 The degree of myocardial fibrosis detected by Sirius red staining (×200)

表3 各组血清IL-18、IL-6、TGF-β1及心肌组织miR-128-3p水平比较（n=12,$\bar{x}±s$）

Tab.3 Comparison of serum levels of IL-18, IL-6, TGF-β1 and expression of miR-128-3p in myocardial tissue between the six groups of rats

组别	IL-18 （µg/L）	IL-6 （µg/L）	TGF-β1 （ng/L）	miR-128-3p
对照组	0.39±0.04	1.32±0.24	120.68±26.83	1.01±0.24
模型组	1.42±0.13^a	14.29±2.65^a	638.15±65.38^a	0.56±0.07^a
MIAT组	0.43±0.09^b	1.97±0.43^b	125.49±28.16^b	0.96±0.19^b
miR-128-3p组	2.38±0.22^b	25.32±3.17^b	931.27±86.67^b	0.31±0.05^b
MIAT+miR- 128-3p组	1.39±0.18^cd	13.14±3.01^cd	629.82±70.54^cd	0.59±0.08^cd
空载质粒组	1.44±0.26	14.75±2.82	635.91±72.08	0.55±0.06
F	228.185^＊＊	170.296^＊＊	321.152^＊＊	46.911^＊＊

图2 TargetScan Release7.2的生物信息学预测LncRNA MIAT和miR-128-3p之间的结合位点

Fig.2 Bioinformatics of TargetScan Release7.2 predicts the binding site between lncRNA MIAT and miR-128-3p

图3 各组细胞相对荧光素酶活性值 A：miR-128-3p无义+空载组;B：miR-128-3p无义+过表达组;C：miR-128-3p野生+空载组;D：miR-128-3p野生+过表达组;E：miR-128-3p突变+空载组;F：miR-128-3p突变+过表达组;n=6,F=25.212,P<0.01;a与miR-128-3p野生+空载组比较,P<0.05。

Fig.3 Relative luciferase activity of cells in each group

参考文献 20

[1]	SEPEHRI SHAMLOO A, DAGRES N, HINDRICKS G. 2020 ESC guidelines on atrial fibrillation:Summary of the most relevant recommendations and innovations[J]. Herz, 2021, 46(1):28-37. doi: 10.1007/s00059-020-05005-y.
[2]	NADEMANEE K. Atrial fibrillation ablation in the 21st century:Almost no stroke risk?[J]. Heart Rhythm, 2020, 17(12):2100-2101. doi: 10.1016/j.hrthm.2020.08.008.
[3]	SHARMA G, GHATI N, SHARIQUE M, et al. Role of inflammation in initiation and maintenance of atrial fibrillation in rheumatic mitral stenosis-An analytical cross-sectional study[J]. J Arrhythm, 2020, 36(6):1007-1015. doi: 10.1002/joa3.12428.
[4]	XIAO L, SALEM J E, CLAUSS S, et al. Ibrutinib-mediated atrial fibrillation attributable to inhibition of C-terminal Src kinase[J]. Circulation, 2020, 142(25):2443-2455. doi: 10.1161/CIRCULATION AHA.120.049210.
[5]	FARSANGI S J, ROSTAMZADEH F, SHEIKHOLESLAMI M, et al. Modulation of the expression of long non-coding RNAs H19,GAS5,and MIAT by endurance exercise in the hearts of rats with myocardial infarction[J]. Cardiovasc Toxicol, 2021, 21(2):162-168. doi: 10.1007/s12012-020-09607-0.
[6]	YANG L, DENG J, MA W, et al. Ablation of lncRNA Miat attenuates pathological hypertrophy and heart failure[J]. Theranostics, 2021, 11(16):7995-8007. doi: 10.7150/thno.50990.
[7]	YAO L, ZHOU B, YOU L, et al. LncRNA MIAT/miR-133a-3p axis regulates atrial fibrillation and atrial fibrillation-induced myocardial fibrosis[J]. Mol Biol Rep, 2020, 47(4):2605-2617. doi: 10.1007/s11033-020-05347-0.
[8]	LI F, LI H, LI S, et al. Long non-coding RNA MIAT mediates non-small cell lung cancer development through regulating the miR-128-3p/PELI3 axis[J]. Biochem Genet, 2020, 58(6):867-882. doi: 10.1007/s10528-020-09979-6.
[9]	YANG P, HAN J, LI S, et al. miR-128-3p inhibits apoptosis and inflammation in LPS-induced sepsis by targeting TGFBR2[J]. Open Med (Wars), 2021, 16(1):274-283. doi: 10.1515/med-2021-0222.
[10]	CAO F, LI Z, DING W M, et al. LncRNA PVT1 regulates atrial fibrosis via miR-128-3p-SP1-TGF-β1-Smad axis in atrial fibrillation[J]. Mol Med, 2019, 25(1):7-17. doi: 10.1186/s10020-019-0074-5.
[11]	焦华琛, 郑书敏, 李运伦, 等. 心AF动大鼠心房肌HCN2,HCN4表达及青山健心片的干预作用[J]. 中西医结合心脑血管病杂志, 2020, 18(14):2230-2233.
	JIAO H C, ZHENG S M, LI Y L, et al. The expression of HCN2,HCN4 in atrial fibrillation rats and the intervention effect of Qingshan Jianxin Tablet[J]. Chinese Journal of Integrative Medicine on Cardio/Cerebrovascular Disease, 2020, 18(14):2230-2233. doi: 10.12102/j.issn.1672-1349.2020.14.008.
[12]	SEARS S F, ANTHONY S, NANIWADEKAR A, et al. Modern atrial fibrillation care:Becoming a pro at using PROs[J]. J Cardiovasc Electrophysiol, 2020, 31(12):3196-3198. doi: 10.1111/jce.14793.
[13]	WANG Q C, WANG Z Y. Big data and atrial fibrillation:current understanding and new opportunities[J]. J Cardiovasc Transl Res, 2020, 13(6):944-952. doi: 10.1007/s12265-020-10008-5.
[14]	NSO N, BOOKANI K R, METZL M, et al. Role of inflammation in atrial fibrillation:A comprehensive review of current knowledge[J]. J Arrhythm, 2020, 37(1):1-10. doi: 10.1002/joa3.12473.
[15]	HARADA M, NATTEL S. Implications of inflammation and fibrosis in atrial fibrillation pathophysiology[J]. Card Electrophysiol Clin, 2021, 13(1):25-35. doi: 10.1016/j.ccep.2020.11.002.
[16]	ZHAO X, REN Y, REN H, et al. The mechanism of myocardial fibrosis is ameliorated by myocardial infarction-associated transcript through the PI3K/Akt signaling pathway to relieve heart failure[J]. J Int Med Res, 2021, 49(7):3000605211031433-3000605211031443. doi: 10.1177/03000605211031433.
[17]	CHEN Y, CHEN X, LI H, et al. Serum extracellular vesicles containing MIAT induces atrial fibrosis,inflammation and oxidative stress to promote atrial remodeling and atrial fibrillation via blockade of miR-485-5p-mediated CXCL10 inhibition[J]. Clin Transl Med, 2021, 11(8):e482-e500. doi: 10.1002/ctm2.482.
[18]	LIU J, WANG S, ZHANG Q, et al. Selenomethionine alleviates LPS-induced chicken myocardial inflammation by regulating the miR-128-3p-p38 MAPK axis and oxidative stress[J]. Metallomics, 2020, 12(1):54-64. doi: 10.1039/c9mt00216b.
[19]	MA H, CHEN P, SANG C, et al. Modulation of apoptosis-related microRNAs following myocardial infarction in fat-1 transgenic mice vs wild-type mice[J]. J Cell Mol Med, 2018, 22(11):5698-5707. doi: 10.1111/jcmm.13846.
[20]	ZHANG C, XIE L, LIANG H, et al. LncRNA MIAT facilitates osteosarcoma progression by regulating mir-128-3p/VEGFC axis[J]. IUBMB Life, 2019, 71(7):845-853. doi: 10.1002/iub.2001.

LncRNA MIAT靶向调节miR-128-3p对心房颤动大鼠心室重构和心肌纤维化的影响

Effects of lncRNA MIAT on ventricular remodeling and myocardial fibrosis in rats with atrial fibrillation through targeting regulation of miR-128-3p

引用本文

RichHTML

PDF (PC)

可视化

摘要/Abstract

使用本文

图/表 6

参考文献 20

相关文章 15

编辑推荐

Metrics

本文评价

[1]	高攀, 谢冰歆, 周赞东, 刘彤. 慢性肾脏病循环中FGF23对心房纤维化的促进作用[J]. 天津医药, 2024, 52(9): 917-923.
[2]	张明龙, 方媛媛, 隋晓鹏, 陈欣欣, 李留东, 王海涛. Peguero-Lo-Presti指数诊断的左心室肥厚与阵发性心房颤动射频导管消融术后复发的关系[J]. 天津医药, 2024, 52(2): 210-214.
[3]	李少儒, 李燕, 刘珊, 户瑞丽. LncRNA SNHG11通过抑制miR-184/CARM1信号轴促进卵巢癌生长[J]. 天津医药, 2023, 51(6): 561-567.
[4]	周梦竹, 张海凤, 张雪, 张跃, 程立君, 刘彤, 刘长乐. NLRP3-CAMKⅡ-IRE-1α通路激活诱导氧化应激增强对糖尿病大鼠心室重构的影响[J]. 天津医药, 2023, 51(6): 580-585.
[5]	宁寅宽, 刘林志, 陈贤平. BMP2重组慢病毒转染兔BMSCs后细胞矿化的实验研究[J]. 天津医药, 2023, 51(5): 454-459.
[6]	郑月, 马运婷, 赵晓莹, 赵新湘. 高血压患者同型半胱氨酸与左心室心肌纤维化的相关性[J]. 天津医药, 2023, 51(4): 395-399.
[7]	杨国红, 余芳芳, 蔡伟, 牛秀珑, 张芯, 赵季红, 李玉明, 陈少伯. VEGFR-3⁺单核细胞对高血压小鼠左心室重塑的影响[J]. 天津医药, 2023, 51(2): 144-148.
[8]	彭明, 李玉凯, 王岚, 黄粮, 成忠, 肖杰. 基于钙超载探讨自主神经调控对HFpEF大鼠心肌结构重塑、电重塑及纤维化的影响[J]. 天津医药, 2023, 51(1): 30-34.
[9]	霍立巍, 刘军, 郑彬彬, 毕学娜. 血清FGF-23对心房颤动患者射频消融术后复发的预测价值[J]. 天津医药, 2023, 51(1): 74-77.
[10]	贺宏梅, 孙健, 王欢欢, 马书静, 张海燕, 邹玉安, 薛茜, 宋爱霞. 非瓣膜性心房颤动患者血管性痴呆危险因素的病例对照研究[J]. 天津医药, 2022, 50(8): 840-843.
[11]	张续腾, 刘芳, 陈军, 刘晓萌, 靳乐, 高红梅△. PI3K/AKT信号通路在心房颤动中的研究进展[J]. 天津医药, 2022, 50(5): 556-560.
[12]	翟亚君, 杨涵, 陈万里, 刘玥, 魏丽萍, 刘克强, 齐新△. 阿利沙坦酯对自发性高血压大鼠稳定降压及心脏保护作用的研究#br#[J]. 天津医药, 2022, 50(5): 481-486.
[13]	霍宁, 詹晓萍, 周梦竹, 张跃, 梁雪, 李广平, 刘长乐. 格列本脲对糖尿病大鼠心房重构及心房颤动诱发的影响[J]. 天津医药, 2022, 50(3): 253-258.
[14]	卢洁, 张芯, 王涛, 杨宁. 环状RNA对房颤患者的影响及其作用机制研究[J]. 天津医药, 2022, 50(3): 333-336.
[15]	李瑞龄, 赵志强. 松弛素在心房颤动中的应用研究进展[J]. 天津医药, 2021, 49(4): 441-444.