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

doi:10.11958/20220015

Abstract

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

CLC Number:

R541.7

Figures/Tables 6

References 20

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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

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

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^＊＊

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^＊＊

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^＊＊