益心定悸方对缺血性心律失常大鼠微RNA-1及其调控蛋白缝隙连接蛋白43的影响

doi:10.20047/j.issn1673-7210.2023.15.06

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摘要目的探究益心定悸方对缺血性心律失常大鼠微RNA-1（miRNA-1）及其调控蛋白缝隙连接蛋白43（Cx43）的影响。方法将60只雄性SPF级10周龄体重（250±20）g的SD大鼠采用随机数字表法分为六组，假手术组，模型组，普萘洛尔对照组，益心定悸方低、中、高剂量组，每组10只。动物喂养1周后，实施灌胃处理。益心定悸方低、中、高剂量组给予13.95、27.90、55.80 g/（kg·d）益心定悸方，普萘洛尔对照组给予普萘洛尔（9.5 mg/7 ml），每天灌胃5 mg/kg，模型组和假手术组灌胃等量生理盐水，l次/d，每组均处理2周。除假手术组外，其余各组灌胃结束后立即造模，通过左冠状动脉前降支结扎，构建大鼠缺血性心律失常模型，假手术组只开胸不结扎。造模后采集大鼠心肌组织，采用定量逆转录聚合酶链反应和Western blot检测miRNA-1与Cx43的表达量及Cx43在Ser262和Ser368位点的磷酸化水平。结果与假手术组比较，模型组miRNA-1表达量升高，Cx43表达量减少（P＜0.05）；与模型组比较，益心定悸方低、中、高剂量组miRNA-1表达量降低，益心定悸方中、高剂量组Cx43表达量升高（P＜0.05）；与益心定悸方低、中剂量组比较，益心定悸方高剂量组miRNA-1表达量降低，Cx43表达量升高（P＜0.05）。与假手术组比较，模型组的Cx43在位点Ser262和Ser368的磷酸化修饰水平降低（P＜0.05）；与模型组比较，益心定悸方高剂量组的Cx43在位点Ser262和Ser368的磷酸化修饰水平升高（P＜0.05）；与益心定悸方低剂量组比较，益心定悸方高剂量组的Cx43在位点Ser262和Ser368的磷酸化修饰水平升高（P＜0.05）；益心定悸方高剂量组的Cx43在位点Ser262和Ser368的磷酸化修饰水平与益心定悸方中剂量组比较，差异无统计学意义（P＞0.05）。苏木精-伊红结果显示，益心定悸方高剂量组可减轻缺血再灌注引起的心肌细胞损伤。结论益心定悸方可能通过下调心肌组织miRNA-1表达量，减弱Cx43的转录后抑制程度，从而有效纠正Cx43的表达。本研究对益心定悸方的药理学做了初步探索，对缺血性心律失常的有效防治提供了理论支撑。

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	杜晔1 张怡清2 张晶3 刘莉2 马学淼4 杨颖2

关键词 ：益心定悸方, 缺血性心律失常, 微RNA-1, 缝隙连接蛋白43

Abstract：Objective To explore the effects of Yixin Dingji Decoction on microRNA-1 (miRNA-1) and its regulatory protein connexin43 (Cx43) in rats with ischemic arrhythmia. Methods Sixty SPF 10-week-old male SD rats with a body weight of (250±20) g were divided into six groups by random number table method: sham operation group, model group, Propranolol control group, low, medium, and high dose groups of Yixin Dingji Decoction, with ten rats in each group. After feeding the animals for one week, gavage was administered. Low, medium, and high dose groups of Yixin Dingji Decoction were given 13.95, 27.90, 55.80 g/(kg·d) Yixin Dingji Decoction. Propranolol control group was given Propranolol (9.5 mg/7 ml) by gavage of 5 mg/kg a day. Model group and sham operation group were given the same amount of normal saline intragastric administration, one time a day. Each group was treated for two weeks. Except for sham operation group, the other groups were constructed immediately after the end of gavage, and the ischemic arrhythmia model of rats was established by ligation of the anterior descending branch of the left coronary artery. Sham operation group only thoracotomy without ligation. After modeling, the myocardial tissue of rats was collected, and the expression levels of miRNA-1 and Cx43 as well as the phosphorylation levels of Cx43 at Ser262 and Ser368 were detected by quantitative reverse transcription polymerase chain reaction and Western blot. Results Compared with sham operation group, the expression of miRNA-1 was increased and Cx43 was decreased in model group (P＜0.05). Compared with model group, miRNA-1 expression was decreased in low, medium, and high dose groups of Yixin Dingji Decoction, while Cx43 expression was increased in medium and high dose groups of Yixin Dingji Decoction (P＜0.05). Compared with low and medium dose groups of Yixin Dingji Decoction, miRNA-1 expression was decreased and Cx43 expression was increased in high dose group of Yixin Dingji Decoction (P＜0.05). Compared with the sham operation group, the phosp- horylation of Cx43 at sites Ser262 and Ser368 was decreased in the model group (P＜0.05). Compared with model group, the phosphorylation of Cx43 at sites Ser262 and Ser368 in high dose group of Yixin Dingji Decoction was increased (P＜0.05). Compared with low dose group of Yixin Dingji Decoction, the phosphorylation of Cx43 at sites Ser262 and Ser368 in high dose group of Yixin Dingji Decoction was increased (P＜0.05). The phosphorylation of Cx43 at sites Ser262 and Ser368 in high dose group of Yixin Dingji Decoction was not significantly different from that in medium dose group of Yixin Dingji Decoction (P＞0.05). Hematoxylin and eosin results showed that high dose group of Yixin Dingji Decoction could alleviate myocardial cell injury caused by ischemia reperfusion. Conclusion Yixin Dingji Decoction may effectively correct the expression of Cx43 by down-regulating the expression of miRNA-1 in myocardial tissue and weakening the post-transcriptional inhibition of Cx43. The pharmacology of Yixin Dingji Decoction is preliminarily explored in this study, which provided theoretical support for the effective prevention and treatment of ischemic arrhythmia.

Key words： Yixin Dingji Decoction Ischemic arrhythmia microRNA-1 Connexin43

基金资助:黑龙江省自然科学基金项目（LH2019H114）。

通讯作者: 杨颖（1965.11-），女，硕士，主任医师，黑龙江中医药大学附属第一医院心血管病四科副主任；主要从事中西医结合临床心血管方向研究工作。

作者简介: 杜晔（1990.11-），女，黑龙江中医药大学临床医学院2020级中西医结合临床专业在读博士研究生；主要从事心血管方向及中西医结合疗法在冠心病、心律失常等方向的应用与机制研究工作。

引用本文:

杜晔1 张怡清2 张晶3 刘莉2 马学淼4 杨颖2. 益心定悸方对缺血性心律失常大鼠微RNA-1及其调控蛋白缝隙连接蛋白43的影响[J]. 中国医药导报, 2023, 20(15): 35-39.
DU Ye1 ZHANG Yiqing2 ZHANG Jing3 LIU Li2 MA Xuemiao4 YANG Ying2. Effects of Yixin Dingji Decoction on microRNA-1 and its regulatory protein connexin43 in rats with ischemic arrhythmia. 中国医药导报, 2023, 20(15): 35-39.

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