[1]石晓欣,邬亦华,昌　菁,等.过表达微RNA-590-3p对大鼠心肌缺血再灌注损伤的保护作用及机制[J].新乡医学院学报,2021,38(7):601-606.[doi:10.7683/xxyxyxb.2021.07.001]
　SHI Xiaoxin,WU Yihua,CHANG Jing,et al.Protective effect and mechanism of microRNA-590-3p overexpression on myocardial ischemia-reperfusion injury in rats[J].Journal of Xinxiang Medical University,2021,38(7):601-606.[doi:10.7683/xxyxyxb.2021.07.001]
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过表达微RNA-590-3p对大鼠心肌缺血再灌注损伤的保护作用及机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年7
页码:
601-606
栏目:
基础研究
出版日期:
2021-07-05

文章信息/Info

Title:
Protective effect and mechanism of microRNA-590-3p overexpression on myocardial ischemia-reperfusion injury in rats
作者:
石晓欣1; 邬亦华2; 昌　菁1; 王瑞良1
（1.上海交通大学医学院附属新华医院老年医学科，上海　200092；2.上海国际医学中心内分泌科，上海　200120）
Author(s):
SHI Xiaoxin1; WU Yihua2; CHANG Jing1; WANG Ruiliang1
(1.Department of Geriatrics，Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine，Shanghai 200092,China；2.Department of Endocrinology，Shanghai International Medical Center，Shanghai 200120，China）
关键词:
微RNA-590-3p; 心肌缺血再灌注; 细胞凋亡; 炎症反应; 氧化应激
Keywords:
microRNA-590-3p; myocardial ischemia-reperfusion; apoptosis; inflammatory reaction; oxidative stress
分类号:
R542.22
DOI:
10.7683/xxyxyxb.2021.07.001
文献标志码:
A
摘要:
目的　探讨微RNA-590-3p（miR-590-3p）过表达对大鼠心肌缺血再灌注损伤（MIRI）的保护作用及其机制。方法　将24只Sprague Dawley大鼠按随机数字表法分为假手术组、模型组、阴性对照（NC）组和agomir-miR-590-3p组，每组6只。模型组、NC组和agomir-miR-590-3p组大鼠通过左冠状动脉结扎法制备MIRI模型，agomir-miR-590-3p组大鼠于结扎前经左心室前壁注入5 nmol agomir-miR-590-3p，NC组大鼠于结扎前经左心室前壁注入5 nmol agomir NC，模型组大鼠注入等量生理盐水；假手术组大鼠仅做开胸和心脏分离不进行结扎，并经左心室前壁注入等量生理盐水。模型制备后4组大鼠经腹主动脉取血，分离血清保存备用；断颈法处死大鼠，取心肌组织用于后续实验。采用实时荧光定量聚合酶链反应法检测大鼠心肌组织中miR-590-3p相对表达量，苏木精-伊红染色法观察大鼠心肌组织病理改变，酶联免疫吸附实验检测大鼠血清中肌酸激酶（CK）、乳酸脱氢酶（LDH）、丙二醛（MDA）、超氧化物歧化酶（SOD）、白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）水平，脱氧核糖核苷酸末端转移酶介导的缺口末端标记法检测大鼠心肌细胞凋亡率，Western blot法检测大鼠心肌组织中B淋巴细胞瘤-2(Bcl-2)基因、Bcl-2相关X(Bax)蛋白表达水平。结果　agomir-miR-590-3p组大鼠心肌组织中miR-590-3p相对表达量显著高于假手术组、模型组、NC组（P＜0.05）；模型组、NC组大鼠心肌组织中miR-590-3p相对表达量显著低于假手术组（P＜0.05）；模型组与NC组大鼠心肌组织中miR-590-3p相对表达量比较差异无统计学意义（P>0.05）。假手术组大鼠心肌纤维结构完整，细胞排列整齐；模型组和NC组大鼠心肌纤维断裂，细胞排列较乱，部分细胞出现坏死；与模型组和NC组相比，agomir-miR-590-3p组大鼠心肌纤维断裂和心肌细胞排列紊乱情况得到缓解。模型组、NC组和agomir-miR-590-3p组大鼠血清IL-1β、TNF-α、CK、LDH、MDA水平显著高于假手术组，SOD水平显著低于假手术组（P＜0.05）；agomir-miR-590-3p组大鼠IL-1β、TNF-α、CK、LDH、MDA水平显著低于模型组和NC组，SOD水平显著高于模型组和NC组（P＜0.05）；模型组与NC组大鼠血清IL-1β、TNF-α、CK、LDH、MDA、SOD水平比较差异无统计学意义（P>0.05）。模型组、NC组和agomir-miR-590-3p组大鼠心肌细胞凋亡率、心肌组织 Bax蛋白表达水平显著高于假手术组，Bcl-2 蛋白表达水平显著低于假手术组（P＜0.05）；agomir-miR-590-3p组大鼠心肌细胞凋亡率、心肌组织 Bax蛋白表达水平显著低于模型组和NC组，Bcl-2蛋白表达水平显著高于模型组和NC组（P＜0.05）；模型组和NC组大鼠心肌细胞凋亡率、心肌组织 Bax和Bcl-2蛋白表达水平比较差异无统计学意义（P>0.05）。结论　过表达miR-590-3p能够改善MIRI，该作用可能与其减少氧化应激及炎症反应、抑制心肌细胞凋亡有关。
Abstract:
Objective　To investigate the protective effect and mechanism of microRNA（miR）-590-3p overexpression on myocardial ischemia-reperfusion injury(MIRI) in rats.Methods　Twenty-four Sprague Dawley rats were randomly divided into the sham operation group，model group，negative control(NC) group and agomir-miR-590-3p group,with 6 rats in each group.MIRI model was established by ligation of left coronary artery of rats in the model group，NC group and agomir-miR-590-3p groupthe rats in the agomir-miR-590-3p group were injected with 5 nmol of agomir-miR-590-3p through the left ventricular anterior wall before ligation，the rats in the NC group were injected with 5 nmol of agomir NC，and the rats in the model group were injected with the same amount of normal saline.In the sham operation group，the rats were only subjected to thoracotomy and heart separation without ligation，and the same amount of normal saline was injected through the anterior wall of left ventricle.After MIRI models preparation，blood was taken from abdominal aorta of rats in each group，and serum was separated and stored for future using.The rats were killed by breaking the neck，and the myocardial tissue was taken for the follow-up experiment.The relative expression level of miR-590-3p in the myocardial tissues of rats were detected by quantitative real-time polymerase chain reaction.Pathological changes of myocardial tissue of rats were observed by hematoxylin-eosin staining.The levels of serum creatine kinase (CK)，lactate dehydrogenase (LDH)，malondialdehyde (MDA)，superoxide dismutase (SOD)，interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) of rats were detected by enzyme linked immunosorbent assay.Apoptosis rate of myocardial cells of rats was detected by TdT mediated dUTP nick end labeling method.The expression levels of B-cell lymphoma-2(Bcl-2) and Bcl-2 associated X protein(Bax) protein in myocardial tissues of rats were detected by Western blot.Results　The relative expression of miR-590-3p in myocardial tissue of rats in the agomir-miR-590-3p group was significantly higher than that in the sham operation group，model group and NC group (P＜0.05)the relative expression of miR-590-3p in myocardial tissue of rats in the model group and NC group was significantly lower than that in the sham operation group (P＜0.05)there was no significant difference in the relative expression of miR-590-3p in myocardial tissue of rats between the model group and NC group (P>0.05).In the sham operation group，the myocardial fiber structure of rats was intact and the myocardial cells were arranged in order.In the model group and NC group，myocardial fibers of rats were broken，the myocardial cells were disordered，and some myocardial cells were necrotic.Compared with the model group and NC group，the myocardial fiber breakage and the disorder of myocardial cell arrangement of rats in the agomir-miR-590-3p group were alleviated.The levels of serum IL-1β，TNF-α，CK，LDH and MDA of rats in the model group，NC group and agomir-miR-590-3p group were significantly higher than those in the sham operation group，the level of SOD was significantly lower than that in the sham operation group(P＜0.05).The levels of serum IL-1β，TNF-α，CK，LDH and MDA of rats in the agomir-miR-590-3p group were significantly lower than those in the model group and NC group(P＜0.05)，the level of SOD was significantly higher than that in the model group and NC group(P＜0.05)there was no significant difference in the levels of serum IL-1β，TNF-α，CK，LDH，MDA and SOD of rats between the model group and NC group (P>0.05).The apoptosis rate of myocardial cells and Bax protein expression in myocardial tissue of rats in the model group，NC group and agomir-miR-590-3p group were significantly higher than those in the sham operation group(P＜0.05)，Bcl-2 protein expression was significantly lower than that in the sham operation group(P＜0.05)the apoptosis rate of myocardial cells and Bax protein expression in myocardial tissue of rats in the agomir-miR-590-3p group was significantly lower than those in the model group and NC group(P＜0.05)，Bcl-2 protein expression in myocardial tissue of rats was significantly higher than that in the model group and NC group(P＜0.05)there was no significant difference in the apoptosis rate of myocardial cells and Bax，Bcl-2 protein expression in myocardial tissue of rats between the model group and NC group (P>0.05).Conclusion　Overexpression of miR-590-3p can improve MIRI，which may be related to reducing oxidative stress and inflammatory response,inhibiting apoptosis of myocardial cells.

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更新日期/Last Update: 2021-07-05