沉默miR-26a减轻缺血/再灌注大鼠心肌损伤

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (9): 1285-1290.

沉默miR-26a减轻缺血/再灌注大鼠心肌损伤

宫丹丹¹, 隋春兴¹, 石晨¹, 李月¹, 姜晓东^2*

1.大连市中心医院心内三科, 辽宁大连 116033;
2.大连医科大学附属第二医院重症医学科,辽宁大连 116027

收稿日期:2021-03-15 修回日期:2021-06-23 出版日期:2021-09-05 发布日期:2021-09-02
通讯作者: *jiangxiaodong@live.cn
基金资助:
辽宁省科学技术计划(20180550232);大连市医学科学研究计划(1712036)

Silencing miR-26a alleviates myocardial ischemia-reperfusion injury in rats

GONG Dan-dan¹, SUI Chun-xing¹, SHI Chen¹, LI Yue¹, JIANG Xiao-dong^2*

1. Department of Cardiology, Dalian Municipal Central Hospital, Dalian 116033;
2.Intensive Care Unit, the Second Hospital of Dalian Medical University, Dalian 116027,China

Received:2021-03-15 Revised:2021-06-23 Online:2021-09-05 Published:2021-09-02
Contact: *jiangxiaodong@live.cn

摘要/Abstract

摘要： 目的探讨微核糖核酸(miR-26a)对大鼠心肌缺血/再灌注损伤(I/R)的影响。方法将大鼠随机分为对照组(n=20)、I/R组(n=20)和siRNA组(n=20)。建立大鼠I/R模型前7 d,尾静脉注射miR-26a siRNA沉默内源性miR-26a。超声心动图检测心脏射血分数(EF%)和缩短分数(FS%);2,3,5-三苯基四唑氯化物(TTC)染色测定心肌梗死面积; HE染色法检测心肌细胞形态;TUNEL法检测心肌细胞凋亡水平;Western blot检测心肌组织中促凋亡蛋白Bcl-2相关X蛋白(Bax)和活性半胱天冬酶-3(caspase-3)和Rho/RhoA信号通路相关蛋白表达。结果 I/R组心肌组织中miR-26a的表达明显高于对照组(P＜0.05)。siRNA组心功能不全显著改善,大鼠的EF%和FS%均提高(P＜0.05)。此外,siRNA组I/R所致的心肌梗死程度明显减轻,心肌梗死面积百分比由43.08%±2.43%减小到21.54%±1.82%(P＜0.05)。siRNA组肌丝排列较整齐,心肌坏死程度较低,细胞水肿较I/R组明显减轻。siRNA组大鼠心肌细胞凋亡水平明显低于I/R组(P＜0.05),Bax和caspase-3表达水平也明显下降(P＜0.05)。miR-26a抑制可以显著降低I/R时Rho/RhoA信号通路蛋白表达(P＜0.05)。结论 miR-26a沉默可能通过降低Rho/RhoA信号通路蛋白表达抑制I/R所致心肌细胞凋亡。

关键词: 心肌缺血/再灌注损伤, miR-26a, 心肌细胞, 凋亡

Abstract: Objective To investigate the effect of miR-26a on myocardial ischemia-reperfusion (I/R) injury in rats. Methods Rats were randomly divided into control group (n=20),I/R group (n=20) and siRNA group(n=20). Seven days before the establishment of I/R model, miR-26a siRNA was injected into tail vein to silence endogenous miR-26a. Ejection fraction (EF%) and shortening fraction (FS%) were measured by echocardiography.The infarct size was measured by TTC.The morphology of cardiomyocytes was detected by HE staining.TUNEL assay was used to detect the level of cardiomyocyte apoptosis.Western blot was used to detect the expression of Bcl-2 related X protein (Bax), caspase-3 and Rho/RhoA signaling pathway related proteins. Results The expression of miR-26a in I/R group was significantly higher than that in control group (P＜0.05).The EF% and FS% of siRNA group were significantly improved (P＜0.05).In siRNA group, the severity of myocardial infarction caused by I/R was significantly reduced, and the percentage of myocardial infarction area was increased from 43.08%±2.43% to 21.54%±1.82% (P＜0.05).In siRNA group, the myofilaments were arranged orderly, the degree of myocardial necrosis was low, and the cell edema was significantly alleviated compared with I/R group.The level of cardiomyocyte apoptosis in siRNA group was significantly lower than that in I/R group (P＜0.05),and the expression levels of Bax and caspase-3 were also significantly decreased (P＜0.05).Inhibition of miR-26a could significantly reduce the expression of Rho/RhoA signaling pathway protein during I/R (P＜0.05). Conclusions miR-26a silencing may inhibit I/R-induced cardiomyocyte apoptosis by reducing expression of Rho/RhoA signaling pathway protein.

Key words: myocardial ischemia-reperfusion injury, miR-26a, myocardial cells, apoptosis

中图分类号:

R364.1

宫丹丹, 隋春兴, 石晨, 李月, 姜晓东. 沉默miR-26a减轻缺血/再灌注大鼠心肌损伤[J]. 基础医学与临床, 2021, 41(9): 1285-1290.

GONG Dan-dan, SUI Chun-xing, SHI Chen, LI Yue, JIANG Xiao-dong. Silencing miR-26a alleviates myocardial ischemia-reperfusion injury in rats[J]. Basic & Clinical Medicine, 2021, 41(9): 1285-1290.

参考文献

[1]Cadenas S. ROS and redox signaling in myocardial ischemia-reperfusion injury and cardioprotection[J]. Free Radic Biol Med, 2018, 117:76-89.
[2]Xu Z, Mcelhanon KE, Beck EX, et al. A Murine model of myocardial ischemia-reperfusion injury[J]. Methods Mol Biol, 2018, 1717:145-153.
[3]Zhao WJ, Zhang HF, Su JY. Downregulation of microRNA-195 promotes angiogenesis induced by cerebral infarction via targeting VEGFA[J]. Mol Med Rep, 2017, 16:5434-5440.
[4]Zhao D, Jian Y, Yang L. Insights for oxidative stress and mTOR signaling in myocardial ischemia/reperfusion injury under diabetes[J]. Oxid Med Cell Longev, 2017, 2017:6437467.doi:10.1155/2017/6437467.
[5]Chen Y, Ba L, Huang W, et al. Role of carvacrol in cardioprotection against myocardial ischemia/reperfusion injury in rats through activation of MAPK/ERK and Akt/eNOS signaling pathways[J]. Eur J Pharmacol, 2017, 796:90-100.
[6]宁娜, 侯晓鸿, 李杨,等. 自噬降低激活β1-AA诱导的心肌细胞内质网应激相关凋亡途径[J].基础医学与临床, 2019, 39: 1-6.
[7]Wang X, Yang L, Kang L, et al. Metformin attenuates myocardial ischemia-reperfusion injury via up-regulation of antioxidant enzymes[J]. PLoS One, 2017, 12:e0182777. doi:10.1371/journal.pone.0182777.
[8]Li D,Wang X,Huang Q, et al. Cardioprotection of CAPE-o NO₂ against myocardial ischemia/reperfusion induced ROS generation via regulating the SIRT1/eNOS/NF-κB pathway in vivo and in vitro[J]. Redox Biol, 2018, 15:62-73.
[9]Guo W,Liu X,Li J,et al. Prdx1 alleviates cardiomyocyte apoptosis through ROS-activated MAPK pathway during myocardial ischemia/reperfusion injury[J]. Int J Biol Macromol,2018,112:608-615.
[10]Ashkenazi A, Fairbrother WJ, Leverson JD, et al. From basic apoptosis discoveries to advanced selective BCL-2 family inhibitors[J]. Nat Rev Drug Discov, 2017, 16:273.
[11]王磊, 朱自强, 袁丽霞,. 丙泊酚减轻H₂O₂诱导的心肌细胞系H9C2损伤[J].基础医学与临床, 2019, 39: 552-557.
[12]Suh JH, Choi E, Cha MJ, et al. Up-regulation of miR-26a promotes apoptosis of hypoxic rat neonatal cardiomyocytes by repressing GSK-3β protein expression[J]. Biochem Biophysi Res Commun, 2012, 423:404-410.

[1]	张敏. 七氟醚对蛛网膜下腔出血大鼠脑组织Wnt/β-catenin信号通路的影响[J]. 基础医学与临床, 2022, 42(8): 1206-1212.
[2]	江宁郭钧. 莪术醇通过Wnt/β-catenin信号通路对人骨肉瘤细胞增殖、凋亡作用[J]. 基础医学与临床, 2022, 42(8): 120-1205.
[3]	薛晋芳宋海飞陈国兵. 程序性细胞死亡在脓毒症发生发展中的研究进展[J]. 基础医学与临床, 2022, 42(8): 1284-1287.
[4]	韩雪莹, 崔丰, 李隽. 乳腺癌缺失因子1(DBC1)影响乳腺癌细胞系对依托泊苷的敏感性[J]. 基础医学与临床, 2022, 42(7): 1060-1066.
[5]	沙务嘎, 李隽. 高通量筛选NMNAT1抑制剂及其肿瘤杀伤作用[J]. 基础医学与临床, 2022, 42(6): 933-939.
[6]	郭婧, 李亚洁, 牟寒霜. LncRNA FGD5-AS1靶向miR-106a-5p减轻ox-LDL诱导的HUVECs损伤[J]. 基础医学与临床, 2022, 42(5): 768-775.
[7]	陈思, 杨富国, 彭红军, 曹卫乐, 王振宇. 丙泊酚减轻缺氧/复氧诱导的心肌细胞损伤[J]. 基础医学与临床, 2022, 42(4): 583-589.
[8]	黄云, 许喜生, 陈凯, 何秀. 下调lncRNA-H19表达促进人瘢痕疙瘩成纤维细胞凋亡和自噬[J]. 基础医学与临床, 2022, 42(4): 633-639.
[9]	陈秋霞, 杨黎星, 马瑞, 赵永晶, 王钰铖, 鞠瑞, 郭磊. 羧胺三唑乳清酸盐抑制人胰腺癌吉西他滨耐药细胞株增殖及线粒体能量代谢[J]. 基础医学与临床, 2022, 42(4): 570-576.
[10]	王帅, 王建军, 孙秀红, 邬鹏宇, 崔志成, 李占清. 萝卜硫素减轻心脏移植模型大鼠缺血/再灌注损伤[J]. 基础医学与临床, 2022, 42(3): 411-416.
[11]	张健, 张吉炯. LncRNA UNC5B-AS1靶向miR-339-5p调控人肺腺癌细胞系A549增殖和凋亡[J]. 基础医学与临床, 2022, 42(3): 454-460.
[12]	赵霞, 蔡庆春, 丁瑞敏, 张倩. 华蟾素抑制人鼻咽癌细胞系CNE2和HONE1增殖[J]. 基础医学与临床, 2022, 42(3): 436-440.
[13]	李明皓, 范亮亮, 项荣. 内质网应激在丙肝病毒感染发病学中作用的研究进展[J]. 基础医学与临床, 2022, 42(2): 316-320.
[14]	陈胜, 王春明, 严雪飞, 毛渊青. 特异性坏死性凋亡抑制剂-1(Nec-1)减轻脊髓损伤模型大鼠胶质瘢痕的形成[J]. 基础医学与临床, 2022, 42(1): 94-99.
[15]	李勇晓, 孙燕, 张瑞生. 丙泊酚抑制人子宫内膜癌细胞系RL95-2增殖[J]. 基础医学与临床, 2022, 42(1): 114-119.