摘要
目的探究微小RNA(miR)-124-3p对自发性高血压(SHR)大鼠心肌线粒体功能和PI3K/AKT信号通路的调控作用。方法45只SHR大鼠分为SHR组、SHR+miR‑124-3p激动剂(agomir)和SHR+miR‑124-3p拮抗剂(antagomir)组,每组各15只,另选15只健康大鼠为对照组。通过尾静脉注射miR-124-3p agomir和miR-124-3p antagomir(300μg)以提高和抑制心肌细胞中miR-124-5p的水平。4周后检测各组大鼠心肌酶指标、心肌组织中miR-124-3p、细胞凋亡水平、线粒体凋亡标志蛋白[细胞色素C(Cyt C)、caspase9、cleaved caspase3]、PI3K/AKT通路水平。双荧光素酶报告实验验证miR-124-3p和PI3K的靶向关系。将心肌细胞分为miR-124-3p NC组和miR-124-3p mimic组,细胞分别转染miR-124-3p NC和miR-124-3p mimic,检测各组细胞中miR-124-3p和PI3K蛋白水平。结果对照组、SHR组、SHR+miR-124-3p agomir和SHR+miR-124-3p antagomir组大鼠心肌组织中的miR-124-3p水平分别为(0.87±0.12)、(2.56±0.38)、(4.51±0.86)和(1.13±0.16)。四组大鼠各项指标比较差异有统计学意义(P<0.05)。SHR组的乳酸脱氢酶[(LDH,1982.35±207.44)IU/L]、肌酸激酶同工酶[(CK-MB,942.55±107.62)IU/L]、凋亡细胞数目[(27.18±2.10)个]、Cyt C、caspase9和cleaved-caspase3蛋白表达量显著高于对照组,PI3K和AKT mRNA和蛋白表达量显著低于对照组(P<0.05)。SHR+miR-124-3p agomir组的LDH[(2371.92±244.26)IU/L]、CK-MB[(1301.42±135.49)IU/L]、凋亡细胞数目[(52.76±3.56)]个、Cyt C、caspase9和cleaved-caspase3蛋白表达量显著高于SHR组,PI3K和AKT mRNA和蛋白表达量显著低于SHR组(P<0.05)。SHR+miR-124-3p antagomir组的LDH[(1624.43±176.31)IU/L]、CK-MB[(722.41±78.43)IU/L]、凋亡细胞数目[(11.85±0.97)个]、Cyt C、caspase9和cleaved-caspase3蛋白表达量显著低于SHR组,PI3K和AKT mRNA和蛋白表达量显著高于SHR组(P<0.05)。双荧光素酶报告结果表明miR-124-3p与PI3K靶向结合。MiR-124-3p mimic组的miR-124-3p水平显著高于miR-124-3p NC组,PI3K mRNA和蛋白的�
Objective To study the regulatory effects of miR-124-3p on myocardial mitochondrial function and PI3K/AKT signaling pathway in spontaneous hypertension rats(SHR).Methods SHR rats(n=45)were divided into SHR group,SHR+miR‑124-3p agomir group and SHR+miR‑124-3p antagomir group(each n=15),and other healthy rats(n=15)were chosen into control group.MiR-124-3p agomir and miR-124-3p antagomir(300μg)were injected through tail vein to increase and inhibit miR-124-5p level in cardiomyocytes.After 4 weeks,the myocardial enzyme indexes,and levels of miR-124-3p in myocardial tissue,apoptosis,mitochondrial apoptosis marker protein(Cyt C,caspase9,cleaved caspase3)and PI3K/AKT pathway were detected in all groups.The targeting relationship between miR-124-3p and PI3K was verified by using dual-luciferase reporter experiment.The cardiomyocytes were divided into miR-124-3p NC group and miR-124-3p mimic group,and the cells were transfected with miR-124-3p NC and miR-124-3p mimic,respectively.The protein levels of miR-124-3p and PI3K were detected in 2 groups.Results The level of miR-124-3p was(0.87±0.12)in control group,(2.56±0.38)in SHR group,(4.51±0.86)in SHR+miR‑124-3p agomir group and(1.13±0.16)in SHR+miR‑124-3p antagomir group(P<0.05).The levels of lactic dehydrogenase[LDH,(1982.35±207.44)IU/L]and CK-MB[(942.55±107.62)IU/L],number of apoptosis cells(27.18±2.10)and protein expressions of Cyt C,caspase9 and cleaved-caspase3 were significantly higher,and mRNA and protein expressions of PI3K and AKT were significantly lower in SHR group than those in control group(P<0.05).The levels of LDH[(2371.92±244.26)IU/L]and CK-MB[(1301.42±135.49)IU/L],number of apoptosis cells(52.76±3.56)and protein expressions of Cyt C,caspase9 and cleaved-caspase3 were significantly higher,and mRNA and protein expressions of PI3K and AKT were significantly lower in SHR+miR-124-3p agomir group than those in SHR group(P<0.05).The levels of LDH[(1624.43±176.31)IU/L]and CK-MB[(722.41±78.43)IU/L],number of apoptosis cells(11.85±0.97)and pro
作者
吴哲
邹彩霞
廖锋
Wu Zhe;Zou Caixia;Liao Feng(Department of Cardiovascular Medicine,West Area,Qingdao Municipal Hospital,Qingdao 266000,China;不详)
出处
《中国循证心血管医学杂志》
2021年第6期747-751,共5页
Chinese Journal of Evidence-Based Cardiovascular Medicine
