微小RNA-323通过前列腺凋亡反应蛋白-4/cleaved caspase-3通路缓解心肌缺血再灌注损伤  

MicroRNA-323 alleviates myocardial ischemia-reperfusion injury by modulating the prostate apoptosis response-4/cleaved caspase-3 pathway

作　　者：宋佶 黄碧珍[1] 孙常青[1] 刘祖恒 郑武扬[1] SONG Ji;HUANG Bizhen;SUN Changqing;LIU zuheng;ZHENG Wuyang(Department of Cardiovascular,The First Affiliated Hospital of Xiamen University,Xiamen,Fujian 361003;Department of Cardiovascular and Neurology,Genertec Medical Chengfei Hospital)

机构地区：[1]厦门大学附属第一医院心血管内科,福建厦门361003 [2]通用医疗成都成飞医院心内神内科

出　　处：《中华高血压杂志（中英文）》2024年第11期1049-1056,共8页Chinese Journal of Hypertension

基　　金：厦门市科技计划项目(3502Z20209020)。

摘　　要：目的探讨微小RNA-323(miR-323)对缺血再灌注诱导心肌细胞损伤的影响。方法建立H9C2缺氧/复氧(HO)损伤细胞模型与小鼠缺血/再灌注(I/R)模型,采用四甲基偶氮唑盐微量酶反应(MTT)比色法检测心肌细胞活力,采用乳酸脱氢酶(LDH)释放试验检测心肌细胞损伤;采用小动物超声检测模型小鼠心功能情况,四唑红-伊文思蓝(TTC-EB)双染色法测量心肌梗死面积。HO处理前后采用Western-blot法检测前列腺凋亡反应蛋白-4(PAR4)、cleaved caspase-3的蛋白水平。结果在细胞模型中,与对照组比较,缺氧/复氧-miR-323(HO-miR-323)组细胞活力改善(t=8.50,P<0.01),LDH释放降低(t=13.65,P<0.01);与miR-323-抑制剂(miR-323-inhibitor)组比较,HO-miR-323-抑制剂(HO-miR-323-inhibitor)组中细胞活力下降(t=8.46,P<0.01),LDH释放增加(t=22.75,P<0.01)。在动物模型中,与对照组相比,miR-323-激动剂(miR-323-agomiR)组心脏功能改善[左室射血分数(LVEF):(39.9±4.4)%比(32.0±3.2)%,t=3.54,P<0.01;左心室缩短分数(LVFS):(19.2±2.0)%比(15.4±2.1)%,t=3.17,P=0.01],miR-323-拮抗剂(miR-323-antagomiR)组心功能降低[LVEF:(29.2±3.7)%比(34.0±4.1)%,t=2.20,P=0.04;LVFS:(11.9±1.7)%比(14.9±1.9)%,t=2.74,P=0.01]。与对照组相比,miR-323-agomiR组心肌梗死面积[(47.2±9.1)%比(57.7±5.9)%,t=2.38,P=0.04]和危险区相对面积[(30.2±4.2)%比(44.9±6.4)%,t=4.70,P<0.01]减少,miR-323-antagomiR组梗死面积[(65.1±7.8)%比(53.2±6.7)%,t=2.83,P=0.02]与危险区相对面积[(53.5±4.5)%比(44.3±7.5)%,t=2.59,P=0.03]增加。转染miR-323可抑制H9C2细胞PAR4的表达,同时减少cleaved caspase-3蛋白表达。结论miR-323通过靶向PAR4调节cleaved caspase-3,从而改善缺血再灌注造成的心肌细胞损伤和改善心功能障碍。Objective To investigate the impact of microRNA-323(miR-323)on cardiomyocyte injury induced by ischemia-reperfusion.Methods The H9C2 cell models for hypoxia/reoxygenation(HO)injury and mice models for ischemia/reperfusion(I/R)were established.Cell viability was assessed through methyl thiazolyl tetrazolium(MTT)colorimetry,while cardiomyocyte injury was determined using lactate dehydrogenase(LDH)release assays.Cardiac function was evaluated via ultrasound and the myocardial infarction area was measured through 2,3,5-triphenyl-2H-tetrazolium chloride-evans blue(TTC-EB)double staining.The protein levels of prostate apoptosis response-4(PAR4)and cleaved caspase-3 were assessed via Western-blot in H9C2 cell models both pre-and post-hypoxia/reoxygenation.Results In the cellular model,after hypoxia/reoxygenation,the HO-miR-323 group demonstrated a substantially reduced LDH content(t=13.65,P<0.01)compared to the control group,concurrently leading to a noteworthy amelioration in cell viability(t=8.50,P<0.01).In contrast,the HO-miR-323-inhibitor group manifested a notable increase in LDH content(t=22.75,P<0.01)and a significant reduction in cell viability(t=8.46,P<0.01).In the animal experiment,miR-323-agomiR significantly improved cardiac function compared to the negative control(NC)group[left ventricular ejection fraction(LVEF):(39.9±4.4)%vs(32.0±3.2)%,t=3.54,P<0.01;left ventricular fractional shortening(LVFS):(19.2±2.0)%vs(15.4±2.1)%,t=3.17,P=0.01].But miR-323-antagomiR reversed this improvement[LVEF:(29.2±3.7)%vs(34.0±4.1)%,t=2.20,P=0.04;LVFS:(11.9±1.7)%vs(14.9±1.9)%,t=2.74,P=0.01].Furthermore,compared to the control group,the miR-323-agomiR group exhibited a significant reduction in the infarct size percentage[(47.2±9.1)%vs(57.7±5.9)%,t=2.38,P=0.04]and relative risk area[(30.2±4.2)%vs(44.9±6.4)%,t=4.70,P<0.01].Conversely,the miR-323-antagomiR group demonstrated an opposite pattern,with an increase in infarction area[(65.1±7.8)%vs(53.2±6.7)%,t=2.83,P=0.02]and the relative area at risk[(53.5±4.5)%vs(44.3�

关 键 词：微小RNA-323 缺血再灌注损伤 心肌细胞 前列腺凋亡反应蛋白-4 凋亡 

分 类 号：R542.22[医药卫生—心血管疾病]