丹参素通过抑制miR-199a-5p对心肌缺血再灌注损伤大鼠的保护作用  

Protection of Danshensu on myocardium against ischemia-reperfusion injury through inhibiting miR-199a-5p

作　　者：史云[1] 顾秀峰[1] 李玉明[2] 李辉[2] 李焕明[1] SHI Yun;GU Xiu-feng;LI Yu-ming;LI Hui;LI Huan-ming(Department of Cardiology,Tianjin Fourth Central Hospital,Tianjin 300140,China;Heart Research Institute,Affiliated Hospital of Logistics College of PAP,Tianjin 300162,China)

机构地区：[1]天津市第四中心医院心内科,天津300140 [2]武警后勤学院附属医院心研所,天津300162

出　　处：《现代药物与临床》2021年第6期1104-1111,共8页Drugs & Clinic

基　　金：国家重点研发项目(2017YFC1307602);天津市科技计划项目(15ZXLCSY00040)。

摘　　要：目的评估丹参素对心肌缺血/再灌注(MI/R)损伤的保护作用及其可能机制。方法32只大鼠随机分为假手术组、模型组组、丹参素30 mg/kg组和丹参素60 mg/kg组,丹参素组在再灌注开始后5 min内iv相应剂量丹参素,假手术组和模型组注射等量生理盐水;再灌注3 h后,向冠状动脉中注入2 mL的3%伊文思蓝染料;取大鼠动脉血和心脏,通过组织化学测定心肌梗死面积大小,通过ELISA测定血清中的肌酸激酶-同工酶(CK-MB)和心肌肌钙蛋白(c TnI)水平。通过氧糖剥夺/恢复(OGD/R)H9c2心肌细胞模型评估丹参素预处理对细胞的直接作用,使用MTT、ELISA和流式细胞术分别测定丹参素对细胞活力、乳酸脱氢酶(LDH)释放和细胞凋亡的影响,使用RT-PCR和Western blotting免疫印迹测定miR-199a-5p及其下游蛋白Akt和ERK1/2的表达。结果丹参素治疗可显著降低心肌梗死面积以及血清中CK-MB和cTnI的水平(P<0.05);丹参素预处理显著改善OGD/R心肌细胞活力恢复能力,减少细胞凋亡水平,显著提高Akt和ERK1/2的m RNA和蛋白表达水平,降低miR-199a-5p的m RNA表达水平(P<0.05);上调miR-199a-5p显著降低Akt和ERK1/2的m RNA和蛋白表达水平(P<0.05),而下调miR-199a-5p可显著提高Akt和ERK1/2的m RNA和蛋白表达水平(P<0.05)。结论丹参素可以通过抑制miR-199a-5p增加Akt和ERK1/2表达来发挥心脏保护作用。Objective To evaluate the protective effects of Danshensu,the effective component of Salvia miltiorrhiza(Danshen),against myocardial ischemia/reperfusion(MI/R)injury in cardiomyocytes and its possible underlying mechanism.Methods 32 rats were randomly divided into four groups:sham operation group,model group,Danshensu 30 mg/kg group and Danshensu 60 mg/kg group.Danshensu 30 mg/kg group and Danshensu 60 mg/kg group were given all drugs intravenously within 5 min after reperfusion,sham operation group and model group were injected with the same amount of physiological saline.Three hours after reperfusion,2 mL of 3%Evans was injected into the coronary artery.The size of myocardial infarction was determined by histochemistry.The levels of creatine kinase MB(CK-MB)and cardiac troponin(cTnI)in serum were measured by ELISA.In addition,the direct toxicity of Danshensu on cardiomyocytes was evaluated by oxygen glucose deprivation/recovery(OGD/R)model.MTT,ELISA and flow cytometry were used to determine the effects of Danshensu on cell viability,lactate dehydrogenase(LDH)release and apoptosis.RT-PCR and Western blotting were used to detect the effects of Danshensu on cell viability,LDH release and apoptosis.The expression of miR-199 a-5 p and its downstream proteins Akt and ERK1/2 were detected by Western blotting.Results Danshensu treatment significantly reduced myocardial infarction area and serum CK-MB and cTnI levels(P<0.05);Danshensu pretreatment significantly improved the viability of OGD/R myocardial cells,reduced the level of apoptosis,significantly increased the mRNA and protein expression levels of Akt and ERK1/2,and decreased the mRNA expression level of miR-199 a-5 p(P<0.05).Up regulation of mi R-199 a-5 p significantly decreased the mRNA and protein expression levels of Akt and ERK1/2(P<0.05),and down-regulation of mi R-199 a-5 p significantly increased the mRNA and protein expression levels of Akt and ERK1/2(P<0.05).Conclusion Danshensu provides the cardioprotective effects against MI/R injury via increasing A

关 键 词：丹参素 心肌缺血再灌注损伤 微小RNA 氧糖剥夺/恢复 miR-199a-5p 

分 类 号：R285.5[医药卫生—中药学]