摘要
目的探究白藜芦醇(Resveratrol,Res)对心肌缺血再灌注(Ischemia Reperfusion,I/R)损伤大鼠模型心肌细胞铁死亡的影响,并探讨其相关机制。方法选取清洁级健康SD大鼠72只,并随机分组为假手术组(Sham组,n=12)、心肌缺血再灌注组(I/R组,n=12)、心肌缺血再灌注+Res低、中、高、剂量组(I/R+ResL组,I/R+ResM组,I/R+ResH组,n=12)、促红细胞生成素组(EPO组,n=12)6组。Sham组行假手术,其余各组通过结扎冠状动脉左前降支后恢复血流灌注进行心肌缺血再灌注损伤模型的制备。I/R+Res 3组在I/R模型建立前分别连续7 d向大鼠腹腔注射RES 0.5 mg/100 g、1.5mg/100 g、3 mg/100 g,EPO组注射促红细胞生成素300 u/100 g,Sham组与I/R组大鼠腹腔注射等容量的生理盐水。再灌注后通过TTC染色观察心肌梗死水平,HE染色观察心肌组织病理状态,动脉血测心肌损伤相关标志物乳酸脱氢酶(LDH)和肌酸激酶同工酶(CK-MB)的浓度,逆转录-聚合酶链反应(RT-PCR)方法检测心肌组织中铁代谢相关的转铁蛋白受体1(TFRl)、铁反应元件结合蛋白2(IREB2)以及铁蛋白重链(FTH)的mRNA的表达水平,Western blot法测定心肌组织中TFRl、IREB2和FTH的蛋白表达,铁检测试剂盒检测心肌细胞内铁沉积情况,流式细胞术检测细胞内ROS荧光强度。结果TTC结果显示,I/R+ResM组、I/R+ResH组与EPO组心肌梗死区域明显低于I/R组;HE染色结果显示,与Sham组相比,I/R+ResM组心肌损伤情况较I/R组明显改善;I/R+Res组和I/R组LDH和CK-MB浓度明显升高(P<0.05);与I/R组相比,I/R+Res组大鼠心肌组织中TFRl、IREB2以及FTH的mRNA及蛋白表达水平均明显下降(P<0.05);组织铁检测试剂盒结果显示I/R+Res组心肌细胞铁含量低于I/R组(P<0.05);流式细胞术结果显示I/R+Res组心肌细胞ROS强度低于I/R组。结论Res能够通过抑制心肌细胞的铁摄取、促进铁的代谢以及铁蛋白的自噬进而降低心肌细胞内铁离子的含量,避免氧化应激损伤,可能以
Objective To explore the effect of Resveratrol(Res)on the iron death of myocardial cells in a rat model of myocardial ischemia-reperfusion(I/R)injury,and to explore its related mechanisms.Methods 72 clean grade healthy SD rats were randomly divided into Sham group(n=12),I/R group(n=12),I/R+Res Low dose group(I/R+ResL),I/R+Res Medium dose group(I/R+ResM),I/R+Res High dose group(I/R+ResH)and EPO group(n=12).Sham operation was performed in sham group,and blood perfusion was restored after ligation of left anterior descending coronary artery in other groups to establish myocardial ischemia-reperfusion injury model.Rats in three I/R+Res groups were intraperitoneally injected with RES 0.5 mg/100 g,1.5 mg/100 g and 3 mg/100 g for 7 consecutive days before the establishment of I/R model,and rats in the EPO group were injected with erythropoietin 300 u/100 g.Rats in Sham group and I/R group were intraperitoneally injected with the same volume of normal saline.After reperfusion,TTC staining was used to observe the level of myocardial infarction,HE staining was used to observe the pathological state of myocardial tissue,and the concentrations of myocardial injury related markers LDH and CK-MB were measured in arterial blood.RT-PCR was used to detect the mRNA expression levels of TFRl,IREB2 and FTH in myocardial tissue.Western blot was used to determine the protein expression of TFRl,IREB2 and FTH in myocardial tissue.Iron deposition in cardiomyocytes was detected by iron detection kit,and intracellular ROS fluorescence intensity was detected by flow cytometry.Results TTC results showed that the area of myocardial infarction in I/R+ResM group,I/R+ResH group and EPO group was significantly lower than that in I/R group.HE staining results showed that compared with Sham group,myocardial injury in I/R+ResM group was significantly improved.Compared with rats in the Sham group,the concentrations of LDH and CK-MB in the I/R+Res group and the I/R group were significantly increased(P<0.05).RT-PCR and WB Results showed that,compared w
作者
孙加燕
任煜
刘婷婷
SUN Jia-yan(Anqing Medical College Pharmacy,Anqing 246052,China)
出处
《牡丹江医学院学报》
2022年第6期14-18,27,共6页
Journal of Mudanjiang Medical University
基金
安徽省高校自然研究项目(KJ2021A1292)。
关键词
白藜芦醇
心肌缺血再灌注
心肌细胞
铁死亡
resveratrol
myocardial ischemia-reperfusion
myocardial cells
ferroptosis
