铁死亡在高脂高糖心肌细胞缺氧复氧损伤中的作用  

Role of ferroptosis in hypoxia-reoxygenation injury in cardiomyocytes cultured in high-fat high-glucose medium

作　　者：黄琴[1] 田立群[1] 赵晓帅 夏中元[1] Huang Qin;Tian Liqun;Zhao Xiaoshuai;Xia Zhongyuan(Department of Anesthesiology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院麻醉科,武汉430060

出　　处：《中华麻醉学杂志》2022年第1期82-87,共6页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81970722)。

摘　　要：目的评价铁死亡在高脂高糖心肌细胞缺氧复氧损伤中的作用。方法正常培养的H9c2心肌细胞,采用随机数字表法分为3组(n=20):对照组(C组)、高脂高糖-缺氧复氧组(HFHG+H/R组)和Ferrostatin-1+高脂高糖-缺氧复氧组(Fer-1+HFHG+H/R组)。采用高脂高糖处理12 h,随后缺氧4 h,复氧2 h的方法制备H9c2心肌细胞缺氧复氧损伤模型。Fer-1+HFHG+H/R组在高脂高糖处理同时加入铁死亡抑制剂Ferrostatin-1,终浓度10μmol/L。于复氧2 h时,采用CCK-8法检测细胞活力,2,4二硝基苯肼显色法检测上清液LDH活性,荧光探针DCFH-DA流式细胞术检测ROS活性,Western blot法检测心肌细胞长链脂酰辅酶A合成酶4(ACSL4)、核受体共激活因子4(NCOA4)和谷胱甘肽过氧化物酶4(GPX4)的表达。结果与C组比较,HFHG+H/R组细胞活力降低,LDH活性、ROS活性、ACSL4和NCOA4表达水平升高(P<0.05),GPX4表达差异无统计学意义(P>0.05);与HFHG+H/R组比较,Fer-1+HFHG+H/R组细胞活力升高,LDH活性、ROS活性、ACSL4和NCOA4表达水平降低(P<0.05),GPX4表达差异无统计学意义(P>0.05)。结论铁死亡参与了高脂高糖心肌细胞缺氧复氧损伤的过程。Objective To evaluate the role of ferroptosis in hypoxia-reoxygenation(H/R)injury in cardiomyocytes cultured in high-fat high-glucose(HFHG)medium.Methods Cardiomyocytes H9c2 cells were commonly cultured and divided into 3 groups(n=20 each)using a random number table method:control group(C group),HFHG-H/R group and Ferrostatin-1(Fer-1)plus HFHG-H/R group(Fer-1+HFHG+H/R group).H9c2 cells were cultured in a HFHG medium for 12 h and then exposed to 1%O2-5%CO2-94%N2 for 4 h,followed by 2 h reoxygenation in a cell incubator.Fer-1 at a final concentration of 10μmol/L was added while the cells were cultured in the HFHG medium in group Fer-1+HFHG+H/R.At 2 h of reoxygenation,the cell viability was measured using CCK-8 assay,the activity of lactate dehydrogenase(LDH)in the supernatant was measured using 2,4-dinitrophenylhydrazine color method,the activity of reactive oxygen species(ROS)was measured by fluorescent probe DCFH-DA flow cytometry,and the expression of acyl-CoA synthetase long-chain family member 4(ACSL4),nuclear receptor coactivator 4(NCOA4),and glutathione peroxidase 4(GPX4)was detected by Western blot.Results Compared with group C,the cell viability was significantly decreased,the activities of LDH release and ROS were increased,and the expression of ACSL4 and NCOA4 was up-regulated(P<0.05),and no significant change was found in the expression of GPX4 in group HFHG+H/R(P>0.05).Compared with group HFHG+H/R,the cell activity was significantly increased,the activities of LDH and ROS were decreased,and the expression of ACSL4 and NCOA4 was down-regulated(P<0.05),and no significant change was found in the expression of GPX4 in Fer-1+HFHG+H/R group(P>0.05).Conclusions Ferroptosis is involved in the process of H/R injury in cardiomyocytes cultured in HFHG medium.

关 键 词：铁 高脂血症 糖尿病 肌细胞 心脏 低氧 

分 类 号：R614[医药卫生—麻醉学]