甘草查尔酮A调控miR-142-3p/FOXO1表达抑制H_(2)O_(2)诱导的心肌细胞凋亡  

Licochalcone A regulates miR-142-3p/FOXO1 expression and inhibits H_(2)O_(2)-induced cardiomyocyte apoptosis

作　　者：马竞 武国利[2] 王潇伟 许金鹏[1] MA Jing;WU Guoli;WANG Xiaowei;XU Jinpeng(Department of Cardiovascular Medicine,Affiliated Hospital of Hebei University,Baoding 071000,China;Intensive Care Unit,Baoding First Central Hospital,Baoding 071000,China)

机构地区：[1]河北大学附属医院心血管内科,河北保定071000 [2]保定市第一中心医院重症医学科,河北保定071000

出　　处：《沈阳药科大学学报》2021年第10期1076-1083,共8页Journal of Shenyang Pharmaceutical University

摘　　要：目的探讨甘草查尔酮A(licochalcone A,LA)对H_(2)O_(2)诱导的心肌细胞凋亡的影响及其机制。方法将体外培养的大鼠心肌H9C2细胞分为Control组、H_(2)O_(2)组(100μmol·L^(-1)H_(2)O_(2)刺激24 h)和H_(2)O_(2)+low dose LA组、H_(2)O_(2)+middle dose LA组、H_(2)O_(2)+high dose LA组(10、20和40μmol·L^(-1)LA孵育4 h后,给予H_(2)O_(2)刺激24 h),采用流式细胞仪检测各组细胞凋亡率,比色法检测细胞Caspase-3活性,RT-PCR检测细胞中miR-142-3p表达,Western blotting检测细胞中FOXO1蛋白表达;采用双荧光素酶报告基因实验和Western blotting检测miR-142-3p与FOXO1靶向调控关系;将miR-142-3p inhibitor或pcDNA3.1-FOXO1过表达载体质粒转染至H9C2细胞后,观察miR-142-3p低表达或FOXO1过表达对40μmol·L^(-1)LA作用下的H9C2细胞凋亡的影响。结果与对照组比较,H_(2)O_(2)组H9C2细胞凋亡率、Caspase-3活性和细胞中FOXO1蛋白表达水平均明显升高,而细胞中miR-142-3p表达水平明显降低(P<0.05);与H_(2)O_(2)组比较,中、高剂量LA可呈剂量依赖性抑制H_(2)O_(2)诱导的H9C2细胞凋亡和Caspase-3活性,并上调miR-142-3p和下调FOXO1蛋白表达(P<0.05)。双荧光素酶报告基因实验和Western blotting实验证实FOXO1是miR-142-3p的靶基因,miR-142-3p可负向调控FOXO1蛋白表达。miR-142-3p低表达或FOXO1过表达可逆转LA对H9C2细胞凋亡的抑制作用。结论LA可通过调控miR-142-3p/FOXO1表达抑制H_(2)O_(2)诱导的心肌细胞凋亡。Objective To investigate the effect of licochalcone A(LA)on H_(2)O_(2)-induced myocardial cell apoptosis and its mechanism.Methods Cultured rat myocardial H9C2 cells in vitro were divided into Control group,H_(2)O_(2)group(100μmol·L^(-1)H_(2)O_(2)stimulation for 24 hours)and H_(2)O_(2)+low dose LA group,middle dose LA group and high dose LA group(10,20 and 40μmol·L^(-1)LA were incubated for 4 hours,and then H_(2)O_(2)was given for 24 hours).The cell apoptosis rate was measured by flow cytometry;Caspase-3 activity was measured by colorimetric method;miR-142-3p expression was detected by RT-PCR,and FOXO1 protein expression in cells were detected by Western blotting.Dual-luciferase reporter gene experiment and Western blotting were used to detect the targeted regulation of miR-142-3p and FOXO1.After transfecting miR-142-3p inhibitor or pcDNA3.1-FOXO1 overexpression vector plasmid into H9C2 cells,the effects of miR-142-3p low expression or FOXO1 overexpression on the apoptosis of H9C2 cells under the action of 40μmol·L^(-1)LA were observed.Results Compared with the Control group,the apoptosis rate,Caspase-3 activity,and FOXO1 protein expression levels in the H_(2)O_(2)group were significantly increased,while the expression level of miR-142-3p in the cells was significantly reduced(P<0.05);Compared with H_(2)O_(2)group,H_(2)O_(2)+middle dose LA group and H_(2)O_(2)+high dose LA group could dose-dependently inhibit H_(2)O_(2)-induced H9C2 apoptosis and Caspase-3 activity,and up-regulate miR-142-3p and down-regulate FOXO1 protein expression(P<0.05).Double luciferase reporter gene experiments and Western blotting experiments confirmed that FOXO1 was a target gene of miR-142-3p,and miR-142-3p could negatively regulate FOXO1 protein expression.Low miR-142-3p expression or FOXO1 overexpression could reverse the inhibitory effect of LA on H9C2 apoptosis.Conclusion LA can inhibit H_(2)O_(2)-induced cardiomyocyte apoptosis by regulating miR-142-3p/FOXO1 expression.

关 键 词：心肌细胞 甘草查尔酮A 细胞凋亡 miR-142-3p FOXO1 

分 类 号：R96[医药卫生—药理学]