CPU0213激活JAK2/STAT3通路减轻大鼠心肌缺血再灌注及氧化应激损伤  

Activation of JAK2/STAT3 pathway by CPU0213 alleviates myocardial ischemia-reperfusion and oxidative stress injury in rats

作　　者：刘亚丽[1] 李雅丽[2] 李春彦[3] 王静[1] 许晓伟[1] 李新军[1] LIU Yali;LI Yali;LI Chunyan;WANG Jing;XU Xiaowei;LI Xinjun(First Department of Cardiology,Second Affiliated Hospital of Hebei North University,Zhangjiakou 075100,China;Comprehensive Ward,Second Affiliated Hospital of Hebei North University,Zhangjiakou 075100,China;Second Department of Cardiology,Second Affiliated Hospital of Hebei North University,Zhangjiakou 075100,China)

机构地区：[1]河北北方学院附属第二医院心内一科,河北张家口075100 [2]河北北方学院附属第二医院综合病区,河北张家口075100 [3]河北北方学院附属第二医院心内二科,河北张家口075100

出　　处：《细胞与分子免疫学杂志》2020年第11期1009-1015,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：2020年度河北省医学科学课题研究计划(20200498)。

摘　　要：目的研究内皮素受体拮抗剂CPU0213在心肌缺血再灌注(I/R)损伤和氧化应激损伤中发挥的作用及其机制。方法为考察CPU0213在I/R中的作用,将SD大鼠随机分为假手术组、缺血再灌注损伤(I/R)组、I/R后CPU0213处理组、I/R后经CPU0213和Janus激酶2(JAK2)特异性抑制剂AG490处理组;为考察CPU0213在氧化应激损伤中的作用,将分离鉴定后培养的心肌细胞分为对照组、H2O2氧化应激组(H2O2组)、氧化应激损伤经CPU0213处理组、氧化应激损伤后经CPU0213和AG490处理组。构建大鼠心肌I/R模型,按实验要求给予大鼠和分离的心肌细胞不同处理后,取大鼠心脏采用氯化三苯基四氮唑(TTC)染色观察心肌梗死面积,测定乳酸脱氢酶(LDH)和肌酸激酶(CK)活性,流式细胞术检测心肌细胞凋亡率,CCK-8法测定细胞生长活力,Western blot法检测Bcl2、JAK2、磷酸化JAK2(p-JAK2)、胱天蛋白酶3(caspase-3)及caspase-9,信号转导子与转录激活子3(STAT3)和磷酸化STAT3(p-STAT3)的表达。结果I/R后经CPU0213处理可降低心肌梗死面积、LDH、CK活性以及细胞凋亡率,并提高JAK2和STAT3磷酸化水平;与I/R联合CPU0213相比,I/R联合CPU0213和AG490组caspase-3、caspase-9表达量上升,Bcl2表达量明显下降,细胞活力明显下降。氧化应激损伤经CPU0213处理后降低LDH、CK活性和细胞凋亡率,提高JAK2和STAT3磷酸化水平;氧化应激损伤经CPU0213和AG490处理后caspase-3、caspase-9表达量上升,Bcl2表达量明显下降,细胞活力明显下降。结论CPU0213激活JAK2/STAT3通路能够抵抗I/R及氧化应激损伤心肌细胞的凋亡。Objective To investigate the role and mechanism of endothelin receptor antagonist CPU0213 in myocardial ischemia-reperfusion(I/R)injury and oxidative stress injury.Methods In order to investigate the role of CPU0213 in I/R,SD rats were randomly divided into sham operation group,ischemia reperfusion injury(I/R)group,CPU0213 treatment group after I/R,CPU0213 and Janus kinase 2(JAK2)specific inhibitor AG490 treatment group after I/R.In order to investigate the role of CPU0213 in oxidative stress damage,the isolated and characterized cardiomyocytes were cultured and divided into control group,H2O2 oxidative stress group(H2O2 group),oxidative stress damaged group treated with CPU0213,and oxidative stress damaged group treated with CPU0213 and AG490.The rat myocardial I/R models were constructed,and the rats and cardiomyocytes were treated with different treatments according to the experimental requirements.The rat heart was stained with triphenyltetrazolium chloride(TTC)to observe the area of myocardial infarction and the lactate dehydrogenase(LDH)and creatine kinase(CK)activity,flow cytometry to detect the apoptosis rate of cardiomyocytes,CCK-8 method to detect cell growth viability,Western blotting to detect the expression of Bcl2,JAK2,phosphorylated JAK2(p-JAK2),caspase-3 and caspase-9,STAT3 and phosphorylated STAT3(p-STAT3).Results After I/R injury in mice,CPU0213 treatment reduced myocardial infarction area,LDH,CK activity and apoptosis rate,but increased the phosphorylation level of JAK2 and STAT3.Compared with I/R combined with CPU0213,I/R combined with CPU0213 and AG490 increased the expression of caspase-3 and caspase-9,decreased significantly the expression of Bcl2 and the cell viability.After oxidative stress damage to cardiomyocytes,CPU0213 treatment reduced LDH,CK activity and cell apoptosis rate,and increased the phosphorylation level of JAK2 and STAT3.In the oxidative stress damaged group treated with CPU0213 and AG490,caspase-3 and caspase-9 expression increased,Bcl2 expression dropped significantly,ce

关 键 词：CPU0213 Janus激酶2(JAK2) 信号转导子与转录激活子3(STAT3) 心肌缺血再灌注损伤 氧化应激损伤 心肌保护 

分 类 号：R965[医药卫生—药理学] R542.22[医药卫生—药学]