胺碘酮通过激活Keap1/Nrf2通路减轻大鼠心脏缺血再灌注损伤
Amiodarone Alleviates Myocardial Ischemia-Reperfusion Injury in Rats by Ac-tivating Keap1/Nrf2 Pathway
刘金江 1赵径 1王娇 1张伟洁 1刘炫2
作者信息
- 1. 四川绵阳四0四医院麻醉科,四川绵阳 621000
- 2. 绵阳市中心医院,四川绵阳 621000
- 折叠
摘要
目的 探讨胺碘酮对心肌缺血大鼠的治疗作用及机制.方法 SD大鼠被随机分为对照组、缺血再灌注(ischemia-reperfusion,IR)组、胺碘酮+IR组,每组8只,采用左冠状动脉前降支夹闭构建心肌IR损伤模型.TTC染色检测梗死心肌面积、苏木素伊红(hematoxylin eosin,HE)染色检测心肌组织病理变化;测定大鼠心脏功能;测定大鼠血清心肌损伤标志物[肌酸激酶同工酶(creatine kinase isoenzyme,CK-MB)、乳酸脱氢酶(lactate dehy-drogenase,LDH)、肌钙蛋白Ⅰ(cardiac troponin Ⅰ,cTnI)]及氧化应激指标[丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)、超氧化物歧化酶(superoxide dismutase,SOD)];蛋白免疫印迹(Western blot,WB)测定Kelch样ECH关联蛋白 1(Kelch like ECH associated protein 1,Keap1)、核因子E2相关因子 2(nuclear factor ery-throid 2-related factor 2,Nrf2)、血红素加氧酶-1(heme oxygenase-1,HO-1)、SOD、醌氧化还原酶[NAD(P)H:quinine oxidoreductase 1,NQO1]的蛋白表达浓度;实时聚合酶链反应(real time polymerase chain reaction,RT-PCR)测定HO-1、SOD、NQO1的mRNA表达水平.结果 与对照组比较,IR组大鼠心肌出现明显梗死、心肌组织呈明显病理损伤,胺碘酮+IR组心肌梗死面积明显减少、心肌损伤明显改善;胺碘酮+IR组大鼠心功能、血清CK-MB、LDH、cTnI浓度明显低于IR组,差异有统计学意义(P<0.05);胺碘酮+IR组大鼠心肌细胞活性氧自由基(reactive oxygen species,ROS)、血清MDA浓度明显低于IR组,而血清GSH、SOD浓度明显高于IR组,差异均有统计学意义(P<0.05);与IR组相比较,胺碘酮+IR组大鼠Nrf2明显升高、Keap1明显下降;胺碘酮+IR组大鼠HO-1、SOD2、NQO1的mRNA、蛋白表达均明显高于IR组,差异均有统计学意义(P<0.05).结论 胺碘酮对心肌IR损伤大鼠具有保护作用,其机制可能与激活Keap1/Nrf2通路抑制氧化应激反应相关.
Abstract
Objectives To explore the protective effect of amiodarone on myocardial ischemia-reperfusion(IR)injury in rats by activating Kelch like ECH associated protein 1(Keap1)/nuclear factor erythroid 2-related factor 2(Nrf2)pathway.Methods SD rats were randomly acted as sham group,IR group and amiodarone+IR group.The myocardial IR injury model was established by clamping the anterior descending branch of the left coronary artery.Then,TTC stain-ing was used to detect the infarcted myocardial area,and hematoxylin eosin(HE)staining was used to detect the patho-logical changes of myocardium.Serum concentrations of creatine kinase isoenzyme(CK-MB),lactate dehydrogenase(LDH),cardiac troponin Ⅰ(cTnI),malondialdehyde(MDA),glutathione(GSH),superoxide dismutase(SOD)were detected.The cardiac function in rats were measured.The protein expressions of Keap1,Nrf2,HO-1,SOD and NQO1 were measured by Western blot(WB).The mRNA expressions of heme oxygenase-1(HO-1),SOD and NAD(P)H:quinine oxidoreductase 1(NQO1)were measured by real time polymerase chain reaction(RT-PCR).Results Compared with sham group,the myocardial tissue of IR group showed obvious myocardial infarction and pathological damage,and the myocardial infarction and damage of IR+amiodarone group was significantly improved.Serum concentrations of CK-MB,LDH and cTnI in IR+amiodarone group were significantly lower than those in IR group(P<0.05).The cardiac func-tion of rats in IR+amiodarone group were improved.The reactive oxygen species(ROS)of myocardial cells and serum concentration of MDA in IR+amiodarone group were significantly lower than those in sham group,but serum concentra-tions of GSH and SOD in IR+amiodarone group were significantly higher than those in sham group(P<0.05).Compared with IR group,Nrf2 in IR+amiodarone group increased significantly and Keap1 decreased significantly(P<0.05).The mRNA and protein expressions of HO-1,SOD and NQO1 in IR+amiodarone group were significantly higher than those in IR group(P<0.05).Conclusions Amiodarone can alleviates myocardial IR injury in rats,and its mechanism may be related to inhibition of oxidative stress response via activating Keap1/Nrf2 pathway.
关键词
胺碘酮/心肌缺血再灌注损伤/Kelch样ECH关联蛋白1/核因子E2相关因子2Key words
amiodarone/myocardial ischemia reperfusion injury/Kelch like ECH associated protein 1/nuclear factor erythroid 2-related factor 2引用本文复制引用
基金项目
四川省医学会科研项目(Q21025)
绵阳市卫健委科研项目(202225)
出版年
2024
NETL
NSTL
万方数据