青藤碱通过调节Nrf2通路保护心肌细胞免受缺氧复氧损伤  

Sinomenine Protects Cardiomyocytes from Hypoxic Reoxygenation Injury by Regulating Nrf2 Pathway

作　　者：崔玉祥 李晨阳 朱丹 张桂娟 CUI Yuxiang;LI Chenyang;ZHU Dan;ZHANG Guijuan(Tangshan Hongci Hospital,Tangshan,Hebei,063000,China)

机构地区：[1]唐山弘慈医院,河北省唐山市063000

出　　处：《医学分子生物学杂志》2021年第6期415-421,共7页Journal of Medical Molecular Biology

基　　金：2018年度中国金属学会冶金安全与健康分会健康卫生科研项目(No.jkws201810)。

摘　　要：目的探究青藤碱通过调节Nrf2通路对心肌细胞缺氧复氧损伤的影响。方法建立缺氧复氧损伤细胞模型,CCK-8法检测细胞存活率;乳酸脱氢酶试剂盒检测LDH水平;肌酸激酶同工酶测试盒检测CK-MB水平;肌钙蛋白测定测试盒检测cTnⅠ水平;DCFH-DA探针检测ROS含量;总超氧化物歧化酶测定试剂盒检测SOD含量;测定试剂盒检测MDA含量;过氧化氢酶测定试剂盒检测CAT含量;谷胱甘肽过氧化物酶测试盒检测GSH-Px含量。ELISA检测TNF-α、IL-6和IL-1β水平;Hoechst染色检测细胞凋亡率;Western印迹检测Cleaved caspase-3、Cleaved caspase-9、Keap1、Nrf2、p-Nrf2、HO-1、NQO1蛋白表达水平;利用siRNA敲减Nrf2低表达Nrf2后再次检测上述指标变化。结果与OGD/R组比较,各剂量组细胞存活率降低,LDH、CK-MB、cTnⅠ水平降低,ROS、MDA含量降低,SOD、CAT和GSH-Px含量升高,TNF-α、IL-6和IL-1β水平降低,细胞凋亡率降低,Cleaved caspase-3、Cleaved caspase-9、Keap1蛋白水平降低,pNrf2/Nrf2、HO-1、NQO1蛋白水平升高。通过siRNA敲减Nrf2低表达Nrf2后,氧化应激,炎性反应和相关蛋白表达被逆转。结论青藤碱通过调节Nrf2通路保护心肌细胞免受缺氧复氧损伤。Objective To investigate the effect of sinomenine on myocardial hypoxia and reoxygenation injury by regulating Nrf2 pathway.Methods The cell model of hypoxia and reoxygenation damage was established,and the cell survival rate was detected by CCK-8 method.Lactate dehydrogenase kit was used to detect LDH levels.CK-MB level was detected by creatine kinase isoenzyme test kit.The level of cTnⅠwas measured by troponin test kit.ROS content was detected by DCFHDA probe.Total superoxide dismutase assay kit was used to detect SOD content.MDA content was detected by malondialdehyde assay kit,CAT by catalase assay kit and GSH-Px by glutathione peroxidase assay kit,and the levels of TNF-α,IL-6 and IL-1βby ELISA.Hoechst staining was used to detect cell apoptosis rate,and Western blotting to detect expression levels of cleaved caspase-3,cleaved caspase-9,Keap1,Nrf2,p-Nrf2,HO-1,and NQO1 protein.siRNA was used to knock down Nrf2.Results The cell survival rate,LDH,CK-MB and cTnⅠlevels,ROS and MDA contents,SOD,CAT and GSH-Px contents,TNF-α,IL-6 and IL-1βlevels,apoptosis rate were decreased in all dose groups when compared with those in the OGD/R group.Cleaved caspase-3,cleaved caspase-9,and Keap1 protein levels were decreased,and p-Nrf2/Nrf2,HO-1,and NQO1 protein levels were increased in all dose groups.Oxidative stress,inflammation and related protein expression were reversed after Nrf2 expression was knocked down by siRNA.Conclusion Sinomenine protects cardiomyocytes from hypoxia and reoxygenation by regulating Nrf2 pathway.

关 键 词：缺血性心脏病 青藤碱 氧化应激 Nrf2通路 

分 类 号：R542.[医药卫生—心血管疾病]