氢溴酸加兰他敏介导AMPKα1/Nrf2/HO-1通路对大鼠心肌缺血再灌注损伤的保护作用  

The Regulation of AMPKα1/Nrf2/HO-1 Pathway Mediated by Galantamine Hydrobromide Lycoremine in Myocardial Ischemia Reperfusion Rats

作　　者：曾菲[1] 李强[2] 曾昪 李希 黄克力[1] 于涛[1] 谭今[1] ZENG Fei;LI Qiang;ZENG Bian;LI Xi;HUANG Ke-li;YU Tao;TAN Jin(Center of Cardiac Surgery,Sichuan Academy of Medical Sciences&Sichuan Provincial People’s Hospital,Chengdu 610072,China;Department of Anesthesiology,the Third People's Hospital of Chengdu,Chengdu 610072,China;Rehabilitation Medicine Department,Sichuan Academy of Medical Sciences&Sichuan Provincial People’s Hospital,Chengdu 610072,China)

机构地区：[1]四川省医学科学院·四川省人民医院心脏外科中心,成都610072 [2]成都市第三人民医院麻醉科,成都610072 [3]四川省医学科学院·四川省人民医院康复医学科,成都610072

出　　处：《四川大学学报（医学版）》2020年第3期337-343,共7页Journal of Sichuan University(Medical Sciences)

基　　金：四川省卫生和计划生育委员会科研课题(No.16PJ297)资助。

摘　　要：目的研究氢溴酸加兰他敏(galantamine hydrobromide lycoremine,Gal)介导腺苷酸激活蛋白激酶(AMPK)α1/Nrf2/血红素加氧酶(heme oxygenase-1,HO-1)通路对心肌缺血再灌注(I/R)大鼠内质网应激型凋亡、心肌凋亡和纤维化的影响。方法构建心肌I/R损伤大鼠模型,将大鼠随机分为5组:对照组、I/R模型组、Gal 1 mg/kg组、Gal 2 mg/kg组、Gal 4 mg/kg组进行后续实验。多普勒超声检测各组大鼠左室射血分数(LVEF)、左心室舒张末期容积(LVEDV)、左心室收缩末期容积(LVESV)、左心室壁厚度(LVWT)、左室短轴缩短率(FS);HE染色检测心肌组织病理损伤程度;免疫组化检测Caspase-3阳性表达率;RT-PCR检测心肌Caspase-3 mRNA表达;蛋白免疫印迹检测内质网应激因子CCAAT/增强子结合蛋白同源蛋白(CCAAT/enhancer-binding protein homologous protein,CHOP)、cleaved Caspase-12、生长抑制和DNA损伤诱导蛋白(growth arrest and DNA damageinducible protein 34,GADD34)、免疫球蛋白重链结合蛋白(immunoglobulin heavy-chainbinding protein,BiP)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、CollagenⅠ、AMPKα1、Nrf2、HO-1蛋白表达水平;并加入AMPK的抑制剂Compound C进行验证。结果与I/R模型组比较,Gal各组心肌组织病理损伤程度明显改善,cleaved Caspase-3阳性表达率和Caspase-3 mRNA水平表达降低(P<0.05);Gal 2 mg/kg、Gal 4 mg/kg组LVWT、FS、LVEF升高(P<0.05),LVEDV、LVESV降低(P<0.05),CHOP、cleaved Caspase-12、α-SMA、CollagenⅠ、AMPKα1、Nrf2、HO-1蛋白表达降低(P<0.05),GADD34、BiP蛋白表达升高(P<0.05)。加入AMPK抑制剂Compound C后,与I/R模型组相比较,CC组AMPKα1、Nrf2、HO-1、BiP蛋白水平降低(P<0.05),CHOP、α-SMA、CollagenⅠ蛋白水平升高(P<0.05),LVESV、FS降低(P<0.05),Caspase-3 mRNA水平升高(P<0.05)。与CC组比较,CC+Gal 4mg/kg组AMPKα1、Nrf2、HO-1、BiP蛋白水平升高(P<0.05),CHOP、α-SMA、CollagenⅠ蛋白水平降低(P<0.05),LVESV、FS升高,Caspase-3 mRNA水平降低(PObjective To investigate the effects of AMPKα1/Nrf2/heme oxygenase-1(HO-1)pathway mediated by galantamine hydrobromide lycoremine(Gal)on endoplasmic reticulum stress apoptosis,myocardial apoptosis and fibrosis in rats with myocardial ischemia reperfusion(I/R).Methods A myocardial ischemia reperfusion injury rat model was established,and the rats were randomly divided into 5 groups:Control group,I/R model group,Gal 1 mg/kg group,Gal 2 mg/kg group and Gal 4 mg/kg group.Left ventricular ejection fraction(LVEF),left ventricular end-diastolic volume(LVEDV),left ventricular end-systolic volume(LVESV),left ventricular wall thickness(LVWT),and left ventricular short-axis shortening rate(FS)were detected by doppler ultrasound.Hematoxylin eosin staining was used to detect the pathological damage of myocardial tissue.The expression of Caspase-3 was detected by immunohistochemistry.Protein expression levels of CCAAT/enhancer-binding protein homologous protein(CHOP),cleaved Caspase-12,growth arrest and DNA damageinducible protein 34(GADD34),immunoglobulin heavy-chain-binding protein(BiP),α-smooth muscle actin(α-SMA),CollagenⅠ,AMPKα1,Nrf2,and HO-1 were measured by western blot,and AMPK inhibitor Compound C was added for verification.Results Compared with the I/R model group,the grade of pathological damage of myocardial tissue in each group of Gal was improved,and cleaved Caspase-3 positive expression rate and Caspase-3 mRNA level were significantly reduced(P<0.05)as well.The results showed that LVWT,FS and LVEF in Gal 2 mg/kg and Gal 4 mg/kg groups were significantly increased(P<0.05),LVEDV and LVESV were significantly reduced(P<0.05)compared with I/R model group.CHOP,cleaved Caspase-12,α-SMA,CollagenⅠ,AMPKα1,Nrf2,HO-1 protein levels were significantly reduced(P<0.05),and GADD34 and BiP protein levels were significantly increased(P<0.05)in Gal 2 mg/kg and Gal 4 mg/kg groups.Conclusion The regulation of AMPKα1/Nrf2/HO-1 pathway mediated by Gal on endoplasmic reticulum stress apoptosis,myocardial apoptosis and fibros

关 键 词：氢溴酸加兰他敏 心肌缺血再灌注 内质网应激 AMPKα1/Nrf2/HO-1通路 

分 类 号：R965[医药卫生—药理学]