山莨菪碱对心肌缺血再灌注大鼠氧化应激、心肌损伤及线粒体凋亡通路的影响  

Effects of anisodamine on oxidative stress,myocardial injury and mitochondrial apoptosis pathway in rats with myocardial ischemia reperfusion

作　　者：宋艳玲[1] 麦华德 林芸芸[1] 陈明慧 王雅纯[1] 顾申红[1] SONG Yan-Ling;MAI Hua-De;LIN Yun-Yun;CHEN Ming-Hui;WANG Ya-Chun;GU Shen-Hong(Department of Geriatrics,the First Affiliated Hospital of Hainan Medical College,Haikou 570000,China)

机构地区：[1]海南医学院第一附属医院老年病科

出　　处：《中国免疫学杂志》2019年第24期2958-2963,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.81560054);2016海南省卫生计生行业科研项目(No.1601320241A2004)资助

摘　　要：目的:探究山莨菪碱(Anis)对心肌缺血再灌注大鼠氧化应激、心肌损伤及线粒体凋亡通路的影响。方法:选取45只雄性SD大鼠,体重200~300 g,随机平分为5组,分别为健康组、模型组、低剂量山莨菪碱组、中剂量山莨菪碱组和高剂量山莨菪碱组;使用冠状动脉结扎术建立缺血再灌注模型,给药组分别静脉注射1、2、4 mg/kg的Anis,健康组和模型组大鼠静脉注射等剂量的0.9%氯化钠溶液。使用HE染色观察大鼠心肌细胞;检测平均动脉压(MAP)和左室收缩压(LVSP)水平;使用试剂盒检测心肌细胞丙二醛(MDA)、超氧化物歧化酶(SOD)、肌酸激酶同工酶(CK-MB)、肌红蛋白(Mb)的水平;使用ELISA法检测各组大鼠血清IL-6、诱生型一氧化氮合酶(iNOS)的含量;使用免疫组化和蛋白质印迹法检测Caspase-3的表达;使用蛋白质印迹法检测线粒体凋亡通路相关蛋白Bak、Bcl2、Apaf1的表达情况。结果:HE染色结果发现,使用Anis处理后各组心肌组织肌纤维断裂溶解弯曲显著减少;相比健康组,模型组大鼠血清MDA、CK-MB、Mb、IL-6、iNOS和心肌组织Caspase-3、Bak、Bcl2、Apaf1蛋白表达水平显著升高,LVSP、MAP和血清SOD水平显著降低(P<0.05);相比模型组,使用Anis理后各组大鼠血清MDA、CK-MB、Mb、IL-6、iNOS和心肌组织Caspase-3、Bak、Bcl2、Apaf1蛋白表达水平显著降低,LVSP、MAP和血清SOD水平显著升高(P<0.05),且随着使用山莨菪碱处理剂量的增加呈剂量依赖性。结论:Anis可通过降低氧化应激、抑制凋亡相关蛋白表达及线粒体凋亡通路实现缓解心肌缺血再灌注损伤。Objective:To investigate the effects of anisodamine(Anis) on oxidative stress,myocardial injury and mitochondrial apoptosis pathway in rats with myocardial ischemia reperfusion.Methods:45 male SD rats were enrolled,weighting 200-300 g.They were randomly equally divided into 5 groups:healthy group,model group,low-dose Anis group,middle-dose Anis group and high-dose Anis group.The ischemia-reperfusion models were established by coronary artery ligation.The drug-administered groups were intravenously injected with 1,2 and 4 mg/kg Anis.The healthy group and model group were intravenously injected with an equal dose of 0.9% sodium chloride solution.Rat cardiomyocytes were observed by HE staining.The levels of mean arterial pressure(MAP) and left ventricular systolic pressure(LVSP)were measured.The levels of cardiomyocytes malondialdehyde(MDA),superoxide dismutase(SOD),creatine kinase MB(CK-MB) and myohemoglobin(Mb) were detected by kit.The contents of interleukin-6(IL-6) and inducible nitric oxide synthase(iNOS) were detected by ELISA.The expression of Caspase-3 was detected by immunohistochemistry and Western blot.The expression of Bak,Bcl2,Apaf1 of mitochondrial apoptosis pathway related protein was detected by Western blot.Results:HE staining results found that fracture,dissolution and bending of myocardial tissue muscle fibers were significantly reduced in groups after treated with Anis.Compared with healthy group,serum MDA,CK-MB,Mb,IL-6,iNOS and expression levels of Caspase-3,Bak,Bcl2 and Apaf1 protein were significantly increased in model group,while levels of LVSP,MAP and serum SOD were significantly decreased(P<0.05).Compared with model group,serum MDA,CK-MB,Mb,IL-6,iNOS and expression levels of Caspase-3,Bak,Bcl2 and Apaf1 protein were significantly decreased in groups treated with Anis,while levels of LVSP,MAP and serum SOD were significantly increased(P<0.05),showing dose-dependence with increase of Anis usage dose.Conclusion:Anis can alleviate myocardial ischemia-reperfusion injury by reducing oxidative st

关 键 词：心肌缺血再灌注 氧化应激 山莨菪碱 线粒体凋亡 

分 类 号：R34[医药卫生—基础医学]