薏苡附子散对小鼠心肌缺血和血管内皮功能损伤的保护作用及其机制  

Protective effect of Yiyi Fuzisan on myocardium ischemia and vascular endothelial function injury in mice and its mechanism

作　　者：李畅 马梓珊 黄汕梅 银帮巧 陈枝凡 聂莎[1,2] 张子谦 李力 刘鹰[1] 唐耀平 LI Chang;MA Zishan;HUANG Shanmei;YIN Bangqiao;CHEN Zhifan;NIE Sha;ZHANG Ziqian;LI Li;LIU Ying;TANG Yaoping(Graduate School,Guangxi University of Traditional Chinese Medicine,Nanning 530200,China;Affiliated Ruikang Hospital,Guangxi University of Traditional Chinese Medicine,Nanning 530011,China;Center of Scientific Research,Guangxi University of Traditional Chinese Medicine,Nanning 530200,China;Key Laboratory of Translational Medicine for Treating High-Incidence Infectious Diseases with Integrative Medicine,Guangxi Zhuang Autonomous Region,Nanning 530200,China)

机构地区：[1]广西中医药大学研究生院,广西南宁530200 [2]广西中医药大学附属瑞康医院心血管内科,广西南宁530200 [3]广西中医药大学科学实验中心,广西南宁530200 [4]广西壮族自治区高发传染病中西医结合转化医学重点实验室,广西南宁530200

出　　处：《吉林大学学报（医学版）》2023年第3期580-589,共10页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金项目(81774115,82160856);广西中医药大学博士科研启动基金项目(2020BS035);广西高校中青年教师科研基础能力提升项目(2022KY0303)。

摘　　要：目的:探讨薏苡附子散对小鼠心肌缺血和血管内皮功能损伤的保护作用,阐明其可能的作用机制。方法:60只雄性SPF级C57BL/6J小鼠,随机分为空白组、假手术组、急性心肌缺血(AMI)组、低剂量薏苡附子散组、中剂量薏苡附子散组和高剂量薏苡附子散组,每组10只;另取40只C57BL/6J小鼠随机分为生理盐水组、低剂量薏苡附子散组、中剂量薏苡附子散组和高剂量薏苡附子散组,每组10只。采用冠脉左前降支(LAD)结扎法建立AMI小鼠模型后进行相应的药物干预处理28 d。采用Western blotting法检测小鼠心肌组织中一氧化氮合酶(eNOS)蛋白表达水平,动脉张力检测法检测各组小鼠离体胸主动脉血管张力和舒张率,总一氧化氮(NO)检测试剂盒检测各组小鼠血清中NO水平,氯化三苯基四氮唑(TTC)染色观察各组小鼠心肌组织缺血面积,HE染色观察各组小鼠心肌组织病理形态表现,观察各组小鼠术后存活率。人冠脉内皮细胞(HCAECs)随机分为空白组、缺氧组、缺氧+低剂量薏苡附子散组、缺氧+中剂量薏苡附子散组和缺氧+高剂量薏苡附子散组,除空白组外,其余各组细胞采用三气培养箱培养24 h建立缺氧模型。Griess实验检测各组HCAECs中NO水平,荧光染色法检测各组HCAECs中线粒体膜电位(MMP)和活性氧(ROS)水平。结果:与建立AMI模型前比较,建立AMI模型后小鼠心电图ST段明显抬高。Western blotting法检测,与空白组比较,注射N-硝基-L-精氨酸甲酯(L-NAME)后假手术组、AMI组和低、中及高剂量薏苡附子散组小鼠心肌组织中eNOS蛋白表达水平降低(P<0.05)。与生理盐水组比较,低、中和高剂量薏苡附子散组小鼠胸主动脉舒张率升高(P<0.05)。与空白组和假手术组比较,AMI组小鼠血清中NO水平降低(P<0.05),高剂量薏苡附子散组小鼠血清中NO水平升高(P<0.05);与AMI组比较,低、中和高剂量薏苡附子散组小鼠血清中NO水平升高(P<0.05)。TTC染�Objective:To discuss the protective effect of Yiyi Fuzisan on the myocardium ischemia and vascular endothelial dysfunction of the mice,and to clarify its possible mechanism.Methods:Sixty male SPF grade C57BL/6J mice were randomly divided into blank group,sham operation group,acute myocardial ischemia(AMI)group,and low,medium and high doses of Yiyi Fuzisan groups,and there were 10 mice in each group;another forty C57BL/6J mice were randomly divided into normal saline group and low,medium,and high doses of Yiyi Fuzisan groups,and there were 10 mice in each group.The left anterior descending coronary artery(LAD)ligation method was used to establish the AMI mouse model,and corresponding drug intervention treatment was performed for 28 d.Western blotting method was used to detect the expression levels of endothelial nitric oxide synthase(eNOS)protein in myocardium tissue of the mice in various groups;arterial tension detection method was used to detect the tension of isolated aortic and relaxation rates of the mice in various groups;total nitric oxide(NO)detection kit was used to detect the levels of NO in serum of the mice in various groups;triphenyltetra zolium chloride(TTC)staining was used to observe the ischemia areas of myocardium tissue of the mice in various groups;HE staining was used to observe the pathomorphology of myocardium tissue of the mice in various groups;the postoperative survival rates of the mice in various groups were observed.The human coronary endothelial cells(HCAECs)were randomly divided into blank group,hypoxia group,hypoxia+low dose of Yiyi Fuzisan group,hypoxia+medium dose of Yiyi Fuzisan group,and hypoxia+high dose of Yiyi Fuzisan group.Except for blank group,the cells in the other groups were cultured in the three gas incubator for 24 h to establish the hypoxia models.Griess test was used to detect the levels of NO in the HCAECs in various groups;fluorescence staining was used to detect the levels of mitochondrial membrane potential(MMP)and reactive oxygen species(ROS)in the HCAECs in v

关 键 词：薏苡附子散 一氧化氮 心肌缺血 人冠脉内皮细胞 线粒体 

分 类 号：R541[医药卫生—心血管疾病]