二甲双胍通过激活自噬改善野百合碱诱导的肺动脉高压大鼠的肺动脉重塑和功能  

Metformin attenuates the remodeling and function of pulmonary artery by activating autophagy in monocrotaline-induced pulmonary arterial hypertension in rats

作　　者：杜阿胖 林立建 练桂丽 黄邦邦 庄伟[4] 谢良地 韩英[1,3] DU A-pang;LIN Li-jian;LIAN Gui-li;HUANG Bang-bang;ZHUANG Wei;XIE Liang-di;HAN Ying(Fujian Hypertension Research Institute,The First Affiliated Hospital of Fujian Medical University,Fuzhou,Fujian 350005,China;Cardiovascular Departement,The Second Affiliated Hospital of Fujian Medical University;Department of Geriatrics,The First Affiliated Hospital of Fujian Medical University;Department of Cardiology,The First Affiliated Hospital of Fujian Medical University)

机构地区：[1]福建医科大学附属第一医院,福建省高血压研究所,福建福州350005 [2]福建医科大学附属第二医院心血管内科三区 [3]福建医科大学附属第一医院老年医学科 [4]福建医科大学附属第一医院心血管内科

出　　处：《中华高血压杂志》2021年第11期1069-1077,共9页Chinese Journal of Hypertension

基　　金：国家自然科学基金面上项目(81873537);福建省教育厅中青年教师教育科研项目(JT18092)。

摘　　要：目的观察二甲双胍对野百合碱诱导的肺动脉高压(PAH)大鼠的肺动脉压及肺小动脉结构和功能的影响,探讨二甲双胍改善PAH的可能机制。方法8周龄雄性SD大鼠48只随机分为正常对照组(n=16)、PAH组(n=16)、二甲双胍组(n=16)。PAH组给予腹腔注射野百合碱(20 mg/kg,1次/周,持续两周)建立大鼠PAH模型;二甲双胍组在野百合碱造模同时予二甲双胍[100 mg/(kg·d)]灌胃,对照组给予等量的生理盐水灌胃。给药4周后测定大鼠的平均肺动脉压(MPAP)、右心室肥厚指数(RVHI)、肺小动脉管壁厚度指数(WT%)及肺小动脉管壁面积指数(WA%)、内皮依赖性血管舒张功能(EDdR)、非内皮依赖性血管舒张功能(EDiR)和血管收缩功能;免疫组织化学法观察肺组织微管相关蛋白1轻链3Ⅱ/Ⅰ(LC3Ⅱ/Ⅰ)、增殖细胞核抗原(PCNA)表达水平;Western blot法检测肺组织自噬蛋白Beclin1、LC3Ⅱ/Ⅰ及腺苷酸活化蛋白激酶(AMPK)和磷酸化腺苷酸活化蛋白激酶(P-AMPK)的表达。结果与对照组相比,PAH组和二甲双胍组的MPAP[(34.43±2.78)比(27.76±1.98)比(20.79±1.92)mm Hg,F=72.692,P<0.001]、RVHI[(48.77±7.06)%比(31.96±6.08)%比(20.88±3.44)%,F=40.121,P<0.001]、WA%[(79.79±3.06)%比(61.59±7.16)%比(51.13±4.33)%,F=64.062,P<0.001]、WT%[(55.96±3.87)%比(40.17±6.17)%比(30.00±3.35)%,F=63.899,P<0.001]水平增高,但二甲双胍组低于PAH组。与对照组相比,PAH组及二甲双胍组的EDdR和EDiR降低,但二甲双胍组较PAH组高。野百合碱腹腔注射和二甲双胍治疗后血管收缩功能无明显变化。与对照组相比,PAH组和二甲双胍组大鼠肺组织P-AMPK/AMPK及自噬标志性蛋白Beclin1、LC3Ⅱ/Ⅰ表达增高,但二甲双胍组高于PAH组(均P<0.05)。PAH组大鼠支气管壁、肺小血管和肺泡周围PCNA表达明显增多,而二甲双胍治疗后有明显下降。结论二甲双胍能改善野百合碱诱导的PAH大鼠肺动脉重塑和功能,其机制可能与自噬激活有关。Objective To observe the effects of metformin on pulmonary arterial pressure,pulmonary arterial structure and function in monocrotaline(MCT)-induced pulmonary arterial hypertension(PAH)rats,and to explore its possible mechanism.Methods Forty-eight male Sprague-Dawley rats were randomly divided into three groups:normal control group(CTRL,n=16),PAH group(n=16)and metformin group(MET,n=16).Rats in PAH and MET groups received intraperitoneal injection of MCT(20 mg/kg)at the first day of the first and second week to establish PAH model.Rats in MET group were given daily intragastric metformin[100 mg/(kg·d)],while rats in CTRL group were administrated with equal volume of 0.9% sodium chloride solution by intragastric administration for 4 weeks.Mean pulmonary arterial pressure(MPAP),right ventricular hypertrophy index(RVHI),ratio of wall/total thickness index(WT%),ratio of wall/total area index(WA%),vascular endothelium-dependent relaxation(EDdR),and endothelium-independent relaxation(EDiR)were measured at the end of the 4 th week after MCT injection.The expression of microtubule associated protein 1 light chain 3(LC3Ⅱ/Ⅰ)in lung tissue was detected by immunohistochemistry.The levels of protein Beclin1,LC3Ⅱ/Ⅰ,adenosine 5-monophosphate-activated protein kinase(AMPK)and phosphorylated AMPK(P-AMPK)in the lung tissue were detected by Western blot.Results Compared with CTRL group,MPAP[(34.43±2.78)vs(27.76±1.98)vs(20.79±1.92)mm Hg,F=72.692,P<0.001],RVHI[(48.77±7.06)%vs(31.96±6.08)%vs(20.88±3.44)%,F=40.121,P<0.001],WA%[(79.79±3.06)%vs(61.59±7.16)%vs(51.13±4.33)%,F=64.062,P<0.001]and WT%[(55.96±3.87)%vs(40.17±6.17)%vs(30.00±3.35)%,F=63.899,P<0.001]were significantly increased in both PAH group and MET group,which were lower in MET group(all P<0.05).Compared with CTRL group,EDdR and EDiR decreased in both MET and PAH group,which were higher in MET group.However,there was no significant difference in pulmonary vascular constriction among three groups.Compared with CTRL group,the expressions of P-AMPK/AMPK,Becl

关 键 词：二甲双胍 野百合碱 肺动脉高压 血管功能 自噬 

分 类 号：R544.1[医药卫生—心血管疾病] R-332[医药卫生—内科学]