胡桃醌对野百合碱诱导肺动脉高压大鼠内皮间充质转化的影响  

Effects of juglone on endothelial-to-mesenchymal transition in murine pulmonary arterial hypertension induced by monocrotaline

作　　者：刘盼盼 高帅 泮思林[1] 罗刚[1] 纪志娴[1] Liu Panpan;Gao Shuai;Pan Silin;Luo Gang;Ji Zhixian(Heart Center,Women and Children’s Hospital,Qingdao University,Qingdao 266034,China)

机构地区：[1]青岛大学附属妇女儿童医院心脏中心,266034

出　　处：《中华实验外科杂志》2022年第8期1472-1475,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81770316、81970249);山东省泰山学者工程资助(2018);山东省医药卫生科技发展计划项目(2018WS370);青岛市科技惠民示范引导专项(20-3-4-47-nsh)。

摘　　要：目的观察胡桃醌(Juglone)对野百合碱(MCT)诱导肺动脉高压(PAH)大鼠内皮间充质转化(EndoMT)的影响。方法采用随机数字表法将30只Sprague Dawley (SD)大鼠分为3组:正常对照组(Con), PAH组, Juglone组, 每组10只。通过MCT建立PAH模型。Juglone组于MCT干预2周后每天给予Juglone腹腔内注射;4周后, 检测右心室收缩压(RVSP)及右心肥厚指数(RVHI), 并用苏木素-伊红(HE)染色观察各组大鼠肺血管形态的改变;免疫荧光法测定大鼠肺小动脉壁中α-肌动蛋白(α-SMA)和血管性血友病因子(vWF)的表达;蛋白印迹法(Western blot)检测各组大鼠肺组织EndoMT相关蛋白的表达水平。多组间均数比较采用单因素方差分析。结果 PAH组大鼠体重低于Con组[(273.00±39.12) g比(408.90±17.63) g, q=15.774, P<0.05], 而RVSP及RVHI则高于Con组[(39.66±5.26) mmHg、(44.02±5.47)%比(18.71±1.72) mmHg、(22.96±3.50)%, q=17.255、16.126, P<0.05];Juglone组大鼠体重高于PAH组[(346.30±16.12) g比(273.00±39.12) g, q=8.320, P<0.05], RVSP及RVHI则低于PAH组[(23.16±3.29) mmHg、(34.03±2.04)%比(39.66±5.26) mmHg、(44.02±5.47)%, q=13.289、7.480, P<0.05]。HE染色示, PAH组肺小动脉管壁增厚;内皮细胞大部分脱落, 部分空泡变性。Juglone干预后管壁明显变薄;内皮细胞分布均匀。免疫荧光染色示Con组内皮层vWF表达为强阳性, 未见α-SMA表达;PAH组内皮层vWF表达减弱, α-SMA为表达加强, 出现vWF和α-SMA的双阳性染色, Juglone组内皮层vWF表达增强, α-SMA表达减弱, 双阳性染色细胞明显减少。Western blot示, PAH组中vWF, 血管内皮钙黏蛋白(VE-cadherin)相对表达量低于Con组(0.54±0.17比0.83±0.01、0.61±0.16比1.06±0.03, q=4.517, 8.231, P<0.05);Juglone组vWF, VE-cadherin蛋白相对表达量高于PAH组(0.78±0.09比0.54±0.17、0.84±0.02比0.61±0.16, q=3.738、4.207, P<0.05);PAH组中波形蛋白(Vimentin)、α-SMA、脯氨酰顺反异构酶1(Pin1)相对表达量高于Con组(1.10±0.05比0.70±0.09、0.94Objective To observe the effect of juglone on endothelial-to-mesenchymal transition(EndoMT)in murine pulmonary arterial hypertension(PAH)induced by monocrotaline(MCT).Methods Totally,30 Sprague Dawley(SD)rats were divided into 3 groups:normal control group(Con),PAH group and Juglone group,10 per group.PAH model was established by MCT.In Juglone group,Juglone was injected intraperitoneally after MCT intervention for 2 weeks.Four weeks later,right ventricle systolic pressure(RVSP)and right ventricular hypertrophy index(RVHI)were measured,pulmonary vascular morphology was observed by hematoxylin-eosin(HE)staining,the expression of alpha smooth muscle Actin(α-SMA)and von Willebrand factor(vWF)in pulmonary arterioles was determined by immunofluorescence,and the expression of EndoMT-related proteins in the lung tissue was detected by Western blotting.The means of multiple groups were compared by single factor analysis of variance.Results The rat weight in PAH group was lower than that in Con group[(273.00±39.12)g vs.(408.90±17.63)g,q=15.774,P<0.05],and that in RVSP and RVHI group was higher than that in Con group[(39.66±5.26)mmHg,(44.02±5.47)%vs.(18.71±1.72)mmHg,(22.96±3.50)%,q=17.255,16.126,P<0.05].The body weight of rats in Juglone group was greater than that in PAH group[(346.30±16.12)g vs.(273.00±39.12)g,q=8.320,P<0.05],and that in RVSP and RVHI group was lower than that in PAH group[(23.16±3.29)mmHg,(34.03±2.04)%vs.(39.66±5.26)mmHg,(44.02±5.47)%,q=13.289,7.480,P<0.05].HE staining showed that in PAH group,the wall of pulmonary arterioles thickened,most of the endothelial cells fell off and some vacuoles degenerated.After the intervention of Juglone,the wall of the vessel became thinner,and the endothelial cells were evenly distributed.Immunofluorescence staining showed that the expression of vWF in endodermis of Con group was strongly positive,but no expression ofα-SMA was found,while in PAH group,the expression of vWF was decreased,while that ofα-SMA was enhanced,and double positive staining of vWF

关 键 词：胡桃醌 肺动脉高压 内皮间充质转化 肺血管重构 

分 类 号：R544.1[医药卫生—心血管疾病]