脯氨酰顺反异构酶1及其抑制剂对大鼠肺动脉高压的影响及其机制  

Function and mechanism of peptidyl-prolyl Cis-trans isomerase NIMA-interacting1 and its inhibitor in murine pulmonary arterial hypertension

作　　者：高帅 杜占慧 刘盼盼 唐雅琪 泮思林[1] Gao Shuai;Du Zhanhui;Liu Panpan;Tang Yaqi;Pan Silin(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3期480-483,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：目的探讨脯氨酰顺反异构酶1(Pin1)及其抑制剂胡桃醌(Juglone)对肺动脉高压(PAH)的影响及作用机制。方法将8周龄雄性SD大鼠按随机数目表法分配原则分为3组:正常对照组(Con)、PAH组、Juglone组,每组8只。野百合碱建立大鼠PAH模型。Juglone组在造模14 d后开始给予Juglone干预,4周后测定右心室收缩压(RVSP),处死动物并取材,计算右心肥厚指数(RVHI)。苏木精-伊红(HE)染色观察肺血管病理学改变;免疫荧光染色检测Pin1在PAH中的表达;蛋白印迹法(Western blot)检测腺苷酸活化蛋白激酶(AMPK)及S期激酶相关蛋白(Skp2)/p27通路蛋白的表达。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[(303.11±20.41)g比(418.83±26.77)g,q=12.912,P<0.01],而RVSP及RVHI则高于Con组[(39.83±2.31)mmHg、(40.49±3.12)%比(17.03±2.12)mmHg、(23.52±1.61)%,q=28.291、16.105,P<0.01];Juglone组大鼠体重高于PAH组[(349.73±21.96)g比(303.11±20.41)g,q=5.204,P<0.01],RVSP及RVHI则低于PAH组[(26.89±1.92)mmHg、(31.40±3.76)%比(39.83±2.31)mmHg、(40.49±3.12)%,q=16.063、8.637,P<0.01]。免疫荧光染色结果显示PAH组Pin1表达高于Con组[(8.39±0.19)%比(3.32±0.26)%,t=13.000,P<0.01]。HE染色结果显示,PAH组肺小动脉管壁增厚,Juglone干预后肺小动脉壁厚度减轻。Western blot结果显示,Con组α-SMA和Pin1的表达低于PAH组(1.02±0.04、1.00±0.07比2.21±0.07、3.37±0.04,t=14.976、27.427,P<0.01)。PAH组p-AMPK表达低于Con组(0.52±0.02比1.00±0.04,F=25.771,P<0.01),Juglone组p-AMPK表达高于PAH组(0.65±0.04比0.52±0.02,F=6.972,P<0.01);PAH组Skp2表达高于Con组(4.05±0.12比1.00±0.07,F=51.213,P<0.01),Juglone组Skp2表达低于PAH组(2.91±0.08比4.05±0.12,F=19.093,P<0.01);PAH组p27表达低于Con组(0.55±0.03比1.00±0.02,F=28.937,P<0.01),Juglone组p27表达高于PAH组(0.75±0.03比0.55±0.03,F=13.051,P<0.01)。结论Pin1在PAH肺组织中过表达,并参与肺血管重构过程。Pin1抑制剂胡桃醌通过激活AMPK从�Objective To investigate the effect of Peptidyl-prolyl cis-trans isomerase NIMA-interacting1(Pin1)and its inhibitor Juglone on pulmonary arterial hypertension(PAH)and its possible mechanism.Methods Totally,258-weeks old male Sprague Dawley rats were randomly divided into 3 groups:control group(Con),PAH group and Juglone group(8 rats each).Monocrotaline was intraperitoneally injected to establish PAH model,and additional Juglone was given in the Juglone group 14 days later.At 4th week,right ventricular systolic pressure(RVSP)and right ventricular hypertrophy index(RVHI)were measured,then animals were sacrificed and lung tissues were extracted.Hematoxylin and eosin(HE)staining was used to testify the pathological changes of lungs,as well as immunofluorescence staining was performed to dectect the expression of Pin1 in PAH.Moerover,the expression of adenosine monophosphate activatedproteinkinase(AMPK)and S-phase kinase-associated protein 2(Skp2)/p27 pathway proteins was revealed by Western blotting.The means of multiple groups were compared by single factor analysis of variance.Results The weight in PAH group was slighter than Con[(303.11±20.41)g vs.(418.83±26.77)g,q=12.912,P<0.01],while RVSP and RVHI in PAH group were higher than Con[(39.83±2.31)mmHg,(40.49±3.12)%vs.(17.03±2.12)mmHg,(23.52±1.61)%,q=28.291,16.105,P<0.01].The weight in Juglone group was heavier than PAH group[(349.73±21.96)g vs.(303.11±20.41)g,q=5.204,P<0.01],but RVSP and RVHI in Juglone group were lower than PAH group[(26.89±1.92)mmHg,(31.40±3.76)%vs.(39.83±2.31)mmHg,(40.49±3.12)%,q=16.063,8.637,P<0.01].Immunofluorescence staining showed that the expression of Pin1 in PAH group was higher than Con[(8.39±0.19)%vs.(3.32±0.26)%,t=13.000,P<0.01].HE staining showed that the pulmonary arteriole wall was thickened in PAH group,while was reduced via Juglone intervention.Western blotting showed that the expression levels of alpha smooth muscle actin(α-SMA)and Pin1 in Con were lower than in PAH group(1.02±0.04,1.00±0.07 vs.2.21±0.07,3.37±0.

关 键 词：脯氨酰顺反异构酶1 胡桃醌 肺动脉高压 肺血管重构 

分 类 号：R544.1[医药卫生—心血管疾病]