摘要
目的观察先天性心脏病肺动脉高压患儿肺组织中芳香烃受体核转位蛋白(ARNT)水平及其对肺动脉平滑肌细胞增殖和迁移的影响,探讨ARNT在先天性心脏病肺动脉高压发病中的作用机制。方法选择我院2018年1月至2018年12月先天性心脏病室间隔缺损相关性肺动脉高压患者60例作为先天性心脏病-肺动脉高压组(CHD-PAH)和先天性心脏病室间隔缺损无肺动脉高压患儿60例作为先天性心脏病组(CHD组)。将人肺动脉平滑肌细胞(HPASMC)细胞分为常氧组和缺氧组,分别给予常氧和缺氧诱导。将HPASMC细胞分为空白对照组(BC组)、阴性对照组(NC组)和siR-ARNT组,siR-ARNT组细胞转染ARNT-shRNA慢病毒,NC组细胞转染阴性对照慢病毒,BC组细胞不转染。采用蛋白质印迹法(Western blot)和反转录-聚合酶连反应(RT-PCR)测定肺组织细胞中ARNT蛋白和mRNA水平。细胞计数试剂盒(CCK-8)测定细胞增殖能力。Transwell和划痕实验测定细胞迁移能力,应用SPSS 20.0统计软件分析。结果CHD-PAH组患儿肺组织ARNT蛋白和mRNA水平(0.53±0.06、2.36±0.17)均高于CHD组(0.09±0.02、1.00±0.12,t=53.889、50.626,P<0.05),差异有统计学意义。缺氧组HPASMC细胞中ARNT蛋白和mRNA水平(0.47±0.09、2.15±0.13)均高于常氧组(0.14±0.06、1.00±0.09,t=8.072、19.243,P<0.05),差异有统计学意义。与BC组[0.65±0.13、1.00±0.08、0.43±0.09、0.81±0.12、1.13±0.14、(95.46±11.37)个、(85.17±6.42)μm]和NC组[0.62±0.14、0.98±0.09、0.41±0.07、0.79±0.13、1.14±0.15、(97.43±12.62)个、(83.91±6.58)μm]比较,siR-ARNT组HPASMC细胞中ARNT蛋白和mRNA水平(0.17±0.06、0.37±0.05)、吸光度(A)值(0.33±0.05、0.56±0.10、0.72±0.13)、迁移细胞数[(72.15±10.94)个]和迁移距离[(32.27±6.12)μm]均降低(F=37.863、158.406、3.794、9.814、20.442、10.171、156.873,P<0.05,差异有统计学意义);BC组和NC组HPASMC细胞中各指标差异无统计学意义(P>0.05)。结论先天性心脏病肺动脉高
Objective To observe the levels of aryl hydrocarbon receptor nuclear translocator(ARNT)in lung tissue of children with congenital heart disease(CHD)pulmonary artery hypertension(PAH)and its effect on proliferation and migration of human pulmonary artery smooth muscle cells(HPASMCs),and to investigate the action mechanism of ARNT in the pathogenesis of CHD-PAH.Methods Sixty patients with CHD ventricular septal defect-associated PAH were selected as CHD-PAH group and 60 children with CHD ventricular septal defect without PAH were selected as CHD group(CHD group)in our hospital.There were no significant difference in age and gender between the two groups.HPASMCs were divided into normoxia group and hypoxia group,which were induced by normoxia and hypoxia respectively.HPASMCs were divided into blank control group(BC group),negative control group(NC group)and siR-ARNT group.The cells in the siR-ARNT group were transfected with ARNT-shRNA lentivirus,and those in the NC group were transfected with negative control lentivirus.The cells in the BC group were not transfected.The expression levels of ARNT protein and mRNA in lung tissue and cells were detected by Western blotting and reverse transcription-polymerase chain reaction(RT-PCR).The cell proliferation ability was determined by the cell counting kit-8(CCK-8)method.Transwell and scratch assays were used to measure cell migration ability.SPSS 20.0 software was used for analysis.The test methods were t test and analysis of variance.Results The expression levels of ARNT protein and mRNA in lung tissue of CHD-PAH group(0.53±0.06 and 2.36±0.17)were significantly higher than those in CHD group(0.09±0.02,1.00±0.12,t=53.889 and 50.626,P<0.05).The ARNT protein and mRNA expression levels in HPASMCs in hypoxia group(0.47±0.09 and 2.15±0.13)were significantly higher than those in normoxia group(0.14±0.06 and 1.00±0.09,t=8.072 and 19.243,P<0.05).As compared with BC group[0.65±0.13,1.00±0.08,0.43±0.09,0.81±0.12,1.13±0.14,(95.46±11.37)cells,(85.17±6.42)μm]and NC gro
作者
张岩伟
彭帮田
刘琳
艾峰
郑家永
胡小松
Zhang Yanwei;Peng Bangtian;Liu Lin;Ai Feng;Zheng Jiayong;Hu Xiaosong(Children’s Heart Center,Fuwai Central Cardiovascular Hospital,Zhengzhou 451464,China;Children’s Heart Center,Heart Center of Henan Provincial People’s Hospital,Zhengzhou 450003,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2020年第1期158-161,共4页
Chinese Journal of Experimental Surgery
基金
河南省医学科技攻关计划普通项目(201702184)
河南省医学科技攻关计划省部共建项目(SB201901099)。
关键词
先天性心脏病
肺动脉高压
芳香烃受体核转位蛋白
肺动脉平滑肌细胞
增殖
迁移
Congenital heart disease
Pulmonary hypertension
Aryl hydrocarbon receptor nuclear translocation protein
Pulmonary artery smooth muscle cells
Proliferation
Migration
