ARC基因沉默对缺氧诱导持续性肺动脉高压小鼠的影响  

Effect of ARC gene silencing on the mniiccee wwiitthh hhyyppooxia-induced persistent pulmonary hypertension

作　　者：张婕 吴素玲 盛文彬 管明 ZHANG Jie;WU Suling;SHENG Wenbin(Department of Respiratory,Hangzhou Children's Hospital,Hangzhou 310041,CHINA)

机构地区：[1]杭州市儿童医院呼吸科,浙江310041 [2]浙江大学医学院附属杭州市第一人民医院耳鼻咽喉科

出　　处：《江苏医药》2023年第8期757-760,F0002,共5页Jiangsu Medical Journal

基　　金：浙江省自然科学基金(LQ20H040001、LY20H130003)。

摘　　要：目的探讨带有Caspases富集功能域的凋亡抑制因子(ARC)基因沉默对缺氧诱导持续性肺动脉高压(PPH)小鼠的影响。方法18只新生C57BL/6小鼠随机分为对照组、PPH组和ARC沉默组,每组6只。PPH组和ARC沉默组采用12%O_(2)诱导构建PPH模型后,ARC沉默组小鼠尾静脉注射短发夹RNA-ARC重组腺相关病毒100μL,PPH组小鼠尾静脉注射生理盐水100μL,每天1次,注射14 d。测定小鼠右心室收缩压(RVSP)和右心室肥厚指数(RVHI),HE染色观察肺血管形态,计算血管壁厚度占血管外径百分比(WT%),TUNEL染色观察肺动脉壁细胞凋亡情况,实时荧光定量PCR检测凋亡因子的表达情况。结果与对照组比较,PPH组和ARC沉默组RVSP、RVHI、WT%升高(P<0.05),肺动脉壁细胞凋亡数减少(P<0.05),Caspase-3和Bcl-2基因相关启动子mRNA表达降低(P<0.05),Bcl-2 mRNA表达增加(P<0.05);与PPH组比较,ARC沉默组RVSP、RVHI和WT%降低(P<0.05),肺动脉壁细胞凋亡数增多(P<0.05),Caspase-3和Fas相关死亡域蛋白mRNA表达增加(P<0.05),Bcl-2 mRNA表达下降(P<0.05)。结论ARC基因沉默能够抑制缺氧诱导PPH小鼠肺血管重塑,促进肺动脉壁细胞凋亡。Objective To investigate the effect of apoptosis repressor with Caspases recruitment domain(ARC)gene silencing on the mice with hypoxia-induced persistent pulmonary hypertension(PPH).Methods Eighteen newborn C57BL/6 mice were randomly divided into three groups with six mice each.Group C was taken as the blank control.PPH model was established by inhalation of 12%O_(2) in groups of A and B.Group A was injected with shRNA-ARC recombinant adeno-associated virus 100μL through caudal vein and group B was injected with normal saline once a day for 14 days.The right ventricular systolic pressures(RVSP)and right ventricular hypertrophy index(RVHI)were detected.The pulmonary vascular morphology was observed by HE staining and the percentage of vascular wall thickness in vascular outer diameter(WT%)was calculated.The apoptosis of pulmonary artery wall cells was observed by TUNEL staining.Real-time fluorescence quantitative PCR was used to detect the expressions of apoptotic factors.Results Compared with group C,RVSP,RVHI and WT%were higher,the number of apoptotic cells in pulmonary artery wall was less,the mRNA expressions of Caspase-3 and Bcl-2 associated death promoter were lower,and Bcl-2 mRNA expression was higher in groups of A and B(P<0.05).Compared with group B,RVSP,RVHI and WT%were lower,the number of apoptotic cells in pulmonary artery wall was higher,the mRNA expressions of Caspase-3 and Fas-associated death domain proteins were higher and Bcl-2 mRNA expression was lower in group A(P<0.05).Conclusion ARC gene silencing can inhibit pulmonary vascular remodeling and promote the apoptosis of pulmonary artery wall in the mice with hypoxia-induced PPH.

关 键 词：肺动脉高压 缺氧 带有半胱天冬氨酸蛋白酶富集功能域的凋亡抑制因子 凋亡 

分 类 号：R722[医药卫生—儿科]