17β-雌二醇/微小核糖核酸-21信号通路抑制低氧性肺动脉高压肺血管重构的机制研究  

17β-estradiol Attenuates Hypoxia-induced Pulmonary Vascular Remodeling via Downregulating miRNA-21 Signaling Pathway

作　　者：沈丽晓 袁博云 王丽[1] 袁雅冬[1] SHEN Li-xiao;YUAN Bo-yun;WANG Li;YUAN Ya-dong(Department of Respiratory and Critical Care Medicine,The Second Hospital,Hebei Medical University,Shijiazhuang（050000）,Hebei,China)

机构地区：[1]河北医科大学第二医院呼吸与危重症医学科,河北省石家庄市050000

出　　处：《中国循环杂志》2018年第11期1118-1123,共6页Chinese Circulation Journal

基　　金：河北省卫生厅重点科技研究计划(20160127)

摘　　要：目的:探讨17β-雌二醇(E2)对低氧性肺动脉高压(HPH)的保护作用是否通过下调微小核糖核酸(mi RNA)-21(mi R-21)表达进而抑制肺动脉平滑肌细胞(PASMC)增殖而实现。方法:(1)动物水平:32只健康雌性SD大鼠行卵巢切除术后随机分入常氧组、常氧+E2组、低氧组、低氧+E2组,每组8只。两个E2干预组大鼠每日皮下注射E2 20μg/kg,余组大鼠皮下注射等量生理盐水。两个低氧组大鼠在低氧环境下饲养,两个常氧组大鼠呼吸正常空气,连续饲养8周建立HPH模型。观察各组大鼠肺血管形态、平均肺动脉压(mPAP)及右心室肥厚指数(RVHI)变化,用实时聚合酶链式反应(PCR)及免疫印迹法测定肺动脉中mi R-21、增殖细胞核抗体(PCNA)表达变化。(2)细胞水平:将体外培养的人PASMC随机分为3组:常氧组、低氧组、低氧+E2组,24 h后用四甲基偶氮唑蓝比色法检测各组细胞增殖情况,用实时PCR及免疫印迹法检测细胞中mi R-21和PCNA表达变化。结果:(1)动物水平:低氧组较常氧组肺小动脉厚度和RVHI均明显增加,m PAP升高,肺动脉中mi R-21及PCNA表达明显上升(P均<0.01);常氧+E2组上述指标较常氧组无明显变化(P均>0.05);低氧+E2组上述三项指标明显低于低氧组(P均<0.01)。(2)细胞水平:与常氧组相比,低氧组细胞增殖明显,mi R-21和PCNA表达明显上升(P均<0.01);与低氧组相比,低氧+E2组细胞增殖程度明显减轻,mi R-21和PCNA表达明显下降(P均<0.01)。结论:E2能够改善HPH大鼠肺血管重构、肺动脉压力、右心室肥厚程度,其保护作用可能是通过下调mi R-21和PCNA表达从而抑制PASMC增殖而实现的。Objectives： To explore the effect of 17β-estradiol （E2） on hypoxic pulmonary hypertension （HPH） and explore if the effects were mediated through suppressing pulmonary artery smooth muscle cells （PASMCs） proliferation by targeting miRNA-21 （miR-21）. Methods： Animal experiment： A total of 32 healthy female SD rats with castrated surgery were randomly divided into 4 groups： normoxia group, normoxia＋E2 group, hypoxia group, hypoxia＋E2 group （n=8 each）. The rats in normoxia＋E2 group and hypoxia＋E2 group received subcutaneous injection of E2 20 ~tg/kg/d, and the rest groups received subcutaneous injection of equal volume saline. The hypoxic groups were placed in the hypoxic chamber （24 hours per day for 8 weeks） to establish HPH model and normoxic groups were kept in the room air. The pulmonary artery remodeling, mean pulmonary artery pressure （mPAP）, right ventricle hypertrophy index （RVHI） were observed. Real-time PCR and Western blot were used to detect the levels of proliferation cell nuclear antibody （PCNA） and miR-21 expression in pulmonary artery. In vitro： human pulmonary artery smooth muscle cells （hPASMCs） were randomly divided into 3 groups： normoxia group, hypoxia group,hypoxia＋E2 group. The levels of cell proliferation in each group were tested by MTT after 24 hours. Real-time PCR and Western blot were used to detect the levels of PCNA and miR-21 in cells. Results： Animal experiment： compared with normoxia group, the hypoxia group showed obviously thickened pulmonary artery wall, increased mPAP and RVHI, and significantly increased expression of miR-21 and PCNA （P〈0.01）; above changed were significantly attenuated in hypoxia＋E2 group （P〈0.01）. In vitro： compared with normoxia group, the hypoxia group showed obvious proliferation and significantly increased expression of miR-21 and PCNA （P〈0.01）; compared with hypoxia group, the proliferation of hPASMCs and expression of miR-21 and PCNA were obviously reduced in hypox

关 键 词：低氧性肺动脉高压 肺动脉平滑肌细胞 17Β-雌二醇 微小核糖核酸 

分 类 号：R54[医药卫生—心血管疾病]