摘要
目的探讨缺氧诱导因子-1α(Hypoxia-inducible factor-1α,HIF-1α)调控内皮-间充质转化(Endothelial to mesenchymal transition,EndoMT)在缺氧性肺动脉高压(Hypoxia induced pulmonary hypertension,HPH)新生大鼠肺血管重塑(Pulmonary vascular remodeling,PVR)中的作用机制。方法96只Wistar新生大鼠随机分为常氧组、缺氧组、缺氧+YC-1(HIF-1α抑制剂)组和缺氧+HIF-1α组,每组再按不同观察时间点随机分为3、7、14 d亚组,每个亚组8只新生大鼠。常氧组给予空病毒尾静脉注射1 d后,每日给予生理盐水腹腔内注射;缺氧组给予空病毒尾静脉注射1 d后,每日生理盐水腹腔注射后开始8 h缺氧处理;缺氧+YC-1组给予空病毒尾静脉注射1 d后,每日YC-1(10 mg/kg)腹腔注射后开始8 h缺氧处理;缺氧+HIF-1α组尾静脉注射8×10^(9)PFU/mL的腺病毒(标有绿色荧光信号,携带HIF-1α的重组腺病毒载体)1 d后,每日生理盐水腹腔注射后开始8 h缺氧处理。在干预处理后3、7、14 d时,用直接测压法测量右心室收缩压(Right ventricular systolic pressure,RVSP);荧光显微镜观察腺病毒转染情况;苏木精-伊红染色观察肺血管形态,计算肺血管重塑指标肺小动脉中层血管壁厚度占血管外径的百分比(MT%)和肺小动脉中层截面积的百分比(MA%);间接免疫荧光双标法检测肺组织中内皮标志物CD31和α平滑肌肌动蛋白(αsmooth muscle actin,αSMA)的表达水平;实时荧光定量聚合酶链反应检测CD31、αSMA和HIF-1αmRNA表达水平。结果与常氧组相比,干预处理后3、7、14 d时,通过缺氧处理的各组RVSP均升高,MA%、MT%增加(P<0.05),CD31表达降低,而αSMA表达升高,HIF-1αmRNA表达升高(P<0.05);与缺氧组相比,干预后7 d、14 d时,缺氧+YC-1组RVSP降低,MA%、MT%减少,CD31表达升高,αSMA表达降低,HIF-1αmRNA表达降低(P<0.05);干预后7 d、14 d时,缺氧+HIF-1α组RVSP进一步升高,MA%、MT%增加,CD31表达降低,αSMA表达升高,HIF-1αmRNA表达升高(P<0.05)。结论缺氧及HIF-1α的抑制及过表达影响了EndoMT的过程,HIF-1α可能通过调控EndoMT促进HPH新生大鼠的PVR。
Objective To investigate the mechanism of hypoxia-inducible factor-1α(HIF-1α)regulation of endothelial to mesenchymal transition(EndoMT)in pulmonary vascular remodeling(PVR)in neonatal rats with hypoxic pulmonary hypertension(HPH).Methods 96 Wistar neonatal rats were randomly di-vided into normoxia group,hypoxia group,hypoxia+YC-1(HIF-1αinhibitor)group and hypoxia+HIF-1αgroup,and each group was randomly divided into 3 d,7 d and 14 d subgroups according to different obser-vation time points,with 8 neonatal rats in each subgroup.The normoxia group was given an empty virus via tail vein injection for 1 day,followed by daily intraperitoneal injection of normal saline.The hypoxia group was given an empty virus via tail vein injection for 1 day,followed by 8-hour hypoxia treatment after daily intraperitoneal injection of normal saline.The hypoxia+YC-1 group was given an empty virus via tail vein injection for 1 day,followed by daily intraperitoneal injection of YC-1(10 mg/kg)and then 8-hour hypoxia treatment.The hypoxia+HIF-1αgroup was given 8×10^(9) PFU/mL adenovirus(labeled with green fluorescence signal,carrying HIF-1αrecombinant adenovirus vector)via tail vein injection for 1 day,followed by daily intraperitoneal injection of normal saline and then 8-hour hypoxia treatment.At 3,7 and 14 days after the intervention treatment,the right ventricular systolic pressure(RVSP)measured by di-rect manometry.Fluorescence microscopy was used to observe adenovirus transfection.Hematoxylin-eo-sin staining was used to observe the morphology of pulmonary blood vessels,and the percentage of the thickness of the middle layer of the pulmonary arteriole to the outer diameter of the blood vessel(MT%)and the percentage of the median cross-sectional area of the pulmonary arteriole(MA%)were calculated.The expression levels of endothelial markers CD31 andαsmooth muscle actin(αSMA)in lung tissues were detected by indirect immunofluorescence double-standard method.Real-time quantitative PCR was used to detect the mRNA expression levels of CD31,αSMA and HIF-1α.Results Compared with normoxia group,RVSP was increased,MA%and MT%was increased in each group after hypoxia on the 3 d,7 d and 14 d of hypoxia(P<0.05),the expression of CD31 was decreased,while the expression ofαSMA and HIF-1αmRNA was increased(P<0.05).Compared with hypoxia group,at 7 d and 14 d of hypoxia,the RVSP,MA%and MT%in the hypoxia+YC-1 group was decreased,the expression of CD31 was in-creased,the expression ofαSMA was decreased,and the expression of HIF-1αmRNA was decreased(P<0.05).On the 7 d and 14 d of hypoxia,RVSP in the hypoxia+HIF-1αgroup further was increased,MA%and MT%was increased significantly,the expression of CD31 was decreased,the expression ofαSMA was increased,and the expression of HIF-1αmRNA was increased(P<0.05).Conclusion Hypoxia and the inhibition and overexpression of HIF-1αaffect the process of EndoMT.HIF-1αmay promote the PVR process in HPH neonatal rats by modulating EndoMT.
作者
李珊珊
王乐
谢鸥
罗洋
赵婷
LI Shanshan;WANG Le;XIE Ou;LUO Yang;ZHAO Ting(School of Pediatrics,Xinjiang Medical University,Urumqi 830017,China;Department of Neonatology,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)
出处
《新疆医科大学学报》
2025年第7期911-917,共7页
Journal of Xinjiang Medical University
基金
国家自然科学基金项目(82060287)。
关键词
缺氧性肺动脉高压
内皮-间充质转化
缺氧诱导因子-1Α
新生大鼠
hypoxia induced pulmonary hypertension
endothelial to mesenchymal transition(EndoMT)
hypoxia-inducible factor-1α(HIF-1α)
neonatal rat
