摘要
目的探讨缺氧诱导因子-1α(HIF-1α)调控经典型瞬时受体电位通道6(TRPC6)在新生大鼠缺氧性肺动脉高压(HPH)肺血管重塑中的作用机制。方法将32只Wistar新生大鼠随机分为常氧组、HPH组、过表达HIF-1α+HPH组(HIF-1α组)、HIF-1α抑制剂+HPH组(2ME组)。常氧组在常氧环境下饲养,其余3组建立HPH模型,其中HIF-1α组大鼠通过尾静脉注射携带HIF-1α基因的腺病毒载体,2ME组每日皮下注射2-甲氧基雌二醇(2ME),各组以干预处理后14 d作为实验终点。直接测压法测量右心室收缩压(RVSP);称重计算右心室肥厚指数(RVHI);苏木精-伊红染色观察肺远端小动脉形态,计算血管重塑指标(MT%、MA%)进行定量分析;免疫组化法检测肺组织增殖细胞核抗原(PCNA)表达以评估肺血管平滑肌细胞增殖水平,同时检测HIF-1α、TRPC6蛋白在肺组织中的表达水平;实时荧光定量-聚合酶链反应法检测HIF-1α、TRPC6、PCNA的转录水平。结果与常氧组比较,HPH组新生大鼠RVSP、RVHI、MT%、MA%升高(P<0.05),肺组织HIF-1α、TRPC6、PCNA的蛋白表达和mRNA水平升高(P<0.05);与HPH组比较,HIF-1α组新生大鼠RVSP、RVHI、MT%、MA%升高(P<0.05),肺组织HIF-1α、TRPC6、PCNA的蛋白表达和mRNA水平升高(P<0.05);与HPH组比较,2ME组RVSP、RVHI、MT%、MA%降低(P<0.05),肺组织HIF-1α、TRPC6、PCNA蛋白表达水平降低(P<0.05),TRPC6、PCNA mRNA水平降低(P<0.05);与HIF-1α组比较,2ME组RVSP、RVHI、MT%、MA%降低(P<0.05),肺组织HIF-1α、TRPC6、PCNA蛋白表达和mRNA水平降低(P<0.05)。结论慢性缺氧刺激下HIF-1α可能通过上调TRPC6,促进新生大鼠HPH肺血管重塑,该研究可能为新生儿HPH提供新的治疗靶点。
Objective To investigate the mechanism by which hypoxia-inducible factor-1α(HIF-1α)regulates the classical transient receptor potential channel 6(TRPC6)to mediate pulmonary vascular remode-ling in a neonatal rat model of hypoxic pulmonary hypertension(HPH).Methods A total of 32 neonatal Wistar rats were randomly divided into normoxia group,HPH group,overexpression of HIF-1α+HPH group(HIF-1αgroup)and HIF-1αinhibitor+HPH group(2ME group).The rats in the normoxia group were fed in normoxia environment,and the other 3 groups were established HPH model,among which the rats in the HIF-1αgroup were injected with adenovirus vector carrying HIF-1αgene through tail vein,and the rats in the 2ME group were injected 2ME every day.The experimental end point was 14 days after in-tervention.Right ventricular systolic pressure(RVSP)was measured using direct catheterization.Right ventricular hypertrophy index(RVHI)was calculated based on ventricular weight measurements.Mor-phology of distal pulmonary arterioles was examined by hematoxylin-eosin(HE)staining.Vascular re-modeling indices(medial wall thickness percentage(MT%)and medial wall area percentage(MA%)were calculated for quantitative assessment.Immunohistochemistry was used to detect the expression of PCNA in lung tissues to evaluate the proliferation level of pulmonary vascular smooth muscle cells,and the ex-pression levels of HIF-1αand TRPC6 proteins in lung tissue were also detected.Transcriptional levels of HIF-1α,TRPC6 and PCNA were quantified using quantitative real-time polymerase chain reaction(qRT-PCR).Results Compared with normoxia group,the RVSP,RVHI,MT%and MA%of neonatal rats in the HPH group increased(P<0.05),and the protein expression and mRNA levels of HIF-1α,TRPC6 and PCNA in lung tissue increased(P<0.05).Compared with the HPH group,the RVSP,RVHI,MT%and MA%of neonatal rats in the HIF-1αgroup increased(P<0.05),and the protein expression and mRNA levels of HIF-1α,TRPC6 and PCNA in lung tissue increased(P<0.05).Compared with the HPH group,the RVSP,RVHI,MT%and MA%of neonatal rats in the 2ME group decreased(P<0.05),and the protein expression levels of HIF-1α,TRPC6 and PCNA in lung tissue decreased(P<0.05),and the mRNA levels of TRPC6 and PCNA decreased(P<0.05).Compared with the HIF-1αgroup,the RVSP,RVHI,MT%and MA%of neonatal rats in the 2ME group decreased(P<0.05),and the protein expression and mRNA levels of HIF-1α,TRPC6 and PCNA in lung tissue decreased(P<0.05).Conclu-sion HIF-1αmay promote pulmonary vascular remodeling in neonatal rats with HPH by up-regulating TRPC6 under chronic hypoxia stimulation,which may provide a new therapeutic target for neonatal HPH.
作者
谢鸥
李珊珊
罗洋
王乐
XIE Ou;LI Shanshan;LUO Yang;WANG Le(Department 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918-923,930,共7页
Journal of Xinjiang Medical University
基金
国家自然科学基金项目(82060287)。
关键词
缺氧诱导因子-1Α
经典型瞬时受体电位通道6
缺氧性肺动脉高压
新生大鼠
hypoxia inducible factor-1α(HIF-1α)
classical transient receptor potential channel 6(TRPC6)
hypoxic pulmonary hypertension(HPH)
neonatal rats
