YC-1对低氧诱导的大鼠肺动脉外膜成纤维细胞增殖及胶原合成的影响  

Effect of YC-1 on hypoxia-induced vascular adventitial fibroblast proliferation and collagen synthesis in vitro

作　　者：刘海潮 任娟 李龙[1] 徐瑞娟 曾群丽 胡振红[1] LIU Haichao;REN Juan;LI Long;XU Ruijuan;ZENG Qunli;HU Zhenhong(Department of Respiratory Medicine,Wuhan General Hospital of Chinese PLA,Wuhan,Hubei 430070,P.R.China)

机构地区：[1]中国人民解放军武汉总医院呼吸内科,湖北武汉430070

出　　处：《中国呼吸与危重监护杂志》2018年第5期504-508,共5页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：湖北省自然科学基金(2017CFC876)

摘　　要：目的初步探讨低氧诱导因子抑制剂3-(5'-羟基甲基-2'-呋喃基)-1-苯基吲哚(YC-1)对低氧诱导的大鼠肺动脉外膜成纤维细胞增殖和胶原生成的影响,以及可能的分子机制。方法在低氧条件下培养的大鼠肺动脉成纤维细胞,将细胞随机分为常氧组、低氧组、低氧+YC-1(0.01、0.05和0.1 mmol/L)组。分别采用MTT法检测细胞增殖情况,3~H-脯氨酸掺入法测定胶原合成,蛋白免疫印迹法检测低氧诱导因子-1α(HIF-1α)蛋白的表达,逆转录-聚合酶链反应检测转化生长因子-β1(TGF-β1)mRNA表达情况。结果低氧组的成纤维细胞增殖及3~H-脯氨酸掺入量显著高于常氧组(均P<0.01)。YC-1组的成纤维细胞增殖及3~H-脯氨酸掺入量显著低于低氧组,但仍高于常氧组;低氧组HIF-1α蛋白、TGF-β1 mRNA的表达明显高于常氧组(P<0.01),YC-1呈剂量依赖性抑制缺氧刺激的HIF-1α蛋白、TGF-β1 mRNA表达,其中YC-1 0.1 mmol/L组中HIF-1α蛋白及TGF-β1 mRNA的表达分别抑制了65%和61%(均P<0.01)。结论 YC-1能抑制低氧诱导的大鼠肺动脉外膜成纤维细胞增殖及胶原合成,并呈剂量依赖关系,其作用可能与下调HIF-1α、TGF-β1 mRNA表达有关。Objective To investigate the effects of 3-（5＇-hydroxymethyl-2＇-fnryl）-1-benzylindazole （YC-1）, a hypoxia-inducible factor-lct （HIF-1 a） inhibitor, on hypoxia induced rat pulmonary arterial adventitial fibroblasts （AFs） proliferation and collagen synthesis, and explore the molecular mechanism. Methods Under hypoxic condition, rat AFs were cultured in DMEM medium supplemented with 10% fetal bovine serum in vitro. The cells were divided into five groups, ie. a normoxia group, a hypoxia group and three hypoxia＋YC-1 groups （treated with YC-1 at concentration of 0.01, 0.05 and 0.1 mmol/L, respectively）. The ceils proliferation was determined by MTT method. Collagen synthesis of AFs was measured by 3H-proline incorporation assay. The expression of HIF- 1α in AFs in different conditions was measured by Western blot, and the mRNA expression of transforming growth factor-β1 （TGF-β1） was measured by reverse-transcription polymerase chain reaction. Results The proliferation rate and the incorporation data of 3H-proline in the hypoxia group were significantly increased as compared with those in the control group （both P〈0.01）. YC-1 significantly reduced the proliferation rate and incorporation data of 3H-proline induced by hypoxia in a dose-dependent manner. YC-1 could also down-regulate the expressions of HIF-1α and TGF- β1 mRNA significantly （both P〈0.01）. Compared with the hypoxia group, the expressions of HIF-1α and TGF-β1 mRNA decreased respectively by 65% and 61% in the hypoxia＋YC-1 （0.1 mmol/L） group （both P〈0.01）. Conclusions YC-1 can inhibit hypoxia-induced AFs proliferation and collagen synthesis in a dose-dependent manner. The mechanism may relate to YC-1＇s inhibitory effect on expressions of HIF-1α and TGF-β1 mRNA.

关 键 词：3-(5'-羟基甲基-2'-呋喃基)-1-苯基吲哚 低氧诱导因子抑制剂 外膜成纤维细胞 增殖 胶原合成 低氧诱导因子-1Α 转化生长因子-β1 

分 类 号：R544.1[医药卫生—心血管疾病]