缺氧诱导因子1α与其脯氨酰羟化酶相互调控对大鼠缺氧性肺动脉高压的作用  

Reciprocal regulation between hypoxia-inducible factor-1α and its prolyl hydroxylases in hypoxic pulmonary hypertension rats

作　　者：陈云荣 戴爱国 胡瑞成 

机构地区：[1]湖南省老年医院-湖南省老年医学研究所呼吸疾病研究室,长沙410001

出　　处：《中华结核和呼吸杂志》2006年第10期668-673,共6页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金(30270581;30570815);教育部科学技术研究重点项目(03091);中国博士后科学基金(2003033436);湖南省自然科学基金(05JJ30073)

摘　　要：目的探讨慢性缺氧性肺动脉高压(HPH)形成过程中缺氧诱导因子1α亚基(HIF-1α)及脯氨酰羟化酶(PHD1、PHD2、PHD3)在肺内的动态表达及相互调控作用。方法 40只成年雄性 SD 大鼠随机分为常氧对照组(C 组)和缺氧3 d、7 d、14 d、21 d 组(H_3、H_7、H_(14)、H_(21)组),每组8只,常压缺氧复制慢性 HPH 大鼠模型。测各组大鼠平均肺动脉压(mPAP)、右心室肥大指数(RVHI)、血管形态学指标;原位杂交、逆转录-聚合酶链反应(RT-PCR)检测肺内 HIF-1α及 PHD1、PHD2、PHD3的mRNA 表达水平,免疫组化、Western blot 检测其蛋白表达水平。结果(1)H_7组大鼠 mPAP 为(21.7±2.4)mm Hg(1mm Hg=0.133 kPa),管壁面积与血管面积比值为(43.9±5.3)%,肺小血管中膜厚度分别为(10.0±0.7)μm,与 C 组[mPAP 为(16.6±1.6)mm Hg,管壁面积与血管面积比值为(36.3±4.8)%,肺小血管中膜厚度为(8.5±1.3)μm]比较差异均有统计学意义(q 分别5.591、4.082、2.929,P 均<0.05),H_(14)组稳定于高水平;H_(14)组 RVHI 为(27.6±1.4)%,与 C 组[(23.6±2.9)%]比较差异有统计学意义(q=5.817,P<0.05);(2)HIF-1α蛋白在 C 组肺小动脉表达不明显(0.080±0.009),H_3组表达均开始升高(0.196±0.018,与 C 组比较 q=18.864,P<0.05),H_7组达高峰(0.203±0.022),H_(14)和 H_(21)组表达稍下降(0.174±0.020、0.156±0.016)。HIF-1α mRNA 在 C 组肺小动脉(0.139±0.017)和肺组织表达阳性,H_(14)组表达略升高(0.176±0.019,与 C 组比较 q=5.401,P<0.05);(3)C 组 PHD1、PHD2 mRNA(0.260±0.031、0.196±0.023)和蛋白(0.244±0.030、0.205±0.025)呈阳性表达,PHD3 mRNA、蛋白表达相对不明显(0.110±0.013、0.153±0.019)。PHD1 mRNA 缺氧后无显著变化,H_3组 PHD2、PHD3 mRNA 表达升高(0.246±0.023、0.262±0.025,与 C 组比较 q 值分别为5.268、15.831,P 均<0.05),PHD3 mRNA 升高更明显。PHD1蛋白缺氧14 d下降(0.210±0.023,与 C 组比较 q=3.885,P<0.05),缺氧21 d 保持较低水平,H_(14)组 PHD3蛋白显著升高(0.259±0.024,与 C 组比较 q=1Objective To investigate the interaction between hypoxia-inducible factors-let subunit （HIF-1α） and its three prolyl hydroxylases（ PHD1, PHD2 and PHD3） during the development of rat hypoxic pulmonary hypertension. Methods Forty male SD rats were randomly divided into 5 groups and exposed to normoxia（ C group） or exposed to hypoxia for 3,7,14 or 21 d （ H3 , H7, H14 , H21 group） , respectively. Mean pulmonary arterial pressure（mPAP） , vessel morphometry and right ventricle hypertrophy index（RVHI） were measured. Reverse transcriptase-polymerase chain reaction（RT-PCR） and in situ hybridization were used to determine the expression of mRNA. Immunohistochemistry and Western blot were used to determine the expression of mRNA. Results The level of mPAP[ （21.7 ± 2. 4） mm Hg, 1 mm Hg = 0. 133 kPa] ,the ratio of vascular wall thickness to external diameter [ WA%, （43.9 ± 5.3 ）% ] and pulmonary artery media thickness[ PAMT, （ 10.0 ±0. 7） μm] were significantly higher in H7 group than those in C group[ （ 16. 6 ± 1.6）mm Hg, （36.3±4.8）% and （8.5 ± 1.3）μm respectively, q value were 5.591,4.082,2.929, respectively,all P 〈0. 05 ]. These parameters reached a high level and remained stable on H14 group, and RVHI was significantly higher in H14 group[ （27.6 ± 1.4） % ] than in C group[ （23.6 ±2.9） % ,q = 5.817, P 〈 0. 05 ]. HIF-1 α protein was barely positive in C group （ 0. 080 ± 0. 009 ）, but markedly up-regulated in H3 group（0.196 ± 0. 018, compared with C group q = 18. 864, P 〈 0.05 ） , reaching its peak in H7 group （0. 203 ±0. 022）,and then declined slightly in Hi4 and H21 group. HIF-1α mRNA increased marginally in H14 group（O. 176 sO. 019 ,compared with C group q =5. 401 ,P 〈0.05,0. 139 ±0. 017）. PHD1 and PHD2 mRNA （0. 260 ± 0. 031,0. 196 ± 0. 023 ） and protein （ 0. 244 ± 0. 030, 0. 205 ± 0. 025 ） were positive in C group. PHD2 mRNA and protein were up-regulated in H3 group（0. 246 ± 0. 023,0. 235±0. 025, compared

关 键 词：缺氧诱导因子 α亚基 脯氨酰羟化酶 高血压 肺性 羟基化 

分 类 号：R543[医药卫生—心血管疾病]