大鼠缺氧性肺动脉高压时三种缺氧诱导因子α亚基在肺动脉中的差异表达  

Differential expression of the three hypoxia-inducible factor α subunits in pulmonary artery of rats with hypoxia-induced pulmonary hypertension

作　　者：李启芳 戴爱国 

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2期113-117,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金资助项目(30270581;30570815);湖南省教育厅重点科研基金资助项目(02A047);中国博士后科学基金资助项目(2003033436)

摘　　要：目的区分大鼠缺氧性肺动脉高压时肺血管壁3种缺氧诱导因子α(HIFα)亚基(HIF1α、HIF2α、HIF3α)的基因表达特征。方法40只雄性Wistar大鼠按随机数字表法分为常氧0d组(H0组)、缺氧3d组(H3组)、7d组(H7组)、14d组(H14组)和21d组(H21组),每组8只。用(10.0±0.5)%的氧浓度每天间断缺氧8h,测各组大鼠平均肺动脉压(mPAP)、肺动脉管壁面积(WA%)、肺动脉中膜厚度(PAMT)、右室肥厚指数(RVHI%);用原位杂交、免疫印迹和免疫组化法检测HIF1α、HIF2α和HIF3α基因表达。结果H7组mPAP为(18.40±0.40)mmHg(1mmHg=0.133kPa),H0组为(14.40±0.40)mmHg,H14组为(21.20±0.20)mmHg,H7组与H0组、H14组比较差异有统计学意义(P均<0.05);血管形态学显示,H7组WA%、PAMT、RVHI%分别为(47.8±0.8)%、(12.3±0.5)μm、(24.0±1.0)%,H14组分别为(60.3±0.4)%、(15.0±0.3)μm、(25.0±1.8)%,H0组分别为(35.5±1.3)%、(11.9±0.6)μm、(23.6±0.5)%,H21组分别为(65.0±0.7)%、(23.0±0.8)μm、(27.7±1.0)%,WA%H7组与H0组、H14组与H0组、H21组与H14组间比较差异均有统计学意义(P均<0.05)。原位杂交显示,H14组HIF1α、HIF2α、HIF3αmRNA水平的吸光度(A)值分别为0.200±0.020、0.080±0.010、0.170±0.010;H7组分别为0.050±0.020、0.160±0.020、0.160±0.020;H0组分别为0.050±0.010、0.140±0.020、0.060±0.010;H14组与H0组三者、H7组与H0组仅HIF3α比较差异有统计学意义(P均<0.05)。免疫组化表明,H3组HIF1α,HIF2α和HIF3α蛋白表达分别为0.200±0.020、0.020±0.010、0.050±0.010;H14组分别为0.160±0.010、0.100±0.020、0.160±0.010;H7组分别为0.220±0.020、0.030±0.010、0.180±0.020;H0组分别为0.050±0.010、0.020±0.010、0.040±0.010,H3组HIF1α蛋白、H14组HIF2α蛋白、H7组、H3组HIF3α蛋白分别与H0组比较差异有统计学意义(P均<0.05)。H7组免疫印迹显示在肺组织中HIF3α表达较H0组显著减弱,H3组HIF2α、HIF3α蛋白表达较H0组增强,随着缺氧时间延长,H14组较H7组更明显。结论3种HIFα表达�Objective To differentiate the expression patterns of all hypoxia-inducible factor α （HIF-α） subunits （HIF-1α, HIF-2α and HIF-3α） in pulmonary artery of rats undergoing systemic hypoxic. Methods Forty male healthy wistar rats were assigned randomly to 5 groups, 8 rats per group, then exposed to hypoxia [O2,（10.0±0.5）%] for 0 d （H0）, 3 d （H3）, 7 d （H7）, 14 d （H14） and 21 d （ H21 ） respectively, 8 h per day intermittently. Mean pulmonary arterial pressure（ mPAP）, arterial wall area （WA, μm）, pulmonary artery medium thickness（ PAMT% ） and right ventricle hypertrophy index （RVHI） were measured. HIF-1α, HIF-2α and HIF-3α gene expression were determined by immunohistochemistry, in sire hybridization and Western blot. Results mPAPs in H7, H0 and H14 groups were [ （ 18.40 ±0. 40） mmHg, 1 mm Hg =0.133 kPa], [（14.40 ± 0.40） mm Hg] and [（21.20 ±0.20） mm Hg], respectively, statistically different when H7 group was compared with H0 and H14 groups （all P 〈 0. 05）. Arterial morphology showed that WA%, PAMT and RVHI% in H7 group were （47. 8 ±0. 8）%, （ 12. 3 ± 0. 5） p,m, （24.0 ± 1.0）%, respectively; in H0 group were （35. 5 ± 1.3）%, （ 11.9 ±0. 6）%, （23.6 ± 0.5） μm, respectively; in H21 group were （65.0 ±0.7）%, （23.0 ±0.8） μm, （27.7 ± 1.0）%, respectively. When H7 group was compared with H0 group, only WA% was statistically different; when H14 group was compared with H0 group, all the three parameters were statistically different （P 〈0.05 ）. In situ hybridization demonstrated that the mRNA levels （absorbance, A） of HIF-1α, HIF-2α, and HIF-3α in H14 group were 0. 200 ±0. 020, 0. 080 ±0. 010, 0. 170 ±0. 010, respectively; in H7 group were 0. 050 ±0. 020, 0. 160 ± 0. 020, 0. 160 ± 0. 020, respectively; in H0 group were 0. 050 ± 0. 010, 0. 140 ± 0. 020, 0. 060 ± 0. 010, respectively. When H7 group was compared with H0 group, only HIF-3α was statistically different; when H14 group was compar

关 键 词：缺氧诱导因子α 缺氧 高血压 肺性 基因表达 

分 类 号：R543.2[医药卫生—心血管疾病] R-332[医药卫生—内科学]