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表达差异可能在缺氧性肺动脉高压发病中发挥作用。
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 compared with H0 group, all the three genes were significantly different （P 〈0. 05）. Immunohistochemistry showed that HIF-1α, HIF-2α and HIF-3α protein levels in H3 group were 0. 200 ± 0. 020, 0. 020 ±0. 010, 0. 050 ± 0. 010, respectively; in H14 group were 0. 160 ± 0. 010, 0. 100 ± 0. 020, 0.160 ± 0.010, respectively; in H7 group were 0.220 ± 0.020, 0.030 ± 0.010, 0.180 ± 0.020, respectively; in H0 group were 0. 050 ±0. 010, 0. 020 ±0. 010, 0. 040 ±0. 010, respectively. HIF-1α in H3 group, HIF-2α in H14 group, HIF-3α in H7 and H3 group were statistically different with that of H0 group （P 〈0. 05）. Protein bands of HIF-α subunits in lung tissue, measured by Western blot, showed that HIF-3αdecreased in H7 group as compared to H0 group, but the other two proteins showed a marked increase in H3 group as compared to H0 group, and increased further corresponding to the duration of hypoxia and peaked in H14 group as compared to H7 group. Conclusion The differential expression of the three HIF-α subunits may play a role in the development of hypoxia-induced pulmonary hypertension.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
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)
关键词
缺氧诱导因子α
缺氧
高血压
肺性
基因表达
Hypoxia-inducible factor α
Hypoxia
Hypertension, pulmonary
Ceneexpression
作者简介
通讯作者：戴爱国
