母体缺氧上调胎兔颈动脉生长因子及细胞外基质表达  

Maternal hypoxia upregulates the expression of growth factors and extracellular matrix in carotid arteries from fetal rabbit

作　　者：陈幼芳 林惠通[2] 吕国荣 杜心清[2] 

机构地区：[1]福建省泉州医学高等专科学校临床医学院内科学教研室,福建泉州362011 [2]福建医科大学附属第二临床学院

出　　处：《中华高血压杂志》2016年第11期1070-1074,共5页Chinese Journal of Hypertension

基　　金：泉州市科委重点项目(2012Z72);福建省卫计委中青年骨干人才培养项目(2013-ZQN-JC-28)

摘　　要：目的研究母体缺氧对新西兰胎兔颈动脉生长因子、细胞外基质表达及血管形态学的影响。方法同期怀孕的新西兰母兔[足月(30±1)d]随机分为对照组(21%O_2,n=5)、缺氧组(12%O_2,n=5),缺氧组于妊娠第10天开始采用缺氧箱进行缺氧暴露,每天2次,上、下午各4h,至妊娠第29天结束。妊娠第29天,母兔麻醉,剖宫取出胎兔,分离胎兔颈动脉,液氮中速冻后转移至-70℃保存备检,另保存胎兔颈动脉用于免疫组织化学检查和Masson染色。用免疫印迹检测缺氧诱导因子1α(HIF-1α)、转化生长因子β_1(TGF-β_1)、Smad3和结缔组织生长因子(CTGF)蛋白表达;实时聚合酶链反应(Real-time PCR)检测HIF-1α、TGF-β_1和CTGF mRNA表达水平;免疫组织化学技术检测颈动脉TGF-β_1、CTGF、Ⅰ型胶原蛋白(CollⅠ)、Ⅲ型胶原蛋白(CollⅢ)表达;Masson染色检测颈动脉胶原纤维含量及内中膜厚度。结果与对照组比较,母体缺氧组胎兔颈动脉HIF-1α(0.26±0.05比0.07±0.04)、TGF-β_1(0.68±0.08比0.51±0.11)、Smad3(0.47±0.04比0.26±0.02)和CTGF(0.81±0.09比0.53±0.12)蛋白表达增多,HIF-1α(1.62±0.37比1.00±0.18)、TGF-β_1(1.90±0.35比1.00±0.12)和CTGF(1.82±0.42比1.00±0.21)mRNA水平升高,及CollⅠ(0.35±0.02比0.27±0.02)、CollⅢ(0.26±0.02比0.22±0.03)表达增多(P<0.05或P<0.01);而Masson染色结果提示:胶原纤维含量、内膜和中膜厚度在两组之间的差异无统计学意义(均P>0.05)。结论母体缺氧上调胎兔颈动脉TGF-β_1、CTGF、CollⅠ、CollⅢ的表达。Objective To investigate the effects of maternal hypoxia on the expression of growth factors, extracellu- lar matrix and morphological changes of carotid arteries from fetal rabbit. Methods Ten New Zealand White rab- bits were randomly divided into 2 groups ： perinatal hypoxia （ 12 %O2 , n = 5） and normoxic control （ 21% O2 , n = 5 ）. Rabbits in hypoxia group were put into the hypoxia box for low oxygen exposure, twice a day（morning and affer- noon） and four hours per time, from pregnancy day 10 to 29. At the end of pregnancy 129-day gestation）, Western blot was used to determine hypoxia-inducible factor-1（HIF-1α）, transforming growth factor β1 （TGF-β1 ）, Smad3 and connective tissue growth factor（CTGF） expression in carotid arteries of fetuses, and real-time polymerase chain reac- tion（RT-PCR） was performed to evaluate HIF-1α, TGF-β1 and CTGF mRNA concentrations. Masson＇s trichrome staining was employed to examine collagen or elastin content, and immunohistochemistry was performed to detect TGF-β1, CTGF, Coil Ⅰ and CollⅢ protein expression in the isolated rings from arteries of fetuses respectively. Results Notably, hypoxia treatment induced a significant increase in protein level in HIF-1α（0. 26±0. 05 vs 0. 07± 0.04）, TGF-β1 （0. 68±0.08 vs 0. 51±0.11） , Smad3（0. 47±0. 04 vs 0.26±0.02） and CTGF（0. 81±0. 09 vs 0. 53± 0.12）, and mRNA level in HIF-1α[1. 62±0. 37 vs 1.00±0.18）, TGF-β1 （1. 904±0. 35 vs 1. 00±0.12） and CTGF （1. 82±0.42 vs 1. 00±0.21）, which were associated with the elevation of TGF-β1, CTGF, collagenⅠ （0. 35±0.02 vs 0.27 ± 0.02） and Ⅲ （0.26 ± 0.02 vs 0.22 ± 0.03） in fetal carotid compared to normoxic control. Maternal hypoxia resulted in an increase in total collagen content of the carotid in the hypoxic animals compared to normoxic animals, but there was no statistical difference between the two groups. Conclusion These results reveal that maternal hypoxia upregulates the expression of TGF-β1 , CT

关 键 词：母体缺氧 转化生长因子Β1 结缔组织生长因子 Ⅰ型胶原蛋白 Ⅲ型胶原蛋白 动脉 

分 类 号：R54[医药卫生—心血管疾病]