国际儿科学杂志
國際兒科學雜誌
국제인과학잡지
INTERNATIONAL JOURNAL OF PEDIATRICS
2014年
3期
305-309
,共5页
李君%富建华%薛辛东%杜延娜%耿海芹
李君%富建華%薛辛東%杜延娜%耿海芹
리군%부건화%설신동%두연나%경해근
低氧诱导因子-1α%血管重塑%肺动脉高压%新生鼠
低氧誘導因子-1α%血管重塑%肺動脈高壓%新生鼠
저양유도인자-1α%혈관중소%폐동맥고압%신생서
Hypoxia-inducible factor-1α%Vascular remodeling%Pulmonary hypertension%Neonatal rats
目的 研究低氧诱导因子-1α(HIF-1α)基因和蛋白在新生鼠肺动脉高压(hypoxiic pulmonary hypertension,HPH)发生、发展中动态表达规律,阐明HIF-1α在肺高压肺小血管重塑中的意义.方法 将64只新生SD大鼠分为模型组(32只)和对照组(32只),建立慢性低氧诱导新生SD大鼠HPH动物模型,对照组置于空气中,分别于实验后第3、7、10、14天利用右心室插管测定右心室收缩压力(mRVSP),采集心脏组织测定右心肥厚指数(RVHI)、肺小动脉中层壁厚占肺小动脉的外径百分比(Mr％)和肺小动脉管壁中层横截面积占肺小动脉总横截面积的百分比(MA％),并分别应用免疫组织化学方法观察HIF-1α,RT-PCR和Western Blot分别测定HIF-1α在基因和蛋白的表达水平.结果 与对照组比较,模型组在第3天出现mRVSP升高,并持续至第14天(P＜0.01);RVHI、MT％、MA％于第7天开始增加,并持续至第14天(P＜0.01);对照组HIF-1α仅有微弱表达,而模型组主要在肺动脉内皮细胞和平滑肌细胞内表达;模型组HIF-1α基因表达水平于第3、7天高于对照组(P＜0.05);蛋白表达水平在第7、10、14天显著高于对照组(P＜0.01).结论 新生鼠暴露于低氧早期,即出现右心室收缩压力升高,继之发生右心室肥厚及肺小动脉结构改变,而HIF-1α不论基因还是蛋白,在PPHN的肺小血管重塑阶段均持续高水平表达,提示HIE-1α可能是促成PPHN肺血管重塑的因子之一.
目的 研究低氧誘導因子-1α(HIF-1α)基因和蛋白在新生鼠肺動脈高壓(hypoxiic pulmonary hypertension,HPH)髮生、髮展中動態錶達規律,闡明HIF-1α在肺高壓肺小血管重塑中的意義.方法 將64隻新生SD大鼠分為模型組(32隻)和對照組(32隻),建立慢性低氧誘導新生SD大鼠HPH動物模型,對照組置于空氣中,分彆于實驗後第3、7、10、14天利用右心室插管測定右心室收縮壓力(mRVSP),採集心髒組織測定右心肥厚指數(RVHI)、肺小動脈中層壁厚佔肺小動脈的外徑百分比(Mr％)和肺小動脈管壁中層橫截麵積佔肺小動脈總橫截麵積的百分比(MA％),併分彆應用免疫組織化學方法觀察HIF-1α,RT-PCR和Western Blot分彆測定HIF-1α在基因和蛋白的錶達水平.結果 與對照組比較,模型組在第3天齣現mRVSP升高,併持續至第14天(P＜0.01);RVHI、MT％、MA％于第7天開始增加,併持續至第14天(P＜0.01);對照組HIF-1α僅有微弱錶達,而模型組主要在肺動脈內皮細胞和平滑肌細胞內錶達;模型組HIF-1α基因錶達水平于第3、7天高于對照組(P＜0.05);蛋白錶達水平在第7、10、14天顯著高于對照組(P＜0.01).結論 新生鼠暴露于低氧早期,即齣現右心室收縮壓力升高,繼之髮生右心室肥厚及肺小動脈結構改變,而HIF-1α不論基因還是蛋白,在PPHN的肺小血管重塑階段均持續高水平錶達,提示HIE-1α可能是促成PPHN肺血管重塑的因子之一.
목적 연구저양유도인자-1α(HIF-1α)기인화단백재신생서폐동맥고압(hypoxiic pulmonary hypertension,HPH)발생、발전중동태표체규률,천명HIF-1α재폐고압폐소혈관중소중적의의.방법 장64지신생SD대서분위모형조(32지)화대조조(32지),건립만성저양유도신생SD대서HPH동물모형,대조조치우공기중,분별우실험후제3、7、10、14천이용우심실삽관측정우심실수축압력(mRVSP),채집심장조직측정우심비후지수(RVHI)、폐소동맥중층벽후점폐소동맥적외경백분비(Mr％)화폐소동맥관벽중층횡절면적점폐소동맥총횡절면적적백분비(MA％),병분별응용면역조직화학방법관찰HIF-1α,RT-PCR화Western Blot분별측정HIF-1α재기인화단백적표체수평.결과 여대조조비교,모형조재제3천출현mRVSP승고,병지속지제14천(P＜0.01);RVHI、MT％、MA％우제7천개시증가,병지속지제14천(P＜0.01);대조조HIF-1α부유미약표체,이모형조주요재폐동맥내피세포화평활기세포내표체;모형조HIF-1α기인표체수평우제3、7천고우대조조(P＜0.05);단백표체수평재제7、10、14천현저고우대조조(P＜0.01).결론 신생서폭로우저양조기,즉출현우심실수축압력승고,계지발생우심실비후급폐소동맥결구개변,이HIF-1α불론기인환시단백,재PPHN적폐소혈관중소계단균지속고수평표체,제시HIE-1α가능시촉성PPHN폐혈관중소적인자지일.
Objective To investigate the dynamic expression from gene and protein levels of hypoxia inducible factor-1α(HIF-1α) during the development of hypoxic pulmonary hypertension (HPH) in neonatal rats.Methods A neonatal rat model of HPH was established,normal neonatal rats were enrolled as the control group.At the 3th 、7th、10th and 14th days,we measured the mRVSP through catheterization of right ventricule,collected hearts to figure out the right ventricular hypertrophy index(RVHI),evaluated vascular remodeling by the percentage of medial thickness to outer diameter of the small pulmonary arteries (MT％) and the percentage of medial cross section on area to the total cross section area of the pulmonary small arteries (MA％),observed the expression of HIF-1α by immunochemistry,and measured the expression of HIF-1α in mRNA and protein by RT-PCR and Western blot respectively.Results The mRVSP increased at the 3th day,and sustained until the 14th day (P ＜ 0.01).RVHI MT％ and MA％ increased at the 7th day,and sustained until the 14th day (P ＜0.01).HIF-1α mainly expressed in endothelium and smooth muscle cells in the CHPH group and the HIF-1αmRNA increased significantly on the 3th and 7th days (P ＜ 0.05),and the HIF-1α protein increased significantly on the 7th、10th and 14th days in the CHPH group compared with the control group(P ＜ 0.01).Conclusion The mRVSP increased at the early stage after hypoxic exposure in neonatal rats,followed by vascular remodeling and right ventricular hypertrophy.Both mRNA and protein levels of HIF-1α sustained higher than control group during the vascular remodeling stage,indicating that HIF-1α might be a important factor contributing to the vascular remodeling.
