In vivo transfection of hepatocyte growth factor gene induces pulmonary angiogenesis in rabbits with hyperkinetic pulmonary hypertension
10.3321/j.issn:0253-3758.2008.12.014
- VernacularTitle:肝细胞生长基因转染诱导肺高压模型动物肺血管生成的实验研究
- Author:
Wei WANG
1
;
Shu-Ming WU
;
Zhong-Ming ZHANG
;
Yi-Qian ZHANG
Author Information
1. 徐州医学院附属医院
- Keywords:
Hypertension,pulmonary;
Hepatocyte growth factor;
Neovascularization,phusiologic;
Genes
- From:
Chinese Journal of Cardiology
2008;36(12):1110-1114
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of adenoviral-mediated exogenous HGF(Ad-HGF)gene transfer on lung angiogenesis in the rabbit lung in rabbits with hyperkinetic pulmonary artery hypertension.Methods A thoracotomy wag performed through a midsternal incision in 1-month-old immature rabbit and an anastomosis between the left inominate artery and the pulmonary trunk was made to establish a chronic patent left to right shunt.Three mortths later,animals were randomly assigned to receive either Ad-HGF(2×109Pfu in 0.2 ml PBS,H1 group),repeated administration of Ad-HGF after one week (H 2 group),Ad-GFP(2×109 Pfu in 0.2 ml PBS,G group),or PBS(0.2 ml,C group)by the intratraeheal method of gene transfection.After two weeks,reverse transcription-polymerase chain reaction (RT-PCR)and immunohistochemical examination was performed to identify HGFmRNA and HGF protein expression.The capillary density and small pulmonary artery density were determined by immunostained with antibodies against factor Ⅷ and a-SMA.After 1 month,the collateral vessels were evaluated by angiogram under digital subtraction angiography(DSA).Results Two weeks after Ad-HGF transfection,484 bp bands could be found by RT-PCR in H l and H2 groups,but not in other groups.The expression of HGF protein could be detected on alveolag epithelium and pulmonary vessel endothelium by immunohistochemistry examination.The number of factor Ⅷ-positive pulmonary capillaries was also significantly increased in the H1 and H2 groups compared with the C and G groups(P<0.05).The capillary density reached(17.0±3.3)mm2 and (19.7±2.8)mm2 in the H1 and H2 group,respectively,whereas it remained(13.2±3.2)mm2 in the C group and(13.5±2.4)mm2 in the G group(P<0.05).One month after Ad-HGF transfeetion.the numbeT of small pulmonary arteries was significantly increased in Hl and H2 group compared with control groups(P<0.05).The collateral vessels were more abundant in HGF transfection groups than that in the two control groups reviewed by angiogram under digital subtraction angiography (DSA).Conclusion In vivo gene transfection with HGF by means of the intra-tracheal injection could induce pulmonary angiogensis in the early stage and small pulmonary arterial angiogensis in later stage.
