Secretory expression of PR39 following adeno-associated viral-encoding fusion gene transfer induces angiogenesis in hypoxia chick embryo
10.3760/cma.j.issn.0253-3758.2009.08.017
- VernacularTitle:分泌表达PR39重组腺相关病毒对缺氧鸡胚促血管生成的影响
- Author:
Yue-Wen HAO
1
;
Li-Jun SUN
;
Ying LIU
;
Quan-Ying WANG
;
Guang-Xiao YANG
Author Information
1. 第四军医大学西京医院
- Keywords:
Neovascularization,physiologic;
THERAPY;
Genes;
Anoxia;
PR39
- From:
Chinese Journal of Cardiology
2009;37(8):746-749
- CountryChina
- Language:Chinese
-
Abstract:
Objectives To investigate the impact of AAV-encoding NT4-TAT-His-PR39 fusion gene expression on HIF-1α level in ECV304 cultured under hypoxic condition ( 1% O2) and on angiogenesis in hypoxic chick embryo. Methods PR39 cDNA was connected with NT4, TAT, 6 x His cDNA by molecular biology methods. The recombinant AAV vector was obtained by three plasmid co-transfection in 293 cells. Then ECV304 were respectively infected with AAV-NT4-TAT-His-PR39, 6 x His expression and HIF-la level in ECV304 were detected by immunocytochemistry. The chicken embryos were randomized into the AAV-PR39, EV and PBS groups ( n = 10 each) subject to hypoxia (5% O2, n = 15 ) or normoxia environments (n = 15) , the vessel density of the chicken chorioallantoic membrane (CAM) were measured by Image Pro Plus (IPP) software. Results The expression of 6 x His protein was detected in AAV-PR39 infected ECV304 cells. HIF-1α protein activity was significantly increased in AAV-PR39 infected ECV304 underwent hypoxia compared to PBS and non-infected ECV304 groups ( P < 0. 05). The vessel density of chicken CAM in hypoxia environment but not in normoxia environment was also significantly higher in AAV-PR39 group than in EV group and PBS group (all P <0. 05). Conclusion AAV-encoding NT4-TAT-His-PR39 fusion gene expression significantly increased HIF-la level in ECV304 exposed to hypoxia and promoted angiogenesis in hypoxic chicken embryo.
