Study on the expression of ?_2-adrenergic receptor gene and enhanced green fluorescent protein gene in cardiomyocytes transferred by adeno associated virus vector
- VernacularTitle:腺病毒相关病毒载体介导人?_2肾上腺素能受体、增强型绿色荧光蛋白基因导入心肌细胞的表达
- Author:
Wenqian GAO
;
Xiaoying LI
;
Xiaobing WU
- Publication Type:Journal Article
- Keywords:
Adenoviruses, human;
Receptors, adrenergic;
Gene;
Tracings, flaorescent protein
- From:
Chinese Journal of Geriatrics
2000;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the way of gene therapy on congestive heart failure and other heart diseases. Methods Recombinant adeno associated viruses (AAV) encoding the human ? 2 adrenergic receptor(AR) and enhanced green fluorescent protein (EGFP) were constructed and the ? 2 AR and EGFP cDNA were transfected in vitro to the neonate rat ventricular myocytes. The expression of ? 2 AR and EGFP on cultured neonate rat ventricular myocytes and the ability of ? 2 AR to potentiate the beta adrenergic signaling were assessed. Results The presence of human ? 2 AR mRNA was demonstrated by Northern blot, the expression of the ? 2 AR gene was demonstrated by Westernimmuno blot. The density of ? AR in the myocytes was greater than that in the control 〔(204.0 ? 6.4) fmol/mg protein vs (76.0 ? 2.8) fmol/mg protein〕 according to a ligand binding assay. Human ? 2 AR gene significantly increased isoproterenol stimulated cAMP as compared with the control〔(116.2?5.8) pmol/10 6 cell vs (58.4 ? 4.6) pmol/10 6 cell〕. EGFP was also expressed in the transfected cardiomyocytes. Conclusions Human ? 2 AR gene could be transfected into cardiomyocytes by AAV vector and could potentiate the beta adrenergic signaling.