低氧对rAAV-2介导的hVEGF165基因在大鼠心肌细胞中表达的影响

Effect of Hypoxia on Expression of Human VEGF_(165) Gene Transferred by Recombinant Adeno-associated Viral Vector2 in Rat Cardiomyocytes in Vitro

目的探讨低氧对人血管内皮生长因子165(hVEGF165)基因在心肌细胞中表达的影响。方法以重组2型腺相关病毒(rAAV-2)为载体,构建rAAV2/hVEGF165,转染大鼠原代心肌细胞。对心肌细胞进行低氧培养,并于培养的0、24、48、72h,分别应用RT-PCR技术检测VEGF165 mRNA表达、ELISA法检测hVEGF165蛋白的水平,并与对照组(予以常氧培养)进行比较。结果rAAV2/hVEGF165体外转染后,低氧培养的心肌细胞在24、48、72hhVEGF165 mRNA转录水平均明显高于对照组,有显著性差异(P<0.05);在低氧培养的24、48、72h其培养上清中hVEGF165蛋白含量亦明显高于对照组,差异具有显著性(P<0.05)。结论低氧可促进重组2型腺相关病毒介导的人血管内皮生长因子(hVEGF165)基因在心肌细胞中表达。

Objective To Study the effect of hypoxia on the expression of human VEGF165 gene in rat cardiomyoctes in vitro. Methods The rAAV2/hVEGF165 was transferred into rat cardiomyocytes. The VEGF mRNA and protein expression in rAAV2/hVEGF165 -tranduced cardiomyocytes were examined by RT- POR and ELISA at different time （0, 24, 48 and 72 hours） under hypoxic condition, and the normoxic culture group was used as control. Results The VEGF mRNA expression level in rAAV2/hVEGF165 - tranduced cardiomyocytes was signiftcantly higher than that in control group at different exposure time under hypoxic condition （ P〈 0.0.5 ） ; the hVEGFm65 protein expression level was also significantly higher （ P〈 0.0.5 ） . Conclusion Hypoxia strongly promotes the expression and secretion of VEGF in cultured cardiomyocytes transfected by rAAV2/hVEGF165.