The Study of the Transfer and Expression of Angiotensin Ⅱ Type 2 Recepto r Gene in Rat Vascular Smooth Muscle Cells Mdiated by Recombinant Adenovirus Ve ctor / 中华高血压杂志

ABSTRACT

Objective To constructed the cell model transferred angiotensin Ⅱ (AngⅡ ) type 2 receptor (AT2R) in vascular smooth muscle cells(VSMC) Method The VSMCs, isolated from the aorta of rat, were cultured by routine method. Recombinant adenoviral vector, AdCMV-AT2R, containing rat AT2 receptor gene was constructed by homologous recombination, and then it was used to transfer AT2 receptor gene to VSMC in vitro. The rate of AT2R expression in VSMC was analysed by flow cytometry. The expression of mRNA, protein were detected by RT-PCR, Western blot and immunohistochemistry respectively. The angiotensin Ⅱ (AngⅡ) type 1 receptor (A T1R) was determined as well. Result The expression rate of AT2R in VSMC was increased signific antly after transferred by AdCMV-AT2R with time, and the peak value detected by flow cytometry was about 89.51% at 48 hours. RT-PCR, Western blot and immunohistochemistry showed that the expression of AT2R mRNA and protein were increased obviously in transferred VSMC. There were no significantly change of AT1R expression during AT2R expression. Conclusion Our study indicates that AdCMV-AT2R did generate high level expression of AT2 receptor and its expression did not affect AT1R exp ression in cultured VSMC. The VSMCs transferred AT2R gene may be used as a good model to study the effect of AT2R on their biological action such as proliferation, migration and apoptosis.