The Study of the Transfer and Expression of Angiotensin II Type 2 Receptor Gene in Rat Vascular Smooth Muscle Cells Mdiated by Recombinant Adenovirus Vector

Objective To constructed the cell model transferred angiotensin Ⅱ (AngⅡ) type 2 receptor (AT\-2R) in vascular smooth muscle cells(VSMC) Method The VSMCs, isolated from the aorta of rat, were cultured by routine method. Recombinant adenoviral vector, AdCMV\|AT\-2R, containing rat AT\-2 receptor gene was constructed by homologous recombination, and then it was used to transfer AT\-2 receptor gene to VSMC in vitro. The rate of AT\-2R 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 (AT\-1R) was determined as well. Result The expression rate of AT\-2R in VSMC was increased significantly after transferred by AdCMV\|AT\-2R 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 AT\-2R mRNA and protein were increased obviously in transferred VSMC. There were no significantly change of AT\-1R expression during AT\-2R expression. Conclusion Our study indicates that AdCMV\|AT\-2R did generate high level expression of AT\-2 receptor and its expression did not affect AT\-1R expression in cultured VSMC. The VSMCs transferred AT\-2R gene may be used as a good model to study the effect of AT\-2R on their biological action such as proliferation, migration and apoptosis.