Involvement of phosphatidylinositol 3-kinase and mitogen-activated protein kinase pathways in AII-mediated enhanced expression of Gi proteins in vascular smooth muscle cells.
We have previously demonstrated that angiotensin II increased Gialpha-2 and Gialpha-3 expression at both protein and mRNA levels in vascular smooth muscle cell (VSMC). The present study was undertaken to investigate the mechanisms responsible for AII-induced enhanced expression of Gi proteins. The levels of Gi protein were determinated by immunoblotting techniques using specific antibodies against Gialpha-2 and Gialpha-3. AII treatment of VSMC increased the levels of Gialpha-2 and Gialpha-3 proteins and actinomycin D, an inhibitor of RNA synthesis attenuated the AII-evoked enhanced expression of Gialpha-2 and Gialpha-3 proteins. In addition, wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI-3-K), rapamycin, an inhibitor of p70(S6K) and PD 098059, an inhibitor of mitogen-activated protein kinase (MAPK) kinase were able to inhibit AII-induced enhanced expression of Gialpha-2 and Gialpha-3 to various degrees. The attenuation of AII-evoked enhanced levels of Gialpha-2 and Gialpha-3 by PD 098059 was concentration dependent. At 50 microM, PD 098059 was able to completely attenuate the enhanced levels of Gialpha-2 and Gialpha-3 caused by AII treatment. These data suggest that the enhanced expression of Gi-proteins by AII treatment may be attributed to increased RNA synthesis of Gi-proteins, and MAPK kinase, PI-3-Kinase and p70(S6K) may be involved in AII-mediated increased expression of Gi-proteins in VSMC.