目的探讨受体组分蛋白(RCP)在静压力促血管平滑肌细胞(VSMC)增殖信号跨膜转导中的作用。方法以鼠源性A10血管平滑肌细胞(A10VSMC)作为研究对象,用静压力处理细胞,噻唑蓝比色法(MTT)检测细胞的活性;Western blot检测增殖细胞核抗原(PCNA)、RCP和G蛋白的表达;RT-PCR检测RCP mRNA表达水平;免疫共沉淀检测G蛋白和RCP间相互作用。结果静压力呈压力和时间依赖性地增加A10VSMC活性、PCNA蛋白表达和RCP表达,并在120 mm Hg静压力和6 h达到最大。静压力在增加细胞pAkt表达水平(P〈0.05)的同时,减少RCP与Gαs的相互作用,增加RCP与Gβ的作用,但对Gγ不明确。结论静压力可使VSMC增殖和RCP表达增加,其作用可能涉及到细胞G蛋白信号转导机制。
Aim To explore the effect of receptor component protein( RCP) in the signal transduction of vascular smooth muscle cell( VSMC) proliferation induced by static pressure. Methods The mouse-derived vascular smooth muscle cell line( A10VSMC)was employed in the experiment. Cells were exposed to static pressure,and MTT assay was used to detect the cell viability. Western blot was used to determine the expressions of PCNA,RCP and p-Akt,RCP mRNA was tested by RT-PCR,and co-immunoprecipitation was used to test the interaction between RCP and G proteins. Results The cell viability,expressions of PCNA and RCP increased with the elevation of static pressure and reached their peaks at 120 mm Hg,and after 6 hours they got a plateau. The static pressure significantly increased the level of p-Akt,meanwhile,the binding of RCP and Gαs significantly decreased.However,the binding of RCP and Gβ increased in response to static pressure after stimuli of static pressure,but Gγ was obscure. Conclusion Static pressure can induce VSMC proliferation and expression of RCP,which may involve G protein signal transduction model.