The Regulatory Role Of Dual Ligand Receptors In Cell Adhesion Molecules Expression On Huvec Cells

Aims:I use LPC and protons to stimulate the receptor G2A on HUVEC cells, then explore its effect in the expression of ICAM-1、VCAM-1, and detect the cell signaling pathways.Contents:I cultured HUVEC, then transfected HUVEC with shRNA-G2A plasmid, and selected cells by G418to get cell lines which plasmid expressed steadily. I used PCR to observe the G2A expression in G2A interferenced HUVEC and normal HUVEC. After cells stimulated by LPC and protons, I detected the calcium concentration and the expression of ICAM-1and VCAM-1on both HUVEC and G2A interference HUVEC, and detected Gi protein inhibitor PTX’s role in this process.Results:I got the G2A interference HUVEC. The G2A expression was reduced after stimulated by protons compared to control. LPC stimulation increased calcium concentration in the HUVEC. Protons decreased calcium concentration which reponse to LPC, and PTX decreased calcium concentration which reponse to LPC too. ICAM-1、VCAM-1was much higher in normal HUVEC than in G2A interference HUVEC after stimulated by LPC. LPC stimulation decreased ICAM-1、VCAM-1in G2A interference HUVEC; stimulated by both LPC and protons, ICAM-1、VCAM-1didn’t change in G2A interference HUVEC.Conclusion:Protons decrease the expression of G2A in HUVEC. G2A is the major receptor which related to the expression of ICAM-1and VCAM-1in HUVEC. LPC stimulated cellular Ca2+mobilization was mediated by G2A coupled with Gi protein and the LPC induced action was antagonilazed by it’s proton ligand. Our results suggest that dual ligand receptors play an important regulatary role in arteriosclerotic process.