Molecular Mechanisms For Resveratrol Against Atherosclerosis

Background: resveratrol is one of the most important substance which has the cardioprotections in red wine.The anti-atherosclerosis effect of resveratrol derives from its powerful anti-inflammatory effect. CD40 pathway, as an inflammatory pathway, involves in the regulation of inflammatory reaction in many cells of atherosclerosis and plays an important part in the occurrence and formation of atherosclerosis.Objective: To investigate the effect of resveratrol on Chemotactic Factors of Monocyte,the Plaque Stabilization of atheromatosis and the effect of antioxidation and the relationship with CD40 Pathway. And to probe the molecular mechanism for resveratrol of action on anti-atherosclerosis .Methods: Divide the cultivated monocytes(THP-1) randomly into groups as the control, 1μmol/LRes,10μmol/LRes,50μmol/LRes. CCR2 mRNA expression were investigated by RT-PCR after THP-1 cells were incubated with resveratrol.THP-1 cells were differentiated to macrophages induced by PMA. TNF-αand sCD40L were used to stimulated the incubated HUVEC and Macrophages which differerntiate from monocytes. Divide the HUVEC and Macrophages randomly into five groups as the control, TNF-α(10μg/L) , (1μmol/L,10μmol/L, 50μmol/L)Res+TNF-α(10μg/L) or sCD40L(10μg/ml), (1μmol/L, 10μmol/L,50μmol/L)Res+sCD40L(10μg/ ml). The empirical sequence of medication group is TNF-αor sCD40L followed by the preincubation of resveratrol 2 hours. MMP-9 mRNA and TIMP-1 mRNA expression were investigated by RT-PCR; MMP-9 and TIMP-1 proteins were detected by western blot; MMP-9 activity was analyzed by gelatin zymography; MCP-1 which were secreted by HUVEC were detected by ELISA, and ROS generate in HUVEC were investigated by immunofluorescence.Results: After the stimulation of TNF-αor sCD40L, the expression of MCP-1 and ROS increases significantly than the control in HUVEC(P<0.05). The preincubation of resveratrol(10μmol/L,50μmol/L)can inhibit the expression of CCR2 gene(P<0.01) in a dose-dependent manner , and expression of MCP-1 ((P<0.05)) and ROS(P<0.01)induced by TNF-αor sCD40L. The expressions of MMP-9/TIMP-1 gene and protein were higher in Macrophages which were differentiated to macrophages induced by PMA than THP-1 cells. Resveratrol(10μmol/L,50μmol/L)can inhibit the expression of MMP-9 genes(P<0.01)and the secretion of MMP-9 protein (P<0.01)which were induced by sCD40L, and degrade the activity of MMP-9(P<0.05) in a dose-dependent manner, and can increase the expression of TIMP-1 genes(P<0.05)and the secretion of TIMP-1 proteins (P<0.05).Conclusion: Resveratrol can inhibit CCR2-gene's expression of mononuclear cells and expressing of HUVEC MCP-1 induced by TNF-αor sCD40L,which can prevent mononuclear cells from migrating into the inflammation region. Meanwhile, resveratrol also inhibits the generating of ROS in active HUVEC, then to diminish oxidative damage. Moreover, resveratrol can inhibit the effect of MMP-9 by CD40 pathway and regulate activity of MMP-9 by stimulates TIMP-1. All above results indicate that resveratrol can suppress the inflammation in the earlier period of atherosclerosis, protect blood vessel endothelium, stabilize the atheromatous plaques and can be seen as a specific inhibitor of CD40 pathway.