Effects Of TEPV1 On M2 Macrophages And The Role Of Evodiamine In The TRPV1-mediated Regulation Of M2 Macrophages

Background:Several studies have showen that inflammation is the most common pathophysiologic characteristics in the development of different stages in cardiovascular diseases such as hypertension and coronary heart disease. Cardiovascular diseases are usually characterized by the dysfun-ction of endothelial cells and chronic inflammation associated with monoc-yte/macrophage infiltration.Therefore, an intervention for inhibiting infla-meematory response will be a potential therapy for cardiovascular diseases. Several studies suggest that the transient receptor potential vanilloid type 1 (TRPV1) channel plays a protective role in inflammation, but the mechan-isms undelying TRPV1-mediated anti-inflammation are still unclear. Evodi-ae has been used in treating cardiovascular disease for hundreds of years. Several studies indicate that Evodiamine extracted from Evodiae may be a TRPV1 agonist. To explore the effects of TRPV1 on M2 macrophages and the role of Evodiamine in the process will provide a basis for future clinical application.Objectives:To determine the mechanisms undrelying TRPV1 and Evod-iamine-mediated anti-inflammation, we explored the effects of TRPV1 on IL-4-induced M2 macrophage phenotypes in the cultured human monocytic leukemia cell line (THP-1), we also determined the role of TRPV1 in Evod-iamine-mediated anti-inflammation.Methods:1. Experimental groups①vehicle group (5% DMSO)②IL-4-treated group (10ng/ml)③CAP+IL-4- treated group (Capsincin 10umol/L, IL-4 10ng/ml)④CAPZ+CAP+IL-4- treated group (CapsazepinelOumol/L, Capsincin 10umol/L, IL-4 10ng/ml)⑤EVO+IL-4-treated group (Evodiamine 10umol/L, IL-4 10ng/ml)⑥APZ+EVO+IL-4- treated group (Capsazepine10umol/L, Evodiamine 10umol/L, IL-4 10ng/ml)2. Determination of parameters①The production of TGF-β 1 was determined by ELISA.②The expression of arginase-1、mannose receptor mRNA was detected by RT-PCR.③The expression of arginase-1、mannose receptor protein was detected by Western blot.Results:1.The production of TGF-β 1 was determined by ELISA.After IL-4 stimulation, the production of TGF-β1 were significantly increased (P<0.05), and activation of TRPV1 significantly reduced the production of TGF-β1-induced by IL-4 (P<0.05). The effects were reverse-d by blocking TRPV1 (P<0.05).2. The expression of arginase-1、mannose receptor mRNA was detected by RT-PCR.After IL-4 stimulation, the expression of arginase-1、mannose receptor mRNA was significantly increased (P<0.05), and activation of TRPV1 obviously reduced the expression of arginase-1、mannose receptor mRNA (P<0.05), the effects were reversed by blocking TRPV1 (P<0.05).3. The expression of arginase-1、mannose receptor protein was detected by Western blot.After IL-4 stimulation, the expression of arginase-1、mannose receptor protein was significantly enhanced (P<0.05), and TRPV1 activation signif-icantly reduced the expression of arginase-1、mannose receptor protein (P<0.05). The effects were reversed by blocking TRPV1(P<0.05).4. Evodiamine treatment significantly inhibited M2 macrophage phenotypes induced by IL-4, as reflected by the decreased production of TGF-β1 and expression of arginase-1、mannose receptor mRNA and protein(P<0.05). Moreover, the Evodiamine-induced effects were reversed by a specific TRPV1 antagonist, CAPZ.Conclusions:1. The IL-4-induced expression of M2 macrophages phenotypes was inhibited by the activation of TRPV1, and the effects were reversed by the TRPV1 antagonist. The results suggest that TRPV1 can regulate the funct-ion of M2 macrophages.2. The IL-4-induced expression of M2 macrophages phenotypes was attenuated by Evodiamine, and the effects were reversed by the TRPV1 antagonist. The data suggest that Evodiamine regulates the function of M2 macrophages via the activation of TRPV1.