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Evidence have shown that endothelial cells and adventitial fibroblasts interact under certain conditions. In this study, we investigated the influence of endothelial cells on the phenotypic modulation of adventitial fibroblasts. Endothelial cells and adventitial fibroblasts from rat thoracic aorta were cultivated in a co\u2010culture system and adventitial fibroblasts were induced with angiotensin II (Ang II). Collagen I and alpha smooth muscle actin (\u03b1\u2010SMA) expression and migration of adventitial fibroblasts were analyzed. Ang II upregulated the expression of collagen I and \u03b1\u2010SMA and the migration of adventitial fibroblasts. Adventitial fibroblasts\u2013endothelial cells co\u2010culturing attenuated the effects of Ang II. Homocysteine\u2010treated endothelial cells, which are functionally impaired, were less inhibitory of the phenotypic modulation of adventitial fibroblasts. Supplementation of endothelial cells with L\u2010arginine (L\u2010Arg) or 8\u2010bromoguanosine 3\u2032,5\u2032\u2010cyclic monophosphate (8\u2010BrcGMP) enhanced the trends, while with L\u2010NG\u2010nitroarginine methyl ester (L\u2010NAME) or 1H\u2010[1,2,4]Oxadiazolo[4,3\u2010a]quinoxalin\u20101\u2010one (ODQ) the opposite effect was observed. Under the influence of Ang II, adventitial fibroblasts were prone to undergo phenotypic modulation, which was closely related to vascular remodeling. Our study showed that endothelial cells influenced fibroblast phenotypic transformation and such effect would be mediated through the nitric oxide (NO)\/cGMP signaling pathway. J. Cell. 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