Objective-To examine the role of perivascular adipose tissue (PVAT)-derived factors in the regulation of adventitial fibroblast (AF) function in vitro and in vivo. Methods and Results-PVAT is an active component of blood vessels. Bioactive substances released from PVAT play regulatory roles in vascular function. However, their effects on vascular AFs remain unclear. PVAT-conditioned medium stimulated AF migration using a transwell technique, and differentiation was evaluated by ␣-smooth muscle-actin induction. We identified the secretome of PVAT by liquid chromatography-tandem mass spectrometry. One of the major secretory proteins in PVAT is complement 3 (C3). The C3 antagonist and neutralizing antibody attenuated PVAT-conditioned medium-induced AF migration and differentiation. Similar to PVAT-conditioned medium, C3 recombinant protein stimulated AF migration and differentiation. We demonstrated that the effects of PVAT-derived C3 were mediated by the c-Jun N-terminal kinase pathway. Moreover, we found morphological changes in perivascular adipocytes and increased expression of C3 in PVAT that was tightly associated with adventitial thickening and myofibroblast clustering around PVAT in deoxycorticosterone acetate-salt hypertensive rats. Key Words: perivascular adipose tissue Ⅲ migration Ⅲ ␣-smooth muscle actin Ⅲ complement 3 Ⅲ hypertensive rats Ⅲ adventitial fibroblasts A lmost all blood vessels are surrounded by perivascular adipose tissue (PVAT), which has been considered merely a structural support for vasculature. Recent studies revealed that PVAT produced and released various bioactive substances, playing a dual role in the regulation of vascular function. 1 Adipocyte-derived relaxing factors released from PVAT have attenuated the contractile response of rat aorta responding to norepinephrine. 2,3 However, PVAT also promoted vasoconstriction through production of superoxide and inflammatory cytokines, such as tumor necrosis factor ␣ (TNF␣) and interleukin 6. 4,5 In particular, the conditioned medium of PVAT induced vascular smooth muscle cell (VSMC) proliferation through activation of mitogen-activated protein kinase (MAPK) pathways via paracrine adipokines. 6,7 In pathophysiologic processes of vascular remodeling, phenotypic changes in perivascular cells correlated with upregulation of proinflammatory adipocytokines, which may contribute to vascular dysfunction. 8,9 The list of bioactive factors released from PVAT is increasing, and the role of these adipocytokines in the regulation of vascular function is being unraveled. 10,11 Multiple lines of evidence suggest that vascular adventitial fibroblasts (AFs), the major cell component of adventitia, play a vital role in regulating the structure and function of blood vessels. 12,13 Previous studies 14,15 indicated that numerous active cytokines induce AF migration, which may be associated with the pathogenesis of neointimal formation. Although AFs are in closer proximity to PVAT compared with VSMCs, little attention has been devoted to the role of PVAT-der...