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Objectives: The differentiation of vascular adventitial fibroblasts (AFs) tomyofibroblasts (MFs) is a key step invascular remodeling. Our previous study demonstrated that TGF-β1 and angiotension II (Ang II)were able to induce this differentiation invitro. The aimof the studywas to identify the moleculeswhichmight be responsible for the cell phenotypic change and investigated the role of complement C3, one of the related molecules, in differentiation of MFs. Design and methods: Cultured rat AFs were treated with TGF-beta 1 (10 ng/ml) and Ang II (10−7M). TGF-beta 1induced gene expression profiling was studied using Affimetrix oligonucleotidemicroarrays. Expressionprofile of complement C3was verifiedby real-time RT-PCR. Then, the role of complement C3 in differentiation of AFs to MFs induced by Ang II was investigated using Western-blot, MTT array and Thymidine-Incorporation Assay. Results: Microarray analysis identified 2121 genes with a 2-fold change or above post-TGF-β1 stimulation. Among 1231 genes with known function, the gene expression profile of secreted phosphoprotein 1(APP1) and Rho-associated coiled-coil forming kinase 2 (ROCK2) was same as α-smooth muscleactin, a MF differentiation mark, and genes of potassium voltage gated channel, Shal-related family and member 2(KCND2) were up-regulated. Furthermore, endothelin 1(EDN1), complement C3, NADPH oxidase 4 (NOX4) andNAD(P)Hdehydrogenase, quinone 1 (Nqo1)were also found to be involved inMFdifferentiation. These resultswere confirmedby realtime quantitative RT-PCR. For the very first time, complement C3 was found to express in vascular adventitial fibroblasts fromSHR,WKYand SD rats and increased by angiotension II. Inhibition of complement C3 decreased the expression of α-smooth muscle-actin and reduced the proliferation of AFs from SD rats and WKY rats post treatment of Ang II. Exogenous C3 stimulated the growth of adventitial fibroblasts. Conclusions: Complement C3 contributes to adventitial fibroblast phenotypic differentiation and may be a target of vascular remodeling therapy. |
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