Gutiérrez-Cañas I, Juarranz Y, Santiago B, Arranz A, Martinez C, Galindo M, Payá M, Gomariz R P, Pablos J L
Servicio de Reumatología, Hospital 12 de Octubre, Avda. de Córdoba s/n 28041, Madrid, Spain.
Rheumatology (Oxford). 2006 May;45(5):527-32. doi: 10.1093/rheumatology/kei219. Epub 2005 Nov 30.
Vasoactive intestinal peptide (VIP) has demonstrated therapeutic effects in arthritis by inhibiting both innate and acquired immune responses. We investigated the potential effects of VIP in the regulation of Toll-like receptor (TLR) expression and function in synovial fibroblasts from patients with rheumatoid arthritis (RA) and osteoarthritis (OA).
Cultured fibroblast-like synoviocytes (FLS) were obtained from patients with RA and OA. The effects of VIP on basal or TNF-alpha or lipopolysaccharide (LPS)-induced TLR2, TLR4 and MyD88 expression and its effects on TLR4-mediated CCL2 and CXCL8 chemokine production were studied by reverse transcription-polymerase chain reaction, western blotting and enzyme-linked immunosorbent assay.
TLR2, TLR4 and MyD88 mRNA expression was increased in RA FLS compared with OA FLS. The largest increase was observed for TLR4 and there was also overexpression at the protein level in RA FLS. TLR4 and MyD88 mRNA and proteins were induced by LPS and TNF-alpha in RA FLS. VIP down-regulated the induced but not the constitutive expression of TLR4 and MyD88 in RA FLS. VIP treatment decreased CCL2 and CXCL8 chemokine production in response to TLR4 activation with LPS in RA FLS.
We demonstrate that VIP down-regulates LPS and TNF-alpha activation of TLR4 expression and the TLR4 functional response in terms of proinflammatory chemokine production. These studies suggest that the pleiotropic anti-inflammatory actions of VIP involve inhibitory effects on TLR4 expression and signalling.
血管活性肠肽（VIP）已通过抑制先天性和获得性免疫反应在关节炎中显示出治疗作用。我们研究了VIP对类风湿性关节炎（RA）和骨关节炎（OA）患者滑膜成纤维细胞中Toll样受体（TLR）表达和功能的潜在影响。
从RA和OA患者中获取培养的成纤维样滑膜细胞（FLS）。通过逆转录-聚合酶链反应、蛋白质印迹法和酶联免疫吸附测定法研究VIP对基础或肿瘤坏死因子-α（TNF-α）或脂多糖（LPS）诱导的TLR2、TLR4和髓样分化因子88（MyD88）表达的影响及其对TLR4介导的趋化因子CCL2和CXCL8产生的影响。
与OA FLS相比，RA FLS中TLR2、TLR4和MyD88 mRNA表达增加。TLR4的增加最为明显，并且在RA FLS的蛋白质水平上也存在过表达。LPS和TNF-α在RA FLS中诱导TLR4和MyD88 mRNA及蛋白质表达。VIP下调RA FLS中TLR4和MyD88的诱导表达而非组成型表达。VIP处理降低了RA FLS中LPS激活TLR4后CCL2和CXCL8趋化因子的产生。
我们证明，就促炎趋化因子的产生而言，VIP下调LPS和TNF-α对TLR4表达的激活以及TLR4功能反应。这些研究表明，VIP的多效抗炎作用涉及对TLR4表达和信号传导的抑制作用。
