Osta Bilal, Roux Jean-Paul, Lavocat Fabien, Pierre Marlène, Ndongo-Thiam Ndieme, Boivin Georges, Miossec Pierre
Immunogenomics and Inflammation Research Unit EA 4130, Department of Clinical Immunology and Rheumatology, Edouard Herriot Hospital, University of Lyon 1 , Lyon , France.
UMR 1033, INSERM, UFR de Médecine Lyon Est , Lyon , France.
Front Immunol. 2015 Apr 7;6:151. doi: 10.3389/fimmu.2015.00151. eCollection 2015.
TNF-α and IL-17A act on fibroblast-like synoviocytes (FLS) and contribute to cytokine production, inflammation, and tissue destruction in rheumatoid arthritis (RA). The aim of this study was to compare their effects on osteogenic differentiation of isolated FLS and on whole bone explants from RA and osteoarthritis (OA) patients.
Fibroblast-like synoviocytes and bone explants were cultured in the presence or absence of TNF-α and/or IL-17A. Mineralization of extracellular matrix of FLS was measured by alizarin red and alkaline phosphatase activity (ALP). mRNA expression was analyzed by qRT-PCR for Wnt5a, BMP2, and RUNX2, key genes associated with osteogenesis. IL-6 and IL-8 levels were measured by enzyme-linked immunosorbent assays. Bone explant structure was quantified by histomorphometry.
In isolated OA and RA FLS, the combination of TNF-α and IL-17A induced matrix mineralization, increased ALP activity and expression of the osteogenesis-associated genes Wnt5a, BMP2, and Runx2, indicating an osteogenic differentiation. Wnt5a levels increased with TNF-α alone and in combination with IL-17A. BMP2 expression decreased with IL-17A and TNF-α after 12 h with OA FLS and 24 h with RA FLS. Runx2 expression decreased only with combination of TNF-α and IL-17A in OA FLS and with cytokines alone and combined in RA FLS. IL-6 and IL-8 production increased with IL-17A and/or TNF-α in both FLS and bone samples, especially from RA. Treatment of bone explants with cytokine combination increased ALP in OA but not RA samples. A decrease in bone volume was seen with cytokine combination, especially with RA explants.
Differences were observed for the effects of IL-17A and TNF-α on osteogenic differentiation. In isolated FLS, increased osteoblastogenesis was observed, contrasting with the inhibitory effect in whole bone, specifically in RA. The net effect of IL-17A and TNF-α appears to depend on the disease state and the presence of other cells.
肿瘤坏死因子-α（TNF-α）和白细胞介素-17A（IL-17A）作用于成纤维样滑膜细胞（FLS），并参与类风湿关节炎（RA）中的细胞因子产生、炎症反应及组织破坏。本研究旨在比较它们对分离的RA和骨关节炎（OA）患者FLS成骨分化以及对全骨外植体的影响。
在有或无TNF-α和/或IL-17A存在的情况下培养成纤维样滑膜细胞和骨外植体。通过茜素红和碱性磷酸酶活性（ALP）检测FLS细胞外基质的矿化情况。采用qRT-PCR分析与成骨相关的关键基因Wnt5a、骨形态发生蛋白2（BMP2）和Runx2的mRNA表达。通过酶联免疫吸附测定法检测白细胞介素-6（IL-6）和白细胞介素-8（IL-8）水平。通过组织形态计量学对骨外植体结构进行定量分析。
在分离的OA和RA FLS中，TNF-α和IL-17A联合作用诱导基质矿化，增加ALP活性以及成骨相关基因Wnt5a、BMP2和Runx2的表达，表明发生了成骨分化。单独使用TNF-α以及与IL-17A联合使用时，Wnt5a水平升高。OA FLS培养12小时、RA FLS培养24小时后，IL-17A和TNF-α联合作用使BMP2表达降低。OA FLS中Runx2表达仅在TNF-α和IL-17A联合作用时降低，而在RA FLS中，单独及联合使用细胞因子时均降低。在FLS和骨样本中，IL-17A和/或TNF-α均使IL-6和IL-8产生增加，尤其是RA样本。细胞因子联合处理骨外植体可使OA样本中的ALP增加，但RA样本中未增加。细胞因子联合作用使骨体积减少，尤其是RA外植体。
观察到IL-17A和TNF-α对成骨分化的影响存在差异。在分离的FLS中，观察到成骨细胞生成增加，这与对全骨（特别是RA）的抑制作用形成对比。IL-17A和TNF-α的净效应似乎取决于疾病状态和其他细胞的存在。
Zotero 插件
