Expression and function of visfatin (Nampt), an adipokine-enzyme involved in inflammatory pathways of osteoarthritis.
Expression and function of visfatin (Nampt), an adipokine-enzyme involved in inflammatory pathways of osteoarthritis.
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DOI：
10.1186/ar4467
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发表时间：
2014-01-31
影响因子：
4.9
通讯作者：
Sellam J
中科院分区：
文献类型：
--
作者：
Laiguillon MC;Houard X;Bougault C;Gosset M;Nourissat G;Sautet A;Jacques C;Berenbaum F;Sellam J
Visfatin 是一种脂肪因子，可能参与骨关节炎 (OA) 的组织间关节通讯。具有同二聚体构象，它发挥烟酰胺磷酸核糖基转移酶 (Nampt) 酶活性，这对于烟酰胺腺嘌呤二核苷酸生物合成至关重要。我们检查了人 OA 关节中 visfatin/Nampt 的组织起源和构象，并通过研究 Nampt 酶活性研究了 visfatin/Nampt 在软骨细胞和成骨细胞中的作用。使用人OA关节的滑膜、软骨和软骨下骨进行蛋白质提取或在无血清培养基（条件培养基）中培养24小时，并从OA患者获得滑液。分别通过蛋白质印迹和酶联免疫吸附测定（ELISA）评估组织提取物和条件培养基中的 Visfatin/Nampt 表达。通过比色测定评估 OA 滑膜中的 Nampt 活性。用visfatin/Nampt刺激小鼠软骨细胞和成骨细胞的原代培养物，并用或不用APO866（一种Nampt活性的药理学抑制剂）预处理。通过定量逆转录酶聚合酶链反应和/或ELISA检查对细胞因子、趋化因子、生长因子和肥大标志物表达的影响。在组织外植体、条件培养基和滑液中，发现内脂素/Nampt为同二聚体，对应于酶活性构象。所有人类OA关节组织均释放visfatin/Nampt（滑膜：628±106 ng/g组织；软骨下骨：195±26 ng/g组织；软骨：152±±46 ng/g组织），其中滑膜水平显着较高（P <0.0005）。在滑膜中离体鉴定 Nampt 活性。在体外，visfatin/Nampt 显着诱导软骨细胞和成骨细胞中白细胞介素 6、角质形成细胞趋化蛋白和单核细胞趋化蛋白 1 的表达。 APO866 降低了两种细胞类型中这些促炎细胞因子的 mRNA 和蛋白质水平（抑制率分别高达 94% 和 63%）。治疗后生长因子（血管内皮生长因子、转化生长因子β）和肥大基因的水平没有变化。 Visfatin/Nampt 由所有人类 OA 组织以二聚体酶促活性构象释放，并且主要由具有 Nampt 活性的滑膜释放。内脏脂肪素的 Nampt 活性参与软骨细胞和成骨细胞的激活，因此针对这种酶活性来破坏关节组织相互作用可能是 OA 治疗中的新颖方法。
Visfatin is an adipokine that may be involved in intertissular joint communication in osteoarthritis (OA). With a homodimeric conformation, it exerts nicotinamide phosphoribosyltransferase (Nampt) enzymatic activity, essential for nicotinamide adenine dinucleotide biosynthesis. We examined the tissular origin and conformation of visfatin/Nampt in human OA joints and investigated the role of visfatin/Nampt in chondrocytes and osteoblasts by studying Nampt enzymatic activity. Synovium, cartilage and subchondral bone from human OA joints were used for protein extraction or incubated for 24 hours in serum-free media (conditioned media), and synovial fluid was obtained from OA patients. Visfatin/Nampt expression in tissular extracts and conditioned media was evaluated by western blot and enzyme-linked immunosorbent assay (ELISA), respectively. Nampt activity was assessed in OA synovium by colorimetric assay. Primary cultures of murine chondrocytes and osteoblasts were stimulated with visfatin/Nampt and pretreated or not with APO866, a pharmacologic inhibitor of Nampt activity. The effect on cytokines, chemokines, growth factors and hypertrophic markers expression was examined by quantitative reverse transcriptase polymerase chain reaction and/or ELISA. In tissular explants, conditioned media and synovial fluid, visfatin/Nampt was found as a homodimer, corresponding to the enzymatically active conformation. All human OA joint tissues released visfatin/Nampt (synovium: 628 ± 106 ng/g tissue; subchondral bone: 195 ± 26 ng/g tissue; cartilage: 152 ± 46 ng/g tissue), with significantly higher level for synovium (P <0.0005). Nampt activity was identified ex vivo in synovium. In vitro, visfatin/Nampt significantly induced the expression of interleukin 6, keratinocyte chemoattractant and monocyte chemoattractant protein 1 in chondrocytes and osteoblasts. APO866 decreased the mRNA and protein levels of these pro-inflammatory cytokines in the two cell types (up to 94% and 63% inhibition, respectively). Levels of growth factors (vascular endothelial growth factor, transforming growth factor β) and hypertrophic genes were unchanged with treatment. Visfatin/Nampt is released by all human OA tissues in a dimeric enzymatically active conformation and mostly by the synovium, which displays Nampt activity. The Nampt activity of visfatin is involved in chondrocyte and osteoblast activation, so targeting this enzymatic activity to disrupt joint tissue interactions may be novel in OA therapy.
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影响因子：
3.5
作者：
Kitani, T;Okuno, S;Fujisawa, H
通讯作者：
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影响因子：
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DOI：
10.1016/j.bbrc.2009.11.066
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2010-01-01
影响因子：
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作者：
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通讯作者：
Gebhardt, R.
影响因子：
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Mahjoub, M.;Berenbaum, F.;Houard, X.
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