Afif Hassan, Benderdour Mohamed, Mfuna-Endam Leandra, Martel-Pelletier Johanne, Pelletier Jean-Pierre, Duval Nicholas, Fahmi Hassan
Osteoarthritis Research Unit, Centre Hospitalier de l'Université de Montréal, Montreal, 1560 Sherbrooke East, Pavillon J,A DeSève, Y-2628, Montreal, QC, H2L 4M1, Canada.
Arthritis Res Ther. 2007;9(2):R31. doi: 10.1186/ar2151.
Peroxisome proliferator-activated receptor gamma (PPARgamma) is a nuclear receptor involved in the regulation of many cellular processes. We and others have previously shown that PPARgamma activators display anti-inflammatory and chondroprotective properties in vitro and improve the clinical course and histopathological features in an experimental animal model of osteoarthritis (OA). However, the expression and regulation of PPARgamma expression in cartilage are poorly defined. This study was undertaken to investigate the quantitative expression and distribution of PPARgamma in normal and OA cartilage and to evaluate the effect of IL-1beta, a prominent cytokine in OA, on PPARgamma expression in cultured chondrocytes. Immunohistochemical analysis revealed that the levels of PPARgamma protein expression were significantly lower in OA cartilage than in normal cartilage. Using real-time RT-PCR, we demonstrated that PPARgamma1 mRNA levels were about 10-fold higher than PPARgamma2 mRNA levels, and that only PPARgamma1 was differentially expressed: its levels in OA cartilage was 2.4-fold lower than in normal cartilage (p < 0.001). IL-1 treatment of OA chondrocytes downregulated PPARgamma1 expression in a dose- and time-dependent manner. This effect probably occurred at the transcriptional level, because IL-1 decreases both PPARgamma1 mRNA expression and PPARgamma1 promoter activity. TNF-alpha, IL-17, and prostaglandin E2 (PGE2), which are involved in the pathogenesis of OA, also downregulated PPARgamma1 expression. Specific inhibitors of the mitogen-activated protein kinases (MAPKs) p38 (SB203580) and c-Jun N-terminal kinase (SP600125), but not of extracellular signal-regulated kinase (PD98059), prevented IL-1-induced downregulation of PPARgamma1 expression. Similarly, inhibitors of NF-kappaB signaling (pyrrolidine dithiocarbamate, MG-132, and SN-50) abolished the suppressive effect of IL-1. Thus, our study demonstrated that PPARgamma1 is downregulated in OA cartilage. The pro-inflammatory cytokine IL-1 may be responsible for this downregulation via a mechanism involving activation of the MAPKs (p38 and JNK) and NF-kappaB signaling pathways. The IL-1-induced downregulation of PPARgamma expression might be a new and additional important process by which IL-1 promotes articular inflammation and cartilage degradation.
过氧化物酶体增殖物激活受体γ(PPARγ)是一种参与调控多种细胞过程的核受体。我们和其他研究人员之前已经表明,PPARγ激活剂在体外具有抗炎和软骨保护特性,并能改善骨关节炎(OA)实验动物模型的临床病程和组织病理学特征。然而,软骨中PPARγ的表达及其调控机制尚不清楚。本研究旨在调查PPARγ在正常和OA软骨中的定量表达及分布情况,并评估OA中一种重要细胞因子白细胞介素-1β(IL-1β)对培养软骨细胞中PPARγ表达的影响。免疫组织化学分析显示,OA软骨中PPARγ蛋白表达水平显著低于正常软骨。通过实时逆转录-聚合酶链反应(RT-PCR),我们发现PPARγ1 mRNA水平比PPARγ2 mRNA水平高约10倍,且只有PPARγ1存在差异表达:其在OA软骨中的水平比正常软骨低2.4倍(p < 0.001)。用IL-1处理OA软骨细胞会以剂量和时间依赖性方式下调PPARγ1表达。这种效应可能发生在转录水平,因为IL-1会降低PPARγ1 mRNA表达和PPARγ1启动子活性。参与OA发病机制的肿瘤坏死因子-α(TNF-α)、IL-17和前列腺素E2(PGE2)也会下调PPARγ1表达。丝裂原活化蛋白激酶(MAPK)p38(SB203580)和c-Jun氨基末端激酶(SP600125)的特异性抑制剂,而非细胞外信号调节激酶(PD98059)的抑制剂,可阻止IL-1诱导的PPARγ1表达下调。同样,核因子κB(NF-κB)信号通路抑制剂(吡咯烷二硫代氨基甲酸盐、MG-132和SN-50)可消除IL-1的抑制作用。因此,我们的研究表明OA软骨中PPARγ1表达下调。促炎细胞因子IL-1可能通过激活MAPKs(p38和JNK)和NF-κB信号通路导致这种下调。IL-1诱导的PPARγ表达下调可能是IL-1促进关节炎症和软骨降解的一个新的重要过程。