Effect of interleukin-1beta on osteogenic protein 1-induced signaling in adult human articular chondrocytes.
Effect of interleukin-1beta on osteogenic protein 1-induced signaling in adult human articular chondrocytes.
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DOI：
10.1002/art.24151
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发表时间：
2009-01
影响因子：
--
通讯作者：
Chubinskaya, Susan
中科院分区：
文献类型：
--
作者：
Elshaier, Arriel M.;Hakimiyan, Arnavaz A.;Rappoport, Lev;Rueger, David C.;Chubinskaya, Susan
目的探讨白细胞介素1β（IL 1β）对成人关节软骨细胞成骨蛋白1（OP 1）信号转导的影响。我们在IL-1β诱导的软骨退变模型中研究了两种主要的受体激活Smad（R-Smad）信号通路：BMP和MAPK。将26例正常人踝关节软骨细胞在无血清培养基中高密度单层培养。采用RT-PCR和流式细胞术研究IL-1β对BMP受体的影响。在用IL-1β（10 ng/ml）、OP-1（100 ng/ml）或组合的IL-1β和OP-1处理的细胞中测试R-Smads的磷酸化。通过蛋白质印迹用针对两个R-Smad磷酸化位点（BMP和MAPK）的多克隆抗体或总的非磷酸化R-Smad作为对照来分析细胞裂解物。为了鉴定IL-1β通过何种MAPK激活接头区，将软骨细胞与特异性MAPK抑制剂（MAP/ERK的PD 98059; JNK的SP 600125;和p38的SB 203580）预孵育。我们发现IL-1β减少ALK-2和ALK-3受体的数量，抑制Smad 1和Smad 6的表达，延迟和过早终止OP-1介导的R-Smad磷酸化的发生，并影响R-Smad/Smad 4复合物的核转位。我们发现，通过MAPK途径（主要是p38和JNK）在连接区的R-Smad的替代磷酸化可能是IL-1β抵消OP-1信号传导和对OP-1应答的可能机制。相反，发现OP-1直接抑制p38的磷酸化。我们的研究结果描述了软骨细胞中OP-1和IL-1β之间相互作用的新机制。该研究还确定了治疗软骨退行性过程的治疗干预的潜在靶点。
To investigate the effect of interleukin-1β (IL-1β) on osteogenic protein-1 (OP-1) signaling in human adult articular chondrocytes. We examined two major receptor-activated Smad (R-Smad) signaling pathways, the BMP and MAPK, in the model of IL-1β-induced cartilage degeneration. Ankle chondrocytes from 26 normal human donors were cultured in high density monolayers in a serum-free media. Effect of IL-1β on BMP receptors was studied by RT-PCR and flow cytometry. Phosphorylation of R-Smads was tested in cells treated with IL-1β (10ng/ml), OP-1 (100ng/ml) or combined IL-1β and OP-1. Cell lysates were analyzed by Western blots with polyclonal antibodies against two R-Smads phosphorylated sites (BMP and MAPK) or total, non-phosphorylated R-Smad as a control. To identify through which MAPK IL-1β activates linker region, chondrocytes were pre-incubated with specific MAPK inhibitors (PD98059 for MAP/ERK; SP600125 for JNK; and SB203580 for p38). We found that IL-1β reduced the number of ALK-2 and ALK-3 receptors, inhibited Smad1 and Smad6 expression, delayed and prematurely terminated the onset of OP-1 mediated R-Smad phosphorylation and affected nuclear translocation of R-Smad/Smad4 complexes. We discovered that the alternative phosphorylation of R-Smad in the linker region via the MAPK pathway (primarily p38 and JNK) could be a possible mechanism through which IL-1β offsets OP-1 signaling and responses to OP-1. Conversely, OP-1 was found to directly inhibit phosphorylation of p38. Our findings describe new mechanisms of the cross-talk between OP-1 and IL-1β in chondrocytes. The study also identifies potential targets for therapeutic interventions in the treatment of cartilage degenerative processes.
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影响因子：
56.9
作者：
HAN, J;LEE, JD;ULEVITCH, RJ
通讯作者：
ULEVITCH, RJ
影响因子：
64.5
作者：
FRESHNEY, NW;RAWLINSON, L;SAKLATVALA, J
通讯作者：
SAKLATVALA, J
影响因子：
64.8
作者：
Imamura, T;Takase, M;Miyazono, K
通讯作者：
Miyazono, K
影响因子：
3.3
作者：
Dore, JJE;Yao, DY;Leof, EB
通讯作者：
Leof, EB
影响因子：
4.8
作者：
Hu, MC;Wasserman, D;Rosenblum, ND
通讯作者：
Rosenblum, ND