p38 MAPK inhibition modulates rabbit nucleus pulposus cell response to IL-1
p38 MAPK inhibition modulates rabbit nucleus pulposus cell response to IL-1
复制标题
DOI：
10.1002/jor.20604
复制
发表时间：
2008-07-01
影响因子：
2.8
通讯作者：
Kang, James D.
中科院分区：
文献类型：
--
作者：
Studer, Rebecca K.;Gilbertson, Lars G.;Kang, James D.
椎间盘基因表达的分析暗示IL-1在兔刺伤模型中椎间盘退变（IDD）的发展中。这些研究的目的是确定p38丝裂原活化蛋白激酶（p38 MAPK）信号传导在髓核细胞对IL-1的反应中的作用，并将兔髓核（rNP）细胞对IL-1活化的反应与椎间盘退变的针刺模型中的反应进行比较。在有或没有p38 MAPK抑制的情况下，将藻酸盐珠培养物中维持的NP细胞暴露于IL-1。分离RNA用于基因表达的逆转录聚合酶链反应（RT-PCR）分析，分析条件培养基中一氧化氮（NO）和前列腺素E-2（PGE-2）的积累，并在10天后测量蛋白聚糖合成。IL-1对环氧化酶-2（考克斯-2）、基质金属蛋白酶-3（MMP-3）、IL-1和IL-6的mRNA的上调被p38抑制减弱，而基质蛋白（胶原I、胶原II、聚集蛋白聚糖）和胰岛素样生长因子I（IFG-1）的下调也被逆转。基质金属蛋白酶组织抑制剂-1（TIMP-1）的mRNA被IL-1适度增加，而转化生长因子-P（TGF-β）SOX-9和多功能蛋白聚糖的mRNA保持不变。阻断p38 MAPK可减少IL-1诱导的NO和PGE-2的积累，并部分恢复蛋白多糖的合成。在对照细胞中，p38 MAPK抑制使基质蛋白（聚集蛋白聚糖、胶原II、多功能蛋白聚糖、胶原1）和合成代谢因子（IGF-1、TGF和SOX-9）的mRNA从50%增加到120%，降低基础PGE-2积累，但对TIMP-1、MMP-3或考克斯-2的信息没有影响。抑制p38 MAPK激活椎间盘细胞钝化基因表达和生产与炎症，疼痛，椎间盘基质catalysts相关的因素，同时逆转IL-1下调基质蛋白基因表达和蛋白多糖合成。这些结果支持了这样的假设，即在椎间盘退变的针刺模型中，IL-1可能是兔NP中所见的许多mRNA变化的原因，并支持了这样的概念，即阻断p38 MAPK的分子技术的发展可以提供一种治疗方法来减缓椎间盘退变的过程。(C)2008骨科研究学会。出版社：Wiley Periodicals，Inc.
Analysis of disc gene expression implicated IL-1 in the development of intervertebral disc degeneration (IDD) in a rabbit stab model. The purpose of these studies is to determine the role of p38 Mitogen Activated Protein Kinase (p38 MAPK) signaling in nucleus pulposus cell response to IL-1, and to compare rabbit nucleus pulposus (rNP) cell responses to IL-1 activation with those in a stab model of disc degeneration. NP cells maintained in alginate bead culture were exposed to IL-1, with or without p38 MAPK inhibition. RNA was isolated for reverse transcription polymerase chain reaction (RT-PCR) analysis of gene expression, conditioned media analyzed for accumulation of nitric oxide (NO) and prostaglandin E-2 (PGE-2), and proteoglycan synthesis measured after 10 days. IL-1 upregulation of mRNA for cycloxygenase-2 (COX-2), matrix metalloproteinase-3 (MMP-3), IL-1, and IL-6, was blunted by p38 inhibition while downregulation of matrix proteins (collagen I, collagen II, aggrecan) and insulin-like-growth-factor I (IFG-1) was also reversed. mRNA for tissue inhibitor of matrixmetalloproteinase-1 (TIMP-1) was modestly increased by IL-1, while those for Transforming Growth Factor-P (TGF-beta) SOX-9, and versican remained unchanged. Blocking p38 MAPK reduced IL-1 induced NO and PGE-2 accumulation and partially restored proteoglycan synthesis. p38 MAPK inhibition in control cells increased mRNA for matrix proteins (aggrecan, collagen II, versican, collagen 1) and anabolic factors (IGF-1, TGF, and SOX-9) from 50% to 120%, decreased basal PGE-2 accumulation, but had no effect on message for TIMP-1, MMP-3, or COX-2. Inhibition of p38 MAPK in cytokine-activated disc cells blunts gene expression and production of factors associated with inflammation, pain, and disc matrix catabolism while reversing IL-1 downregulation of matrix protein gene expression and proteoglycan synthesis. The results support the hypothesis that IL-1 could be responsible for many of the mRNA changes seen in rabbit NP in the stab model of disc degeneration, and uphold the concept that development of molecular techniques to block p38 MAPK could provide a therapeutic approach to slow the course of intervertebral disc degeneration. (C) 2008 Orthopaedic Research Society. *Published by Wiley Periodicals, Inc.