探討葡萄糖胺在人類初代滑液膜纖維母細胞抑制間質分解酵素及趨化激素的分子機制

Study of inhibitory molecular mechanisms of glucosamine on the MMPs and chemokines expression in human primary synovial fibroblast

林雅婷(Ya-Ting Lin)

指導教授 ： 陳建和 ； 許明照

臺北醫學大學/醫學院/醫學檢驗生物技術學研究所/碩士(2006年)

摘要

退化性關節炎，被認為是關節軟骨的正常功能喪失，而導致軟骨細胞的合成及分解功能失去平衡所引起的。而間質分解酵素能降解細胞外間質，故間質分解酵素在退化性關節炎病理中扮演重要角色。IL-1β 是一種細胞激素，存在於退化性關節炎軟骨及滑液膜中。IL-1β 是一個重要的調解者，其會參與基質降解及減少醣蛋白合成。IL-1β 會來調解代謝過程，包括了調控酵素的分泌，例如一氧化氮合成?、間質分解酵素及ADAMAs 。葡萄醣胺可能會刺激醣蛋白合成、抑制醣蛋白降解作用並且在體內刺激軟骨中的醣蛋白再生。在本篇碩士論文中，glucosamine 加入滑液膜纖維母細胞中，發現glucosamine 會調控MMP-1、MMP-3、MMP-13 和chemokines，例如IL-8, MCP-1 and RANTES 的protein以及 mRNA 表現。為了了解glucosamine 如何調節MMPs 及chemokines 的訊息傳遞路徑，因此探討一些蛋白激?磷酸化的訊號傳遞。從結果上可以看到glucosamine HCl 活化經由IL-1β 所誘導的MAKP pathway 中的JNK 及ERK 的磷酸化，除此之外，glucosamine 會抑制p-IκBα 磷酸化及IκBα 的分解，所以NF-κB 無法進入核內。從以上的結果證明glucosamine 在人類初代滑液膜纖維母細胞中，glucosamine會經由NF-κB這條路徑中抑制p-IκBα 磷酸化，而來抑制IL-1β活化啟動下游基因MMP-1、MMP-3、MMP-13以及chemokine的表現。

關鍵字

退化性關節炎 ； IL-1β ； 間質分解酵素 ； 趨化激素 ； 滑液膜纖維母細胞 ； 葡萄醣胺

並列摘要

Osteoarthritis is characterized by loss of the functional integrity of articular cartilage due to an imbalance between catabolic and anabolic chondrocyte activity. Degradation of the collagenous extracellular matrix by matrix metalloproteinases plays an important role in the pathogenesis of osteoarthritis. IL-1β, a cytokine presents which in elevated in osteoarthritis cartilage and synovium. IL-1β is an important mediator of increase matrix degradation and reduced proteoglycan synthesis. IL-1β mediates this process by a number of metabolic processes including up-regulating enzymes such as nitric oxide synthase, matrix metalloproteinases and ADAMDs. Glucosamine is capable of stimulating proteoglycan synthesis, inhibiting the degradation of proteoglycans, and stimulating the regeneration of cartilage in vivo. It showed that the expression of MMP-1, MMP-3, MMP-13 and chemokines such as IL-8, MCP-1 and RANTES, was down-regulated protein and mRNA production by treatment of glucosamine in human primary synovial fibroblast. To examine which signal pathways are involved in the matrix metalloproteinases and chemokines expression by glucosamine, several kinase pathways are investigated the phosphorylation status. The results showed that glucosamine HCl activated ERK and JNK pathway. Beside, glucosamine could inhibited p-IκBα phosphoylation and IκBα degradation. So that NF-κB could not ?translocated to nucleus. These results provide evidence of glucosamine inhibit IL-1β induced NF-κB pathway via p-IκBα phosphorylation down-regulation matrix metalloproteinases and chemokines in human primary synovial fibroblast.

並列關鍵字

osteoarthritis ； IL-1β ； matrix metalloproteinases ； Chemokines ； synovial fibroblast ； glucosamine

參考文獻

1. Kuettner, K.E. and A.A. Cole, Cartilage degeneration in different human joints. Osteoarthritis Cartilage, 2005. 13(2): p. 93-103.

2. McAlindon, T., Glusoamine for osteoarthritis: dawn of a new era? Lancet, 2001. 357(9252): p. 247-8.

3. Reginster, J.Y., et al., Long-term effects of glucosamine sulphate on osteoarthritis progression: a randomised, placebo-controlled clinical trial. Lancet, 2001. 357(9252): p. 251-6.

4. Pacifici, M., E. Koyama, and M. Iwamoto, Mechanisms of synovial joint and articular cartilage formation: recent advances, but many lingering mysteries. Birth Defects Res C Embryo Today, 2005. 75(3): p. 237-48.

5. Mor, A., S.B. Abramson, and M.H. Pillinger, The fibroblast-like synovial cell in rheumatoid arthritis: a key player in inflammation and joint destruction. Clin Immunol, 2005. 115(2): p. 118-28.