探討高分子量玻尿酸與低分子量玻尿酸在退化性關節炎中之分子機制

Investigating the mechanism of high molecular-HA and oligosaccharide-HA on pathogenesis of Osteoarthritis

陳奕璇(Yi-Hsuan Chen)

指導教授 ： 陳建和

共同指導教授 ： 梁有志

臺北醫學大學/醫學院/醫學科學研究所/碩士(2008年)

https://doi.org/10.6831/TMU.2008.00098

摘要

退化性關節炎為好發於年紀大的老年人，其特徵是因軟骨發炎導致軟骨的結構破壞，進而瓦解軟骨產生關節疼痛感。近來，對於退化性關節炎的治療包括服用非類固醇抗發炎藥物或服用葡萄糖胺來減緩疼痛，此外亦有使用外科關節置換手術。然而有些病患對於藥物反應不佳，且又礙於年紀而無法做關節置換手術，因此發展新的治療方式頗為重要。近年來研究指出，注射玻尿酸至關節腔內，不僅可潤滑關節腔及包覆軟骨，並且能修復軟骨細胞外基質成份如aggrecan、typeⅡ collagen等保護作用。先前文獻指出高分子量玻尿酸具有降低由IL-1β誘發所引起的TNF-α、MMPs 及 iNOS等，與退化性關節炎相關的發炎物質基因表現。而在本研究中加入高分子量玻尿酸至軟骨肉瘤細胞的結果發現，高分子量玻尿酸可抑制環氧化酶-2 (COX-2)及血紅素氧化酶1 (HO-1)的表現，證實了HO-1在不同組織的特異性。除此之外，高分子量玻尿酸也會提升軟骨肉瘤細胞核接受器 peroxisome proliferator-activated receptor (PPARγ)的表現。由於PPARγ的增加，表示此藥物有著抗發炎的作用，亦即證實高分子量玻尿酸具抗發炎的作用。深入探討發現，高分子量玻尿酸是經由抑制JNK磷酸化或經由提升Akt的磷酸化而抑制了NF-κB的訊息傳遞路徑來抑制這些發炎物質的產生。本研究中，我們也同時探討低分子量玻尿酸的作用，結果發現低分子量玻尿酸的作用卻是增加環氧化酶-2 ( COX-2 )及血紅素氧化酶1 (HO-1 )的表現，並且抑制PPARγ的表現量。從分子機制證實，低分子量玻尿酸是藉由提升p38及抑制Akt的磷酸化，以提升NF-κB的訊息傳遞路徑，進而增加發炎性物質的產生。

關鍵字

玻尿酸 ； 退化性關節炎

並列摘要

Osteoarthritis (OA), which is a primarily inflammatory and degenerative joint disease characterized by progressive loss of articular cartilage, is the most common form of arthritis occurred in the elder people. Loss of cartilage cushion causes friction between the bones, will lead to pain of joint. Current treatments of OA are taking Non-steroidal anti-inflammatory drugs (NSAIDs), Glucosamine and chondrotin sulfate, or Total Knee Arthroplastry (TKA). Because of poor response to medicine and too older for TKA, a new treatment for OA is important. Previous report showed that injection of Hyaluronic acid in knee joint was helpful for OA, which could provide a backbone for the attachment of aggrecan side chains by link proteins in chondrocyte. Present study aims to investigate the effects of HMW-HA on the gene expression of OA- associated cytokines and enzymes, including IL-1β, TNF-α MMP-1, MMP-13, and inducible NOS (iNOS) in primary chondrocyte from OA patients. In this study, the expression of COX-2 and HO-1 were down regulated by HMW-HA treatment in chondrosarcoma cell (SW1353). Besides, up-regulation of PPARγ could also be found in SW1353 after HMW-HA treatment. It indicated that treatment of HMW-HA had anti-inflammatory effect. Further examination showed that the regulation of MMPs、COX-2、HO-1 and PPARγ expression were through the Akt phosphorylation and NF-κB inhibition. In contrast to HMW-HA, LMW-HA would induced inflammatory mediators, including activated MMPs, COX-2 and HO-1 expression as well as down regulated PPARγ expression. These results were proved lately by in vitro studies that through activation of NF-κB and inhibition of Akt phosphorylation. From all the results obtained from this study, the HMW- and LMW-HA have opposite mechanism on the chondrosarcoma cell, SW1353.

並列關鍵字

HA ； Osteoarthritis

參考文獻

1. Cahue, S., et al., The ratio of type II collagen breakdown to synthesis and its relationship with the progression of knee osteoarthritis. Osteoarthritis and Cartilage, 2007. 15(7): p. 819-823.

3. Homandberg, G.A., Cartilage damage by matrix degradation products: Fibronectin fragments. Clinical Orthopaedics and Related Research, 2001(391 SUPPL.).

4. Ala-Aho, R. and V.M. Ka?ha?ri, Collagenases in cancer. Biochimie, 2005. 87(3-4 SPEC. ISS.): p. 273-286.

5. Sternlicht, M.D. and Z. Werb, How matrix metalloproteinases regulate cell behavior. Annual Review of Cell and Developmental Biology, 2001. 17: p. 463-516.

6. Sounni, N.E. and A. Noel, Membrane type-matrix metalloproteinases and tumor progression. Biochimie, 2005. 87(3-4 SPEC. ISS.): p. 329-342.