- 學位論文
白芨多醣體與白藜蘆醇應用於預防及早期治療退化性關節炎
The Evaluation of Polysaccharide from Bletilla Striata Mixed with Resveratrol for Osteoarthritis Prevention in Early Phase
摘要
退化性關節炎(Osteoarthritis，OA)是一種慢性關節損傷疾病，其漸進性的病程會導致軟骨纖維化或缺損，且軟骨組織受損後不易復原。對於退化性關節炎，當今醫療只能針對症狀進行緩解而無法預防或治癒該疾病，因此退化性關節炎在再生醫學研究方向為持續發展新的治療策略與生醫材料。已有文獻指出白藜蘆醇(Resveratrol) 能顯著抑制退化性關節炎產生之發炎狀態，但白藜蘆醇在進入臨床試驗後其生物藥效率並未達到共識，為了改善此點，本實驗目的為利用傳統中藥白芨多醣體(Polysaccharide Isolated from Bletilla striata)的抗氧化、抗發炎以及促進受損組織細胞修復的功能來加強白藜蘆醇減緩軟骨細胞之發炎情況。 在本研究中，將50 µM白藜蘆醇混合100 µg/ml白芨多醣體的混合物(RB)作用於受脂多醣誘導發炎的豬軟骨細胞實驗模型中，評估RB對細胞之細胞毒性、細胞外基質活性與基因表現。實驗結果顯示萃取之白芨多醣體具有正確之官能基，且RB經由WST-1試驗和 LIVE/DEAD 染色分析證實具有良好生物相容性。RB經由ICC檢測第二型膠原蛋白含量與Alcian blue染色結果顯示能維持細胞外基質的表現量，RB也能有效降低脂多醣刺激軟骨所造成的發炎基因表現量。綜合上述實驗結果白芨多醣體能維持細胞外基質的表現量，白藜蘆醇則能減輕發炎症狀，而白芨多醣體與白藜蘆醇之混合物能維持兩者之優點，因此在早期治療及預防退化性關節炎之症狀具有很大的潛力。
並列摘要
Osteoarthritis (OA) is a chronic joint injury disease. Its progressive course leads to cartilage fibrosis due to tissue nature to not easy to recover after damage. Present aids for OA are primarily focuses for alleviating the symptoms of the disease rather than prevent or cure the disease process. Therefore, development of new therapeutic strategies and biomaterials are always on-demand for OA regenerative medicine. It has been reported that resveratrol can inhibit the inflammatory rate of OA, but bioavailability of resveratrol is still controversy. In order to improve this point, the polysaccharide isolated from Bletilla striata (BSP) is added. Recent studies found that BSP has antioxidative and anti-inflammatory properties, which can strengthen the role of resveratrol to provide an anti-inflammatory effect. In this study, a mixture of 50 μM resveratrol and 100 µg/mL BSP (RB) was tested for cytotoxicity potential, then applied to an experimental model of OA-like chondrocytes induced by lipopolysaccharide (LPS) thus evaluated its ECM activity and gene expression. The experimental results showed that the BSP have the correct functional groups, and RB has been proved to have good biocompatibility through the WST-1 test and LIVE/DEAD staining. ICC type II collagen, Alcian blue staining, and gene expression evaluation showed that RB able to maintained the expression of ECM and inflammatory-related genes caused by LPS-stimulated chondrocyte, could be effectively reduced. To conclude, BSP can significantly maintain the expression of extracellular matrix, while resveratrol can reduce the symptoms of inflammation, and the mixture of BSP and resveratrol has the advantages of both, so it has great potentials for OA early treatment and prevention.