臺灣博碩士論文加值系統

骨關節炎（osteoarthritis, OA）是老年人中最常見的肌肉骨骼疾病，其症狀主要是關節疼痛或僵硬。 OA的最重要原因是隨著時間的推移，骨表面保護性關節軟骨的降解和退化，因此也被稱為退化性關節疾病。先前的許多研究表明，具有多能性和發展潛力的幹細胞，例如胚胎幹細胞（embryonic stem cells, ESCs）和誘導性多能幹細胞（induced pluripotent stem cells, iPSCs），可以用作體外分化軟骨細胞，作為研究OA藥物篩選的合適模型。相比之下，iPSCs可以從體細胞中重新編程，並且在免疫排斥上的問題較小，也較不具有道德爭議。我們實驗室先前產生的iPS細胞沒有致癌基因（c-Myc和Klf4）和病毒感染，在缺氧條件下只轉染了兩個轉錄因子（Oct4和Sox2），稱為iPS-OSH[1]（補充圖 1）。因此，本研究目的是找出一個簡單、有效的方案，以成功地將iPS-OSH分化為軟骨細胞，並用於OA研究。目前，我們成功地將iPS-OSH分化成為軟骨細胞（其幹細胞相關基因Oct4、Nanog表達水平顯著降低，軟骨形成相關基因Col2a1、Acan表達水平顯著上升），而此軟骨細胞使用細胞激素（cytokine）白介素-1α（interleukin-1α, IL-1α）處理，以模擬OA的發炎環境（其發炎反應相關基因Ccl2表達水平略有上升，軟骨分解代謝相關基因Mmp9、Mmp13與Adamts4表達水平也略有上升）。此一模式將可成為藥物篩選的平台。目前我們也已發現純化物5895s可以降低發炎反應相關基因的表達，未來將用於動物模式測試。此研究將可以開發一個OA藥物篩選平台，用於發現可抑制發炎作用的新藥並研究新藥參與的機制，進而減少或預防骨關節炎的發生。

Osteoarthritis (OA) is a most common musculoskeletal disease in the elderly, and the symptoms are mostly joint pain or stiffness. The most important cause of OA is the degradation and degeneration of protective articular cartilage on the bone surface over time, so it is also called degenerative joint disease. Many studies have previously indicated that stem cells with pluripotency and developmental potential, such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), can be used a suitable in vitro model for studying the differentiation of chondrocytes and drug screening of OA. iPSCs can be generated from somatic cells and is less controversial with immune rejection and ethical issues. Our laboratory generated an iPS cell line without oncogenes (c-Myc and Klf4) and viral infection, which transfected only two factors (Oct4 and Sox2) under hypoxic conditions, named iPS-OSH. Therefore, the aim of this study is to find out a simple and effective method to successfully differentiate iPS-OSH into chondrocytes for the subsequent study of OA. We have success differentiated iPS-OSH into chondrocytes. iPSC-derived chondrocytes were treated with cytokine interleukin-1α (IL-1α) to model the inflammatory environment of OA. It will become the platform to drug screening. We observed a novel pure compounds, 5895s, could decrease both the expression of inflammatory related genes, Ccl2, and the expression of catabolic related genes including Mmp9, Mmp13, and Adamts4. We will test 5895s anti-inflammation effect in vivo in the future. This study has established a iPSC-derived drug screening platform for OA and could be used to find out the mechanism of potential drugs and help reduce or prevent the occurrence of osteoarthritis.

中文摘要 I
英文摘要 III
目次 V
圖表目次 VI
縮寫表 VII
第一章 緒論 1
第二章 研究動機與方向 7
第三章 材料與方法 10
第四章 結果 21
第五章 結論 29
第六章 討論 30
第七章 圖表 34
第八章 參考文獻 57

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