【摘要】：膝關(guān)節(jié)骨性關(guān)節(jié)炎(Knee osteoarthritis, KOA)是最常見的關(guān)節(jié)疾患之一,主要表現(xiàn)為關(guān)節(jié)緩慢發(fā)展的疼痛、僵硬、腫大,伴關(guān)節(jié)功能障礙,甚至發(fā)生殘疾。隨著老齡化社會的到來,KOA對人類的健康和生存質(zhì)量影響增大,本病的發(fā)病率日趨升高,對其的研究已成為醫(yī)學(xué)領(lǐng)域中的重要課題。國內(nèi)外公認的是膝關(guān)節(jié)骨關(guān)節(jié)炎是一種長期的、隱匿的慢性疾病。診斷通常是基于臨床癥狀和影像學(xué)變化。然而X線靈敏度、精確度相對比較差,目前臨床上的診斷手段不允許0A的早期診斷或關(guān)節(jié)損傷進展的評估。因此,尋找膝關(guān)節(jié)骨性關(guān)節(jié)炎的生物標志物作為0A早期的診斷標準是目前研究的一個熱門方向。蛋白質(zhì)組學(xué)能通過對全體蛋白質(zhì)表達的規(guī)�；治�,研究生物過程相關(guān)蛋白質(zhì)的動態(tài)表達和功能的改變；對于尋找疾病診斷的特異性標志物以及對疾病的發(fā)病機理的研究來說,是一種有效的高通量的研究方法,可以獲得傳統(tǒng)手段無法得到的蛋白標志物,目前已經(jīng)成為尋找新的腫瘤蛋白標志物的主要方法。本研究我們首先運用蛋白質(zhì)組學(xué)研究方法及手段,通過對膝骨性關(guān)節(jié)炎K-LO、Ⅱ、Ⅳ級的3個亞組的血清進行差異蛋白質(zhì)組學(xué)分析,篩選出了一些有潛在診斷價值的差異蛋白質(zhì),并對其中部分差異蛋白質(zhì)進行了驗證和進一步分析。目的：通過對膝骨性關(guān)節(jié)炎K/L分級的各階段的血清樣本進行ITRAQ定量蛋白組學(xué)分析檢測進行差異蛋白的篩選,以發(fā)現(xiàn)膝骨性關(guān)節(jié)炎各個時期的的潛在分子標志物。方法：收集膝關(guān)節(jié)骨性關(guān)節(jié)炎患者60例,參照Kellgren-Lawrence(K-L)分級標準,分為K-L 0、 Ⅱ、Ⅳ級的亞組,每個亞組隨機選取10例男性與10例女性。去除血清高豐度蛋白,進行穩(wěn)定同位素116、117、118的iTRAQ標記、反相色譜柱分離、質(zhì)譜檢測及Swissport數(shù)據(jù)庫檢索；再利用生物信息學(xué)軟件進行分析。對的部分差異蛋白進行Western Blot驗證。結(jié)果：通過對不同樣品的iTRAQ肽段實驗標記、Q-star質(zhì)譜鑒定和MASCOT搜庫,共篩選出有意義差異蛋白169個,K-L 0級與K-LⅣ級差異蛋白153個K-L 0級與K-L Ⅱ級差異蛋白153個；K-L II級與K-L IV級差異蛋白145個。Western Blot驗證的ADIPOQ、CRP兩個差異蛋白與實驗iTRAQ定量蛋白組學(xué)分析檢測的結(jié)果相符合。結(jié)論：1.應(yīng)用iTRAQ技術(shù)可篩選出與膝關(guān)節(jié)骨性關(guān)節(jié)炎發(fā)生發(fā)展相關(guān)的多個差異蛋白。提示iTRAQ技術(shù)對于膝關(guān)節(jié)骨性關(guān)節(jié)炎蛋白質(zhì)組學(xué)血清生物標記物的研究有很好的應(yīng)用前景。2. Adiponectin可能是診斷膝關(guān)節(jié)骨性關(guān)節(jié)炎早期潛在的血清生物標志物。同時指出脂類代謝紊亂可能才是膝骨性關(guān)節(jié)炎發(fā)病的根本原因。
[Abstract]:Knee osteoarthritis (Knee osteoarthritis, KOA) is one of the most common joint diseases, which is characterized by slow development of joint pain, stiffness, swelling, joint dysfunction and even disability. With the arrival of an aging society, the impact of KOA on human health and quality of life is increasing, the incidence of this disease is increasing, and its research has become an important subject in the field of medicine. Knee osteoarthritis is recognized at home and abroad as a long-term, hidden chronic disease. Diagnosis is usually based on clinical symptoms and imaging changes. However, X-ray sensitivity and accuracy are relatively poor. The current clinical diagnostic methods do not allow the early diagnosis of 0A or the evaluation of the progression of joint injury. Therefore, looking for biomarkers of knee osteoarthritis as an early diagnostic criterion of 0 A is a hot topic. Proteomics can study the dynamic expression and functional changes of proteins associated with biological processes through large-scale analysis of the expression of all proteins; for searching for specific markers for disease diagnosis and for studying the pathogenesis of disease, It is an effective high-throughput research method, which can obtain protein markers that can not be obtained by traditional methods, and has become the main method to find new tumor protein markers. In this study, we first used proteomics research methods and methods to analyze the differential proteomics of three subgroups of knee osteoarthritis (K-LOO, 鈪，

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