【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(huì)的到來,KOA對(duì)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(duì)其的研究已成為醫(yī)學(xué)領(lǐng)域中的重要課題。國(guó)內(nèi)外公認(rèn)的是膝關(guān)節(jié)骨關(guān)節(jié)炎是一種長(zhǎng)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(duì)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(jìn)展的評(píng)估。因此,尋找膝關(guān)節(jié)骨性關(guān)節(jié)炎的生物標(biāo)志物作為0A早期的診斷標(biāo)準(zhǔn)是目前研究的一個(gè)熱門方向。蛋白質(zhì)組學(xué)能通過對(duì)全體蛋白質(zhì)表達(dá)的規(guī)�；治�,研究生物過程相關(guān)蛋白質(zhì)的動(dòng)態(tài)表達(dá)和功能的改變；對(duì)于尋找疾病診斷的特異性標(biāo)志物以及對(duì)疾病的發(fā)病機(jī)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(biāo)志物,目前已經(jīng)成為尋找新的腫瘤蛋白標(biāo)志物的主要方法。本研究我們首先運(yùn)用蛋白質(zhì)組學(xué)研究方法及手段,通過對(duì)膝骨性關(guān)節(jié)炎K-LO、Ⅱ、Ⅳ級(jí)的3個(gè)亞組的血清進(jìn)行差異蛋白質(zhì)組學(xué)分析,篩選出了一些有潛在診斷價(jià)值的差異蛋白質(zhì),并對(duì)其中部分差異蛋白質(zhì)進(jìn)行了驗(yàn)證和進(jìn)一步分析。目的：通過對(duì)膝骨性關(guān)節(jié)炎K/L分級(jí)的各階段的血清樣本進(jìn)行ITRAQ定量蛋白組學(xué)分析檢測(cè)進(jìn)行差異蛋白的篩選,以發(fā)現(xiàn)膝骨性關(guān)節(jié)炎各個(gè)時(shí)期的的潛在分子標(biāo)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(jí)標(biāo)準(zhǔn),分為K-L 0、 Ⅱ、Ⅳ級(jí)的亞組,每個(gè)亞組隨機(jī)選取10例男性與10例女性。去除血清高豐度蛋白,進(jìn)行穩(wěn)定同位素116、117、118的iTRAQ標(biāo)記、反相色譜柱分離、質(zhì)譜檢測(cè)及Swissport數(shù)據(jù)庫檢索；再利用生物信息學(xué)軟件進(jìn)行分析。對(duì)的部分差異蛋白進(jìn)行Western Blot驗(yàn)證。結(jié)果：通過對(duì)不同樣品的iTRAQ肽段實(shí)驗(yàn)標(biāo)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(gè),K-L 0級(jí)與K-LⅣ級(jí)差異蛋白153個(gè)K-L 0級(jí)與K-L Ⅱ級(jí)差異蛋白153個(gè)；K-L II級(jí)與K-L IV級(jí)差異蛋白145個(gè)。Western Blot驗(yàn)證的ADIPOQ、CRP兩個(gè)差異蛋白與實(shí)驗(yàn)iTRAQ定量蛋白組學(xué)分析檢測(cè)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(gè)差異蛋白。提示iTRAQ技術(shù)對(duì)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(biāo)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(biāo)志物。同時(shí)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，

本文編號(hào)：2146067