發(fā)布時(shí)間：2018-07-28 07:26

【摘要】：研究背景類(lèi)風(fēng)濕關(guān)節(jié)炎(RA)是一類(lèi)以長(zhǎng)病程、侵襲性破壞關(guān)節(jié)為特征的自身免疫性疾病,以多關(guān)節(jié)腫痛,滑膜炎為主要特點(diǎn),未經(jīng)正規(guī)治療可導(dǎo)致關(guān)節(jié)殘疾及功能障礙,可累及到各個(gè)臟器受累。其中,滑膜組織最容易受到累及,成纖維細(xì)胞樣滑膜細(xì)胞(FLS)具有“類(lèi)腫瘤”的特征,通過(guò)分泌炎性介質(zhì)、增殖、侵襲及遷移等行為影響了 RA的病程,在關(guān)節(jié)穩(wěn)態(tài)和RA發(fā)病機(jī)制中起重要作用,FLS的增殖、遷移行為受多種細(xì)胞因子及信號(hào)通路影響,而一種新型的抗類(lèi)風(fēng)濕藥艾拉莫德(T-614)能作用于某些共同的細(xì)胞因子及信號(hào)通路。因此本文通過(guò)實(shí)驗(yàn)研究了解艾拉莫德是否能影響FLS的增殖及遷移行為,并通過(guò)臨床觀(guān)察艾拉莫德的療效。研究目的1.體外原代培養(yǎng)RA患者滑膜細(xì)胞,了解艾拉莫德對(duì)FLS增殖及遷移功能的影響。2.觀(guān)察了解不同艾拉莫德用藥方法,并與傳統(tǒng)DMARDs聯(lián)合藥物進(jìn)行比較,了解對(duì)RA患者的臨床血清學(xué)指標(biāo)影響及病情改善情況。研究方法組織塊法培養(yǎng)RA患者滑膜細(xì)胞,無(wú)菌條件下,通過(guò)關(guān)節(jié)鏡手術(shù)選取RA患者滑膜組織,培養(yǎng)皿中并倒置放入培養(yǎng)箱內(nèi),觀(guān)察不同濃度下艾拉莫德(100、200、400、800 ng/ml)作用滑膜細(xì)胞72小時(shí)后的細(xì)胞毒作用,是否引起部分細(xì)胞凋亡,以及觀(guān)察滑膜細(xì)胞是否有遷移行為,完成細(xì)胞凋亡實(shí)驗(yàn)確定引起細(xì)胞凋亡的有效濃度并避免對(duì)隨后實(shí)驗(yàn)的干擾,并通過(guò)Edu細(xì)胞增殖實(shí)驗(yàn)、細(xì)胞遷移Transwell實(shí)驗(yàn)了解艾拉莫德對(duì)RAFLS增殖和遷移的影響,并探討可能的機(jī)制。臨床實(shí)驗(yàn)篩選確診為類(lèi)風(fēng)濕關(guān)節(jié)炎并且病情活動(dòng)的患者90人,其中男性19人,女性71人,年齡21～67歲,平均年齡47.23±15.62歲,平均病程4.75±3.53年),患者隨機(jī)分為3組,聯(lián)合治療組(31例)予艾拉莫德25mg/bid+MTX 10mg/qw,艾拉莫德單藥組(28例)艾拉莫德25mg/bid,對(duì)照組(31例)予甲氨蝶呤(MTX)10mg/qw+來(lái)氟米特(LEF)20mg/qd。分別在用藥0,1,3,6,12月觀(guān)察抗CCP抗體、RF、ESR、CRP、PLT、病人VAS評(píng)分、醫(yī)生VAS評(píng)分、DAS28-ESR、HAQ評(píng)分等指標(biāo)綜合改變情況。觀(guān)察過(guò)程中因患者個(gè)人原因或藥物不良反應(yīng)退出觀(guān)察研究8例,最后同時(shí)符合納入標(biāo)準(zhǔn)、排除標(biāo)準(zhǔn)并完成整個(gè)臨床治療觀(guān)察過(guò)程的患者共82例。本研究中所有定量資料在進(jìn)行統(tǒng)計(jì)學(xué)分析之前均進(jìn)行正態(tài)性檢驗(yàn),正態(tài)分布數(shù)據(jù)采用均數(shù)±標(biāo)準(zhǔn)差表示,非正態(tài)分布采用中位數(shù)M表示。用統(tǒng)計(jì)學(xué)方法分析治療前及治療中各時(shí)間點(diǎn)指標(biāo)情況,是否具統(tǒng)計(jì)學(xué)意義,并比較每個(gè)指標(biāo)在不同治療組隨時(shí)間變化情況,行組間比較,是否具有統(tǒng)計(jì)學(xué)意義。研究結(jié)果1.艾拉莫德(濃度達(dá)到800ng/ml時(shí))能促進(jìn)RAFLS凋亡,當(dāng)濃度≤400ng/ml時(shí)對(duì)凋亡無(wú)明顯影響;艾拉莫德(濃度達(dá)到400ng/ml時(shí))能抑制滑膜RAFLS增殖。艾拉莫德(濃度達(dá)到200ng/ml時(shí))能抑制滑膜RAFLS遷移,該抑制作用呈濃度依賴(lài)性。2.艾拉莫德能降低抗CCP抗體和RF水平,艾拉莫德聯(lián)合MTX在降抗體方面療效較傳統(tǒng)DMARDs聯(lián)合組(MTX+LEF)更有優(yōu)勢(shì)。3.傳統(tǒng)DMARDs聯(lián)合組(MTX+LEF)在血沉、CRP等指標(biāo)的遠(yuǎn)期療效方面較艾拉莫德單用組好。4.艾拉莫德與MTX聯(lián)用有協(xié)同增效作用,具有較好的安全性,是RA治療的一個(gè)值得推薦的方案。研究結(jié)論艾拉莫德能呈濃度依賴(lài)性抑制滑膜FLS增殖及遷移,能降低自身抗體水平,治療RA改善炎癥指標(biāo)及病情,安全有效,與MTX聯(lián)用能協(xié)同增效,值得推廣。
[Abstract]:Background rheumatoid arthritis (RA) is a kind of autoimmune disease characterized by long course of disease and invasive destruction of joints. It is characterized by multiple joint swelling and pain and synovitis. Without regular treatment, joint disability and dysfunction can be caused by involvement of various organs. Among them, synovial tissue is the most vulnerable and fibroblasts. FLS, which has the characteristics of "tumor like", affects the course of RA by secreting inflammatory mediators, proliferation, invasion and migration, and plays an important role in the pathogenesis of joint homeostasis and RA. The proliferation of FLS, migration behavior is influenced by many cytokines and signal pathways, and a new anti rheumatoid drug Ira Maude (T-614 It can act on some common cytokines and signaling pathways. Therefore, this article is to investigate whether Ira Maude can affect the proliferation and migration of FLS and to observe the effect of Ira Maude by clinical observation. Objective 1. primary culture RA patients' synovial cells were cultured in vitro to understand the effect of Ira Maude on the proliferation and migration of FLS.2 To observe and understand the different Ira Maude medication methods, and compare with the traditional DMARDs combined drugs, to understand the effect of the clinical serological index and the condition improvement of the RA patients. The tissue block method was used to train the synovial cells of the patients with RA. Under the aseptic condition, the synovial tissue of the patients with RA was selected by arthroscopy, and the culture Petri dish and the inversion were placed in the culture dish. In the incubator, the cytotoxic effect of Ella mod (100200400800 ng/ml) on the synovial cells after 72 hours was observed, whether some cell apoptosis was caused, and whether the synovial cells had migration behavior, the apoptosis experiment was completed to determine the effective concentration of apoptosis and avoid interference with the subsequent experiment. Edu cell proliferation test, cell migration Transwell experiment to understand the effect of Ira Maude on the proliferation and migration of RAFLS, and explore possible mechanisms. Clinical trials screening 90 patients with rheumatoid arthritis and disease activity, including 19 men, 71 women, 21~67 years old, the average age of 47.23 + 15.62 years, and the average course of 4.75 +. 3.53 years), the patients were randomly divided into 3 groups, and the combined treatment group (31 cases) was given Ira Maude 25mg/bid+MTX 10mg/qw, Ira Maude single drug group (28 cases) Ira Maude 25mg/bid, and the control group (31 cases) was given methotrexate (MTX) 10mg/qw+ leflunomide (LEF) 20mg/qd. to observe the anti CCP antibody in 0,1,3,6,12 month, respectively, RF, ESR, CRP, and the patient's score. DAS28-ESR, HAQ score and other indexes were changed. In the observation, 8 cases were withdrawn due to individual reasons or adverse drug reactions. At the same time, 82 cases were conformed to the inclusion criteria, excluding standards and completed the whole clinical observation process. The quantitative data in this study were all advanced before statistical analysis. The normal distribution of normal distribution data was represented by mean number + standard deviation, and the non normal distribution was represented by median M. Statistical analysis was used to analyze the indexes of each time point before and during the treatment. Whether there were statistical significance, and comparing each index at any time in different treatment groups, whether there was statistical difference between groups, was statistically significant. Results 1. Ella mod (when the concentration reaches 800ng/ml) can promote RAFLS apoptosis. When the concentration is less than 400ng/ml, there is no obvious effect on apoptosis; Ira Maude (when the concentration reaches 400ng/ml) can inhibit the proliferation of RAFLS in the synovial membrane. Ella mod (when the concentration reaches 200ng/ml) can inhibit the RAFLS migration of the synovium, which is a concentration dependent.2. Ella. Mod can reduce the level of anti CCP antibody and RF. The curative effect of Ira Maude combined with MTX in descending antibody is better than that of the traditional DMARDs combined group (MTX+LEF). The long-term effect of.3. traditional DMARDs combined group (MTX+LEF) in the erythrocyte sedimentation rate, CRP and other indexes is more synergistic than the combination of.4. Ella Maude and MTX, which has better safety. Full nature is a recommendable scheme for RA treatment. Conclusion Ira Maude can show the proliferation and migration of FLS in a concentration dependent suppressing synovial membrane. It can reduce the level of autoantibody and improve the inflammatory index and condition of RA. It is safe and effective. It can be synergized with the combination of MTX, and it is worth promoting.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R593.22

【參考文獻(xiàn)】

相關(guān)期刊論文 前6條

1 王衛(wèi)華;王昌正;蔣一;吳本儼;;miR-92b對(duì)人胃癌細(xì)胞SGC7901遷移、粘附和侵襲的影響[J];南方醫(yī)科大學(xué)學(xué)報(bào);2014年12期

2 江維;趙毅;林輝;劉毅;陳進(jìn);;艾拉莫德治療原發(fā)性干燥綜合征療效觀(guān)察[J];西部醫(yī)學(xué);2014年06期

3 曾小峰;朱松林;譚愛(ài)春;謝小平;;我國(guó)類(lèi)風(fēng)濕關(guān)節(jié)炎疾病負(fù)擔(dān)和生存質(zhì)量研究的系統(tǒng)評(píng)價(jià)[J];中國(guó)循證醫(yī)學(xué)雜志;2013年03期

4 周瓊竦;;甲氨蝶呤聯(lián)合用藥治療類(lèi)風(fēng)濕關(guān)節(jié)炎療效觀(guān)察[J];現(xiàn)代中西醫(yī)結(jié)合雜志;2011年04期

5 陳廷生;;來(lái)氟米特聯(lián)合甲氨蝶呤治療難治性類(lèi)風(fēng)濕關(guān)節(jié)炎30例臨床觀(guān)察[J];華夏醫(yī)學(xué);2010年04期

6 孫瑋;劉秀梅;;甲氨蝶呤聯(lián)合來(lái)氟米特或雷公藤多苷治療抗環(huán)瓜氨酸肽抗體陽(yáng)性的早期類(lèi)風(fēng)濕關(guān)節(jié)炎的療效比較[J];山西醫(yī)藥雜志;2010年01期

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