【摘要】：背景:類風(fēng)濕關(guān)節(jié)炎(RA)以慢性和侵蝕性滑膜炎為主要特點(diǎn)的一種自身免疫病,可影響多個(gè)關(guān)節(jié)導(dǎo)致其破壞、變形,并最終造成身體殘疾,給患者帶來(lái)極大的痛苦。RA患病率為0.5%~1%,大量研究表明成纖維樣滑膜細(xì)胞(FLS)的異常增生是RA發(fā)病的關(guān)鍵。粘著斑激酶(FAK)是一種蛋白激酶,參與細(xì)胞內(nèi)的信號(hào)轉(zhuǎn)導(dǎo)。在腫瘤研究領(lǐng)域,FAK能通過(guò)其N-末端的FERM結(jié)構(gòu)域與p53結(jié)合,進(jìn)而抑制細(xì)胞凋亡,即FAK可通過(guò)抑制p53基因的表達(dá)來(lái)阻止腫瘤細(xì)胞凋亡從而導(dǎo)致腫瘤的發(fā)生。研究發(fā)現(xiàn)RA患者FLS與腫瘤細(xì)胞有許多相似的特性,如進(jìn)行性增生、遷移和侵襲,其機(jī)制可能同腫瘤細(xì)胞無(wú)限增殖及轉(zhuǎn)移的機(jī)制相似。我科前期研究表明,FAK在RA滑膜組織中表達(dá)量較正�；そM織顯著升高,且通過(guò)抗風(fēng)濕治療后,可抑制FAK表達(dá),促進(jìn)FLS凋亡,這表明FAK在FLS的過(guò)度增生過(guò)程中起重要作用。還有研究表明,p53缺失可能參與RA滑膜組織增生這一病理過(guò)程。那么在RA的發(fā)病過(guò)程中,FAK是否可與p53相互作用,使p53表達(dá)減少導(dǎo)致FLS過(guò)度增殖,從而參與RA的發(fā)病機(jī)制,值得研究。目的:1、通過(guò)體外研究FAK、p53在RA成纖維滑膜細(xì)胞中的表達(dá),探討RA的發(fā)病機(jī)制;2、探討FAK、p53對(duì)RA成纖維滑膜細(xì)胞增殖的影響,為治療RA提供新靶點(diǎn)。方法:1、取確診的RA患者滑膜組織進(jìn)行體外培養(yǎng);2、對(duì)體外培養(yǎng)的RA患者滑膜細(xì)胞進(jìn)行不同處理,分別加入TNF-α(10ng/ml)及不同濃度的蛋白酶抑制劑(MG-132)(1μM、5μM、10μM、20μM),48h后RT-PCR檢測(cè)各組FLS上FAK及p53 m RNA的水平;3、體外培養(yǎng)的滑膜細(xì)胞分別加入不同濃度的蛋白酶抑制劑(MG-132)(1μM、5μM、10μM、20μM),分別于處理后的24h、48h、72h、96h進(jìn)行細(xì)胞增殖試驗(yàn);4、采用SPSS 17.0統(tǒng)計(jì)軟件分析。結(jié)果:1、TNF-α作用后滑膜細(xì)胞的FAK m RNA水平較對(duì)照組增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05);TNF-α作用后p53 m RNA水平與對(duì)照組比較有所降低,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);2、MG-132作用后滑膜細(xì)胞的p53 m RNA水平較對(duì)照組增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05),以濃度為5μM時(shí)p53 m RNA水平最高;MG-132作用后FAK m RNA水平較對(duì)照組有所降低,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);3、不同濃度的MG-132作用后,FLS的增殖情況與對(duì)照組比較受到抑制,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:1、粘著斑激酶在體外經(jīng)TNF-α作用后其m RNA水平增高,經(jīng)MG-132作用后其m RNA水平略有降低,粘著斑激酶參與類風(fēng)濕關(guān)節(jié)炎發(fā)病過(guò)程;2、p53在體外經(jīng)MG-132作用后其m RNA水平增高,p53可抑制類風(fēng)濕關(guān)節(jié)炎成纖維滑膜細(xì)胞的過(guò)度增殖,在類風(fēng)濕關(guān)節(jié)炎發(fā)病機(jī)制中有重要作用。
[Abstract]:Background: rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic and erosive synovitis. It affects multiple joints and results in destruction, deformation, and ultimately physical disability. The prevalence of RA was 0.5%. A large number of studies showed that the abnormal proliferation of (FLS) in fibroblast synoviocytes was the key to the pathogenesis of RA. Focal adhesion kinase (FAK) is a protein kinase involved in intracellular signal transduction. In the field of tumor research, FAK can bind to p53 through its N-terminal FERM domain, which can inhibit the apoptosis of tumor cells. That is, FAK can inhibit the apoptosis of tumor cells by inhibiting the expression of p53 gene, leading to tumorigenesis. It has been found that FLS in RA patients has many similar characteristics with tumor cells, such as progressive proliferation, migration and invasion, and its mechanism may be similar to that of infinite proliferation and metastasis of tumor cells. Our previous study showed that the expression of FAK in RA synovial tissue was significantly higher than that in normal synovial tissue, and FAK could inhibit the expression of FAK and promote the apoptosis of FLS after anti-rheumatism treatment, which indicated that FAK played an important role in the process of FLS hyperproliferation. It has also been suggested that p53 deficiency may be involved in RA synovial hyperplasia. Therefore, it is worth studying whether FAK can interact with p53 in the pathogenesis of RA, which can reduce the expression of p53 and lead to excessive proliferation of FLS, and thus participate in the pathogenesis of RA. Objective to investigate the expression of FAKP p53 in RA fibroblast synoviocytes in vitro, to explore the pathogenesis of RA and the effect of FAKN p53 on the proliferation of fibroblast synoviocytes, and to provide a new target for the treatment of RA. Methods the synovial tissue of the confirmed RA patients was cultured in vitro, and the synovial cells of the RA patients were treated in different ways. TNF- 偽 (10ng/ml) and different concentrations of protease inhibitor (MG-132) (1 渭 M5 渭 MU 10 渭 MU 20 渭 M) were added to RT-PCR for 48 h. The levels of FAK and p53 m RNA on FLS were detected by RT-PCR. The cultured synovial cells were treated with different concentrations of MG-132 (1 渭 M5 渭 M10 渭 M10 渭 M20 渭 M),) respectively at 24 h, 48 h, 72 h, 96 h after treatment with TNF- 偽 (TNF- 偽) and different concentrations of protease inhibitor (MG-132). Cell proliferation test 4 was analyzed by SPSS 17.0 software. Results the level of FAK m RNA in synovial cells treated with TNF- 偽 was significantly higher than that in control group (P0.05), and the level of p53 m RNA decreased after TNF- 偽 treatment. There was no significant difference (P0.05) the level of p53 m RNA in synovial cells treated with MG-132 was significantly higher than that in control group (P0.05). The highest level of p53 m RNA was found in 5 渭 M and the level of FAK m RNA decreased after MG-132 treatment. There was no significant difference (P0.05), the proliferation of MG-132 was inhibited after different concentrations of MG-132 compared with the control group, the difference was statistically significant (P0.05). Conclusion the level of m RNA increased after TNF- 偽 treatment in vitro, and the level of m RNA decreased slightly after treatment with MG-132. Adhesion kinase participates in the pathogenesis of rheumatoid arthritis (RA) and its m RNA level increases after treated with MG-132 in vitro. P53 can inhibit the excessive proliferation of fibroblasts from rheumatoid arthritis (RA) and play an important role in the pathogenesis of rheumatoid arthritis (RA).
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R593.22

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 蘇劍敏,桂律,周逸平,查錫良;粘著斑激酶在腫瘤中的表達(dá)及其臨床意義[J];中國(guó)腫瘤;2002年11期

2 溫進(jìn)坤;韓梅;程云會(huì);楊長(zhǎng)春;劉智敏;;整合素β_3-粘著斑激酶信號(hào)途徑的活化參與大鼠血管新生內(nèi)膜的形成[J];中國(guó)病理生理雜志;2006年10期

3 尹航 ,汪麗蕙 ,霍勇 ,彭旭 ,夏春芳 ,唐朝樞;粘著斑激酶活化對(duì)平滑肌細(xì)胞粘附和遷移的影響(英文)[J];Chinese Medical Journal;2002年04期

4 梁光波,金惠銘;粘著斑激酶在血管內(nèi)皮細(xì)胞遷移中的作用[J];中國(guó)病理生理雜志;2004年06期

5 傅敏剛;平萍;范志宏;;粘著斑激酶在增生性瘢痕成纖維細(xì)胞中的表達(dá)和意義[J];中國(guó)美容整形外科雜志;2007年02期

6 蔡林;楊樹(shù)旺;李楚彥;;粘著斑激酶在腫瘤治療上的研究進(jìn)展[J];武警醫(yī)學(xué)院學(xué)報(bào);2008年09期

7 尹航,汪麗蕙,彭旭,霍勇,夏春芳,唐朝樞;粘著斑激酶活化對(duì)平滑肌細(xì)胞粘附和遷移的影響[J];中國(guó)病理生理雜志;2002年06期

8 鮑翠玉;程芳洲;柯俊;;粘著斑激酶磷酸化與大鼠血管內(nèi)膜剝脫術(shù)后血管新生內(nèi)膜中細(xì)胞增生程度的相關(guān)性(英文)[J];中國(guó)組織工程研究與臨床康復(fù);2007年49期

9 尹航,汪麗蕙,彭旭,霍勇,夏春芳,唐朝樞;粘著斑激酶反義寡核苷酸對(duì)平滑肌細(xì)胞粘附遷移和凋亡的影響[J];基礎(chǔ)醫(yī)學(xué)與臨床;2001年05期

10 方y(tǒng)N,王麗影,陳伉麗,靳嘉巍,查錫良;粘著斑激酶在TNF-α作用下影響SMMC-7721細(xì)胞PKB的表達(dá)水平[J];中國(guó)生物化學(xué)與分子生物學(xué)報(bào);2001年02期

相關(guān)會(huì)議論文 前1條

1 王華;常立文;李文斌;陳紅兵;;大鼠肺發(fā)育過(guò)程中粘著斑激酶表達(dá)變化[A];中華醫(yī)學(xué)會(huì)第五次全國(guó)圍產(chǎn)醫(yī)學(xué)學(xué)術(shù)會(huì)議論文匯編[C];2005年

相關(guān)碩士學(xué)位論文 前1條

1 張思雨;類風(fēng)濕關(guān)節(jié)炎滑膜成纖維細(xì)胞粘著斑激酶與p53對(duì)細(xì)胞增殖的研究[D];山西醫(yī)科大學(xué);2016年

，

本文編號(hào)：2182973