調(diào)節(jié)性T細(xì)胞在急慢性痛風(fēng)性關(guān)節(jié)炎轉(zhuǎn)換中的作用研究
發(fā)布時(shí)間：2018-12-16 08:16
【摘要】：研究背景痛風(fēng)是以高尿酸血癥和反復(fù)關(guān)節(jié)炎發(fā)作為特征的一種炎癥性疾病。在一些誘發(fā)因素下,尿酸鹽晶體沉積在關(guān)節(jié)部位后觸發(fā)一系列炎性細(xì)胞因子風(fēng)暴,中性粒細(xì)胞聚集,固有免疫系統(tǒng)激活從而導(dǎo)致痛風(fēng)的急性發(fā)作。一般情況下,急性痛風(fēng)發(fā)作時(shí)即使沒(méi)有治療也會(huì)在2周內(nèi)自然緩解,但痛風(fēng)患者反復(fù)發(fā)作后,則容易遷延不愈形成慢性痛風(fēng)。目前痛風(fēng)的緩解機(jī)制主要強(qiáng)調(diào)關(guān)節(jié)局部單核/巨噬細(xì)胞、中性粒細(xì)胞等固有免疫系統(tǒng)自我調(diào)控,但這種機(jī)制不能解釋為什么慢性痛風(fēng)不能自行緩解。主要原因可能是忽視了炎癥反應(yīng)對(duì)適應(yīng)性免疫系統(tǒng)的影響,特別是對(duì)炎癥有抑制作用的調(diào)節(jié)性T細(xì)胞(Regulatory T,Treg)的影響。Treg是一種具有強(qiáng)大的抗炎作用和免疫抑制作用的輔助性T細(xì)胞,在維持自身免疫和炎癥病理的穩(wěn)態(tài)中起著重要作用。關(guān)于Treg細(xì)胞參與免疫抑制作用,在其他一些風(fēng)濕病中(如系統(tǒng)性紅斑狼瘡)研究較多,但痛風(fēng)作為一種自身炎癥性疾病,Treg細(xì)胞是否在痛風(fēng)急性炎癥的緩解中起重要作用?慢性痛風(fēng)的遷移不愈是否與其功能減退有關(guān)?目前國(guó)內(nèi)外尚未有相關(guān)文獻(xiàn)報(bào)道。因此本文主要探討調(diào)節(jié)性T細(xì)胞在不同時(shí)期痛風(fēng)患者體內(nèi)的表達(dá)水平,進(jìn)一步分析其在痛風(fēng)急慢性轉(zhuǎn)換中的作用,為痛風(fēng)的防治提供新的思路。目的探討不同時(shí)期(急性期、慢性期)痛風(fēng)患者外周血淋巴細(xì)胞中CD4~+CD25~+Foxp3~+Treg細(xì)胞的百分率變化以及CD4~+T細(xì)胞中Foxp3 m RNA的相對(duì)表達(dá)量的差異,探討其在痛風(fēng)性關(guān)節(jié)炎急慢性轉(zhuǎn)換中的作用。方法病例選取自2015年4月-2016年12月期間就診于我院風(fēng)濕免疫科門(mén)診和住院部的痛風(fēng)患者,鑒于痛風(fēng)患者以男性為主,選取的病例均為年齡大于18歲的男性,其中痛風(fēng)急性發(fā)作期患者16例,痛風(fēng)慢性期36例,健康對(duì)照25例;(1)使用流式細(xì)胞儀檢測(cè)急性期、慢性期痛風(fēng)患者及健康對(duì)照組外周血CD4~+CD25~+Foxp3~+Treg細(xì)胞百分率。(2)采用磁珠分選的方法分選出CD4~+T細(xì)胞,提取RNA,采用熒光定量PCR(Real-time PCR,RT-PCR)檢測(cè)CD4~+T中Foxp3 m RNA的相對(duì)表達(dá)量水平。使用SPSS 16.0軟件進(jìn)行統(tǒng)計(jì)分析。結(jié)果(1)急性痛風(fēng)組患者外周血CD4~+CD25~+Foxp3~+Treg細(xì)胞百分率(1.659±0.335)%高于慢性痛風(fēng)組(0.435±0.240)%和健康對(duì)照組(1.232±0.290)%;健康對(duì)照組外周血CD4~+CD25~+Foxp3~+Treg細(xì)胞百分率高于慢性痛風(fēng)組,差異均有統(tǒng)計(jì)學(xué)意義(P=0.000)。(2)急性痛風(fēng)組外周血CD4~+T細(xì)胞中Foxp3 m RNA表達(dá)水平[4.54(2.853,17.663)]高于正常對(duì)照組[0.921(0.591,1.769)],差異有統(tǒng)計(jì)學(xué)意義(Z=-3.232,P=0.001);慢性痛風(fēng)組外周血CD4~+T細(xì)胞中Foxp3 m RNA表達(dá)水平高于正常對(duì)照組,差異具有統(tǒng)計(jì)學(xué)意義(Z=-2.310,P=0.021);急性痛風(fēng)組外周血CD4~+T細(xì)胞中Foxp3 m RNA表達(dá)水平和慢性痛風(fēng)組[2.913(1.605,12.233)]間比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(Z=-1.049,P=0.294)。(3)采用Spearman等級(jí)相關(guān)分析,結(jié)果顯示正常對(duì)照組、急性痛風(fēng)組、慢性痛風(fēng)組的Tregs細(xì)胞百分率水平與Foxp3 m RNA表達(dá)量水平均無(wú)顯著相關(guān)性(均P0.05)。結(jié)論急性痛風(fēng)的自限性可能與患者體內(nèi)Treg細(xì)胞增多相關(guān);慢性痛風(fēng)患者體內(nèi)Treg細(xì)胞減少,可能是關(guān)節(jié)炎遷延不愈的主要原因。慢性痛風(fēng)患者Treg細(xì)胞變化與細(xì)胞內(nèi)Foxp3 m RNA表達(dá)不同步,提示Foxp3降解增加所導(dǎo)致的Treg細(xì)胞減少可能是急性痛風(fēng)向慢性痛風(fēng)轉(zhuǎn)換的重要機(jī)制。
[Abstract]:Background of the study gout is an inflammatory disease characterized by hyperuricemia and repeated arthritis. In some of the inducing factors, a series of inflammatory cytokine storms, the aggregation of neutrophils, and the activation of the innate immune system are triggered after the deposition of the urate crystals on the joint site, leading to an acute attack of gout. In general, even if there is no treatment in the acute gout attack, it will be naturally relieved within 2 weeks, but after repeated episodes of the gout, the chronic gout can be easily delayed. At present, the mechanism of the relief of gout mainly emphasizes the self-regulation of the intrinsic immune system such as the local mononuclear/ macrophage and the neutrophils, but the mechanism cannot explain why the chronic gout can not be self-relieved. The primary cause may be to ignore the effects of inflammatory reactions on the adaptive immune system, in particular regulatory T cells (Treg) that have an inhibitory effect on inflammation. Treg is an auxiliary T-cell with strong anti-inflammatory and immunosuppression effects and plays an important role in maintaining the steady state of autoimmune and inflammatory pathologies. As regards the involvement of Treg cells in immunosuppression, there are more studies in other rheumatism, such as systemic lupus erythematosus, but gout is an autoimmune disease, and whether Treg cells play an important role in the response of gout acute inflammation? Whether the migration of chronic gout is related to the decrease of its function? At present, no relevant literature has been reported at home and abroad. Therefore, this paper mainly discusses the expression level of the regulatory T cells in the patients with gout during different times, and further analyzes the role of the regulatory T cells in the acute and chronic conversion of gout, and provides a new idea for the prevention and treatment of gout. Objective To study the changes of the percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells in peripheral blood lymphocytes of patients with gout (acute and chronic) and the relative expression of Foxp3 mRNA in CD4 ~ + T cells. Methods The cases of gout were selected from April 2015 to December 2016 in the patients with gout in the outpatient department and the hospitalization department of the rheumatic and immunocology department of our hospital. In view of the fact that the patients with gout were male, the selected cases were male with age of more than 18 years, of which 16 cases of the patients with acute onset of gout, (1) The percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells in peripheral blood of patients with acute and chronic gout and healthy control group were detected by flow cytometry. (2) CD4 ~ + T cells were selected by magnetic bead sorting, and the relative expression level of Foxp3 mRNA in CD4 ~ + T was detected by fluorescence quantitative PCR (RT-PCR). The statistical analysis was performed using the SPSS 10.0 software. Results (1) The percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells in the peripheral blood of the acute gout group was higher than that of the chronic gout group (0.435-0.240)% and the healthy control group (1.232-0.290)%; the percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells in the peripheral blood of the healthy control group was higher than that of the chronic gout group (P = 0.000). (2) The expression of Foxp3 mRNA in the peripheral blood CD4 ~ + T cells in the acute gout group was higher than that in the normal control group[0.921 (0.591, 1.769)], and the difference was significant (Z =-3.232, P = 0.001); the expression level of Foxp3 mRNA in the peripheral blood CD4 ~ + T cells in the chronic gout group was higher than that of the normal control group (Z =-2.310, The expression of Foxp3 mRNA in the peripheral blood of the acute gout group and the expression of Foxp3 mRNA in the peripheral blood of the acute gout group and the chronic gout group[2. 913 (1. 605, 12. 233)] were not statistically significant (Z =-1.049, P = 0.294). (3) Spearman grade correlation was used to show that the percentage of Tregs in the normal control group, the acute gout group and the chronic gout group had no significant correlation with the level of the expression of Foxp3 mRNA (P0.05). Conclusion The self-infection of acute gout may be related to the increase of Treg cells in the patients. The decrease of Treg cells in patients with chronic gout may be the main cause of the non-recovery of arthritis. The changes of Treg cells in patients with chronic gout were not synchronized with the expression of Foxp3 mRNA in the cells, suggesting that the decrease of Treg cells caused by the increase of Foxp3 could be an important mechanism for acute gout to chronic gout.
【學(xué)位授予單位】：安徽醫(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)】：R589.7
本文編號(hào)：2382045
[Abstract]:Background of the study gout is an inflammatory disease characterized by hyperuricemia and repeated arthritis. In some of the inducing factors, a series of inflammatory cytokine storms, the aggregation of neutrophils, and the activation of the innate immune system are triggered after the deposition of the urate crystals on the joint site, leading to an acute attack of gout. In general, even if there is no treatment in the acute gout attack, it will be naturally relieved within 2 weeks, but after repeated episodes of the gout, the chronic gout can be easily delayed. At present, the mechanism of the relief of gout mainly emphasizes the self-regulation of the intrinsic immune system such as the local mononuclear/ macrophage and the neutrophils, but the mechanism cannot explain why the chronic gout can not be self-relieved. The primary cause may be to ignore the effects of inflammatory reactions on the adaptive immune system, in particular regulatory T cells (Treg) that have an inhibitory effect on inflammation. Treg is an auxiliary T-cell with strong anti-inflammatory and immunosuppression effects and plays an important role in maintaining the steady state of autoimmune and inflammatory pathologies. As regards the involvement of Treg cells in immunosuppression, there are more studies in other rheumatism, such as systemic lupus erythematosus, but gout is an autoimmune disease, and whether Treg cells play an important role in the response of gout acute inflammation? Whether the migration of chronic gout is related to the decrease of its function? At present, no relevant literature has been reported at home and abroad. Therefore, this paper mainly discusses the expression level of the regulatory T cells in the patients with gout during different times, and further analyzes the role of the regulatory T cells in the acute and chronic conversion of gout, and provides a new idea for the prevention and treatment of gout. Objective To study the changes of the percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells in peripheral blood lymphocytes of patients with gout (acute and chronic) and the relative expression of Foxp3 mRNA in CD4 ~ + T cells. Methods The cases of gout were selected from April 2015 to December 2016 in the patients with gout in the outpatient department and the hospitalization department of the rheumatic and immunocology department of our hospital. In view of the fact that the patients with gout were male, the selected cases were male with age of more than 18 years, of which 16 cases of the patients with acute onset of gout, (1) The percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells in peripheral blood of patients with acute and chronic gout and healthy control group were detected by flow cytometry. (2) CD4 ~ + T cells were selected by magnetic bead sorting, and the relative expression level of Foxp3 mRNA in CD4 ~ + T was detected by fluorescence quantitative PCR (RT-PCR). The statistical analysis was performed using the SPSS 10.0 software. Results (1) The percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells in the peripheral blood of the acute gout group was higher than that of the chronic gout group (0.435-0.240)% and the healthy control group (1.232-0.290)%; the percentage of CD4 ~ + CD25 ~ + Foxp3 ~ + Treg cells in the peripheral blood of the healthy control group was higher than that of the chronic gout group (P = 0.000). (2) The expression of Foxp3 mRNA in the peripheral blood CD4 ~ + T cells in the acute gout group was higher than that in the normal control group[0.921 (0.591, 1.769)], and the difference was significant (Z =-3.232, P = 0.001); the expression level of Foxp3 mRNA in the peripheral blood CD4 ~ + T cells in the chronic gout group was higher than that of the normal control group (Z =-2.310, The expression of Foxp3 mRNA in the peripheral blood of the acute gout group and the expression of Foxp3 mRNA in the peripheral blood of the acute gout group and the chronic gout group[2. 913 (1. 605, 12. 233)] were not statistically significant (Z =-1.049, P = 0.294). (3) Spearman grade correlation was used to show that the percentage of Tregs in the normal control group, the acute gout group and the chronic gout group had no significant correlation with the level of the expression of Foxp3 mRNA (P0.05). Conclusion The self-infection of acute gout may be related to the increase of Treg cells in the patients. The decrease of Treg cells in patients with chronic gout may be the main cause of the non-recovery of arthritis. The changes of Treg cells in patients with chronic gout were not synchronized with the expression of Foxp3 mRNA in the cells, suggesting that the decrease of Treg cells caused by the increase of Foxp3 could be an important mechanism for acute gout to chronic gout.
【學(xué)位授予單位】：安徽醫(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)】：R589.7
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 張永;陶金輝;李向培;唐江平;厲小梅;汪國(guó)生;張敏;馬艷;;腺嘌呤核苷三磷酸-嘌呤受體P2X配體門(mén)控離子通道7-白細(xì)胞介素-1β通路參與痛風(fēng)性關(guān)節(jié)炎發(fā)病的初步研究[J];中華風(fēng)濕病學(xué)雜志;2015年05期
，本文編號(hào)：2382045
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