調節(jié)性T細胞在急慢性痛風性關節(jié)炎轉換中的作用研究
發(fā)布時間：2018-12-16 08:16
【摘要】：研究背景痛風是以高尿酸血癥和反復關節(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節(jié)部位后觸發(fā)一系列炎性細胞因子風暴,中性粒細胞聚集,固有免疫系統激活從而導致痛風的急性發(fā)作。一般情況下,急性痛風發(fā)作時即使沒有治療也會在2周內自然緩解,但痛風患者反復發(fā)作后,則容易遷延不愈形成慢性痛風。目前痛風的緩解機制主要強調關節(jié)局部單核/巨噬細胞、中性粒細胞等固有免疫系統自我調控,但這種機制不能解釋為什么慢性痛風不能自行緩解。主要原因可能是忽視了炎癥反應對適應性免疫系統的影響,特別是對炎癥有抑制作用的調節(jié)性T細胞(Regulatory T,Treg)的影響。Treg是一種具有強大的抗炎作用和免疫抑制作用的輔助性T細胞,在維持自身免疫和炎癥病理的穩(wěn)態(tài)中起著重要作用。關于Treg細胞參與免疫抑制作用,在其他一些風濕病中(如系統性紅斑狼瘡)研究較多,但痛風作為一種自身炎癥性疾病,Treg細胞是否在痛風急性炎癥的緩解中起重要作用?慢性痛風的遷移不愈是否與其功能減退有關?目前國內外尚未有相關文獻報道。因此本文主要探討調節(jié)性T細胞在不同時期痛風患者體內的表達水平,進一步分析其在痛風急慢性轉換中的作用,為痛風的防治提供新的思路。目的探討不同時期(急性期、慢性期)痛風患者外周血淋巴細胞中CD4~+CD25~+Foxp3~+Treg細胞的百分率變化以及CD4~+T細胞中Foxp3 m RNA的相對表達量的差異,探討其在痛風性關節(jié)炎急慢性轉換中的作用。方法病例選取自2015年4月-2016年12月期間就診于我院風濕免疫科門診和住院部的痛風患者,鑒于痛風患者以男性為主,選取的病例均為年齡大于18歲的男性,其中痛風急性發(fā)作期患者16例,痛風慢性期36例,健康對照25例;(1)使用流式細胞儀檢測急性期、慢性期痛風患者及健康對照組外周血CD4~+CD25~+Foxp3~+Treg細胞百分率。(2)采用磁珠分選的方法分選出CD4~+T細胞,提取RNA,采用熒光定量PCR(Real-time PCR,RT-PCR)檢測CD4~+T中Foxp3 m RNA的相對表達量水平。使用SPSS 16.0軟件進行統計分析。結果(1)急性痛風組患者外周血CD4~+CD25~+Foxp3~+Treg細胞百分率(1.659±0.335)%高于慢性痛風組(0.435±0.240)%和健康對照組(1.232±0.290)%;健康對照組外周血CD4~+CD25~+Foxp3~+Treg細胞百分率高于慢性痛風組,差異均有統計學意義(P=0.000)。(2)急性痛風組外周血CD4~+T細胞中Foxp3 m RNA表達水平[4.54(2.853,17.663)]高于正常對照組[0.921(0.591,1.769)],差異有統計學意義(Z=-3.232,P=0.001);慢性痛風組外周血CD4~+T細胞中Foxp3 m RNA表達水平高于正常對照組,差異具有統計學意義(Z=-2.310,P=0.021);急性痛風組外周血CD4~+T細胞中Foxp3 m RNA表達水平和慢性痛風組[2.913(1.605,12.233)]間比較,差異無統計學意義(Z=-1.049,P=0.294)。(3)采用Spearman等級相關分析,結果顯示正常對照組、急性痛風組、慢性痛風組的Tregs細胞百分率水平與Foxp3 m RNA表達量水平均無顯著相關性(均P0.05)。結論急性痛風的自限性可能與患者體內Treg細胞增多相關;慢性痛風患者體內Treg細胞減少,可能是關節(jié)炎遷延不愈的主要原因。慢性痛風患者Treg細胞變化與細胞內Foxp3 m RNA表達不同步,提示Foxp3降解增加所導致的Treg細胞減少可能是急性痛風向慢性痛風轉換的重要機制。
[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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R589.7
本文編號：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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R589.7
【參考文獻】
相關期刊論文 前1條
1 張永;陶金輝;李向培;唐江平;厲小梅;汪國生;張敏;馬艷;;腺嘌呤核苷三磷酸-嘌呤受體P2X配體門控離子通道7-白細胞介素-1β通路參與痛風性關節(jié)炎發(fā)病的初步研究[J];中華風濕病學雜志;2015年05期
，本文編號：2382045
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