调节性T细胞在急慢性痛风性关节炎转换中的作用研究
发布时间：2018-12-16 08:16
【摘要】：研究背景痛风是以高尿酸血症和反复关节炎发作为特征的一种炎症性疾病。在一些诱发因素下,尿酸盐晶体沉积在关节部位后触发一系列炎性细胞因子风暴,中性粒细胞聚集,固有免疫系统激活从而导致痛风的急性发作。一般情况下,急性痛风发作时即使没有治疗也会在2周内自然缓解,但痛风患者反复发作后,则容易迁延不愈形成慢性痛风。目前痛风的缓解机制主要强调关节局部单核/巨噬细胞、中性粒细胞等固有免疫系统自我调控,但这种机制不能解释为什么慢性痛风不能自行缓解。主要原因可能是忽视了炎症反应对适应性免疫系统的影响,特别是对炎症有抑制作用的调节性T细胞(Regulatory T,Treg)的影响。Treg是一种具有强大的抗炎作用和免疫抑制作用的辅助性T细胞,在维持自身免疫和炎症病理的稳态中起着重要作用。关于Treg细胞参与免疫抑制作用,在其他一些风湿病中(如系统性红斑狼疮)研究较多,但痛风作为一种自身炎症性疾病,Treg细胞是否在痛风急性炎症的缓解中起重要作用?慢性痛风的迁移不愈是否与其功能减退有关?目前国内外尚未有相关文献报道。因此本文主要探讨调节性T细胞在不同时期痛风患者体内的表达水平,进一步分析其在痛风急慢性转换中的作用,为痛风的防治提供新的思路。目的探讨不同时期(急性期、慢性期)痛风患者外周血淋巴细胞中CD4~+CD25~+Foxp3~+Treg细胞的百分率变化以及CD4~+T细胞中Foxp3 m RNA的相对表达量的差异,探讨其在痛风性关节炎急慢性转换中的作用。方法病例选取自2015年4月-2016年12月期间就诊于我院风湿免疫科门诊和住院部的痛风患者,鉴于痛风患者以男性为主,选取的病例均为年龄大于18岁的男性,其中痛风急性发作期患者16例,痛风慢性期36例,健康对照25例;(1)使用流式细胞仪检测急性期、慢性期痛风患者及健康对照组外周血CD4~+CD25~+Foxp3~+Treg细胞百分率。(2)采用磁珠分选的方法分选出CD4~+T细胞,提取RNA,采用荧光定量PCR(Real-time PCR,RT-PCR)检测CD4~+T中Foxp3 m RNA的相对表达量水平。使用SPSS 16.0软件进行统计分析。结果(1)急性痛风组患者外周血CD4~+CD25~+Foxp3~+Treg细胞百分率(1.659±0.335)%高于慢性痛风组(0.435±0.240)%和健康对照组(1.232±0.290)%;健康对照组外周血CD4~+CD25~+Foxp3~+Treg细胞百分率高于慢性痛风组,差异均有统计学意义(P=0.000)。(2)急性痛风组外周血CD4~+T细胞中Foxp3 m RNA表达水平[4.54(2.853,17.663)]高于正常对照组[0.921(0.591,1.769)],差异有统计学意义(Z=-3.232,P=0.001);慢性痛风组外周血CD4~+T细胞中Foxp3 m RNA表达水平高于正常对照组,差异具有统计学意义(Z=-2.310,P=0.021);急性痛风组外周血CD4~+T细胞中Foxp3 m RNA表达水平和慢性痛风组[2.913(1.605,12.233)]间比较,差异无统计学意义(Z=-1.049,P=0.294)。(3)采用Spearman等级相关分析,结果显示正常对照组、急性痛风组、慢性痛风组的Tregs细胞百分率水平与Foxp3 m RNA表达量水平均无显著相关性(均P0.05)。结论急性痛风的自限性可能与患者体内Treg细胞增多相关;慢性痛风患者体内Treg细胞减少,可能是关节炎迁延不愈的主要原因。慢性痛风患者Treg细胞变化与细胞内Foxp3 m RNA表达不同步,提示Foxp3降解增加所导致的Treg细胞减少可能是急性痛风向慢性痛风转换的重要机制。
[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.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R589.7
本文编号：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.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R589.7
【参考文献】
相关期刊论文 前1条
1 张永;陶金辉;李向培;唐江平;厉小梅;汪国生;张敏;马艳;;腺嘌呤核苷三磷酸-嘌呤受体P2X配体门控离子通道7-白细胞介素-1β通路参与痛风性关节炎发病的初步研究[J];中华风湿病学杂志;2015年05期
，本文编号：2382045
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