摘要
目的探讨磷酸酯酶与张力蛋白同源物(PTEN)类泛素化修饰在类风湿关节炎(RA)调节性T细胞(Treg)免疫功能失调中的作用,以进一步探索RA的发病机制。方法收集2021年9月至2022年12月余姚市人民医院45名健康志愿者和45例RA患者的外周血样本,ELISA检测血清中炎性因子IFN-γ和Treg相关细胞因子IL-10的表达水平,流式细胞术分析评估CD4^(+)CD25^(+)FOXP3^(+)Treg细胞的比例。蛋白印迹法检测分离的Treg细胞中类泛素化-PTEN、脂肪酸合成酶(FASN)、PTEN、泛素化FASN、Neddylation E3酶X连锁凋亡抑制蛋白(XIAP)和泛素化E3酶三分模体包含蛋白质21(TRIM21)表达水平。采用两独立样本t检验或Mann-Whitney U进行组间比较。结果与健康对照组相比,RA患者组的IFN-γ表达明显升高[(599±65)pg/ml与(1066±85)pg/ml,t=-2.06,P<0.001],而IL-10[(601±49)pg/ml与(245±41)pg/ml,t=2.05,P<0.001]和CD4^(+)CD25^(+)FOXP3^(+)Treg细胞比例[(14.3±0.4)%与(6.2±0.6)%],t=19.36,P<0.001)明显下降。RA患者Treg细胞中的FASN明显泛素化,而PTEN类泛素化水平下降。此外,RA患者TRIM21的表达水平上调(0.66±0.03与1.27±0.04,t=-20.85,P<0.001),而PTEN(0.858±0.036与0.377±0.022,t=19.36,P<0.001)、XIAP(1.107±0.065与0.530±0.015,t=15.03,P<0.001)、FASN蛋白表达水平(1.654±0.031与0.858±0.025,t=34.64,P<0.001)的表达水平下调。结论PTEN蛋白的Neddylation修饰水平下降,可能抑制了PTEN蛋白进入细胞核,从而促进了FASN的泛素化降解,抑制了Treg细胞中的脂肪酸合成,影响了Treg细胞的免疫抑制功能稳定性,并可能促进了RA的进展。
Objective To investigate the role of phosphatase and tensin homolog(PTEN)neddylation in the dysregulation of regulatory T cell(Treg)immune function in rheumatoid arthritis(RA)and further explore the pathogenesis of RA.Methods Peripheral blood samples were collected from 45 healthy volunteers and 45 RA patients in the Yuyao People′s Hospital from Sep.2021 to Dec.2022.ELISA was performed to detect the expression levels of the inflammatory factor IFN-γand Treg-related cytokine IL-10 in serum,and flow cytometry was used to assess the proportion of CD4^(+)CD25^(+)FOXP3^(+)Treg cells.Western Blot analysis was conducted to measure the expression levels of Nedd8-PTEN,ubiquitinated FASN,PTEN,FASN,Neddylation E3 enzyme XIAP,and ubiquitination E3 enzyme TRIM21 in isolated Treg cells.Group comparisons were conducted using an independent samples t-test or the Mann-Whitney U test.Results Compared with the healthy control group,the expression of IFN-γwas significantly increased[(599±65)pg/ml vs.(1066±85)pg/ml,t=-2.06,P<0.001],while IL-10 levels[(601±49)pg/ml vs.(245±41)pg/ml,t=2.05,P<0.001]and the proportion of CD4^(+)CD25^(+)FOXP3^(+)Treg cell[(14.3±0.4)%vs.(6.2±0.6)%,t=19.36,P<0.001]were significantly decreased in the RA patient group(respectively).FASN in Treg cells of RA patients was significantly ubiquitinated,while the PTEN Neddylation level decreased.Additionally,TRIM21 expression was significantly upregulated in RA patients(0.66±0.03 vs.1.27±0.04,t=-20.85,P<0.001),whereas the expression levels of PTEN(0.858±0.036 vs.0.377±0.022,t=19.36,P<0.001),XIAP(1.107±0.065 vs.0.530±0.015,t=15.03,P<0.001),and FASN protein(1.654±0.031 vs.0.858±0.025,t=34.64,P<0.001)were significantly downregulated.Conclusion Decreased levels of PTEN protein Neddylation modification may inhibit the entry of PTEN protein into the nucleus,thereby promoting the ubiquitination degradation of FASN,suppressing fatty acid synthesis in Treg cells,affecting the stability of Treg cell immunosuppressive function,and possibly promoting the progression of RA.
作者
沈孟达
魏聪
邹维
朱盈俊
魏巍
Shen Mengda;Wei Cong;Zou Wei;Zhu Yingjun;Wei Wei(Department of Rheumatology and Immunology,Affiliated Yangming Hospital of Ningbo University(Yuyao People′s Hospital),Ningbo 315400,China)
出处
《中华风湿病学杂志》
2025年第9期759-763,I0002,共6页
Chinese Journal of Rheumatology
基金
余姚市医疗卫生科技计划(2022YYB01)。
