类风湿关节炎患者关节滑液及滑膜组织中CCP/AST表达水平及临床意义

Expression Level of CCP/AST in Synovial Fluid and Synovial Tissue of Rheumatoid Arthritis Patients and Its Clinical Significance

目的探究类风湿关节炎(rheumatoid arthritis,RA)患者关节滑液及滑膜组织中环瓜氨酸肽抗原特异性T细胞(cyclic citrullinated peptide antigen-specific T cells,CCP/AST)表达水平及临床意义。方法选取石家庄市中医院2021年1月~12月128例RA患者为RA组,并取同期在该院因关节疼痛需做关节镜检查者50例为对照组。其中RA患者轻度组46例,中度组52例和重度组30例;通过蛋白印迹分析各组受试者滑膜组织类风湿因子(rheumatoid factors,RF)和抗瓜氨酸蛋白抗体(anticitrullinated protein antibodies,ACPA)蛋白表达水平;通过流式细胞仪分析受试者关节滑液中CCP/AST频率;双重免疫荧光染色/激光共聚焦扫描观察滑膜组织中CCP/AST染色强度;Pearson相关性分析评估关节滑液、滑膜组织中CCP/AST表达与RF和ACPA的相关性;采用Logistic回归分析RA发生的危险因素。结果对照组、轻度组、中度组和重度组患者滑膜组织中RF(1.01±0.01,1.53±0.03,2.01±0.08,2.66±0.12kDa)和ACPA蛋白(1.03±0.01,1.61±0.03,2.04±0.10,2.59±0.13 kDa)依次升高,差异具有统计学意义(F=14.207,12.446,均P<0.05)。对照组、轻度组、中度组和重度组患者关节滑液中CCP/AST表达依次升高(8.26%±1.68%,22.46%±3.28%,33.58%±4.37%,46.15%±5.44%),差异具有统计学意义(F=25.306,P<0.05)。对照组、轻度组、中度组和重度组患者滑膜组织CCP/AST染色强度依次升高(1.05±0.26,1.35±0.89,2.04±0.56,2.78±0.15分),差异具有统计学意义(F=70.67,P<0.05)。RA患者关节滑液、滑膜组织中CCP/AST表达与RF呈明显正相关(r=0.861,0.934,均P<0.05),与ACPA呈明显正相关(r=0.854,0.913,均P<0.05)。Logistic回归分析结果显示,高血压(OR=3.241,95%CI:1.491~6.752)、糖尿病(OR=2.565,95%CI:1.126~5.813)以及关节滑液(OR=4.450,95%CI:1.652~11.622)、滑膜组织(OR=5.629,95%CI:2.474~12.390)中CCP/AST表达均为RA发生的独立危险因素(P<0.05)。结论RA患者关节滑液及滑膜组织中CCP/AST呈现高表达,且与疾病活动度及关节破坏有关,可用于评估此类患者临床关节活动度及骨破坏程度。

Objective To investigate the expression level and clinical significance of cyclic citrullinated peptide antigen-specific T cells(CCP/AST)in synovial fluid and synovial tissue of rheumatoid arthritis(RA)patients.Methods A total of 128 RA patients in Shijiazhuang Hospital of Traditional Chinese Medicine from January to December 2021 were selected as the RA group,and 50 patients who needed arthroscopy for joint pain in the hospital during the same period were selected as the control group.Among the RA group,there were 46 cases in the mild group,52 cases in the moderate group,and 30 cases in the severe group.The protein expression levels of rheumatoid factors(RF)and anticitrullinated protein antibodies(ACPA)in synovial tissues of the subjects in each group were analyzed by Western blot.The frequency of CCP/AST in the synovial fluid of the subjects was analyzed by flow cytometry.The intensity of the staining of CCP/AST in synovial tissues was observed by double immunofluorescence staining/laser confocal scanning.Pearson correlation analysis was used to assess the correlation between the CCP/AST expression of synovial fluid and synovial tissue and RF and ACPA.Logistic regression was used to analyze the risk factors for the development of rheumatoid arthritis.Results In the order of control,mild,moderate and severe groups,RF(1.01±0.01,1.53±0.03,2.01±0.08,2.66±0.12 kDa)and ACPA proteins(1.03±0.01,1.61±0.03,2.04±0.10,2.59±0.13 kDa)in synovial tissues of patients were sequentially elevated,and the differences were all statistically significant(F=14.207,12.446,all P<0.05).The expression of CCP/AST in synovial fluid of patients in the control,mild,moderate and severe groups was increased sequentially(8.26%±1.68%,22.46%±3.28%,33.58%±4.37%,46.15%±5.44%),and the difference was statistically significant(F=25.306,P<0.05).Meanwhile,the intensity of CCP/AST staining in synovial tissues of patients in the control,mild,moderate and severe groups was also increased sequentially(1.05±0.26,1.35±0.89,2.04±0.56,2.78±0.15 score),and the difference was statistically significant(F=70.67,P<0.05).The expression of CCP/AST in the synovial fluid and synovial tissues of patients with RA was positively correlated with RF(r=0.861,0.934,all P<0.05)and ACPA in synovial fluid and synovial tissue(r=0.854,0.913,all P<0.05).Logistic regression analysis showed that hypertension(OR=3.241,95%CI:1.491~6.752),diabetes mellitus(OR=2.565,95%CI:1.126~5.813),synovial fluid(OR=4.450,95%CI:1.652~11.622),and CCP/AST expression in synovial tissues(OR=5.629,95%CI:2.474~12.390)were independent risk factors for the development of RA(P<0.05).Conclusion CCP/AST showed high expression in synovial fluid and synovial tissue of RA patients and related to disease activity and joint destruction,which can be used to assess the clinical joint mobility and bone destruction degree in such patients.