类风湿性关节炎患者高凝血状态与核因子κB活化及致炎因子增加有关

Hypercoagulable state is associated with NF-kappa B activation and increased inflammatory factors in patients with rheumatoid arthritis

目的基于核因子κB(NF-κB)信号转导通路探讨类风湿性关节炎(RA)患者高凝血状态的机制。方法选取RA患者35例,健康志愿者20例。ELISA检测两组血清中白细胞介素10(IL-10)、IL-6、IL-4、IL-17、NF-κB激活子1(Act1)、p50、p65、NF-κB抑制蛋白α(IκBα)、血小板激活因子(PAF)、PAF-乙酰水解酶(PAF-AH)、抗环瓜氨酸肽(CCP)抗体水平,全自动血细胞分析仪检测血小板(PLT)数量,魏氏法检测血沉(ESR),全自动生化仪检测C反应蛋白(CRP)、类风湿因子(RF),全自动凝血仪检测D-二聚体(D-D)、纤维蛋白原(FBG)、凝血酶时间(TT)、凝血酶原时间(PT)、部分凝血酶原时间(APTT)水平;同时半定量反转录PCR检测Act1、p65、p50、IκBα、IκB激酶α(IKKα)mRNA水平,Western blot法检测p65、p50、IκBα蛋白水平。采用Spearman分析RA患者外周血中凝血纤溶指标与细胞因子、NF-κB、活动性指标以及临床症状体征之间的相关性。结果与正常组相比,RA患者外周血中D-D、FBG、PLT明显升高,TT缩短,APTT和PT无明显改变;RA患者血清IL-4、IL-10、PAF-AH的水平明显降低,IL-6、IL-17、Act1、p50、p65、IκBα、IKKα,PAF明显升高;相关性分析显示,RA患者外周血中凝血纤溶指标与细胞因子、NF-κB、活动性指标以及临床症状体征均明显相关。结论 RA患者体内普遍存在高凝状态,并与炎性因子、活动性指标及NF-κB异常活化密切相关。

Objective To investigate the mechanism of hypercoagulable state based on nuclear factor κB（ NF-κB） pathway in patients with rheumatoid arthritis（ RA）. Methods Thirty-five RA patients were enrolled as well as 20 healthy volunteers as a control group. Interleukin-10（ IL-10）,IL-6,IL-4,IL-17,NF-κB activator 1（ Act1）,p50,p65,IκBα,platelet activating factor（ PAF）,PAF-acetylhydrolase（ PAF-AH） and anti-cyclic citrullinated peptide（ CCP） were detected using ELISA. The number of platelet（ PLT） was detected using Sysmex XT-2000 i automated hematology analyzer. The levels of D-dimer（ D-D）,fibrinogen（ FBG）,thrombin time（ TT）,prothrombin time（ PT）,and partial thromboplastin time（ APTT） were detected using Sysmex CA-1500 automatic coagulation analyzer. Erythrocyte sedimentation rate（ ESR） was detected using Westergren method. C-reactive protein and rheumatoid factor（ RF） were detected using Hitachi 7060 automatic biochemical analyzer.Meanwhile,the mRNA expressions of Act1,p65,p50,IκBα and IκB kinase α（ IKKα） were detected using semi-quantitative reverse transcription PCR. The expressions of p65,p50 and IκBα proteins were examined using Western blotting. The correlations of the above indexes were analyzed by Spearman correlation test. Results Compared with the normal group,the levels of DD,FBG,PLT significantly increased in the peripheral blood of RA patients,TT decreased,while APTT and PT were not significantly changed. IL-4,IL-10 and PAF-AH were significantly reduced in the sera of RA patients,while IL-6,IL-17,Act1,p50,p65,IκBα,IKKα and PAF were significantly elevated. Spearman correlation analysis showed that coagulant and fibrinolytic indexes were significantly correlated with cytokines,NF-κB,activity indexes and clinical symptoms and signs.Conclusion The hypercoagulable state is common in the peripheral blood of RA patients,and it is closely related to inflammatory factors,activity indexes and abnormal activation of NF-κB.