新风胶囊通过调控miR-126-VEGF/PI3K/AKT信号通路改善类风湿关节炎患者血瘀状态的作用机制研究

Effect of Xifeng Capsule on blood stasis in patients with rheumatoid arthritis by regulating miR-126-VEGF/PI3K/AKT signaling pathway

目的:从miR-126-血管内皮细胞因子(vascular endothelia growth factor,VEGF)/磷脂酰肌醇3-激酶(phosphoinositide 3-kinases,PI3K)/丝-苏氨酸蛋白激酶(serine-threonine kinase,AKT)信号通路角度探讨新风胶囊(XFC)改善类风湿关节炎(RA)患者血瘀状态的作用机制。方法:选取符合诊断标准的60例RA患者,按照随机数字表法将其分为XFC治疗组30例,来氟米特(LEF)对照组30例,治疗组口服新风胶囊(每次3粒,3次/d),对照组口服来氟米特(每次1粒,1次/d)。观察RA患者血瘀症状评分,检测实验室指标血沉(erythrocyte sedimentation rate,ESR)、C反应蛋白(C-reactive protein,CRP)、类风湿因子(rheumatoid factors,RF)、抗环瓜氨酸肽抗体(anti-cyclic citrullinated peptides,CCP)、凝血酶时间(thrombin time,TT)、部分凝血酶原时间(activated partial thromboplastin time,APTT)、凝血酶原时间(prothrombin time,PT)、D二聚体(d-dimer,D-D)、纤维蛋白原(fibrinogen,FBG)、血小板(platelet,PLT)、血小板平均体积(mean platelet volume,MPV)、血小板分布宽度(platelet distribution width,PDW)水平,实时荧光定量PCR法检测miR-126水平,ELISA法检测肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)、IL-35、VEGF、PI3K、AKT水平。采用Spearman方法对RA患者血瘀症状总积分、血瘀相关指标与疾病活动性指标、细胞因子、miR-126、VEGF、PI3K、AKT进行相关性分析。结果:两组治疗后相比,XFC组在改善血瘀症状、疾病活动性指标及降低miR-126、VEGF、PI3K、AKT、TNF-α、IL-6、D-D、FBG、PLT和升高IL-35水平方面明显优于LEF组,差异有统计学意义(P<0.05)。相关性分析显示,RA患者血瘀症状总积分、血瘀相关指标与疾病活动性指标、细胞因子、miR-126、VEGF、PI3K、AKT之间存在一定相关性。结论:RA患者体内存在血瘀状态,XFC可能通过miR-126-VEGF/PI3K/AKT信号通路改善患者细胞因子网络紊乱从而改善RA患者血瘀状态。

Objective:To discuss the mechanism of improving effect of Xinfeng Capsule(XFC)on the blood stasis of rheumatoid arthritis(RA)patients via miR-126-vascular endothelial cytokine(VEGF)/phosphatidylinositol 3-kinase(PI3K)/serthreonine protein kinase(AKT)signaling pathway.Methods:Sixty RA patients meeting the diagnostic criteria were selected and randomly divided into the XFC treatment group(30 cases)and the leflunomide(LEF)control group(30 cases)according to the random number table method.The treatment group took Xinfeng Capsules(3 capsules each time,3 times/d),and the control group took leflunomide(1 capsule each time,1 time/d).The blood stasis symptom scores of RA patients were observed,and the laboratory indicators of erythrocyte sedimentation rate(ESR),C-reactive protein(CRP),rheumatoid factor(RF),anti-cyclic citrullinated peptide antibody(CCP),thrombin time(TT),part prothrombin time(APTT),prothrombin time(PT),D dimer(DD),fibrinogen(FBG),platelets(PLT),mean platelet volume(MPV),and platelet distribution width(PDW)levels were determined.Real-time fluorescent quantitative PCR method was used to detect the level of miR-126,and the ELISA method was used to detect the levels of tumor necrosis factor(TNF-α),interleukin-6(IL-6),IL-35,VEGF,PI3K,and AKT.Spearman method was used to analyze the correlation between the total score of blood stasis symptoms,blood stasis-related indicators and disease activity indicators,cytokines,miR-126,VEGF,PI3K,and AKT in RA patients.Results:The XFC group improved blood stasis symptoms,disease activity indicators,decreased miR-126,VEGF,PI3K,AKT,TNF-α,IL-6,D-D,FBG,PLT,and increased IL-35 level,which was significantly better than the LEF group(P<0.05,P<0.01).Correlation analysis showed that there was a certain correlation between the total score of blood stasis symptoms,blood stasis-related indicators and disease activity indicators,cytokines,miR-126,VEGF,PI3K,and AKT in RA patients.Conclusion:XFC may improve the cytokine disorder of RA patients with blood stasis through miR-126-VEGF/PI3K/AKT signaling pathway,thereby improving the blood stasis status.