新风胶囊对骨关节炎血瘀状态的影响

Effect of Xinfeng Capsule in Improving Blood Stasis State of OA Patients

目的探讨骨关节炎(osteoarthritis,OA)患者的血瘀状态与核因子-κB(nuclear factor-κ-gene binding,NF-κB)通路的活化及miRNA-146之间的关系及新风胶囊对其影响。方法将70例OA患者按随机数字表法分为新风胶囊组(治疗组,每日3次,1次3粒)和氨基葡萄糖组(对照组,每日3次,1次1粒),每组35例,两组疗程均为3个月。采用ELISA检测两组血清中的NF-κB信号通路指标p50、p65、核因子κB抑制因子α(inhibitor of nuclear factorκBα,IκBα)、核因子κB激活剂1(nuclear factor kappa B activator 1,Act1)、转化生长因子激酶1(TGF-β-activated kinase 1,TAK1)和IL-1、IL-17、IL-10、血栓素A2(thromboxane A2,TXA2)及前列腺环素(prostacyclin,PGI2)的含量;荧光定量PCR检测Act1、p65、p50、TAK1mRNA水平;Western blot检测p50、p65蛋白表达水平;一步法PCR反应测定miRNA-146水平。结果 (1)与本组治疗前比较,两组治疗后血瘀积分、血小板(platelet,PLT)、D二聚体(D-dimer,D-D)、TXA2、p50、p65、IL-1、IL-17、IκBα、红细胞沉降率(erythrocyte sedimentation rate,ESR)、超敏C-反应蛋白(high-sensitivity C-reactive protein,hs-CRP)降低,p50、p65、Act1、TAK1mRNA含量降低,p50、p65蛋白表达降低,miRNA-146含量水平降低,活化部分凝血活酶时间(activated partial thromboplastin time,APTT)、凝血酶原时间(prothrombin time,PT)、PGI2、IL-10水平升高,差异均有统计学意义(P<0.05,P<0.01);与对照组比较,治疗组治疗后血瘀积分、PLT、FIB、TXA2、IL-17、hs-CRP、ESR水平降低,p65、TAK1mRNA水平降低,p65蛋白水平降低,PT、PGI2水平升高(P<0.05,P<0.01)。结论新风胶囊可能是通过上调miRNA-146,抑制NF-κB信号通路的激活,调节免疫及恢复细胞因子网络平衡,保护血管内皮细胞,从而改善骨关节炎的血瘀状态。

Objective To observe the relationship between blood stasis state and activation of nuclear factor-κ-gene binding（NF-κB）signaling pathway/miRNA-146 in osteoarthritis（OA）patients and the effect of Xinfeng Capsule（XFC）on them.Methods Totally 70 OA patients were assigned to the treatment group and the control group according to random number table,35 in each group.Patients in the treatment group took XFC,3 pills each time,3times per day,while those in the control group took Glucosamine,1 pill each time,3 times per day.The therapeutic course for all was 3 months.Serum contents of p50,p65,inhibitor of nuclear factorκBα（IκBα）,nuclear factor kappa B activator 1（Act1）,TGF-β-activated kinase 1（TAK1）,IL-1,IL-17,IL-10,and thromboxane A2（TXA2）,prostacycline（PGI2）were detected by ELISA.mRNA levels of Act1,p65,p50,and TAK1 were detected using fluorescent quantitative PCR.The protein levels of p50 and p65 were detected using Western blot.The level of miRNA-146 was detected using in one-step PCR.Results（1）Compared with pre-treatment in the same group,the levels of bloodstasis score,platelets（PLT）,D-dimer（D-D）,TXA2,IL-1,IL-17,high-sensitivity C-reactive protein（hs-CRP）,and erythrocyte sedimentation rate（ESR）all decreased;mRNA levels of p50,p65,Act1,and TAK1 were lowered;protein expressions of p50 and p65 decreased;serum levels of miRNA-146 decreased;activated partial thromboplastin time（APTT）,prothrombin time（PT）,prostaglandin 2（PGI2）,IL-10 increased in the two groups after treatment with statistical difference（P〈0.05,P〈0.01）.Compared with the control group,the levels of blood stasis score,PLT,FIB,TXA2,IL-17,hs-CRP,and ESR were lowered;mRNA expressions of p65 and TAK1 were lowered;protein expressions of p50 decreased;levels of PT and PGI2 increased in the treatment group after treatment（P〈0.05,P〈0.01）.Conclusion XFC could regulate the immunity and restore the equilibrium of cytokine network,and protect vascular endothelial cells possibly by up-regulating miRNA-146 expression and inhibiting activation of NF-κB signaling pathway,thus improving blood stasis state of OA.