新风胶囊通过抑制miR-155/NF-κB信号通路改善强直性脊柱炎活动期患者高凝状态

Xinfeng capsule improves hypercoagulative state by inhibiting miR-155/NF-κB signaling pathway in patients with active ankylosing spondylitis

目的新风胶囊(XFC)对强直性脊柱炎(AS)活动期患者miR-155、核因子κB(NF-κB)信号通路、高凝状态的影响,探讨其可能的机制。方法采用随机数字表法将56例AS活动期患者分为新风胶囊组(XFC组)和柳氮磺吡啶组(SASP组)。实时荧光定量PCR检测miR-155。反转录PCR检测核因子κB激活蛋白1(Act1)、NF-κB抑制蛋白α(IκBα)、IκB激酶β(IKKβ)、NF-κB p65、NF-κB p50 mRNA的水平,Western blot法检测NF-κB P65、NF-κB P50蛋白表达,ELISA检测血清血栓素B2(TXB2)、6-酮-前列环素F1(6-keto-PGF1)、血小板颗粒膜蛋白(GMP140)、血小板活化因子(PAF)、纤溶酶原激活抑制剂(PAI-2)、肿瘤坏死因子α(TNF-α)、白细胞介素4(IL-4)、IL-10、IL-17。同时评价XFC对AS患者临床疗效。结果 XFC组Bath AS疾病活动指数50%达标率(BASDAI50)显著高于SASP组。与SASP组治疗后相比,XFC组治疗后血小板(PLT)、纤维蛋白原(FBG)、D-D二聚体(D-D)、TXB2、GMP140、PAF、PAI-2、IL-17、血沉(ESR)、C反应蛋白(CRP)、视觉模拟评分(VAS)、Bath AS疾病活动指数(BASDAI)、Bath AS功能指数(BASFI)、Bath AS总体指数(BAS-G)降低更明显,且其可明显升高6-keto-PGF1、IL-4、IL-10;与SASP治疗后相比,XFC治疗后IKKβ、IκBα、NF-κB p65、NF-κB p50 mRNA及NF-κB P65、NF-κB P50蛋白、miR-155表达水平更低。结论 XFC能有效改善AS活动期患者的高凝状态,可能与抑制miR-155、抑制NF-κB信号通路活化有关。

Objective To observe the effect of Xinfeng capsule （XFC） on miR-155, nuclear factor kappa B （NF-KB） signal pathway, and indexes related to hypercoagulative state in patients with active ankylosing spondylitis （AS）, and investigate the possible mechanism. Methods Fifty-six cases in active AS were randomly divided into XFC group and sulfasalazine （SASP） group. All cases in the XFC group took three capsules three times daily for twelve consecutive weeks. The ones in the SASP group took four tablets two times daily for twelve consecutive weeks. The expression of miR-155 was detected by real-time PCR. The mRNA expressions of nuclear factor KB activator 1 （Actl）, NF-KB inhibitor-alpha （IKBa）, inhibitor of kappa-B kinase beta （ IKKIS）, NF-KB p65, and NF-KB pS0 were tested by reverse transcription PCR （RT-PCR）. Meanwhile, the protein expressions of NF-KB P65 and NF-KB PS0 were determined by Western blotting. Tumor necrosis factor-alpha （TNF-a）, interleukin （ IL）-4, IL-10, IL-11, thromboxane B2 （TXB2）, 6-ketone-prostaglandin FI （6-keto-PGFl）, platelet granular membrane protein 140 （GMP140）, platelet activation factor （PAF）, and plasminogen activator inhibitor-2 （PAl-2） were determined by ELISA. Clinical efficacy was evaluated in the two groups. Results Compared with the SASP group, 50% Bath ankylosing spondylitis disease activity index （BASDAISO） was significantly higher in the XFC group. Compared with theSASP group after treatment, platelet （PLT）, fibrinogen （FBG） and D-D dimer （D-D）, TXB2, GMPI40, PAF, PAl-2, IL-17, erythrocyte sedimentation rate （ESR）, C-reactive protein （CRP）, visual analog scale （VAS）, BASDAI, BASFI, and BAS-G were reduced more obviously in the XFC group after treatment; meanwhile, 6-keto-PGF1, IL-4, and IL-10 significantly increased. Compared with the SASP group after treatment, the expressions of IKK mRNA, IKBa mRNA, NF-KB 1065 mRNA, NF-KB pSO mRNA, NF-KB P65 protein, NF-KB P50 protein, and miR-155 were lower in the XFC group after treatment. Conclusion XFC could effectively improve hypercoagulative state in active AS patients. The potential mechanism may be associated with the inhibition of miR-155 and NF-KB signal pathway.