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目的基于Act1/NF-κB信号传导通路研究新风胶囊(Xinfeng Capsule,XFC)改善类风湿关节炎患者血瘀状态的机制。方法将76例RA患者按随机数字表法分为2组:XFC组(3粒/次,每日3次)和来氟米特(LEF)组(10 mg/次,每日1次),每组38例,连续治疗3个月。评定两组中医证候临床疗效;ELISA法检测血清白介素-10(Interleukin-10,IL-10)、IL-6、IL-17、核因子κB激活蛋白1抗体(NF-κB activator 1,Act1)、p50、p65,血小板凝聚因子(platelet activating factor,PAF)、血小板活化因子乙酰水解酶(platelet activating factor acetyl hydrolase,PAF-AH);评定血瘀证症状的评分;采用荧光定量PCR法检测Act1、p65、p50mRNA表达;Western blot法检测p50、p65蛋白表达。采用Spearman方法对RA患者外周血凝血指标及血瘀证总积分与IL-10、IL-6、IL-17、Act1、p50、p65之间进行相关性分析。结果 XFC组总有效率为89.5%(34/38);LEF组总有效率为94.7%(36/38),两组总有效率比较,差异无统计学意义(P>0.05)。与本组治疗前比较,两组治疗后D-D、FBG、PLT、PAF、IL-17及IL-6水平下降,Act1、p50、p65mRNA表达及含量水平降低,p50、p65蛋白水平降低,血瘀证各个症状评分均降低,而PAF-AH及IL-10水平上升(P<0.05,P<0.01);与LEF组比较,XFC组治疗后D-D、FBG、PLT、IL-17及IL-6水平下降,Act1、p65mRNA表达及含量水平降低,p 65蛋白表达降低,关节刺痛、舌质、皮下瘀斑及血瘀证总积分的评分均明显降低(P<0.05,P<0.01)。RA外周血D-D与IL-17、IL-6、Act1、p65呈正相关,与IL-10呈负相关(P<0.05,P<0.01);FBG与IL-6呈正相关(P<0.05);PLT与IL-17呈正相关(P<0.05);血瘀证总积分与IL-6、Act1、p65呈正相关,与IL-10呈负相关(P<0.05,P<0.01);PAF与IL-17、IL-6、Act1、p65呈正相关(P<0.05,P<0.01);PAF-AH与p50呈负相关(P<0.05)。结论 RA患者血瘀状态的发生机制及新风胶囊对血瘀状态的影响可能是与Act1/NF-κB信号传导通路密切相关。
Objective To study the mechanism of Xinfeng Capsule (XFC) on improving the blood stasis status of patients with rheumatoid arthritis based on Act1 / NF-κB signaling pathway. Methods 76 patients with RA were divided into two groups according to the random number table: XFC group (3 capsules / time, 3 times daily) and LEF group (10 mg / time, once daily) 38 cases in each group, continuous treatment for 3 months. The clinical efficacy of TCM syndromes was evaluated in both groups. The levels of serum IL-10, IL-6, IL-17 and Act1 were detected by ELISA. , P50, p65, platelet activating factor (PAF) and platelet activating factor acetyl-hydrolase (PAF-AH) were measured. The scores of symptoms of blood stasis were evaluated. Fluorescent quantitative PCR was used to detect the expression of Act1, p65, p50mRNA expression; Western blot detection of p50, p65 protein expression. The Spearman method was used to analyze the correlation between the blood clotting indexes and the total score of blood stasis syndrome and IL-10, IL-6, IL-17, Act1, p50 and p65 in RA patients. Results The total effective rate was 89.5% (34/38) in the XFC group. The total effective rate in the LEF group was 94.7% (36/38). There was no significant difference in the total effective rate between the two groups (P> 0.05). Compared with before treatment, the levels of DD, FBG, PLT, PAF, IL-17 and IL-6 in the two groups decreased, the expression and content of Act1, p50 and p65 mRNA decreased, the levels of p50 and p65 decreased, The levels of PAF-AH and IL-10 in the XFC group were significantly lower than those in the LEF group (P <0.05, P <0.01). The levels of DD, FBG, PLT, IL-17 and IL- The expression of Act1 and p65 mRNA decreased, the expression of p65 protein decreased, the scores of joint pain, tongue, subcutaneous ecchymosis and blood stasis syndrome were significantly decreased (P <0.05, P <0.01). There was a positive correlation between DD and IL-17, IL-6, Act1 and p65 in RA peripheral blood and a negative correlation with IL-10 (P <0.05, P <0.01) (P <0.05, P <0.05). The total score of blood stasis syndrome was positively correlated with IL-6, Act1, p65 and negatively correlated with IL-10 , IL-6, Act1 and p65 (P <0.05, P <0.01). There was a negative correlation between PAF-AH and p50 (P <0.05). Conclusion The mechanism of blood stasis in patients with RA and the effect of Xinfeng capsule on the state of blood stasis may be closely related to the Act1 / NF-κB signaling pathway.