摘要
目的从细胞层面探讨抗风湿中药单体青藤碱(SIN)对辅助性T淋巴细胞(Th)17/调节性T淋巴细胞(Tregs)细胞比例的影响,进一步阐明SIN治疗类风湿关节炎(RA)的机制。方法分别抽取SIN组和双氯酚酸钠组RA患者治疗前、治疗12周后以及对照组健康体检者的外周静脉血20mL,以肝素钠抗凝,采用Ficol1密度梯度离心法分离外周血单核细胞,进行荧光抗体标记,应用流式细胞仪测定Th17(CD4+IL-17+)和Tregs(CD4+CD25+CD127lowFoxP3+)细胞比例,观察SIN组和双氯芬酸钠组治疗前后类风湿因子(RF)、红细胞沉降率(ESR)、C反应蛋白(CRP)、抗环瓜氨酸多肽(CCP)抗体水平和疾病活动性评分(DAS)28评分的变化,比较SIN组和双氯芬酸钠组Th17/Tregs细胞比例,观察SIN的临床疗效。结果 SIN组和双氯酚酸钠组治疗后的CRP、ESR、RF、抗CCP抗体水平均较同组治疗前显著下降(P值分别<0.05、0.01),但两组间治疗前后上述各项指标的差异均无统计学意义(P值均>0.05)。SIN组和双氯酚酸钠组治疗后的DAS28评分均较同组治疗前显著下降(P值均<0.05),双氯芬钠组治疗后的DAS28评分显著高于SIN组(P<0.05)。SIN组和双氯芬酸钠组治疗前的Th17细胞比例均显著高于对照组(P值均<0.05),SIN组治疗后的Th17细胞比例较同组治疗前显著降低(P<0.05)。SIN组和双氯芬酸钠组治疗前的Tregs细胞比例均显著低于对照组(P值均<0.01),SIN组治疗后的Tregs细胞比例较同组治疗前显著升高(P<0.01),双氯芬酸钠组治疗前后Tregs细胞比例的差异无统计学意义(P>0.05)。双氯芬酸钠组治疗后的Tregs细胞比例显著低于SIN组(P<0.01)。结论 SIN可能通过抑制致炎细胞Th17升高,具有免疫抑制作用的Tregs细胞比例,而产生治疗RA的作用。
Objective To investigate the effect of sinomenine on the proportion of T helper cells 17 （Th17） and Regulatory T cells （Tregs） and to explore the mechanism of sinomenine for the treatment of rheumatoid arthritis. Methods Peripheral venous blood （20 mL） was collected from 40 patients with rheumatoid arthritis （20 treated with sinomenine and 20 treated with diclofenac sodium） and 40 healthy volunteers before treatment and at 12 weeks after treatment. Heparin sodium was used for anticoagulation. The peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation and labeled with fluorescent antibodies. The number of Th17 （CD4＋ IL-17＋ ） and Treg （CD4＋ 0D25＋ CD127low FoxP3＋ ） was measured by flow cytometry. Rheumatoid factor （RF）, erythrocyte sedimentation rate （ESR）, C reactive protein （ORP）, cyclic citrulline peptide （CCP） antibody and Disease Activity Score （DAS） were compared between sinomenine group and diclofenac group. The proportion of Th17/Tregs was analyzed. Results The clinical parameters, such as CRP, ESR, RF, and CCP level were all significantly decreased after treatment with sinomenine or diclofenac sodium （P〈0.05, 0.01）. The parameters in the sinomenine group were even lower than those in the diclofenac group, but the difference was not statistically significant （all P〈0.05）. DAS28 score was significantly reduced after treatment with sinomenine or diclofenac sodium （P〈0.05）, and there was significant difference between the two treatment groups （P 〈 0.05）. Before treatment, the proportion of Th17 in the two treatment groups were significantly higher than that in control group （P〈 O, 05）. The proportion of Th17 after treatment with sinomenine or diclofenac sodium was significantly lower than before treatment （ P〈 0.05）, and that in the sinomenine group was even lower than diclofenac group （ P〈 0.05）. Before treatment, the proportion of Treg in the two treatment groups were significantly lower than that in control group （all P〈0.01）. The proportion of Treg was elevated after treatment with sinomenine （P〈0.01）. There was no significant difference in the proportion of Treg before and after treatment with diclofenac sodium （ P〉 0.05）, but there was significant difference between two treatment groups （P〈0.01）. Conclusion The therapeutic effect of sinomenine on rheumatoid arthritis may be related to inhibiting proinflammatory Th17 and promoting immunosuppressive Treg. （Shanghai Med J, 2013, 36： 254-258）
出处
《上海医学》
CAS
CSCD
北大核心
2013年第3期254-258,共5页
Shanghai Medical Journal
基金
上海市卫生局科研计划课题资助项目(20090Y79)
