壮医药线点灸联合甲氨蝶呤对佐剂关节炎大鼠NF-κB/IκB信号通路的影响

Effect of Zhuang's Medicated Thread Moxibustion Combined with Methotrexate on NF-κB/IκB in Rats with Adjuvant Arthritis

目的:从分子生物学角度探究壮医药线点灸联合甲氨蝶呤对佐剂关节炎大鼠NF-κB/IκB信号通路的影响,为壮医药线点灸联合甲氨蝶呤治疗类风湿性关节炎(RA)提供理论依据。方法:建立佐剂关节炎大鼠模型,并分为正常对照组(NC)、关节炎模型组(RA)、甲氨蝶呤组(MTX)和壮医药线点灸联合甲氨蝶呤组(CT)。观察并评估大鼠关节炎指数; ELISA法检测各组大鼠血清中TNF-α和IL-1β的含量; qPCR和Western blotting法检测滑膜组织中P65、I-k Bα、TNF-α、IL-1β和COX-2的表达。结果:与RA组相比,MTX组和CT组均能显著降低关节炎指数(P <0. 05),且CT组显著低于MTX组(P <0. 05);与RA组相比,MTX组和CT组大鼠血清中TNF-α和IL-1β含量均显著下降(P <0. 05),其中CT组下降更为显著(P <0. 05 VS MTX); MTX组和CT组大鼠关节滑膜组织中P65、TNF-α、IL-1β和COX-2的mRNA和蛋白表达显著下调(P <0. 05 VS RA),I-k Bα显著上调(P <0. 05 VS RA),且MTX组和CT组相比差异亦具有显著性差异(P <0. 05)。结论:壮医药线点灸联合甲氨蝶呤干预能够显著改善大鼠关节炎症状,且其作用机制可能是通过调控NF-κB/IκB信号通路,从而抑制炎症因子释放和环氧化酶的表达来实现的。

Objective： To explore the effect of Zhuang＇s medicated thread moxibustion combined with Methotrexate（MTX） on NF -KB/IKB in rats with adjuvant arthritis（AA） based on molecular biology, thus to provide theoretical basis of the therapy for rheumatoid arthritis （RA）. Methods： AA rat model was established and randomly divided into the model group, MTX group and the combination group （Zhuangg medicated thrad moxibustion ＋ MTX ） , and the normal rats were taken as the blank control. The arthritis index was observed and evaluated in rats. The serum contents of TNF -α and IL - 1 β were detected by ELISA method. The expression levels of P65, I - κBα, TNF - α, IL - 1 β and COX - 2 were examined by the methods of qPCR and Western bohting. Results ： Compared to the model group, the arthritis index was significantly reuced in both the MTX group and the combination group （P 〈 0.05 ） ; and the combination group im- proved more than the MTX group（P 〈 0.05）. Compared to the model group, the serum contents of TNF-α and IL - 1 β were significantly decreased in both the MTX group and the combination group （ P 〈 0.05 ） ; and the decrease was more obvious in the combination group than that in MTX group （P 〈 0.05）. The expressions of P65, TNF - α, IL - 1β and COX - 2 in synovial tissues were significanly down - regualted and I - κBα was significantly up - regulated in MTX group and in the combination group compared to those in the model group （P 〈 0.05） ; of which the difference between the two medicated groups was also significant （P 〈 0.05）. Conclusion： Zhuangg medicated thread moxibustion combined with Methotrexate can significantly improve the symptoms of arthritis in rats. The mechanism may be achieved by regula- ting the NF - KB/IκB signaling pathway, thereby inhibiting the release of inflammatory factors and the expression of cy- clooxygenase.