目的:检测痹肿消汤按湿、热、毒、瘀拆方对胶原诱导性关节炎大鼠滑膜细胞外信号调节激酶1/2(ERK1/2)、细胞外信号调节激酶5(ERK5)表达的影响,从湿、热、毒、瘀探讨类风湿性关节炎(RA)的作用机制。方法:采用皮下注射牛Ⅱ型胶原蛋白与完全弗氏佐剂诱导SD大鼠胶原诱导性关节炎(CIA)模型,并设立正常组。采用免疫印迹法(Western blot)同步检测正常组、模型组、痹肿消汤组、清热解毒组(拆方Ⅰ组)、祛湿组(拆方Ⅱ组)、活血组(拆方Ⅲ组)大鼠在不同时间点滑膜ERK1/2、ERK5的磷酸化水平,运用灰度扫描软件对结果进行半定量分析。结果:造模14d后,模型组滑膜ERK1/2、ERK5表达高于正常组(P〈0.05)。随着免疫时间延长,模型组ERK1/2、ERK5表达于21、28d呈递增趋势(P〈0.05),28d达到最高。4用药组治疗后,ERK1/2、ERK5的磷酸化表达水平均低于同期模型组(P〈0.05)。在21、28d水平,表达呈递减趋势(P〈0.05)。痹肿消汤组的ERK1/2、ERK5各时间点的表达水平比拆方3组更低,拆方Ⅱ组的ERK1/2表达的下降幅度比拆方I组、拆方Ⅲ组更明显(P〈0.05)。而拆方I组的ERK5表达的下降幅度比拆方Ⅱ组、拆方Ⅲ组更显著(P〈0.05)。结论:拆方各组可有效地抑制ERK1/2、ERK5转导通路的活化,达到阻抑关节滑膜增殖、骨质侵蚀的作用。但清热解毒、祛湿通络、活血祛瘀各治法对上述通路调节作用优势有不同。
Objective: To elucidate the mechanism of the separated groups from Bizhongxiao decoction(BZXD) in treatment of rheumatoid arthritis(RA) by analyzing the effects of the separated groupⅠ(Antipyretic Detoxicate group),separated group Ⅱ(Dissolving Dampness group) and separated group Ⅲ(Activating Blood Circulation group) from BZXD on ERK1/2 and ERK5 expressions in synovium of CIA rats.Methods: The experiment arthritis rat model was established by subcutaneous injection of Ⅱ collagen and complete Freunds adjuvant.ERK1/2 and ERK5 expressions at different times in five groups were detected by Western blotting,and semi-quantification analysis was performed with gray scale scanning software.Results: ERK1/2,ERK5 expressions of synovium in model group on the 21th day were higher than those in normal group(P〈0.05).ERK1/2,ERK5 expressions of synovium in model group on the 28th day were higher than those on the 21th day(P〈0.05).After separated groupⅠ,separated group Ⅱ and separated group Ⅲ treatments,ERK1/2,ERK5 expressions were sigificantly lower than those in model group at the different timepoints(P〈0.05) and decreased gradually(P〈0.05).ERK1/2 expression of synovium in the separated group Ⅱdecreased more significantly than that in the separated groupⅠand separated group Ⅲ(P〈0.05),ERK5 expression of synovium in the separated group Ⅰdecreased more significantly than that in the separated groupⅡand separated group Ⅲ(P〈0.05).Conclusion: Remove groups could effectively inhibit ERK1/2,ERK5 transduction pathway activation,inhibition of synovial proliferation,bone to erosion.But Qingrejiedu,dampness,promoting blood circulation and removing blood stasis on the treatment to the pathways regulating action advantage had different.