藏药五味甘露药浴颗粒对AA模型大鼠的分子作用机制研究

Molecular mechanism of Wuwei Ganlu Yaoyu Keli on AA model rats

目的:从细胞层面观察五味甘露药浴颗粒对AA模型大鼠滑膜组织中细胞因子的影响以及对蛋白表达的调控作用,并探讨其中可能的分子机制。方法:除正常对照组外,大鼠采用弗氏完全佐剂(FCA)造模,随机分为模型对照组、地塞米松组(0.150 mg·kg^(-1))、五味甘露药浴颗粒高剂量组(11.80 g·L^(-1))、中剂量组(5.90 g·L^(-1))、低剂量组(2.95 g·L^(-1)),药浴组40℃药浴30 min·d^(-1),模型对照组和正常对照组40℃温水浴30 min·d^(-1)。给药四周后采用酶联反应、免疫组化法检测并分析五味甘露药浴颗粒对滑膜组织中细胞因子及蛋白表达的干预效应。结果:五味甘露药浴颗粒组大鼠血清中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、前列腺素E2(PGE2)、基质金属蛋白酶(MMP-3、TIMP-1)的含量均有不同程度的下降。结论:五味甘露药浴颗粒在类风湿关节炎滑膜病理改变的过程中,通过下调致炎因子(TNF-α、IL-1、IL-6)、上调抗炎因子(IL-10)来激活体内免疫系统,抑制成纤维样滑膜细胞分泌各类细胞因子对滑膜的刺激,来控制关节滑膜炎症,达到抑制RA滑膜细胞增殖、促进细胞凋亡、抗炎、消肿、保护关节骨与软骨的目的。

Objective:To observe the effect of Wuwei Ganlu Yaoyu Keli on the cytokines in synovial tissue of AA model rats and the regulation of protein expression,and explore the possible molecular mechanism.Method:Except for the normal control group,the rat model was established by Freund’s complete adjuvant.The rats were randomly divided into adjuvant arthritis group(AA)dexamethasone group(0.150 mg·kg^(-1)),high-dose Wuwei Ganlu Yaoyu Keli group(11.80 g·L^(-1)),medium-dose Wuwei Ganlu Yaoyu Keli group(5.90 g·L^(-1))and low-dose Wuwei Ganlu Yaoyu Keli group(2.95 g·L^(-1)).In the Tibetan medicine-bath group,the bath temperature was 40℃and bathed 30 min·d^(-1).The control group and the AA model group used 40℃warm water and bathed 30 min·d^(-1).After four weeks,the intervention effect of Wuwei Ganlu Yaoyu Keli on the expression of cytokines and proteins in synovial tissue was analyzed by enzyme-linked reaction and immunohistochemistry.Result:The levels of IL-1β,TNF-α,PGE2,MMP-3 and TIMP-1 in serum of rats in Wuwei Ganlu Yaoyu Keli group were decreased in different degrees.Conclusion:In the process of pathological changes of rheumatoid arthritis synovium,Wuwei Ganlu Yaoyu Keli can activate the immune system in vivo by down regulating inflammatory factors(TNF-α,IL-1β,IL-6)and up regulating anti-inflammatory factors(IL-10),inhibit fibroblast like synovial cells to secrete various cytokines to stimulate synovium,so as to control synovium inflammation,inhibit RA synovium cell proliferation and promote cell apoptosis,inhibit the proliferation of RA synovial cells,promote apoptosis,antiinflammatory,detumescence,protect joint bone and cartilage.