补肾祛寒治尪汤对胶原诱导关节炎大鼠抗炎与抗骨破坏的作用及对Wnt/β-catenin通路的影响  

Effects of kidney-supplementing cold-dispelling Zhi Wang Tang decoction on inflammation, bone erosion and Wnt/β-catenin pathway in rats with collagen-induced arthritis

作　　者：徐愿[1] 阎小萍[1] 肖诚[1] 马茹 陈光耀[2] 陶庆文[1] Xu Yuan;Yan Xiaoping;Xiao Cheng;Ma Ru;Chen Guangyao;Tao Qingwen(TCM Department of Rheumatism,Beijing Key Lab of Immune-Mediated Inflammatory Diseases,China-Japan Friendship Hospital,Beijing 100029.China; Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]中日友好医院中医风湿病科,免疫炎性疾病北京市重点实验室,北京100029 [2]北京中医药大学

出　　处：《北京中医药大学学报》2020年第4期289-295,共7页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(No.81603588)。

摘　　要：目的研究补肾祛寒治尪汤改善胶原诱导关节炎(CIA)大鼠炎症和骨破坏的作用及可能机制。方法大鼠分为空白组,模型组,补肾祛寒治尪汤组(25 g/kg灌胃,每天1次),甲氨蝶呤组(3 mg/kg灌胃,3天1次)。除空白组外,其余大鼠分别于饲养1、2周时尾部注射0.1 mL胶原乳剂(含牛Ⅱ型胶原200μg)制备关节炎模型。给药时间为初次免疫1周后开始,连续给药6周,每周记录关节炎指数(AI)。实验第7周处死动物,采集膝关节行HE染色,用免疫组化、免疫印迹法和荧光定量法评估肿瘤坏死因子α(TNF-α)、基质金属蛋白酶13(MMP13)、β-链蛋白(β-catenin)的表达。结果治疗6周后,补肾祛寒治尪汤组AI评分低于模型组(P<0.05);HE染色显示,补肾祛寒治尪汤组骨破坏较模型组明显减轻。与模型组比较,补肾祛寒治尪汤组TNF-α、MMP13、β-catenin蛋白及mRNA的表达减少(P<0.05)。结论补肾祛寒治尪汤能改善CIA大鼠骨破坏与关节炎症状,作用机制可能与调节Wnt/β-catenin通路,抑制TNF-α、MMP13表达有关。Objective To investigatethe effects of kidney-supplementing cold-dispelling Zhi Wang Tang(ZWT)decoction on relieving inflammation and bone erosion and modulating TNF-α, MMP13, β-catenin in rats of collagen-induced arthritis(CIA) models. Methods SD rats were injected at the tails with emulsion(0.1 mL) containing bovine type Ⅱ collagen(200 μg) after 1-2 weeks of orientation feeding to simulate the development of arthritis except those in the blank group.Rats were divided into the following four groups: blank group(n=6),model group(n=7), ZWT group(25 g/kg, oral, once a day, n=7), MTX group(methotrexate, 3 mg/kg, oral, once every three days, n=7).Intervention was administered for 6 continuous weeks starting in the week after immunization. Arthritis indexes(AI) were recorded.All rats were then sacrificed at week 7.Knee joint samples were taken for HE staining. Levels of TNF-a, MMP13 and β-catenin were evaluated with immunohistochemistry, Western blot and qPCR assays. Results At week 6 of intervention, AI scores in the ZWT group were lower than those of model group(P<0.05);HE staining result indicated that bone erosion changes in the ZWT group were also significantly relieved compared to the model group. Expressions of β-catenin, pro-inflammatory cytokines TNF-α and MMP13 were all reduced(P<0.05) compared to the model group. Conclusion ZWT seemed to relieve inflammation and bone erosion in CIA rats;its mechanism maybe associated with modulating Wnt/β-catenin pathway, and inhibiting the expression TNF-α and MMP13.

关 键 词：类风湿关节炎 胶原诱导关节炎 补肾祛寒治尪汤 WNT/Β-CATENIN通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]