Therapeutic Action Mechanism Of Wang-bi Capsule On Collagen-Induced Arthritis Mice

Objective:The study aimed to establish the mice with Collagen Induced Arthritis(CIA)model and explore the anti-inflammatory and osteoprotective effects and action mechanism of Wang-BI Capsule(WB)treating for rheumatoid arthritis(RA)Methods:1.CIA mice model were established,the mice were divided into control group(Control,Con),model group(Model,Mod),methotrexate group(Methotrexate,MTX),WB low-dose group(WB-L),WB middle dose group(WB-M)and WB high-dose group(WB-H),8 mice in each group,were intragastric administered in the volume of 0.1 mL/10 g,two different administration cycles were designed for two weeks and four weeks.We recorded arthritis scores and weight twice a week.H&E staining was used to observe the pathological changes in the ankle joint,and the histology score was performed2.Serum level of IL-1β,IL-6 and TNF-α were detected by ELISA Immunohistochemistry was used to determine the expression levels of IL-1β,IL-6 and TNF-a in the ankle j oint.The expression levels of p-IKKα/β,p-p65 and IκBα in the ankle joint were determined by Western Blot.Observation of destruction of ankle cartilage tissue in mice by using Safranin O-solid green staining.Micro-CT was used to scan,reconstruction and analysis the bone structure.The differentiation and maturation of osteoclasts in the inflammatory joints were observed by TRAP stainingResults:1.Compared with two weeks of administration,the administration of WB for four weeks could effectively reduce the degree of paw swelling and arthritis score in CIA mice,improved histopathological conditions,at the same time,there was no significant effect on liver and kidney function2.ELISA results showed that WB reduced the levels of inflammatory factors IL-1β,IL-6 and TNF-α in serum of CIA mice(P<0.05 or P<0.01).Immunohistochemistry results showed that WB effectively reduced the levels of IL-1β,IL-6 and TNF-α in the ankle joint of CIA mice(P<0.05 or P<0.01).Western Blot results showed that WB could decrease the expression of p-IKKα/β and p-p65(P<0.05 or P<0.01)and increase the expression of IκBαprotein(P<0.05 or P<0.01)3.The results of Safranin O-solid green staining showed that administration of WB for four weeks could protect the structural intergrity of CIA mice cartilage tissue.Micro-CT results showed that WB could reduce the damage of trabecular bone in CIA mice.TRAP staining results showed that WB could reduce the number of osteoclasts at the inflammation joint(P<0.05 or P<0.01).Immunohistochemistry results showed that WB reduced the levels of RANKL in the ankle joint of CIA mice(P<0.05 or P<0.01),and improved the levels of OPG(P<0.05 或 P<0.01)Conclusion:1.The Wang-Bi capsule has anti-inflammatory and osteoprotective effects on CIA mice,compared with two weeks administration,the effect was better when administered for 4 weeks,and there were no obvious signification effect on liver and kidney function2.The action mechanism of Wang-Bi capsule on RA might be related to decreasing the expression of inflammatory factors IL-1β,IL-6 and TNF-α,inhibiting the activation of NF-κB pathway,inhibiting the differentiation and maturation of osteoclasts,and down-regulating the ratio of RANKL/OPG.