Effect of Topical Application of Fushan Rheumatism External Prescription on Inflammatory Cytokines and Notch2 Pathway in Rats with Collagen-Induced Arthritis / 世界科学技术-中医药现代化

ABSTRACT

Objective To explore the effect of topical application of Fushan rheumatism external prescription on inflammatory cytokines and notch2 pathway in rats with collagen-induced arthritis(CIA).Methods 36 female Wistar rats were randomly divided into normal group(n=10)and model group(n=26).CIA model was successfully established in 20 rats,which were randomly divided into model group(n=10)and Fushan rheumatism external prescription group(n= 10).Treatment was initiated on day 14,and 0.4 mL ointment was evenly applied to the ankle joints of rats in Fushan rheumatism external prescription group.Normal group and model group were topical smeared with the same volume of normal saline.Activity was taken twice a day for 42 days.The joint swelling of rats was observed every week and the arthritis index was scored.Thereafter blood was collected from abdominal aorta,and then ankle joint,spleen,liver,and kidney of rats were taken out after the end of the interventions.The severity of arthritis and the pathological changes of ankle joint,liver and kidney were evaluated by HE staining;Inflammatory cytokines expression in serum were detected by ELISA;The expression of Notch2,Delta-like ligand protein 1(delta-like ligand protein-1,DLL1)and nuclear factor-κ Bp65(nuclear factor-κ Bp65,NF-κ Bp65)mRNA and protein in synovium of ankle joints and spleen were detected by qRT-PCR and Western blot,and its positive expression in ankle joints were detected by immunohistochemical method;The changes of liver and kidney function of rats in each group were detected in serum.Results Compared with normal group,the arthritis index score in model group were increased(P<0.01),joint injury and pathological score were increased(P<0.01),the levels of inflammatory cytokines TNF-α,IL-6,IL-17 and IFN-γ in serum were increased(P<0.01),the expression of Notch2,DLL1 and NF-κBp65 mRNA and protein in joints and spleen were increased(P<0.01),and the positive expression in joints were also increased(P<0.01);Compared with model group,the arthritis index score in Fushan rheumatism external prescription group were decreased(P<0.05,P<0.01),joint injury and pathological score were decreased(P<0.01),the levels of inflammatory cytokines TNF-α,IL-6,IL-17 and IFN-γ in serum were decreased(P<0.01),the expression of Notch2,DLL1 and NF-κBp65 mRNA and protein in joints and spleen were decreased(P<0.01),and the positive expression in joints were also decreased(P<0.01).In addition,no noticeable tissue damages were observed in liver and kidney in Fushan rheumatism external prescription group,and the levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),blood urea nitrogen(BUN)and creatinine(Cr)in serum were no changes(P>0.05).Conclusion Fushan rheumatism external prescription relieves arthritis symptoms and joint injury in CIA rats,and has no toxic and side effects on liver and kidney.Its mechanism may be related to the reduction of inflammatory cytokines and down-regulation of Nocth2 pathway.