Anti-arthritic And Mechanistic Studies Of Kadsura Coccinea Stem In Adjuvant-induced Arthritis In Rats

Objective: To evaluate the therapeutic effect of kadsura coccinea in the treatment of adjuvant-induced arthritis(AIA)and explore its mechanism.Methods:1.In animal experiment,60 male SD rats were randomly divided into normal group,model group,MTX group,Kadsura coccinea low-dose group(HLH-L),Kadsura coccinea medium-dose group(HLH-M)and Kadsura coccinea high-dose group(HLH-H),with 10 in each group.Except for the normal group,the other groups were subcutaneously injected with 0.1 m L complete freund adjuvant(CFA)to induce the establishment of AIA model.From the first day to the 36 th day,MTX group was given MTX(1mg/kg)by oral administration once a week.HLH-L,HLH-M,and HLH-H groups were intragastrically administrated once a day,with doses of 200mg/kg,400mg/kg and800mg/kg,respectively.The body weight of the rats was weighed every 3days.From the 9th day after modeling,the paw volume and arthritic score were measured every 3 days.After the treatment,the degree of joint bone destruction was evaluated by CT.The histopathological changes of the ankle joint were observed by HE staining.The expression of the IL-1α,IL-1β,IL-2,IL-4,IL-5,IL-6,IL-7,IL-10,IL-12(p70),IL-13,IL-17 a,IL-18,G-CSF,GM-CSF,GRO/KC,IFN-α,M-CSF,MIP-1α,MIP-3α,RANTES,TNF-α,VEGF,MCP-1 in rat serum were tested by bio-plex kit.The effect of seco-coccinic acid F(SCAF)on the RAW264.7 cell induced by LPS was observed.LPS groups: add the final concentration of 0.1μg/ml LPS;LPS+SCAF group: add the final concentration of 0.1μg/ml LPS and concentration of 2.5,5,10 and 20μM SCAF,testing the cell viability.Results:1.Compared with the model group,the body weight,paw volume and arthritic score of normal group,MTX group,HLH-M and HLH-H were significantly decreased.2.The model group and HLH-L group had serious bone damage.The degree of bone destruction in MTX group,HLH-M and HLH-H group were less.3.Compared with the model group,the expression of IL-1α,IL-1β,IL-2,IL-7,GM-CSF,GRO/KC,MIP-1α,MIP-3α,VEGF,MCP-1,TNF-α,IL-6 in MTX group,HLH-M and HLH-H group were significantly reduced.4.Cell experiment results show that compared with LPS group,LPS+20μM SCAF group could significantly inhibit the growth of RAW264.7 cells induced by LPS,P<0.0001).Conclusion:1.Kadsura coccinea can effectively relieve the degree of paw swelling in AIA model rats,increase body weight,reduce paw volume and arthritic scores,protect bone joints and reduce bone damage.2.The mechanism of Kadsura coccinea in treating RA may caused by a variety of cytokines: Kadsura coccinea can inhibit the release of IL-1α,IL-1β,IL-2,IL-7,GM-CSF,TNF-α and IL-6,thus inhibiting inflammation development;Inhibiting the GRO/KC,MIP-1α,MIP-3α,MCP-1,blocking its chemotaxis activities on the immune cells or other related cells,reduce inflammation,inhibiting osteoclast activation,reduce bone damage;Inhibiting the expression of VEGF,reduce synovial pannus formation.3.Seco-coccinic acid F can inhibit the proliferation and growth of RAW264.7 cell induced by LPS,and it is speculated that it may be one of the anti-inflammatory active components of Kadsura coccinea.