Effect Of Koumine On Spleen T Helper Cells Of Collagen-induced Rheumatoid Arthritis Rats

Objective: To observe the effect of koumine on CIA rats and its regulation on spleen T helper cells in CIA ratsMethods: Male Wistar rats were used to create CIA model. 40 rats were divided in to four groups(model, MTX, km=15mg/kg, km=0.6mg/kg), each group had 10 rats. Another 10 rats were used as control group. We gave koumine to rats by gavage 21 day after first immunization. Once a day for 10 consecutive days; rats in the control and model group were gavaged 0.9% normal saline; rats in MTX group were gavaged once two day. Hind paw value, mechanical pain threshold, arthritis index, serum C-reactive protein(CRP) level and mechanical pain were used as an indicator to observe koumine therapeutic effect on CIA rats. We collected serum of rats in day 0, 21, 24, 27, 30 to detect serum CRP levels of rats by Elisa.Hematoxylin-eosin staining was used to observe joint change of rats; separate spleen to preparate spleen cell suspension to detect the number of Th1, Th2, Th17,Treg by flow cytometry. Spleen cell suspension supernatant inflammatory cytokine IFN-γ, IL-4, IL-17, TGF-βlevels were detect by Elisa.Result: Compare to control group, mechanical pain threshold of CIA rats are significantly lower than control group 14 days after first immunization, and its hind paw value, arthritis index. Serum C-reactive protein(CRP) level are significantly improve 21 days after first immunization. Aankle joints of rats have a large number of inflammatory cell infiltration, we can see joint structural damage under microscope. These results point that koumine can significantly relieve mechanical pain, inflammation and joint destruction. It has a significantly therapeutic effect on CIA. Flow cytometry showed that CIA splenic T cell subsets appear polarization offset. It showed a significantly increased of Th1 / Th2, Th17 / Treg cells proportion. The Elisa tests show a same result as flow cytometry, proportion of IFN-γ / IL-4 and IL-17 A / TGF-β has a significantly improvement.Conclusion: Koumine may have a significant therapeutic effect on rheumatoid arthritis and its ability to decrease the ratios of Th1 / Th2, Th17 / Treg cells in the spleen that maybe relevant to the therapeutic effect.