Therapeutic Effect Of Koumine On Rheumatoid Arthritis In C57Bl/6Mice

Objective: To observe the therapeutic effect of koumine on rheumatoid arthritis in C57BL/6mice through thermal hyperalgesia, hind paw value, arthritis index and the histopathology in ankle, and the relationship with the number of regulatory T cells (Treg cells) from spleen of CIA mice and the serum level of transforming growth factor (TGF-β).Methods: Male C57BL/6mice were injected at the base of tail with100ul emulsion which containing an equal volume of chicken typeⅡcollagen solution and complete Freund’ adjuvant.21days after the first injection, the second injection was repeated in order to establish the CIA mice model. Selected successful CIA mice model were randomly divided into CIA model group, methotrexate group and koumine high, medium and low-dose treatment groups, non-immunized mice served as controls. Koumine high, medium and low treatment groups orally administered koumine with10mg/kg,2mg/kg,0.4mg/kg for10days starting from the23rd after the initial immunization, to observe the features of CIA in paw withdrawal thermal latency, hind paw value, arthritis index. Selected successful CIA mice model were randomly divided into CIA model group, methotrexate group and koumine high and low-dose treatment groups, non-immunized mice served as controls. The same to the scheme intragastric administration, after the last administration, the mice were killed3hours late, the ankle joints of mice were took to investigate the change of pathology. Collecting serum to detect the level of IgG1and TGF-βwith the enzyme-linked immunosorbent assay (ELISA). Removing the spleen to detect the percentages of CD4+CD25+Foxp3+regulatory T cells in CD4+T cells with flow cytometry.Result: The thermal hyperalgesia, hind paw value, arthritis index were reduced and the retrogression of cartilage was relieved by the treatment of Koumine. CIA model group was higher than control group (P<0.05). CD4+CD25+Foxp3+Treg content in peripheral blood of CIA model group was significantly lower than those of control group (P<0.01), However, with the treatment of Koumine, the expression of CD4+CD25+Foxp3+Treg significantly higher than those CIA group mice(P<0.05). ELISA results showed that, the levels of TGF-βwere decreased, compared with the control group. Koumine have antagonistic to the trend, but no statistically significan (P>0.05). Conclusion: Koumine may have a significant therapeutic effect on rheumatoid arthritis and its ability to increase the numbers of Treg cells in the spleen maybe relevant to the therapeutic response.