Research On Immunosuppressive Effects And Action Mechanism Of Active Ingredient Of Elephantopus Scaber

Objective:To study the immunosuppressive effects of the active ingredient of elephantopus scaber DDC-105, reveal the immune mechanism of DDC-105 on rheumatoid arthritis (RA) treatment, and lay a theoretical foundation for new medicine research of the active ingredient of elephantopus scaber DDC-105 on RA treatment.Method:1. Established T cell in vitro activation model induced by mitogen Con A, and tested cell proliferation by adopting MTT method; Tested DDC-105 on ConA induction of generation level of spleen lymphocyte cytokines IL-2, IFN-γ and IL-6 by adopting enzyme linked immunosorbent assay (ELISA); established antigen specific immune response induced by ovalbumin (OVA), and evaluated immunosuppressive effect of in vivo DDC-105 and its mechanism;2. Established Bovine Type Ⅱ DBA/1 Collagen induced arthritis (CIA) of laboratory rats. According to clinical scoring criteria, it is divided into three groups:CIA model set, positive medicine MTX group, DDC-105 drug administration group. Observed pathogenic degree of CIA of laboratory rats by adopting arthritis clinical scoring method; determined pathological damage degree of CIA of laboratory rats by adopting paraffin section H & E dyeing; tested generation level of collagen induced lymphocytes Th1 (IFN-γ), Th2 (IL-4),Th17(IL-17A), Treg cytokines (IL-10) and inflammatory factor (TNF-α) by adopting ELISA method.Results:1. No obvious cytotoxic effects detected when the concentration of DDC-105 is at 30 μM. The concentration dependence inhibits T lymphocyte multiplication induced by mitogen ConA. And the concentration of DDC-105 inhibits the generation level of splenic lymphocyte IL-2, IFN-γ and IL-6 induced by ConA. In vivo oral drug administration of DDC-105 inhibits the generation level of T lymphocyte multiplication and Thl type cytokines induced by DDC-105.2. DDC-105 significantly reduced clinical score of CIA of laboratory rats, improved symptoms of local CIA, and alleviated inflammatory cell infiltration; and reduced CIA collagen specificity immune response (P<0.01), inhibited cytokines expression of CIA splenic lymphocyte IFN-γ and IL-17A (both are P<0.05), and up-regulated the generation level of IL-10 (P<0.01);Conclusion:1. DDC-105 inhibits T cell immune response. Immunosuppressive effect has been revealed within the body. Down-regulation of Thl cytokines is probably one of the mechanism of its immunosuppressive effects.2. DDC-105 has inhibition effect on CIA of laboratory rats. Down-regulation of highly pathogenic Th17 expression and regulation of Th17 cell/Treg may be one of the mechanisms for RA treatment.