Clematichinenoside AR induces immunosuppression involving Treg cells in Peyer's patches of rats with adjuvant induced arthritis

Ethnopharmacological relevance: Clematichinenoside AR (AR) has been defined as a major active ingredient of triterpenoid saponins extracted from Clematidis Radix et Rhizoma, which is a traditional Chinese herbal medicine that has long been used in the treatment of rheumatoid arthritis (RA). To further explore the mechanism of AR in the treatment of RA, we investigated whether its immunomodulatory effects are related to Treg-mediated suppression derived from Peyer's patches (PPs) in adjuvant induced arthritis (AIA) rat model. Materials and methods: AR (8, 16, 32 mg/kg) was orally administered daily from Day 18 to Day 31 after immunization. The effect of AR on AIA rats was evaluated by hind paw swelling and histopathological examination. Percentages of CD4(+)CD25(+)Foxp3(+) T regulatory cells were determined by flow cytometry. Levels of IL-10, TGF-beta(1), IL-17A and TNF-alpha were measured by ELISA. Expressions of Foxp3 and ROR gamma in synovium were detected using immunohistochemical analysis. Results: AR treatment significantly reduced paw swelling of ALA rats, and histopathological analysis confirmed it could suppress severity of established arthritis. AR treatment upregulated the percentages of CD4(+)CD25(+)Foxp3(+) Treg cells among CD4+ T cells in PPs lymphocytes, and increased the levels of IL-10 and TGF-beta(1) secreted from ConA-activated PPs lymphocytes, whereas decreased the levels of IL-17 A and TNE-alpha. Similar tendency of circulating CD4(+)CD25(+)Foxp3(+) Treg cells percentages and serum cytokine levels were observed. Moreover, AR decreased the expression levels of Foxp3 and ROR gamma in joint synovial membrane. Conclusions: In conclusion, these results suggested AR has a potent protective effect on the progression of AIA, probably by augmenting CD4(+)CD25(+)Foxp3(+) Treg cells in PPs to induce immunosuppression, and modulating the balance between Treg cells and Th17 cells systemically. These findings may help to develop AR as a potent immunosuppressive agent for the treatment of RA. (C) 2014 Elsevier Ireland Ltd. All rights reserved.