title: Clematichinenoside AR induces immunosuppression involving Treg cells in Peyer׳s patches of rats with adjuvant induced arthritis.
abstract: BACKGROUND
  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.
  
  
  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-β1, IL-17A and TNF-α were measured by ELISA. Expressions of Foxp3 and RORγ in synovium were detected using immunohistochemical analysis.
  
  
  RESULTS
  AR treatment significantly reduced paw swelling of AIA 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-β1 secreted from ConA-activated PPs lymphocytes, whereas decreased the levels of IL-17 A and TNF-α. 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 RORγ 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.
authors: State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, # 24 TongJiaXiang, Nanjing 210009, PR China.
authors: Ying
authors: Y
authors: Xiong
authors: State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, # 24 TongJiaXiang, Nanjing 210009, PR China.
authors: Yan
authors: Y
authors: Ma
authors: State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, # 24 TongJiaXiang, Nanjing 210009, PR China.
authors: Wang
authors: W
authors: Han
authors: State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, # 24 TongJiaXiang, Nanjing 210009, PR China.
authors: Nandani Darshika
authors: ND
authors: Kodithuwakku
authors: Department of Pharmacognosy, the Key Laboratory of Modern Chinese Medicines (Ministry of Education), China Pharmaceutical University, # 24 TongJiaXiang, Nanjing 210009, PR China.
authors: Li-Fang
authors: LF
authors: Liu
authors: Department of Traditional Chinese Pharmacy, China Pharmaceutical University, # 24 TongJiaXiang, Nanjing 210009, PR China.
authors: Feng-Wen
authors: FW
authors: Li
authors: State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, # 24 TongJiaXiang, Nanjing 210009, PR China. Electronic address: weirongfang@163.com.
authors: Wei-Rong
authors: WR
authors: Fang
authors: State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, # 24 TongJiaXiang, Nanjing 210009, PR China. Electronic address: yunmanlicpu@hotmail.com.
authors: Yun-Man
authors: YM
authors: Li
mesh_terms: D000818:Animals; D001169:Arthritis, Experimental / Q000139:chemically induced / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D018450:Disease Progression; D004305:Dose-Response Relationship, Drug; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D005620:Freund's Adjuvant; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D018836:Inflammation Mediators / Q000276:immunology / Q000378:metabolism; D007596:Joints / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D008297:Male; D010581:Peyer's Patches / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D008517:Phytotherapy; D010946:Plants, Medicinal; D017207:Rats, Sprague-Dawley; D012503:Saponins / Q000494:pharmacology*; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D058504:Th17 Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D013997:Time Factors; D014315:Triterpenes / Q000494:pharmacology*
chemical_list: D016207:Cytokines; D004365:Drugs, Chinese Herbal; D007166:Immunosuppressive Agents; D018836:Inflammation Mediators; D012503:Saponins; D014315:Triterpenes; C000599570:clematichinenoside AR; D005620:Freund's Adjuvant
keywords: Adjuvant induced arthritis; CD4+CD25+Foxp3+Treg cell; Clematichinenoside AR; Immunosuppression; Peyer׳s patch; Rheumatoid arthritis