title: Herbal formula Xian-Fang-Huo-Ming-Yin regulates differentiation of lymphocytes and production of pro-inflammatory cytokines in collagen-induced arthritis mice.
abstract: BACKGROUND
  Xian-Fang-Huo-Ming-Yin (XFHM), a traditional herbal formula, has been used to treat sores and carbuncles for hundreds of years in Asia. Nowadays, its clinical effects in treatment of rheumatoid arthritis (RA) have been validated. In this study, we want to study its possible molecular mechanisms of regulating the differentiation of lymphocytes and production of pro-inflammatory cytokines in collagen-induced arthritis (CIA) mice for RA treatment.
  
  
  METHODS
  A high performance liquid chromatography-electrospray ionization/mass spectrometer (HPLC-ESI/MSn) system was used to analyze the constituents of XFHM granules. An arthritics mouse model was induced by collagen and leflunomide (LEF) was used as a positive control medicine. Pathological changes at the metatarsophalangeal joint were studied through Safranin O and immunohistochemical staining. The differentiation of T, B and NK cells was examined by flow cytometry and pro-inflammatory cytokines were assayed using an Inflammation Antibody Array assay. The expression of key molecules of the nuclear factor κB (NF-κB) and Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathways in spleen were studied by western-blot analysis.
  
  
  RESULTS
  In our study. 21 different dominant chemical constituents were identified in XFHM. Treatment with XFHM suppressed the pathological changes in arthrosis of CIA. Additionally, XFHM down-regulated the proliferation and differentiation of CD3+T cells and CD3-CD19+B cells significantly. However, XFHM had no significant effect on CD3-NK1.1+NK cells. Further study showed that the production of pro-inflammatory cytokines had been suppressed by inhibiting the activation of NF-κB and JAK/STAT signaling.
  
  
  CONCLUSIONS
  XFHM can regulate and maintain the immunologic balance of lymphocytic immunity and inhibit the production of pro-inflammatory cytokines, thus suppressing the pathological changes of RA. Therefore, XFHM may be used as an application of traditional medicine against RA in modern complementary and alternative therapeutics.
authors.affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China.|Department of orthopedics Rheumatology, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China.
authors.forename: Jinyu
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China.
authors.forename: Xue
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Anatomy, School of Medicine, Nankai University, Tianjin, China.
authors.forename: Dashuai
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Nie
authors.affiliation: Department of Rheumatology, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China.
authors.forename: Jingwei
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China.
authors.forename: Lixia
authors.initials: L
authors.lastname: Lou
authors.affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Dong
authors.affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China.
authors.forename: Aiming
authors.initials: A
authors.lastname: Wu
authors.affiliation: Department of Anatomy, School of Medicine, Nankai University, Tianjin, China.
authors.forename: Yongzhe
authors.initials: Y
authors.lastname: Che
authors.affiliation: School of Preclinical Medicine, Beijing University of Chinese Medicine, Beijing, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Chen
authors.affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China.
authors.forename: Lingqun
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Department of orthopedics Rheumatology, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China.
authors.forename: Mingwei
authors.initials: M
authors.lastname: Mu
authors.affiliation: Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China. liminchai@hotmail.com.
authors.forename: Limin
authors.initials: L
authors.lastname: Chai
mesh_terms: D000818:Animals; D001168:Arthritis / Q000188:drug therapy* / Q000235:genetics / Q000276:immunology / Q000503:physiopathology; D002454:Cell Differentiation; D003094:Collagen / Q000009:adverse effects; D016207:Cytokines / Q000235:genetics / Q000276:immunology*; D004195:Disease Models, Animal; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage*; D006801:Humans; D053612:Janus Kinases / Q000235:genetics / Q000276:immunology; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology; D008297:Male; D051379:Mice; D008811:Mice, Inbred DBA; D016328:NF-kappa B / Q000235:genetics / Q000276:immunology
chemical_list: D016207:Cytokines; D004365:Drugs, Chinese Herbal; D016328:NF-kappa B; D003094:Collagen; D053612:Janus Kinases
keywords: Collagen-induced arthritis (CIA); Janus-activated kinase-signal transducer and activator of transcription (JAK/STAT); Lymphocyte; Nuclear factor κB (NF-κB); Pro-inflammatory cytokine; Xian-fang-huo-ming-Yin (XFHM)