title: Dissection of the mechanism of traditional Chinese medical prescription-Yiqihuoxue formula as an effective anti-fibrotic treatment for systemic sclerosis.
abstract: BACKGROUND
  Systemic sclerosis (SSc) is a connective tissue fibrotic disease for which there is no effective treatment. Traditional Chinese Medicine (TCM), such as the Yiqihuoxue formula used in Shanghai TCM-integrated Hospital, has shown the efficacy of anti-fibrosis in clinical applications. This study was aiming to dissect the anti-fibrotic mechanism of Yiqihuoxue treatment for SSc.
  
  
  METHODS
  Bleomycin-induced mice and SSc dermal fibroblasts were treated with Yiqihuoxue decoction; NIH-3T3 fibroblasts were exposed to exogenous TGF-β1, and then cultured with or without Yiqihuoxue decoction. Luciferase reporter gene assay was used to determine the activity of Smad binding element (SBE). Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to examine the mRNA levels of extracellular matrix (ECM) genes. The protein levels of type I collagen, Smad3 and phosphorylated-Smad3 (p-Smad3) were detected by western blotting. Student's t-tests were used to determine the significance of the results.
  
  
  RESULTS
  Bleomycin-induced mice, SSc dermal fibroblasts and TGF-β1-induced NIH/3T3 fibroblasts showed higher levels of ECM gene transcriptions and collagen production. In addition, the phosphorylation level of Smad3 and activity of SBE were significantly increased after exogenous TGF-β1 induction. Whereas, Yiqihuoxue treatment could obviously attenuate fibrosis in bleomycin-induced mice, down regulate ECM gene expressions and collagen production in SSc dermal fibroblasts and TGF-β1-induced NIH/3T3 fibroblasts. Furthermore, the aberrantly high phosphorylation level of Smad3 and activity of SBE in the TGF-β1-induced NIH/3T3 fibroblasts were also dramatically decreased by Yiqihuoxue treatment.
  
  
  CONCLUSIONS
  Yiqihuoxue treatment could effectively reduce collagen production via down-regulating the phosphorylation of Smad3 and then the activity of SBE, which are involved in the TGF-β pathway and constitutively activated in the progression of SSc.
authors.forename: Ting
authors.initials: T
authors.lastname: Wu
authors.forename: Haiyan
authors.initials: H
authors.lastname: Chu
authors.forename: Wenzhen
authors.initials: W
authors.lastname: Tu
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Song
authors.forename: Dongdong
authors.initials: D
authors.lastname: Chen
authors.forename: Jin
authors.initials: J
authors.lastname: Yuan
authors.forename: Ling
authors.initials: L
authors.lastname: Yu
authors.forename: Yanyun
authors.initials: Y
authors.lastname: Ma
authors.forename: Qingmei
authors.initials: Q
authors.lastname: Liu
authors.forename: Li
authors.initials: L
authors.lastname: Jin
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Zhou
authors.forename: Hejian
authors.initials: H
authors.lastname: Zou
authors.affiliation: National Ministry of Education Key Laboratory of Contemporary Anthropology, School of Life Sciences, Fudan University, Shanghai, PR China. wuwenyu@medmail.com.cn.
authors.forename: Wenyu
authors.initials: W
authors.lastname: Wu
authors.forename: Jiucun
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005355:Fibrosis / Q000188:drug therapy / Q000473:pathology; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D041681:NIH 3T3 Cells; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011897:Random Allocation; D012595:Scleroderma, Systemic / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: D004365:Drugs, Chinese Herbal; D016326:Extracellular Matrix Proteins; D012333:RNA, Messenger; C033946:yiki-huoxue