title: Saikosaponin A of Bupleurum chinense (Chaihu) elevates bone morphogenetic protein 4 (BMP-4) during hepatic stellate cell activation.
abstract: BACKGROUND
  Saikosaponin a (SSa) is a compound extracted from a Chinese herb which has been widely used in treating liver diseases such as liver fibrosis. However, the mechanism of SSa in treatment of liver fibrosis still remain unclear. Our previous study demonstrated that BMP4 stimulated the expression of smooth muscle alpha actin (α-SMA) in the liver. Therefore, the current study investigates the effect of SSa on BMP4 expression during hepatic stellate cell activation in a human hepatic stellate cell line.
  
  
  METHODS
  LX-2 cells were cultured in DMEM/F12 with fetal bovine serum and treated with SSa in different times and concentrations. The expression of BMP4 was examined by both RT-PCR and western blot analysis. WST-1 proliferation reagent was used to evaluate cell proliferation. α-SMA and Bax protein expression was determined by western blot analysis.
  
  
  RESULTS
  Both mRNA and protein levels of BMP-4 were significantly inhibited in LX-2 cells after 5 μM SSa treatment. SSa significantly inhibited LX-2 proliferation at the concentration of 5μM while BMP-4 had no effect on LX-2 proliferation. BMP-4 increased α-SMA expression in LX-2 while SSa reduced α-SMA expression. In addition SSa could neutralize the effect of BMP-4 on α-SMA expression. SSd also inhibited BMP4 expression but not NG. Bax protein expression was induced in these cells by 5 μM SSa.
  
  
  CONCLUSIONS
  SSa could down-regulate BMP-4 expression and inhibit hepatic stellate cell activation. Therefore, SSa could be used for treatment of liver disease with elevated BMP-4 expression.
authors: Faculty of Pharmacy, University of Manitoba, Winnipeg, Manitoba, Canada R3E 0T5; Department of TCM Basic Clinical Research, Beijing University of Chinese Medicine, 11 North No. 3 Road East, Chaoyang District, Beijing 100029, China.
authors: Xiaotong
authors: X
authors: Wang
authors: Qingguo
authors: Q
authors: Wang
authors: Frank J
authors: FJ
authors: Burczynski
authors: Weisi
authors: W
authors: Kong
authors: Yuewen
authors: Y
authors: Gong
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics / Q000378:metabolism*; D029972:Bupleurum / Q000737:chemistry*; D002417:Cattle; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004365:Drugs, Chinese Herbal / Q000494:pharmacology / Q000627:therapeutic use; D055166:Hepatic Stellate Cells / Q000187:drug effects*; D006801:Humans; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008103:Liver Cirrhosis / Q000188:drug therapy / Q000378:metabolism*; D009828:Oleanolic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D008517:Phytotherapy; D012333:RNA, Messenger / Q000378:metabolism; D012503:Saponins / Q000494:pharmacology* / Q000627:therapeutic use; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000199:Actins; C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D004365:Drugs, Chinese Herbal; D012333:RNA, Messenger; D012503:Saponins; D051028:bcl-2-Associated X Protein; D009828:Oleanolic Acid; C025759:saikosaponin D
keywords: ALK; BMP-4; Bone morphogenetic protein; Bupleurum chinense; CCl-4; DMEM/F12; Dulbecco's modified Eagle medium/F12; ECL; ECM; FBS; GDFs; HSCs; Hepatic fibrogenesis; Hepatic stellate cells; MAPKs; NG; RT-PCR; SDS; SSa; SSd; Saikosaponins and gene regulation; TBS; TGF-β; Tris-buffered-saline; activin receptor-like kinase; bone morphogenetic protein-4; carbon tetrachloride; enhanced chemiluminescence; extracellular matrix; fetal bovine serum; growth differentiation factors; hepatic stellate cells; mitogen-activated protein kinases; naringin; reverse transcriptase polymerase chain reaction; saikosaponin a; saikosaponin d; smooth muscle alpha actin; sodium dodecyl sulfate; transforming growth factor-β; α-SMA