title: [The role of extracellular signal-regulated kinase in induction of apoptosis with salvia miltiorrhiza monomer IH764-3 in hepatic stellate cells].
abstract: OBJECTIVE
  To explore the effect of Salvia miltiorrhiza monomer IH764-3 on apoptosis in hydrogen peroxide (H2O2)-stimulated hepatic stellate cells (HSCs).
  
  
  METHODS
  HSCs were cultured in medium with different IH764-3 doses (10 mg/L, 20 mg/L, 30 mg/L, 40 mg/L) and without IH764-3. Direct cell count, 3H-thymidine incorporation, Annexin-V/Propidium Iodide double-labeled flow cytometry, TUNEL and transmission electron microscopy were employed to estimate the influence of IH764-3 on proliferation and apoptosis of HSCs. The expression of extracellular signal-regulated kinase 1 (ERK1) mRNA and protein in HSCs were detected using RT-PCR and Western blot respectively.
  
  
  RESULTS
  It was showed that H2O2 could promote HSC proliferation. In contrast, IH764-3 at concentrations of 10 mg/L, 20 mg/L, 30 mg/L and 40 mg/L inhibited its proliferation. The inhibition rates were 7.13%, 28.36%, 53.80% and 73.10% (P < 0.01). And the inhibition rates of IH764-3 at concentrations of 30 mg/L at 12 h, 24 h and 48 h were 22.24%, 40.51% and 61.65%. Furthermore, IH764-3 could also induce the HSC apoptosis in dose-dependent an dtime-dependent manners (P < 0.01). In addition, after exposed of HSCs to IH764-3 for 24 h, ERK production decreased and ERK1 mRNA was down-regulated earlier about 2 h after exposure to IH764-3.
  
  
  CONCLUSIONS
  IH764-3 may inhibit the proliferation and induce apoptosis of HSCs in both dose-dependent and time-dependent manners, which may be related to down-regulation of ERK expression.
authors.affiliation: Department of Gastroenterology, the Second Hospital of Hebei Medical University, Hebei Key Laboratory of Gastroenterology, Hebei Institute of Gastroenterology, Shijiazhuang 050000, China.
authors.forename: Shu-Ming
authors.initials: SM
authors.lastname: Fang
authors.forename: Chun-Sheng
authors.initials: CS
authors.lastname: Li
authors.forename: Jun-Yan
authors.initials: JY
authors.lastname: An
authors.forename: Zhi-Na
authors.initials: ZN
authors.lastname: Dun
authors.forename: Dong-Mei
authors.initials: DM
authors.lastname: Yao
authors.forename: Lei
authors.initials: L
authors.lastname: Liu
authors.forename: Xiao-Lan
authors.initials: XL
authors.lastname: Zhang
mesh_terms: D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D002460:Cell Line; D015536:Down-Regulation / Q000187:drug effects; D004365:Drugs, Chinese Herbal / Q000302:isolation & purification / Q000494:pharmacology*; D055166:Hepatic Stellate Cells / Q000166:cytology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D048052:Mitogen-Activated Protein Kinase 3 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D027561:Salvia miltiorrhiza / Q000737:chemistry*
chemical_list: D004365:Drugs, Chinese Herbal; C079053:IH 764-3; D012333:RNA, Messenger; D006861:Hydrogen Peroxide; D048052:Mitogen-Activated Protein Kinase 3