title: [Activation of hepatocyte growth factor-induced apoptosis in hepatic stellate cells].
abstract: OBJECTIVE
  To determine whether apoptosis is induced in rat hepatic stellate cells (HSCs) in response to activation of the hepatocyte growth factor (HGF) by hepatocyte growth factor activator (HGFA) by using a co-culture system of bone marrow mesenchymal stem cells (BMSCs) and HSCs.
  
  
  METHODS
  In this study, cells were divided into the following five groups: HSC control group: HSCs co-cultured with fibroblast cells; HSCs blank group: HSCs cultured alone; BMSCs blank group: BMSCs cultured alone; Experimental group: BMSCs + HSCs; HGFA intervention group: HSCs treated with 70 ng/mL of HGFA. The culture systems were established in culture plates with transwell inserts, and cells were assessed at 24, 48, and 72 h of growth. Dynamic changes in cell morphology were observed under an inverted phase contrast microscope. The surface markers of BMSCs and the apoptosis rate of HSCs were detected by Annexin-V-FITC/propidium iodide (PI). Expression of a-smooth muscle actin (SMA) in HSCs was evaluated by immunohistochemistry. The presence of activated HGF (HGF-a chain) was determined by immunofluorescent staining. HSC proliferation was measured by MTT assay, and the concentrations of HGF and HGFA were quantified by enzyme-linked immunosorbent assay (ELISA).
  
  
  RESULTS
  MTT results indicated that treatment with HGF alone had no effect on HSC proliferation rate (vs. HSC blank group, P more than 0.05), but that 24 h treatment with HGFA significantly inhibited the proliferation rate (0.26 ± 0.00 vs. blank group: 0.13 ± 0.04, P = 0.02); moreover, this effect was concentration-dependent. Expression of HGF-a was lower in the experimental group than in the HGFA intervention group at 72 h (37.24 ± 1.03 vs. 40.44 ± 0.77, P = 0.04), and both of these groups had higher expression than the control group at all time points examined (P less than 0.05). The apoptosis rate was consistently higher in the experimental group than in the HGFA intervention group, but most robustly at 72 h (40.77 ± 1.16% vs. 33.35 ± 2.04%, P = 0.00); moreover, the apoptosis rate was significantly higher than that in the control group at all time points examined (P less than 0.01). The concentration of HGF in the experimental group and the HGFA intervention group showed a time-dependent reduction, and was consistently lower than that in the HSCs control group (P less than 0.05). Finally, the concentration of HGFA was higher in the experimental group than in the blank group at all time points examined (P less than 0.05).
  
  
  CONCLUSIONS
  The BMSC-HSC co-culture system can promote secretion of HGFA from HSCs and HGF activation, thereby inducing apoptosis of HSCs.
authors.affiliation: Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China.
authors.forename: Yun-chao
authors.initials: YC
authors.lastname: Meng
authors.forename: Hai-xing
authors.initials: HX
authors.lastname: Jiang
authors.forename: Jun-hong
authors.initials: JH
authors.lastname: Zhang
authors.forename: Zheng-feng
authors.initials: ZF
authors.lastname: Lu
authors.forename: Shan-yu
authors.initials: SY
authors.lastname: Qin
authors.forename: Lin
authors.initials: L
authors.lastname: Ning
authors.forename: Wen
authors.initials: W
authors.lastname: Yang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001854:Bone Marrow Cells / Q000166:cytology; D002478:Cells, Cultured; D018920:Coculture Techniques; D055166:Hepatic Stellate Cells / Q000166:cytology*; D017228:Hepatocyte Growth Factor / Q000378:metabolism* / Q000494:pharmacology; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012697:Serine Endopeptidases / Q000378:metabolism
chemical_list: D017228:Hepatocyte Growth Factor; C081092:HGF activator; D012697:Serine Endopeptidases