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Objective To investigate the effect of hepatopoietin Cn(HPPCn) on liver stem cells.Methods In this study, WB-F344 cell line was used, and MTT and flow cytometry assay were conducted to determine cell proliferation and apoptosis.Transwell assay was used to test the migration of WB-F344 cells.A 2AAF-partial hepatectomy(PH) mouse model was used to observe the effect of HPPCn on liver stem cell proliferation in vivo.Results HPPCn enhanced WB-F344 cell proliferation and migration and activated the SphK1, Erk and Stat3 signal pathways.The analysis of the 2AAF-PH mouse model showed that oval cells in the experimental group far outnumbered those in control and the regeneration of the liver was improved post PH.Conclusion HPPCn can increase the liver stem cell proliferation and survival while promoting the regenenation of the liver by augmenting oval cell proliferation.
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AIM:To look for a suitable signal peptide which may effectively conduct hepatopoietin(HPO)secretion,various recombinant eukaryotic expression vectors were constructed.METHODS:Different exogenous signal sequences were spliced with HPO cDNA by PCR,and the spliced genes were cloned into eukaryotic expression plasmids.The different recombinants were respectively tansfected into COS-7 cells by Lipofectamine 2000 method and the secretion of HPO was analyzed by Western blotting.RESULTS:Western blotting analysis indicated that the signal peptides from interleukin-1 receptor antagonist(IL-1ra)and an artifical signal peptide did not secret HPO directly and effectively,but the signal peptide from murine Ig kappa secreted HPO directly with great efficiency.The molecular weight of the secreted HPO was 30 kD,which means that the secreted HPO existed in homodimer.CONCLUSION:Secreted recombinant expression plasmid is successful constructed.The result may pave the way for the gene therapy of hepatic fibrosis.