摘要
Elucidation of the mechanisms of liver fibrogenesis is important to treat liver fibrosis.In this study,we established rat models of liver fibrosis with stages from 0–1,2,and 3–4 to 4 at 2,4,6,and 8 weeks,respectively,by injection of pig serum.Liver fibrogenesis was detected by Masson’s trichrome staining.Rat non-parenchymal cells(NPCs)were enriched 4-fold by Percoll density gradient centrifugation.Protein extracts from NPCs were prepared at 4 and 8 weeks,separated by two-dimensional electrophoresis,and then stained with Coomassie Blue G-250.At 4 weeks,we identified 18 non-redundant differentially expressed proteins of which protein disulfide-isomerase associated protein 3(PDIA3)and NDUV showed consistent expression at protein and mRNA levels from 4 to 8 weeks.PDIA3 was found to be down-regulated by Western blotting in the rat model and immunohistochemically in human liver.Our results revealed important aspects of the pathogenesis/progression of liver fibrosis and demonstrated important changes in protein expression levels of NPCs at various stages of liver fibrosis.
Elucidation of the mechanisms of liver fibrogenesis is important to treat liver fibrosis. In this study, we established rat models of liver fibrosis with stages from 0-1, 2, and 3-4 to 4 at 2, 4, 6, and 8 weeks, respectively, by injection of pig serum. Liver fi- brogenesis was detected by Masson＇s trichrome staining. Rat non-parenchymal cells （NPCs） were enriched 4-fold by Percoll density gradient centrifugation. Protein extracts from NPCs were prepared at 4 and 8 weeks, separated by two-dimensional electrophoresis, and then stained with Coomassie Blue G-250. At 4 weeks, we identified 18 non-redundant differentially ex- pressed proteins of which protein disulfide-isomerase associated protein 3 （PDIA3） and NDUV showed consistent expression at protein and mRNA levels from 4 to 8 weeks. PDIA3 was found to be down-regulated by Western blotting in the rat model and immunohistochemically in human liver. Our results revealed important aspects of the pathogenesis/progression of liver fi- brosis and demonstrated important changes in protein expression levels of NPCs at various stages of liver fibrosis.
基金
supported by the Major New Drug Discovery Science and Technology(2012ZX09303013)
the National Basic Research Program of China(2011CB910700)
National Natural Science Foundation of China(81271834)
China Postdoctoral Science Foundation(20100471238)
China Postdoctoral Science Special Foundation(201104516)
National Key Technology R&D Program of China(2012ZX10001003)
the Postdoctoral Science Foundation of Central South University,Science and Technology Commission of Shanghai Municipality(11DZ2292900)
