| Objective:Preliminary discussion the regularity of hepatic oval cells(Hocs) proliferation in liver proliferation modle. Detected the expression of transforming growth factor beta(TGF beta), active receptor-like kinase5(ALK-5) and active receptor-like kinase 1(ALK-1) in liver tissue and the relation between transforming growth factor beta(TGF beta), active receptor-like kinase 5(ALK-5) and active receptor-like kinase 1(ALK-1) and hepatic oval cells(Hocs).Methods:Using random number table method to divide F344 rats into three groups: sham-operated group(suture incision and do not do any processing on the middle abdominal incision),partial-hepatectomy group(PH group: two-thirds of liver resection and 50% alcohol filled the stomach in postoperative) and the 2-acetaminofluorence group(2-AAF/PH group: two-thirds of liver resection and2-AAF solvent filled the stomach in postoperative). According to the time points, three groups were randomly divided three groups to 9 subgroups: sham-operated group 7d, 14 d and 21d; PH group 7d, 14 d and 21d; 2-AAF group: 7d, 14 d and 21d; every subgroup 4 F344 rats. Each subgroup takes liver tissue accoding to the sent time points in advance.To observe cytology morphology of Hocs in hematoxylin-eosin staining(HE). Using immunohisto–chemical(Streptomyces avidin peroxidase link method: SP)to identificate Hoc. Detected the protein and mRNA expression of TGFbeta, ALK-5 and ALK-1 in liver tissue on 7d, 14 d and 21 d of three groups using the reverse transcription-polymerase chain reaction(RT-PCR) and Western blot.Result:HE: Postoperative visible hepatic lobules are clean and in alignment, liver cells structure is normal in sham-operated group. In PH group, liver cells are edema postoperative and structure also gradually changes even a few liver cells present point neucrosis after postoperative. In 2-AAF/PH group can survey many cells which surrounding clusters or scattered distribution around hepatic portal area. The nucleis of these cells are ovoid, the volume about one third of the normal liver cell and these cells were less cytoplasm.Immunohistochemical: In each time point, it was hard to see Epithelial cell adhesion molecule(EpCAM) and(Thymine-1) Thy-1 positive cells(Hocs) are scarce in sham- operated group. In PH and2-AAF/PH groups, a few Hocs were observed at 7th and migrated to liver parenchyma at 14 th but the positive cells number of later is more. At 21 th, hepatic oval cells are a few in PH group but many in the2-AAF/PH group.RT-PCR and Western blot testing results: expressions of TGFβ mRNA and ptotein in PH group and2-AAF/PH group were higher than in the sham-operated group and the differences were statistically significant(P<0.05), PH group was greatly higher than 2-AAF/PH group and the differences were significant statistically significance(P<0.01).There was no statistically significant differences insham-operated group within three time points(P>0.05), but the differences in PH and 2-AAF/PH group had statistically significance(P<0.05). ALK-5 mRNA and ptotein expression: PH and 2-AAF/PH group were higher than the sham-operated group and the differences was statistically significant(P<0.05); the expression of differences in 2-AAF/PH group was higher than PH group and had remarkable statistics meaning(P<0.01). There time points expression differences of the sham-operated group was no statistically significant(P>0.05) but in PH and 2- AAF/PH group was significant statistically(P<0.01). the expressions of ALK-1 mRNA and ptotein in PH and 2-AAF/PH group were higher than the sham-operated group,which had statistically significant differences(P<0.05), PH group higher than 2-AAF/PH group and the differences was statistically significance(P<0.05). However, the expression of differences of sham-operated group in three time points was no statistically significant(P>0.05), the expression in PH and2- AAF/PH group had statistically significant meaning(P<0.01).Conlusion:1. The method of two-thirds of liver resection and postoperative gastric 2-AAF solvent to inhibite liver cells proliferation can successfully construct hepatic oval cell proliferation model in micro environment of liver proliferation. The expression of EPCAM and Thy-1 was positive in hepatic oval cells in Hoc proliferation model.2. The expression of TGF beta and Hoc proliferation presents the dynamic tendency, but the tendency between the expression of TGF beta and Hoc proliferation was not synchronic in hepatic oval cell proliferation model. Hoc proliferation dynamic tendency synchronizes with ALK- 5 expression of dynamic tendency but different with ALK- 1 dynamic tendency. This phenomenon may be predicted that the functions of regulatory mechanism in Hoc proliferation were various. |
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