Research On The Effect Of Fructus Polygoni Orientalis On Rat Liver Regeneration And Bone Marrow Cell Proliferation

Objective: The aim of this study was to explore the effect of fructus polygoni orientalis on rat liverregeneration and the proliferation of bone marrow cell on the rat partial hepatectomy (PH) model whichwas made as described by Higgins et al, and provide the experimental data for developing the traditionalchinese medicine to treat the liver disease.Methods: Except for the SO group, other adult male wistar rat were operated2/3partial hepatectomyprior to randomly divided into5experimental groups: PH group, CP group and the low,middle,high doseof EFPO groups, respectively. EFPO groups were administrated i.g. with EFPO0.9,1.8,3.6g·kg^-1,respectively. The CP groups were treated i.p. with0.03g·kg^-1CP injection, and PH and SO groups i.g.with0.9%NaCl. The rat were sacrificed7days after administration and liver were removed, and the liverwere fixed in10%neutral formalin and embedded in paraffin. The paraffin section was used to study theexpression of PCNA by immunohistochemistry and histological change of liver by H-E staining; theliver,thymus,spleen of the rats were removed and weighed immediately to calculate organ index; Bonemarrow cells were isolated and the expression of PCNA of bone marrow cells was studied byimmunocytochemistry; Then bone marrow mononuclear cells isolated and purified by density gradientcentrifugation with60%percoll were used to test cell cycle by flow cytometry; Bone marrow biopsyimprint were used to research morphological changes and the proliferation of bone marrow cells by G-Wstaining. The max dosage was used to evaluat the safety of the EFPO.Results:The effects of EFPO on rat liver regeneration:①The immunohistochemistry result showedthat the expression of PCNA of the liver tissue of EFPO3.6g·kg^-1groups were lower than the PH groups(P <0.05).②The hepatocytes of the EFPO groups arranged radially along the central vein, and thehepatic sinusoid cavity was widen than the PH group. EFPO3.6g·kg^-1groups showed obviously widenhepatic sinusoid cavity, several enlarged nucleus, slightly ballooning degeneration of hepatocyte and spottyand focal necrosis were found in portal area, away from the central vein.③The EFPO could inhibite theweight of thymus, spleen and liver of liver regenerating rat, the organ index were significantly lower thanPH groups with a dose-dependent manner (P <0.05). The effects of EFPO on bone marrow cellsproliferation of PH rats:①The PCNA immunocytochemistry staining of bone marrow cells on rat liver regeneration shows that the EFPO1.8g·kg^-1,3.6g·kg^-1could significant inhibit the proliferation of bonemarrow cells (P <0.05).②The content of DNA and cell cycle of bone marrow cells were observed withthe flow cytometer (FCM), EFPO3.6g·kg^-1could decreased the content of DNA of BMCs, and induceBMCs gathered in G₀/G₁phase (P <0.05), and the percentage of BMCs in G₂/M and S phases weredecreased markedly,contrast to the PH group.③The result of G-W staining showed that karyocyte ofEFPO groups obviously decreased and mature erythrocyte increase than that of PH group. EFPO couldinhibit myelosis of liver regenerating rat with a dose-dependent manner (P <0.05), and the EFPO3.6g·kg^-1induced active proliferation, while SO and PH group showed apparent active myeloproliferative. Themax dosage of mice i.g. EFPO was148.8g·kg^-1, which was amount to80higher than that of adult people,so that i.g. with EFPO on mice148.8g·kg^-1was safety.Conclusion:①The EFPO inhibit the rat liver regeneration.②The EFPO inhibit the rat bone marrowcells proliferation.③The max dosage of mice i.g. was148.8g·kg^-1, which was amount to80higher thanthat of adult people, so that i.g. with EFPO on mice148.8g·kg^-1was safety.