Reversal effect of transplantation of bone marrow-derived endothelial progenitor cells from the liver fibrosis environment in rats with liver fibrosis

doi:10.3969/j.issn.2095-4344.2017.13.017

Abstract

Abstract:

BACKGROUND: Theoretically, bone marrow-derived endothelial progenitor cells (EPCs) from liver fibrosis rats could be “filtered” by the pathological environment in vivo. These EPCs would be more adapted to the micro-environment of liver fibrosis, and easier to differentiate into mature endothelial cells participating in the intrahepatic vascular remodeling after transplanted into the liver.
OBJECTIVE: To explore the effectiveness of transplantation of bone marrow-derived EPCs from the liver fibrosis environment in liver fibrosis rats.
METHODS: Twenty-eight Wistar rats were randomly divided into three groups as follows: normal group (n=8) were injected with olive oil, twice per week; model group (n=10) were infused with carbon tetrachloride at a dose of 3 mL/kg body weight (double doses for the first time), twice per week, and infused with normal saline through the tail vein at 2, 3 and 5 weeks; EPCs transplantation group (n=10) were infused with carbon tetrachloride at a dose of 3 mL/kg body weight (double doses for the first time), twice per week, and infused with EPCs suspension through the tail vein at 2, 3 and 5 weeks. Six weeks after final injection, the angiogenesis, hepatocyte proliferation and pathological changes in the liver tissues were observed. The liver function and coagulation function were tested.
RESULTS AND CONCLUSION: (1) The pathological changes of the liver: in the model group, fatty degeneration and hepatocyte necrosis in the liver tissue were serious, inflammatory cells were infiltrated around the portal and central vein, the portal areas expanded, and fibrous tissues overgrew. Compared with the model group, these changes were significantly relieved in the EPCs transplantation group (P < 0.05). (2) The expressions of liver-related proteins: compared with the normal group, the levels of hyaluronic acid, laminin, type III procollagen, vascular endothelial growth factor, epidermal growth factor were significantly increased in the model group (P < 0.05). Compared with the model group, the levels of hyaluronic acid, laminin and type III procollagen were decreased significantly (P < 0.05), and the levels of vascular endothelial growth factor and epidermal growth factor were increased in the EPCs transplantation group (P < 0.05). (3) Liver function and coagulation function: compared with the normal group, the liver function and blood blotting function of rats were seriously damaged in the model group (P < 0.05). Compared with the model group, the liver function and coagulation function were obviously improved in the EPCs transplantation group (P < 0.05). To conclude, transplantation of bone marrow-derived EPCs from the liver fibrosis environment is effective for liver fibrosis in rats. The mechanism may be associated with the promotion of angiogenesis in the liver.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Liver Cirrhosis, Tissue Engineering

CLC Number:

R394.2

Liu Ran, Lan Ling, Liu Bo-wei, Yuan Yuan, Qin Ling-yun, Yu Jing. Reversal effect of transplantation of bone marrow-derived endothelial progenitor cells from the liver fibrosis environment in rats with liver fibrosis[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(13): 2068-2073.

Figures/Tables 2

2.1 内皮祖细胞的细胞形态内皮祖细胞经诱导培养后 4 d内贴壁；6 d呈现首尾相连的直线状条索样结构；7 d可形成集落，呈放射状向外周生长，周边有梭形细胞不断生成，在不同的培养时间可能会呈现出椭圆形、长梭形或纺锤形；10 d起增殖加速，大部分呈多角形，并逐渐分化成具有“铺路石”样结构特征的内皮细胞[8]。 2.2 各组大鼠一般情况同正常组相比，模型组大鼠生长缓慢，体型消瘦，毛色灰暗，部分大鼠出现腹水、黄疸等表现；实验组大鼠体质量增长较快，毛色光泽度接近正常，除有1只大鼠出现腹水和黄疸表现外，余大鼠未见肝硬化体征。最终进入实验统计的大鼠数量如下：正常组8只，模型组8只(2只分别在第3周和第5周皮下注射中死亡)，实验组8只(1只在第2周死亡，1只在第4周死亡)。 2.3 各组肝组织病理比较模型组大鼠肝组织呈广泛而严重的肝细胞脂肪变性和坏死，门静脉和中央静脉周围炎症细胞浸润，汇管区扩大及大量纤维组织增生；与模型组相比，实验组肝细胞变性坏死、炎症细胞浸润及纤维组织增生等均有所减轻(图1)。 2.4 各组肝组织细胞外基质水平的比较肝组织透明质酸、层粘连蛋白和Ⅲ型前胶原水平能反映细胞外基质的沉积程度。模型组大鼠肝组织透明质酸、层粘连蛋白和Ⅲ型前胶原水平较正常组明显升高(P < 0.05)；实验组上述3项指标水平较模型组均显著下降(P < 0.05)，见表1。 2.5 各组肝组织血管内皮生长因子和表皮生长因子水平的比较与正常组相比，模型组肝组织血管内皮生长因子、表皮生长因子水平升高(P < 0.05)；同模型组相比，实验组血管内皮生长因子、表皮生长因子水平均明显升高(P < 0.05)，见表2。 2.6 各组肝功能和凝血功能的比较与正常组相比，模型组大鼠血清谷丙转氨酶、谷草转氨酶和总胆红素水平均显著升高(P < 0.05)，全血凝血酶原时间和活化部分凝血激酶时间明显延长(P < 0.05)；内皮祖细胞移植虽对总胆红素和全血凝血酶原时间水平改善作用不明显，但能显著改善谷丙转氨酶、谷草转氨酶和活化部分凝血激酶时间3项指标水平(P < 0.05)，见表3。"

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