Development of regenerative medicine and therapeutic system for hepatic failure applied by hepatocyte nuclear factor 4

我们将腺病毒介导的HNF-4转染入肝前体细胞，并分析其肝脏特异性功能的表达。腺病毒介导的HNF-4基因转移导致HNF-4、载脂蛋白（Apo）A1、ApoC 3和PXR的mRNA表达增加。然后将HNF-4过表达的肝祖细胞注射到受体小鼠中，受体小鼠用二甲基亚硝胺和30%部分肝切除术治疗。治疗组小鼠的存活时间明显长于对照组小鼠。用HNF-4转染的细胞处理的小鼠的白蛋白、总胆固醇和葡萄糖的血浆水平高于对照小鼠。雄性Wistar大鼠皮下注射2-乙酰氨基芴。7天后，他们接受了70%的部分肝切除术，并感染了携带HNF-4或Lac-Z cDNA的重组腺病毒。在HNF-4处理的大鼠中，卵圆细胞的总数显著减少。在这一点上，HNF-4治疗的大鼠经历了肝细胞增殖的抑制，而HNF-4，载脂蛋白CIII，酪氨酸氨基转移酶和白蛋白，肝细胞分化的标志物，mRNA的表达增加。与此相反，细胞角蛋白-19的mRNA表达，胆管上皮细胞分化的标志物，在HNF-4转移大鼠的肝脏下降。这些结果表明，腺病毒介导的HNF-4转染诱导分化从肝祖细胞肝实质细胞在体外，这些细胞可能是有用的作为一个来源的细胞移植在肝脏疾病。转染HNF-4基因可促进肝卵圆细胞向肝细胞分化。

We transfected adenovirus-mediated HNF-4 into hepatic progenitor cells, and analyzed the expressions of the liver-specific functions. Adenovirus-mediated HNF-4 gene transfer resulted in increases in the expressions of HNF-4, apolipoprotein (Apo) A1 ApoC3, and pregnane X receptor (PXR) mRNA. HNF-4-overexpressing hepatic progenitor cells were then injected into recipient mice, which were treated with dimethylnitrosamine and 30% partial hepatectomy. The treated mice survived significantly longer than the control mice. The plasma levels of albumin, total cholesterol, and glucose were higher in the mice treated with cells transfected by HNF-4 than in the control mice. Male Wistar rats were administered 2-acetylaminofluorene subcutaneously. Seven days later, they received 70% partial hepatectomy and infected with a recombinant adenovirus carrying the cDNA for HNF-4 or Lac-Z. The total numbers of oval cells significantly decreased in the HNF-4 treated rats. At this point, HNF-4-treated rats experienced an inhibition of liver cell proliferation, while mRNA expressions of HNF-4, apolipoprotein CIII, tyrosine amino transferase and albumin, markers for the differentiation of hepatocytes, increased. In contrast, mRNA expression of cytokeratin-19, a marker for the differentiation of biliary epithelial cells, decreased in the livers of HNF-4 transferred rats. These findings demonstrate that adenovirus-mediated HNF-4 transfection induces the differentiation from hepatic progenitor cells to hepatic parenchymal cells in vitro and that these cells may be useful as a source for cell transplantation in liver diseases. The transfer of HNF-4 gene could accelerate the differentiation of hepatic oval cells to hepatocytes.