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目的探讨HNF4α和HNF6在肝发育过程中的作用。方法用RT-PCR和原位杂交检测HNF4αmRNA和HNF6mRNA在胚胎第8、9、13、15、17d(E8、E9、E13、E15、E17)、出生后第1d(P1)以及成年小鼠肝中的表达。结果RT-PCR结果显示,HNF4α在肝中的表达始于E9,并持续到新生,至成年时表达减弱。原位杂交结果显示,在胚胎发育的各个阶段多数肝索细胞呈HNF4α阳性反应,成年时仍有少数HNF4α阳性的肝索细胞,而胆管板及胆管上皮细胞、血管内皮细胞、造血细胞等均为阴性。HNF6mRNA的表达在E9开始出现,E13消失,E15又重新出现,并一直维持到出生。E9、E15多数肝索细胞呈HNF6mRNA阳性反应,E17及P1HNF6mRNA在肝索细胞中的反应减弱,胆管板细胞出现强阳性反应。结论HNF4αmRNA表达与肝芽的形成有关,并参与肝干细胞向肝细胞的增殖、分化的过程,其在正常成年肝中的表达可能与肝细胞正常形态的维持有关。HNF6可能在肝干细胞的形成和肝干细胞向胆管细胞的分化,以及胆管细胞的增殖、分化过程中发挥作用,其在正常成年肝中可能介导维持胆管上皮细胞的正常形态。
Objective To investigate the role of HNF4α and HNF6 in liver development. Methods The expression of HNF4α mRNA and HNF6 mRNA in embryonic day 8, 9, 13, 15 and 17 (E8, E9, E13, E15 and E17), 1st day after birth (P1) expression. Results The results of RT-PCR showed that the expression of HNF4α in the liver started at E9 and continued to the newborn, and the expression was reduced to the adult. In situ hybridization results showed that most hepatic fibroblasts were positive for HNF4α in all stages of embryonic development, and there were still a few HNF4α-positive hepatic cord cells in adulthood. The bile duct epithelium, bile duct epithelial cells, vascular endothelial cells and hematopoietic cells were all negative. HNF6 mRNA expression began in E9, E13 disappeared, E15 reappeared, and has been maintained until birth. E9, E15 majority of hepatic cells showed HNF6 mRNA positive reaction, E17 and P1HNF6 mRNA response in hepatic cord cells weakened, bile duct plate cells strong positive reaction. Conclusions The expression of HNF4α mRNA is related to the formation of liver buds and is involved in the proliferation and differentiation of hepatic stem cells. The expression of HNF4α in normal adult liver may be related to the maintenance of normal morphology of hepatocytes. HNF6 may play a role in the formation of hepatic stem cells and the differentiation of hepatic stem cells into cholangiocytes and the proliferation and differentiation of cholangiocytes, which may mediate the normal morphology of biliary epithelial cells in normal adult liver.