Immunohistochemical analyses of cell cycle progression and gene expression of biliary epithelial cells during liver regeneration after partial hepatectomy of the mouse
Immunohistochemical analyses of cell cycle progression and gene expression of biliary epithelial cells during liver regeneration after partial hepatectomy of the mouse
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DOI：
10.1538/expanim.15-0082
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发表时间：
2016-04-01
影响因子：
2.4
通讯作者：
Shiojiri, Nobuyoshi
中科院分区：
文献类型：
--
作者：
Fukuda, Tatsuya;Fukuchi, Tomokazu;Shiojiri, Nobuyoshi
肝脏具有显著的再生能力，在手术切除其质量块后，剩余的组织通过其组成细胞的补偿性生长进行快速再生。尽管肝细胞在邻近细胞的各种信号分子的调控下同步增殖，但关于肝切除后胆管细胞如何再生的详细分析还很少。本研究旨在通过免疫组织化学和RT-PCR技术对小鼠肝部分切除后胆管细胞的细胞周期进程和基因表达进行广泛的分析，以阐明胆管细胞是如何再生的。免疫组织化学检测肝再生过程中细胞周期标志物增殖细胞核抗原、Ki67抗原、拓扑异构酶11a和磷酸化组蛋白H3的表达，发现肝细胞在切除后48~72h出现S期和M期高峰。而胆管上皮细胞的增殖活性明显低于肝细胞，其S时相的高峰期被推迟。有丝分裂样在胆管细胞中很少检测到。RT-PCR分析胆汁标志物Spp1(骨桥蛋白)、Epcam和HNF1B的基因表达，表明它们在肝再生过程中表达上调。门静脉周围肝细胞表达部分胆汁标志物，包括Spp1mRNA和蛋白。部分门脉周围肝细胞HNF4α和HNF1α表达下调。在肝再生过程中，肝母细胞发育过程中决定细胞命运的Notch信号分子的基因表达上调。Notch信号可能参与胆道再生。
The liver has a remarkable regeneration capacity, and, after surgical removal of its mass, the remaining tissue undergoes rapid regeneration through compensatory growth of its constituent cells. Although hepatocytes synchronously proliferate under the control of various signaling molecules from neighboring cells, there have been few detailed analyses on how biliary cells regenerate for their cell population after liver resection. The present study was undertaken to clarify how biliary cells regenerate after partial hepatectomy of mice through extensive analyses of their cell cycle progression and gene expression using immunohistochemical and RT-PCR techniques. When expression of PCNA, Ki67 antigen, topoisomerase I la and phosphorylated histone H3, which are cell cycle markers, was immunohistochemically examined during liver regeneration, hepatocytes had a peak of the S phase and M phase at 48-72 h after resection. By contrast, biliary epithelial cells had much lower proliferative activity than that of hepatocytes, and their peak of the S phase was delayed. Mitotic figures were rarely detectable in biliary cells. RT-PCR analyses of gene expression of biliary markers such as Spp1 (osteopontin), Epcam and Hnf1b demonstrated that they were upregulated during liver regeneration. Periportal hepatocytes expressed some of biliary markers, including Spp1 mRNA and protein. Some periportal hepatocytes had downregulated expression of HNF4 alpha and HNF1 alpha. Gene expression of Notch signaling molecules responsible for cell fate decision of hepatoblasts to biliary cells during development was upregulated during liver regeneration. Notch signaling may be involved in biliary regeneration.