Kruitwagen H S, Spee B, Viebahn C S, Venema H B, Penning L C, Grinwis G C M, Favier R P, van den Ingh T S G A M, Rothuizen J, Schotanus B A
Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM Utrecht, The Netherlands.
Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands.
Vet J. 2014 Sep;201(3):345-52. doi: 10.1016/j.tvjl.2014.05.024. Epub 2014 May 22.
Hepatic progenitor cells (HPCs) are an adult stem cell compartment in the liver that contributes to liver regeneration when replication of mature hepatocytes is insufficient. In this study, laser microdissection was used to isolate HPC niches from the livers of healthy dogs and dogs with lobular dissecting hepatitis (LDH), in which HPCs are massively activated. Gene expression of HPC, hepatocyte and biliary markers was determined by quantitative reverse transcriptase PCR. Expression and localisation of selected markers were further studied at the protein level by immunohistochemistry and immunofluorescent double staining in samples of normal liver and liver from dogs with LDH, acute and chronic hepatitis, and extrahepatic cholestasis. Activated HPC niches had higher gene expression of the hepatic progenitor markers OPN, FN14, CD29, CD44, CD133, LIF, LIFR and BMI1 compared to HPCs from normal liver. There was lower expression of albumin, but activated HPC niches were positive for the biliary markers SOX9, HNF1β and keratin 19 by immunohistochemistry and immunofluorescence. Laminin, activated stellate cells and macrophages are abundant extracellular matrix and cellular components of the canine HPC niche. This study demonstrates that the molecular and cellular characteristics of canine HPCs are similar to rodent and human HPCs, and that canine HPCs are distinctively activated in different types of liver disease.
肝祖细胞（HPCs）是肝脏中的一种成体干细胞区室，当成熟肝细胞的复制不足时，它有助于肝脏再生。在本研究中，使用激光显微切割技术从健康犬和患有小叶性析离性肝炎（LDH）的犬的肝脏中分离肝祖细胞龛，在LDH中肝祖细胞被大量激活。通过定量逆转录聚合酶链反应测定肝祖细胞、肝细胞和胆管标志物的基因表达。通过免疫组织化学和免疫荧光双重染色，在正常肝脏以及患有LDH、急慢性肝炎和肝外胆汁淤积的犬的肝脏样本中，进一步在蛋白质水平研究选定标志物的表达和定位。与正常肝脏的肝祖细胞相比，激活的肝祖细胞龛中肝祖细胞标志物骨桥蛋白（OPN）、14 kDa 肿瘤坏死因子受体（FN14）、整合素β1（CD29）、细胞表面糖蛋白（CD44）、Prominin-1（CD133）、白血病抑制因子（LIF）、白血病抑制因子受体（LIFR）和多梳蛋白抑制复合物2核心成分之一（BMI1）的基因表达更高。白蛋白表达较低，但通过免疫组织化学和免疫荧光检测，激活的肝祖细胞龛对胆管标志物性别决定区Y框蛋白9（SOX9）、肝细胞核因子1β（HNF1β）和角蛋白19呈阳性。层粘连蛋白、激活的星状细胞和巨噬细胞是犬肝祖细胞龛丰富的细胞外基质和细胞成分。本研究表明，犬肝祖细胞的分子和细胞特征与啮齿动物和人类肝祖细胞相似，并且犬肝祖细胞在不同类型的肝脏疾病中被特异性激活。
