Kizaki Keiichiro, Kageyama Tomomi, Toji Noriyuki, Koshi Katsuo, Sasaki Kouya, Yamagishi Norio, Ishiguro-Oonuma Toshina, Takahashi Toru, Hashizume Kazuyoshi
Cooperative Department of Veterinary Medicine, Iwate University, Morioka, Japan.
United Graduate School of Veterinary Sciences, Gifu University, Gifu, Japan.
Anim Sci J. 2020 Jan-Dec;91(1):e13324. doi: 10.1111/asj.13324. Epub 2019 Dec 20.
Liver performs several important functions; however, predicting its functions is difficult. Methods of analyzing gene expression profiles, for example, microarray, provide functional information of tissues. Liver and peripheral blood leukocytes (PBLs) were collected from Holstein cows subjected to two different physiological conditions (non-pregnant and pregnant), and PBLs were fractionated by gradient cell separation. RNA from PBLs and liver were applied to oligo-DNA microarray and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). It revealed a group of stable bovine liver genes under constant physiological conditions. When they applied to physiological conditions including non-pregnant and pregnant, the profiles of some genes in liver were consistent with those in PBLs. Microarray data subjected to a principal component analysis (PCA) showed that the hepatic gene expression profiles were more consistent with those of granulocytes than mononuclear cells. The relationship of gene profiles in liver with granulocytes was confirmed by RT-qPCR and hierarchical cluster analysis. Gene profiles of granulocytes were more reliable than those of mononuclear cells, which reflected liver functions. These results suggest that the genes expressed in PBLs, particularly granulocytes, may be convenient bioindicators for the diagnosis of clinical disorder and/or detecting aberration of liver functions in cows subjected to different physiological conditions.
肝脏执行多种重要功能;然而,预测其功能却很困难。例如,分析基因表达谱的方法,如微阵列,可提供组织的功能信息。从处于两种不同生理状态(未怀孕和怀孕)的荷斯坦奶牛身上采集肝脏和外周血白细胞(PBLs),并通过梯度细胞分离法对PBLs进行分级分离。将来自PBLs和肝脏的RNA应用于寡核苷酸DNA微阵列和逆转录定量聚合酶链反应(RT-qPCR)。研究揭示了一组在恒定生理条件下稳定的牛肝脏基因。当将它们应用于包括未怀孕和怀孕在内的生理状态时,肝脏中一些基因的表达谱与PBLs中的一致。对微阵列数据进行主成分分析(PCA)表明,肝脏基因表达谱与粒细胞的表达谱比与单核细胞的更一致。通过RT-qPCR和层次聚类分析证实了肝脏基因谱与粒细胞的关系。粒细胞的基因谱比单核细胞的更可靠,能反映肝脏功能。这些结果表明,在PBLs中表达的基因,尤其是粒细胞中的基因,可能是用于诊断奶牛在不同生理条件下临床疾病和/或检测肝功能异常的便捷生物指标。