Clinic for Small Animal Internal Medicine, Vetsuisse Faculty, University of Zürich, Winterthurerstr. 268, 8057 Zürich, Switzerland.
Vet J. 2010 Mar;183(3):287-93. doi: 10.1016/j.tvjl.2008.11.012. Epub 2008 Dec 31.
The cat has recently been proposed as a valuable model for type 2 diabetes mellitus (T2DM), because feline diabetes shares several similarities with the disease in humans. Impaired beta-cell function, decreased beta-cell mass, insulin resistance that is often related to obesity, and pancreatic amyloid deposition, are among these common features. In this study, and to further develop the cat as a model of T2DM, feline pancreatic islets were isolated and real-time PCR quantification of mRNA transcripts of genes central to beta-cell function and survival established. In particular, mRNA quantification systems were determined for insulin, the insulin enhancer pancreatic duodenal homeobox-1 (PDX-1), the insulin suppressor CCAAT/enhancer binding protein-beta (C/EBPbeta), glucose transporter isoform 2 (GLUT2), Fas receptor, the caspase-8 inhibitor FLIP (FLICE [caspase-8]-inhibitory protein) and two chemokines, interleukin (IL)-8 and monocyte chemoattractant protein-1 (MCP-1). Pancreatic islets were isolated by collagenase digestion from healthy cat donors. Partial feline mRNA sequences were determined for PDX-1, C/EBPbeta, GLUT2 and FLIP using primers identified from conserved regions of human, dog and rat mRNA. These novel and the previously available sequences (insulin, Fas receptor, IL-8 and MCP-1) were used to design feline-specific primers suitable for real-time PCR in isolated pancreatic islets. The adopted protocol of collagenase digestion yielded pancreatic islets that were frequently surrounded by acinar cells. Quantification of mRNA transcripts was simple and reproducible in healthy cats. Characterisation of genes related to insulin signalling in cats will prove useful to better understand the pathogenesis of feline diabetes and possibly of human T2DM.
猫最近被提议作为 2 型糖尿病（T2DM）的有价值模型，因为猫糖尿病与人类的糖尿病有几个相似之处。受损的β细胞功能、β细胞数量减少、常与肥胖有关的胰岛素抵抗以及胰腺淀粉样沉积等都是这些共同特征之一。在这项研究中，为了进一步将猫发展为 T2DM 模型，分离了猫的胰岛，并建立了与β细胞功能和存活相关的基因 mRNA 转录本的实时 PCR 定量。特别是，确定了胰岛素、胰岛素增强因子胰腺十二指肠同源盒-1（PDX-1）、胰岛素抑制因子 CCAAT/增强子结合蛋白-β（C/EBPβ）、葡萄糖转运体同工型 2（GLUT2）、Fas 受体、caspase-8 抑制剂 FLIP（FLICE[caspase-8]-inhibitory protein）和两种趋化因子白细胞介素（IL）-8 和单核细胞趋化蛋白-1（MCP-1）的 mRNA 定量系统。通过胶原酶消化从健康的猫供体中分离胰岛。使用从人、狗和大鼠 mRNA 的保守区域鉴定的引物，确定了 PDX-1、C/EBPβ、GLUT2 和 FLIP 的部分猫 mRNA 序列。这些新的和以前可用的序列（胰岛素、Fas 受体、IL-8 和 MCP-1）被用于设计适合分离胰岛实时 PCR 的猫特异性引物。采用的胶原酶消化方案产生的胰岛经常被胰岛包围。在健康猫中，mRNA 转录本的定量简单且可重复。猫中与胰岛素信号相关的基因特征将有助于更好地理解猫糖尿病的发病机制，并可能有助于理解人类 T2DM。
