Igarashi Hirotaka, Ohno Koichi, Maeda Shingo, Kanemoto Hideyuki, Fukushima Kenjiro, Uchida Kazuyuki, Tsujimoto Hajime
Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Science, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Vet Immunol Immunopathol. 2014 May 15;159(1-2):1-10. doi: 10.1016/j.vetimm.2014.03.003. Epub 2014 Mar 12.
Inflammatory colorectal polyps (ICRPs) are commonly seen in miniature dachshund (MD) dogs; typically, multiple polyps form with severe neutrophil infiltration. ICRP is thought to be a novel form of inflammatory bowel disease (IBD), but its etiology has not been investigated. The innate immune system is implicated in the pathogenesis of both human and canine IBD. Therefore, the aim of the current study was to evaluate the messenger RNA (mRNA) expression profiles of pattern recognition receptors (PRRs) and cytokines in ICRPs. Polyp tissues were collected by colonoscopic biopsies from 24 MDs with ICRPs. Non-polypoid colonic mucosa was collected from all MDs with ICRPs and 21 clinically healthy beagles (as the controls). The expression levels of the mRNAs encoding toll-like receptors (TLRs) 1-10; nucleotide-binding oligomerization domain (NOD)-like receptors NOD1 and NOD2; and cytokines IL-1β, IL-6, IL-8/CXCL8, and TNF-α were evaluated by quantitative real-time RT-PCR. Three of the 10 well-known candidate reference genes were selected as housekeeper genes based on analyses from the GeNorm, NormFinder, and BestKeeper programs. Levels of TLR1, TLR2, TLR4, TLR6, TLR7, TLR8, TLR9, TLR10, NOD2, and all cytokines were significantly upregulated in the polyps relative to those in the controls. There was significant decrease in the expression levels of TLR3 and NOD1 in the polyp tissues compared to the non-polypoid colonic mucosa obtained from MDs with ICRPs. All upregulated PRR mRNAs were positively correlated with all proinflammatory cytokine mRNAs. This study demonstrated the dysregulation of PRRs and proinflammatory cytokines in ICRPs of MDs, which may play an important role in the pathogenesis of this disease.
炎症性结肠息肉(ICRPs)常见于迷你腊肠犬(MD);通常会形成多个息肉,并伴有严重的中性粒细胞浸润。ICRP被认为是炎症性肠病(IBD)的一种新形式,但其病因尚未得到研究。先天性免疫系统与人类和犬类IBD的发病机制有关。因此,本研究的目的是评估ICRPs中模式识别受体(PRRs)和细胞因子的信使核糖核酸(mRNA)表达谱。通过结肠镜活检从24只患有ICRPs的MD中收集息肉组织。从所有患有ICRPs的MD和21只临床健康的比格犬(作为对照)中收集非息肉样结肠黏膜。通过定量实时逆转录聚合酶链反应(RT-PCR)评估编码Toll样受体(TLRs)1-10、核苷酸结合寡聚化结构域(NOD)样受体NOD1和NOD2以及细胞因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8/CXC趋化因子配体8(IL-8/CXCL8)和肿瘤坏死因子-α(TNF-α)的mRNA表达水平。根据GeNorm、NormFinder和BestKeeper程序的分析,从10个著名的候选参考基因中选择了3个作为管家基因。与对照组相比,息肉中TLR1、TLR2、TLR4、TLR6、TLR7、TLR8、TLR9、TLR10、NOD2和所有细胞因子的水平均显著上调。与从患有ICRPs的MD中获得的非息肉样结肠黏膜相比,息肉组织中TLR3和NOD1的表达水平显著降低。所有上调的PRR mRNA与所有促炎细胞因子mRNA均呈正相关。本研究证明了MD的ICRPs中PRRs和促炎细胞因子的失调,这可能在该疾病的发病机制中起重要作用。