Department of Veterinary Sciences and Services, Royal Veterinary College, University of London, North Mymms, Hatfield, Hertfordshire, UK; Small Animal Clinic (Internal Medicine), Justus-Liebig University, Giessen, Germany.
Scand J Immunol. 2014 Aug;80(2):85-94. doi: 10.1111/sji.12186.
The composition of the microbiome plays a significant role in the pathogenesis of inflammatory bowel disease (IBD) in humans and chronic enteropathies (CE) in dogs. The administration of probiotic micro-organisms is one way of modulating the microbiome, but experiments elucidating mechanisms of action of probiotics in the intestine of healthy and CE dogs are lacking. The aim of our study was to investigate the effects of different Toll-like receptor (TLR) ligands and Enterococcus faecium (EF) on ex vivo cultured duodenal samples and whole blood (WB) from dogs with food-responsive chronic enteropathy (FRE) when compared to healthy dogs. Biopsy stimulation was performed in 17 FRE and 11 healthy dogs; WB stimulation was performed in 16 FRE and 16 healthy dogs. Expression of TLR2, 4, 5 and 9, IL-17A, IL-22, IFNy, TNFα, IL-4, IL-10, TGFβ and PPARy was determined in biopsies by quantitative polymerase chain reaction (PCR). In addition, production of TNFα, IL-10, IFNy and IL-17A protein in WB and biopsy supernatants was assessed by ELISA. Treatment with individual TLR ligands or EF induced a variety of changes in the expression of different TLRs and cytokines, but not necessarily a consistent change with a single stimulating agent. Even though cytokine protein could not be detected in supernatants from ex vivo stimulated biopsies, we found TNFα protein responses in blood to be opposite of the transcriptional responses seen in the biopsies. Stimulation of canine duodenal biopsies with TLR ligands can potentially induce anti-inflammatory gene expression, especially in healthy tissue, whereas the effects of EF were limited.
微生物组的组成在人类炎症性肠病（IBD）和犬慢性肠炎（CE）的发病机制中起着重要作用。益生菌微生物的给药是调节微生物组的一种方法，但缺乏阐明益生菌在健康和 CE 犬肠道中作用机制的实验。我们的研究目的是研究不同 Toll 样受体（TLR）配体和屎肠球菌（EF）对食物反应性慢性肠炎（FRE）犬与健康犬的离体培养十二指肠样本和全血（WB）的影响。对 17 只 FRE 和 11 只健康犬进行了活检刺激；对 16 只 FRE 和 16 只健康犬进行了 WB 刺激。通过定量聚合酶链反应（PCR）在活检中测定 TLR2、4、5 和 9、IL-17A、IL-22、IFNy、TNFα、IL-4、IL-10、TGFβ和 PPARγ的表达。此外，通过 ELISA 评估 WB 和活检上清液中 TNFα、IL-10、IFNy 和 IL-17A 蛋白的产生。用单个 TLR 配体或 EF 处理诱导了不同 TLR 和细胞因子表达的多种变化，但不一定是单一刺激剂的一致变化。尽管不能在离体刺激的活检上清液中检测到细胞因子蛋白，但我们发现血液中 TNFα 蛋白的反应与活检中观察到的转录反应相反。TLR 配体对犬十二指肠活检的刺激可能会诱导抗炎基因表达，特别是在健康组织中，而 EF 的作用有限。
