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Use of quantitative real-time PCR to determine the local inflammatory response in the intestinal mucosa and muscularis of horses undergoing small intestinal resection.

使用定量实时PCR确定接受小肠切除术的马的肠粘膜和肌层中的局部炎症反应。

  • 影响因子:1.86
  • DOI:10.1111/evj.13429
  • 作者列表:"Lisowski ZM","Lefevre L","Mair TS","Clark EL","Hudson NPH","Hume DA","Pirie RS
  • 发表时间:2021-02-01
Abstract

BACKGROUND:Studies in rodents and humans have demonstrated that intestinal manipulation or surgical trauma initiates an inflammatory response in the intestine which results in leucocyte recruitment to the muscularis externa causing smooth muscle dysfunction. OBJECTIVES:To examine the intestinal inflammatory response in horses undergoing colic surgery by measuring relative differential gene expression in intestinal tissues harvested from surgical colic cases and control horses. STUDY DESIGN:Prospective case-control study. METHODS:Mucosa and muscularis externa were harvested from healthy margins of resected small intestine from horses undergoing colic surgery (n = 12) and from intestine derived from control horses euthanised for reasons unrelated to the gastrointestinal tract (n = 6). Tissue was analysed for genes encoding proteins involved in the inflammatory response: interleukin (IL) 6 and IL1β, C-C motif chemokine ligand 2 (CCL2), tumour necrosis factor (TNF), prostaglandin-endoperoxide synthase 2 (PTGS2) and indoleamine 2,3-dioxygenase (IDO1). Relative expression of these genes was compared between the two groups. Further analysis was applied to the colic cases to determine whether the magnitude of relative gene expression was associated with the subsequent development of post-operative reflux (POR). RESULTS:Samples obtained from colic cases had increased relative expression of IL1β, IL6, CCL2 and TNF in the mucosa and muscularis externa when compared with the control group. There was no difference in relative gene expression between proximal and distal resection margins and no association between duration of colic, age, resection length, short-term survival and the presence of pre-operative reflux and the relative expression of the genes of interest. Horses that developed POR had significantly greater relative gene expression of TNF in the mucosa compared with horses that did not develop POR. MAIN LIMITATIONS:Small sample size per group and variation within the colic cases. CONCLUSIONS:These preliminary data support an upregulation of inflammatory genes in the intestine of horses undergoing colic surgery.

摘要

背景: 对啮齿类动物和人类的研究表明,肠道操作或手术创伤在肠道中引发炎症反应,导致白细胞募集到外肌,引起平滑肌功能障碍。 目的: 通过测量从外科绞痛病例和对照马收获的肠组织中的相对差异基因表达,检查接受绞痛手术的马的肠道炎症反应。 研究设计: 前瞻性病例对照研究。 方法: 从接受结肠手术的马 (n = 12) 切除的小肠的健康边缘和出于与胃肠道无关的原因实施安乐死的对照马 (n = 6) 的肠中收获粘膜和外肌。分析组织中编码参与炎症反应的蛋白质的基因: 白细胞介素 (IL) 6和IL1β 、c-c基序趋化因子配体2 (CCL2) 、肿瘤坏死因子 (TNF) 、前列腺素内过氧化物合酶2 (PTGS2) 和吲哚胺2,3-双加氧酶 (IDO1)。比较两组之间这些基因的相对表达。对结肠病例进行进一步分析,以确定相对基因表达的大小是否与术后反流 (POR) 的后续发展相关。 结果: 与对照组相比,从结肠病例获得的样本在粘膜和外肌中IL1β 、IL6、CCL2和TNF的相对表达增加。近端和远端切缘之间的相对基因表达没有差异,绞痛持续时间、年龄、切除长度、短期存活和术前反流的存在与目的基因的相对表达之间没有关联。与未发生POR的马相比,发生POR的马在粘膜中具有显著更大的TNF相对基因表达。 主要局限性: 每组样本量小,结肠病例内变异。 结论: 这些初步数据支持在接受结肠手术的马的肠道中炎症基因的上调。

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