C57BL/6小鼠炎症性肠病模型的建立与指标评价  

Establishment and Index Evaluation of Inflammatory Bowel Disease Model in C57BL/6 Mice

作　　者：何家宇 杨茗淇 伍娇 杨宇[1] 姚欢 易金娥[1] 孔丽 HE Jiayu;YANG Mingqi;WU Jiao;YANG Yu;YAO Huan;YI Jine;KONG Li(Hunan Engineering Research Center of Livestock and Poultry Health Care,Colleges of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学动物医学院,畜禽保健湖南省工程研究中心,长沙410128

出　　处：《中国畜牧兽医》2024年第9期4153-4163,共11页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(32273084);湖南省研究生科研创新基金(CX20220670);湖南农业大学研究生科研创新基金(2022XC010)。

摘　　要：【目的】通过葡聚糖硫酸钠(dextran sulfate sodium,DSS)或脂多糖(lipopolysaccharide,LPS)刺激构建小鼠炎症性肠病(inflammatory bowel disease,IBD)模型,为IBD研究提供高效经济的模型参考,并为IBD抗炎药物筛选与药效评价提供可靠的方法支撑。【方法】本试验以C57BL/6雄性小鼠为研究对象,分别暴露于不同浓度的DSS(3%、4%和5%)7 d或LPS(5、10和20 mg/kg)4 h。通过测量小鼠结肠长度,HE染色观察肠道形态结构及实时荧光定量PCR检测肠道炎性因子的mRNA表达等对模型进行评价。【结果】DSS诱导小鼠结肠变短、病理出血、腺体萎缩、结构排列絮乱、炎性细胞浸润以及结肠中炎性因子白细胞介素-1β(interleukin-1β,IL-1β)、IL-6、IL-10和肿瘤坏死因子-α(tumour necrosis factor-α,TNF-α)的mRNA表达显著上升(P<0.05),4%DSS即可高效建立IBD模型。同时,LPS引起小鼠肠系膜出血、肠道肿胀和充血,肠绒毛高度降低,隐窝深度升高,小肠中IL-10和IL-1βmRNA水平升高、IL-6 mRNA水平下降,十二指肠和回肠TNF-αmRNA水平上升,空肠TNF-αmRNA水平降低,且以5 mg/kg LPS综合影响最为显著。【结论】4%DSS和5 mg/kg LPS是C57BL/6小鼠IBD模型建立的最适浓度,肠道病理形态改变、肠道炎性细胞浸润及炎性因子IL-1β、IL-6、IL-10和TNF-α的mRNA表达异常是评价IBD模型的重要指标。【Objective】The study was aimed to construct a mouse inflammatory bowel disease(IBD)model by stimulating with dextran sulfate sodium(DSS)or lipopolysaccharide(LPS),providing an efficient and economical model reference for IBD research,and providing reliable methodological support for the screening and efficacy evaluation of IBD anti-inflammatory drugs.【Method】C57BL/6 male mice were chosen and exposed to different concentrations of DSS(3%,4%and 5%)for 7 d or LPS(5,10 and 20 mg/kg)for 4 h in the current study.The model was evaluated by measuring colon length,observing intestinal morphology and structure with HE staining,and detecting mRNA expression of intestinal inflammatory factors by Real-time quantitative PCR.【Result】DSS could decrease the colon length of mice,led to pathological bleeding,atrophy of glands and disordered structure in colon.Moreover,DSS significantly induced inflammatory cell infiltration and the mRNA expression of the inflammatory cytokines interleukin-1β(IL-1β),IL-6,IL-10 and tumor necrosis factor-α(TNF-α)in the colon of mice,which implied that 4%DSS could establish the IBD efficiently.Meanwhile,LPS could cause mesenteric hemorrhage,intestinal swelling and congestion of mice.Moreover,LPS could decrease the villus height and increased the crypt depth in the intestine.In addition,LPS could also upregulate mRNA levels of intestinal inflammatory cytokines IL-10 and IL-1β,downregulate IL-6 mRNA level and elevate TNF-αmRNA level in the duodenum and ileum,and reduce TNF-αmRNA level in the jejunum.It was found that the most significant effect of intestinal damage was 5 mg/kg LPS treatment.【Conclusion】4%DSS or 5 mg/kg LPS treatment were the optimal concentrations to establish IBD model in C57BL/6 mice.Intestinal pathomorphological changes,intestinal inflammatory cell infiltration and inflammatory factors disorder such as the mRNA expression of IL-1β,IL-6,IL-10 and TNF-αwere important indicators to evaluate the models of IBD.

关 键 词：葡聚糖硫酸钠(DSS) 脂多糖(LPS) 炎症性肠病 肠道炎症 C57BL/6小鼠 

分 类 号：S856.4[农业科学—临床兽医学]