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投稿时间：2022-03-23 网络发布日期：2022-06-20
投稿时间：2022-03-23 网络发布日期：2022-06-20
中文摘要: 目的 利用麦醇溶蛋白（gliadin）诱导小鼠乳糜泻模型，通过检测乳糜泻相关指标来评价该模型的可行性。方法 将健康的C57BL/6小鼠随机分成2组：正常组与乳糜泻组（CeD组）。第1天对CeD组的小鼠腹部两侧皮下注射100 μl的 gliadin与完全弗氏佐剂的乳液（含100 μg gliadin），对正常组的小鼠腹部两侧皮下注射100 μl的1×PBS溶液。第8天对CeD组的小鼠腹部两侧皮下注射100 μl的gliadin与不完全弗氏佐剂的乳液（含50 μg gliadin），对正常组的小鼠腹部两侧再次皮下注射100 μl的1×PBS溶液。第14天，测量注射25 μl gliadin溶液（2.5 mg/ml）和DMSO溶液前后的CeD组，小鼠的足底厚度。连续测量灌胃0.2 ml gliadin的0.1 mol/L醋酸溶液的小鼠体重。造模结束后，处死小鼠，测量小肠长度。利用实时荧光定量聚合酶链式反应（RT-qPCR）检测小鼠十二指肠组织中的干扰素（IFN）-γ、白细胞介素-15（IL-15）、肿瘤坏死因子α （TNF-α）、组织转谷氨酰胺酶（TG2）的水平。结果 gliadin溶液刺激足底后，CeD小鼠足底的厚度明显高于刺激前 ［(3.25±0.05)mm vs (2.30±0.05)mm，P<0.01］，CeD组（gliadin）小鼠的足底肿胀程度为（0.95±0.05）mm，明显大于正常组的（0.54±0.08）mm及CeD（DMSO）组的（0.53±0.21）mm（P＜0.01）。相对于对照组，CeD组小鼠出现体重下降、〖JP2〗肠损伤。CeD组的TNF-α，TG2、IL-15 和 IFN-γ这4种乳糜泻发生的标志性产物的含量显著增加（P<0.01）。结论 该造模方法在肠损伤、肠道炎症等方面均能很好的模仿临床上乳糜泻的疾病特征，可为未来乳糜泻治疗的研究提供基础。
Abstract:Objective To evaluate the feasibility of the mouse model of celiac disease (CeD) induced by gliadin through testing the relevant indexes of celiac disease. Methods Healthy C57BL/6 mice were randomly divided into normal group and CeD group. On Day 1, 10 mice in the CeD group were injected subcutaneously with 100 μl complete Freund adjuvant (containing 100 μg gliadin) on both sides of the abdomen, and the mice in the normal group were injected subcutaneously with 100 μl 1× PBS solution on both sides of the abdomen. On Day 8， the CeD group were injected subcutaneously with 100 μl Freund incomplete adjuvant (containing 50 μg gliadin) on both sides of the abdomen, and the normal group were injected subcutaneously with 100 μl 1× PBS solution on both sides of the abdomen again. On Day 14, the plantar thickness of mice in CeD group before and after injection of 25 μl gliadin (2.5 mg/ml) or DMSO was measured. The body weight of mice was continuously measured by gavage of 0.2 ml gliadin in 0.1 mol/L acetic acid solution. After modeling, the mice were executed and the length of small intestine was measured. Real time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the levels of interferon (IFN)-γ, interleukin-15 (IL-15), tumor necrosis factor α (TNF-α) and tissue transglutaminase (TG2) in mouse duodenum. Results After gliadin solution stimulated the plantar, the thickness of plantar was significantly higher than that before stimulation ［(3.25±0.05) mm vs (2.30±0.05) mm，P<0.01］. The degree of plantar swelling in CeD （gliadin） group was (0.95±0.05) mm, which was significantly higher than that in normal group ［(0.54±0.08) mm］ and CeD (DMSO) group ［(0.53±0.21) mm］. Compared with the control group, the mice in the CeD group had weight loss and intestinal injury. The level of TNF-α，TG2, IL-15 and IFN-γ in CeD group was significantly increased (P<0.01). Conclusion This modeling method can well imitate the clinical characteristics of celiac disease in terms of intestinal injury and intestinal inflammation, which provides a basis for the research of celiac disease treatment in the future.
keywords: Gliadin Celiac disease Animal model Delayed hypersensitivity Interferon-γ Interleukin-15 Tumor necrosis factor-α Transglutaminase
文章编号： 中图分类号：R574.4 文献标志码：A
基金项目:江苏省“六大人才高峰”高层次人才项目（WSW-006）；南京医科大学姑苏学院重点项目立项（GSKY20220109）
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引用文本：
殷双双，范文涛，范志宁，赵黎黎，刘莉.常规饲养C57BL/6乳糜泻小鼠模型的构建及其评价[J].中国临床研究,2022,35(6):770-774.
殷双双，范文涛，范志宁，赵黎黎，刘莉.常规饲养C57BL/6乳糜泻小鼠模型的构建及其评价[J].中国临床研究,2022,35(6):770-774.