溃疡性结肠炎模型的构建及其炎症反应机制研究

神经药理学报 ›› 2014, Vol. 4 ›› Issue (5): 15-23.

溃疡性结肠炎模型的构建及其炎症反应机制研究

王倩，田慧，刘靓靓，钟明，梅艳飞，张力

河北北方学院，河北张家口，075000，中国

出版日期:2014-10-26 发布日期:2015-01-20
通讯作者: 张力，男，教授，硕士生导师；研究方向：神经药理学；E-mail：zmczl@hotmail.com
作者简介:王倩，女，硕士；研究方向：神经药理学；E-mail：wangqianpharmacy@126.com
基金资助:
国家自然基金课题(No. 81274005)

Establishment of model about Ulcerative Colitis and the Mechanism of Colitis-related Inflammatory Responses

WANG Qian TIAN Hui LIU Liang-jing ZHONG Ming MEI Yan-fei ZHANG Li

Hebei North University, Zhangjiakou, 075000, China

Online:2014-10-26 Published:2015-01-20
Contact: 张力，男，教授，硕士生导师；研究方向：神经药理学；E-mail：zmczl@hotmail.com
About author:王倩，女，硕士；研究方向：神经药理学；E-mail：wangqianpharmacy@126.com
Supported by:
国家自然基金课题(No. 81274005)

摘要/Abstract

摘要： 目的：构建2,4,6-三硝基苯磺酸(2,4,6-trinitrobenzene sulfonic acid，TNBS)以及恶唑酮(Oxazolone，OXZ)诱导的小鼠溃疡性结肠炎(Ulcerative Colitis，UC)模型。方法：将120只昆明小鼠（♂）随机分为4组（n=30）。Ⅰ组、Ⅱ组参照Morris及Walter的方法制备小鼠TNBS模型：Ⅰ组（TNBS溶剂对照组），50%乙醇0.1ml灌肠；Ⅱ组（TNBS模型组），0.6%TNBS溶液0.1ml灌肠；两组灌肠给药一次后在d 1，d 2，d 3，d 5，d 7每组处死6只。Ⅲ组、Ⅳ组参照Heller方法制备小鼠OXZ模型：Ⅲ组（OXZ溶剂对照组），皮肤涂抹100%乙醇0.1ml，每天一次，连续2d，d 7以50%乙醇0.1ml灌肠；Ⅳ组（OXZ模型组），皮肤涂抹1%OXZ溶液(100%乙醇溶解)0.1ml每天一次，连续2d（致敏），d 7以0.5%OXZ(50%乙醇溶解)0.1ml灌肠；两组灌肠给药一次后在d 1~d 5每天处死6只小鼠。观察Ⅰ组~Ⅳ组小鼠疾病活动指数(Disease Activity Index，DAI)、结肠组织大体损伤指数(Colon Macroscopic Damage Index，CMDI)和病理组织学评分(Histopathological Score，HPS)，并检测小鼠结肠组织中髓过氧化物酶(Myeloperoxidase，MPO)、白细胞介素-4(Interleukin-4，IL-4)、肿瘤坏死因子-α (Tumor Necrosis Factor -α，TNF-α)的水平。结果：两种模型组小鼠DAI，CMDI和HPS均较对照组有明显改变；TNBS和OXZ诱导的结肠炎均可导致MPO明显上升，TNBS结肠炎TNF-α明显上升，OXZ结肠炎IL-4明显下降。结论：TNBS及OXZ均能诱导小鼠溃疡性结肠炎模型。两种模型各有特点，其中TNBS诱导的小鼠溃疡性结肠炎以辅助性T1（helper1，Th1）型炎症反应为主，OXZ诱导的小鼠溃疡性结肠炎以辅助性T2（helper2，Th2）型炎症反应为主。

关键词: 溃疡性结肠炎, 2,4,6-三硝基苯磺酸(TNBS), 恶唑酮(OXZ), 白细胞介素-4(IL-4), 肿瘤坏死因子-&, alpha, (TNF-&, alpha, )

Abstract: Objective： To establish 2,4,6- trinitrobenzene sulfonic acid (TNBS) and oxazolone (OXZ) induced murine models of ulcerative colitis (UC). Methods：Totally 120 mice (♂) were randomly divided into four groups (n = 30). Ⅰ,Ⅱgroup referring to Morris and Walter TNBS model mice were prepared: Ⅰgroup (TNBS solvent control group), 50% ethanol 0.1ml enema; Ⅱgroup (TNBS model group), 0.6% TNBS solution 0.1ml enema; after the two groups enema administered once, on d 1, d 2, d 3, d 5, d 7, 6 mice in each group were sacrificed. Ⅲ, Ⅳ group referring to Heller OXZ model mice were prepared: Ⅲ group (OXZ solvent control group), skin smear 100% ethanol 0.1ml, once a day for 2d, d 7 with 50 percent ethanol 0.1ml enema; Ⅳ group (OXZ model group), skin applicator 1% OXZ solution (100% ethanol solution) 0.1ml once a day for 2d (sensitization), d 7 with 0.5% OXZ (50% ethanol solution) 0.1ml enema; after the two groups enema administered once, on d 1 to d5 6 mice in each group were sacrificed. Observed mice Disease Activity Index (DAI), Colon Macroscopic Damage Index (CMDI) and Histopathological Score (HPS) of group Ⅰ ~ Ⅳ, and detected mice colon tissue Myeloperoxidase (MPO), Interleukin -4 ( IL-4), Tumor Necrosis Factor -α (TNF-α) levels. Results： Both models mice DAI, CMDI and HPS, compared with control group, had significantly changed; TNBS-induced colitis and OXZ can lead to significantly increased MPO, TNBS colitis TNF-α significantly increased, OXZ colitis significantly decreased IL-4 . Conclusion：TNBS and OXZ could induce mouse model of ulcerative colitis, TNBS-induced mouse ulcerative colitis with Th1-type inflammation dominated, OXZ ulcerative colitis in mice induced Th2-type inflammatory response with the main , and so, ulcerative colitis is a result that Th1 and Th2 imbalance adjusted .

Key words: ulcerative colitis , 2,4,6-trinitrobenzene sulfonic acid(TNBS), Oxazolone(OXZ), Interleukin-4(IL-4), Tumor necrosis factor -&, alpha, (TNF-&, alpha, )

中图分类号:

R964

王倩, 田慧, 刘靓靓, 钟明, 梅艳飞, 张力. 溃疡性结肠炎模型的构建及其炎症反应机制研究[J]. 神经药理学报, 2014, 4(5): 15-23.

WANG Qian TIAN Hui LIU Liang-jing ZHONG Ming MEI Yan-fei ZHANG Li. Establishment of model about Ulcerative Colitis and the Mechanism of Colitis-related Inflammatory Responses[J]. ACTA NEUROPHARMACOLOGICA, 2014, 4(5): 15-23.

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