中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (27): 4363-4368.doi: 10.3969/j.issn.2095-4344.1386

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

不同造模次数对 2，4，6-三硝基苯磺酸/乙醇诱导重症溃疡性结肠炎模型大鼠的影响

刘佳丽1，杨  坤1，徐爱玲1，刘一东1，谷雪松1，孙平良2   

  1. (1广西中医药大学研究生院，广西壮族自治区南宁市  530001；2广西中医药大学第一附属医院，广西壮族自治区南宁市  530023)
  • 收稿日期:2019-03-26 出版日期:2019-09-28 发布日期:2019-09-28
  • 通讯作者: 孙平良，博士，主任医师，硕士生导师，广西中医药大学第一附属医院，广西壮族自治区南宁市 530023
  • 作者简介:刘佳丽，女，1993年生，广东省河源市人，汉族，广西中医药大学在读硕士，主要从事肛肠疾病诊治的研究。
  • 基金资助:

    国家自然科学基金项目(81660795)，项目负责人：孙平良；广西自然科学基金项目(2017GXNSFAA198303)，项目负责人：孙平良

Effect of different modeling times on the rat models of severe ulcerative colitis induced by 2,4,6-trinitrobenzene sulfonic acid/ethanol

Liu Jiali1, Yang Kun1, Xu Ailing1, Liu Yidong1, Gu Xuesong1, Sun Pingliang2   

  1. (1Graduate College of Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China; 2the First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530023, Guangxi Zhuang Autonomous Region, China)
  • Received:2019-03-26 Online:2019-09-28 Published:2019-09-28
  • Contact: Sun Pingliang, MD, Chief physician, Master’s supervisor, the First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530023, Guangxi Zhuang Autonomous Region, China
  • About author:Liu Jiali, Master candidate, Graduate College of Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81660795 (to SPL); the Natural Science Foundation of Guangxi Zhuang Autonomous Region, No. 2017GXNSFAA198303 (to SPL)

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摘要：

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文题释义：
重症溃疡性结肠炎：经激素治疗无效的患者，体征稳定，腹泻次数> 6次/d，有明显黏液性脓血便，体温>    37.5 ℃，心率> 90次/min，血红蛋白(Hb)<100 g/L，红细胞沉降率(ESR)>30 mm/h。病理表现为可以破坏黏膜，形成黏膜表面的剥蚀或者向下穿过黏膜肌层。
2，4，6-三硝基苯磺酸(TNBS)/乙醇：三硝基苯磺酸是一种化学品，化学式是C6H3N3O9S，其联合乙醇用于大鼠造模，主要机制为乙醇破坏结肠黏膜屏障，TNBS渗入结肠作为半抗原与大分子物质结合，形成全抗原引起肠壁一系列免疫应答与炎症反应，其发病机制与人类溃疡性结肠炎相似。
摘要
背景：实验采取复合法2，4，6-三硝基苯磺酸(2，4，6-Trinitrobenzenesulfonic acid，TNBS)/乙醇建立溃疡性结肠炎大鼠模型，操作简单，持续时间长且组织学变化与人类溃疡性结肠炎相似，是目前常用的溃疡性结肠炎动物模型之一，但对不同剂量不同造模次数仍存在较大差异。
目的：采用相同剂量TNBS/乙醇不同次数诱导重症溃疡性结肠炎大鼠模型，探索重症溃疡性结肠炎适宜的造模次数。
方法：选用SPF级SD大鼠80只，购自湖南斯莱克景达实验动物有限公司，实验方案经广西中医药大学第一附属医院动物实验伦理委员会批准[批准号为2013(KF)-E-003]。随机将大鼠分为正常对照组、TNBS+乙醇灌肠1次组、TNBS+乙醇灌肠2次组、TNBS+乙醇灌肠3次组。用药组分别使用灌胃针经大鼠直肠注入相同剂量的100 mg/kg TNBS+等体积乙醇1 d、连续2 d、连续3 d。分别于给药后的3，7 d各处死10只大鼠，观察大鼠生理状态、结肠形态、疾病活动指数及组织病理学改变情况。
结果与结论：①造模7 d后大鼠体质量变化，随着造模次数的增加，大鼠体质量呈渐进性下降、症状呈渐进性加重趋势，DAI评分各组呈上升趋势；②TNBS+乙醇灌肠3次组体质量显著低于其他组(P < 0.05)；DAI评分和大体评分均显著高于其他组(P < 0.05)；③病理结果示，TNBS+乙醇灌肠3次组病变位于结肠黏膜肌层，杯状细胞丢失，隐窝形态不规则，排列紊乱，大部分可见溃疡面，较TNBS+乙醇灌肠1次组和TNBS+乙醇灌肠2次组更符合重症溃疡性结肠炎的病理表现；④结果说明，TNBS/乙醇造模中，100 mg/kg TNBS+乙醇灌肠3次连续灌肠更符合重症溃疡性结肠炎模型。

关键词: 重症溃疡性结肠炎, 2, 4, 6-三硝基苯磺酸, 大鼠, 动物模型, 造模次数, 国家自然科学基金

Abstract:

BACKGROUND: Establishing the rat model of ulcerative colitis by 2,4,6-trinitrobenzene sulfonic acid (TNBS)/ethanol method is easy to operate, has long duration and the histological changes are similar to human ulcerative colitis, which is commonly used animal model. However, the optimal dose and modeling times still remain controversial.
OBJECTIVE: To establish the rat model of the severe ulcerative colitis by different times of different doses of TNBS/ethanol, and to explore the optimal modeling times.
METHODS: Eighty Sprague-Dawley rats, SPF level, were provided by Hunan Slack Jingda Experimental Animal Co., Ltd., and the study was approved by the Experimental Animal Ethics Committee of the First Affiliated Hospital of Guangxi University of Chinese Medicine, approval number: 2013(KF)-E-003. The rats were randomly divided into normal control group, and TNBS/ethanol (100 mg/kg once, 100 mg/kg twice, and 100 mg/kg thrice) group. The rats were intragastrically injected with TNBS/ethanol through rectum for 1, 2, and 3 days continuously. Then ten rats were killed at 3 and 7 days after administration, so as to observe the physiological status, colon morphology, disease activity index and histopathological changes.
RESULTS AND CONCLUSION: (1) At 7 days after modeling, the body mass of rats progressively decreased and symptoms gradually exacerbated with the modeling times increased. The disease activity index was on a rise. (2) The body mass in the TNBS/ethanol thrice group was significantly lower than that in the other groups (P < 0.05), and the disease activity index and gross scores were significantly higher than those in the other groups (P < 0.05). (3) The pathological results showed that the TNBS/ethanol thrice group caused the lesions located in the colonic mucosa and muscularis, the goblet cells lost, irregular crypts in shape, disordered arrangement and the most of visible ulcers, which were more consistent with the pathological manifestations of severe ulcerative colitis than the other two groups. (4) In summary, in the TNBS/ethanol modeling, using 100 mg/kg TNBS/ethanol thrice is more consistent with the model of severe ulcerative colitis.

Key words: severe ulcerative colitis, 2,4,6-trinitrobenzene sulfonic acid, rats, animal model, modeling times, the National Natural Science Foundation of China

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