| 张浩浩,马澜婧,许 冰,李文姣,丁美玲,孙丽娟,聂勇战.小鼠急慢性结肠炎模型的建立及结肠上皮组织分离方法的探讨[J].现代生物医学进展英文版,2018,(12):2228-2232. |
| 小鼠急慢性结肠炎模型的建立及结肠上皮组织分离方法的探讨 |
| Establishment of Mouse Models of Acute and Chronic Colitis and a Method for Colonic Epithelial Tissue Isolation in Mouse |
| Received:December 06, 2017 Revised:December 27, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.12.006 |
| 中文关键词: 葡聚糖硫酸钠 急性结肠炎 慢性结肠炎 肠上皮分离 |
| 英文关键词: Dextran Sulfate Sodium Acute colitis model Chronic colitis model Enterocyte isolation |
| 基金项目:国家杰出青年科学基金项目（81225003） |
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| 中文摘要: |
| 摘要 目的：采用葡聚糖硫酸钠(Dextran Sulfate Sodium，DSS)诱导小鼠急慢性结肠炎模型，并探讨小鼠结肠上皮组织分离技术的可行性。方法：将40只8周龄C57小鼠随机分为急性和慢性结肠炎模型DSS组和急慢和性结肠炎对照组，每组10只小鼠。急性结肠炎模型：给予C57小鼠3% DSS自由饮水7天，蒸馏水3天；慢性结肠炎模型：给予C57小鼠2.5% DSS水5天，换蒸馏水自由饮水7天，再给予2.5% DSS水5天后换蒸馏水水7天，重复3个循环，共36天，对照组蒸馏水自由饮水。期间每天观察并记录小鼠体重、便隐血、大便性状并评分。造模完成后对结肠组织行苏木素-伊红(HE)染色评价有无组织学炎性损伤。小鼠肠上皮分离采用运用机械涡旋的方法。结果：与对照组相比，小鼠急性结肠炎模型小鼠体重减轻明显(P<0.001)、便隐血阳性、大便性状发生改变，结肠长度明显缩短(P<0.01)。小鼠慢性结肠炎模型小鼠体重随着DSS和蒸馏水的交替出现下降和上升的变化趋势，HE染色提示急性和慢性小鼠结肠炎模型结肠上皮发生急性和慢性的炎症伤；分离得到的肠上皮组织样本提取总蛋白证实蛋白未发生降解，Actin和GAPDH内参条带清晰。结论：小鼠急性和慢性结肠炎模型的建立成功，本实验采用的小鼠结肠上皮分离方法稳定可行。 |
| 英文摘要: |
| ABSTRACT Objective: To establish acute colitis model and chronic colitis model in mice by using DSS and to explore the efficiency of colonic epithelial isolation from colon of mice. Methods: Forty 8-week old C57 mice were randomly divided into four groups, the acute and chronic colitis model groups and the acute and chronic colitis control groups. the acute colitis model of mice was given 3% DSS wa- ter for 7 days and distilled water for 3 days ad libitum; chronic colitis model of mice were subjected to three cycles of DSS treatment, in which each cycle consisted of 2.5% DSS for 5 days followed by a 7 day recovery period with distilled water. The control group were given distilled water ad libitum. Body weight loss, stool consistency and the presence of occult/gross blood were assessed daily for each mouse, Colonic tissues were evaluated by hematoxylin eosin (HE) staining. The use of the isolation of intestinal epithelium was established by the methed of mechanical vortex. Results: The acute colitis model exhibited progressive body weight loss(P<0.001), colon length shorter, rectal bleeding and change of stool character, the chronic colitis model showed a curve of the mice body weight decreasing and rising with giving DSS and then distilled water. And the Histological analysis by H&E staining suggested that the two groups of mice colitis model mice had acute inflammatory and chronic inflammatory tissue injury; the quality of total protein from isolated colon epithelial tis- sue was stable. Conclusion: Acute and chronic colitis model were established successfully in mice and the quality of total protein from the isolated epithelial tissue of mice colon suggested that our method of epithelial isolation is stable and effective． |
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