目的探讨小鼠巨结肠模型的建立及其生物学特性。方法90只雄性昆明小鼠随机分为3组,实验组以0.1%Benzalkonium chloride(BAC)处理降结肠15min,对照组以生理盐水代替。术后分别于第2、7、14、21、28天通过大体观察,组织学、免疫组织化学及逆转录-聚合酶链反应(RT-PCR)检测观察该模型相关生物学特性。结果自术后第7天开始,实验组HE及免疫组织化学染色可见肌层神经节细胞明显减少至完全消失,平滑肌细胞及黏膜层无明显变化,RT-PCR显示NF舢、ACHE、GFAP表达下调。对照组无明显变化。结论运用0.1%Benzalkonium Chloride(SAC)可以成功构建小鼠巨结肠模型,该模型与先天性巨结肠具有相似的病理特征,为下一步的神经干细胞抑制奠定实验基础。
Objective To establish an aganglionosis mouse model for the study of neural stem cells transplantation and explore the relevant biological characteristics. Methods Ninety male Kunming mice were randomly divided into normal group,control group and treatment group. In treatment group,0. 1% Benzalkonium cldoride (BAC) was applied onto the colon descendens for 15 min and normal saline was used instead of BAC in control group. Mice in normal group received no treatment. On the day 1,3, 5,7,15 and 30 after treatment, general observation, histological examination, immunoldstochemical exami- nation and RT-PCR were performed. Results The results of HE and immunohistochemical staining showed obvious reduction or complete disappearence of ganglion cells in the muscle layer 7 days after treatment. There were no obvious changes in smooth muscle cell mucous layer. RT-PCR revealed down-regulation of NF-200, ACHE and GFAP. Conclusion The aganglionosis mouse model for neural stem cells transplantation was established using 0. 1% BAC.