摘要
目的建立接近于人类先天性巨结肠（HD）的乳鼠模型,并进行模型评价。方法9窝6~7日龄SD乳鼠随机分为实验组、对照组和正常组,每组3窝。实验组以0.1%苯扎氯铵（BAC）、对照组用生理盐水处理降结肠15 min,正常组不做任何处理。各组分别于术后第1、3、5、6和7周行大体解剖结肠形态学观察及组织病理学检查,以SABC法检测各组结肠神经节细胞S-100蛋白和神经细胞特异性烯醇化酶（NSE）表达,c-Kit免疫荧光检测各组结肠Cajal间质细胞（ICC）的分布和表达。结果①术后7周,实验组均出现不同程度的排便减少、腹胀、精神萎靡和消瘦,粪便颗粒较对照组和正常组大而干燥,处死后大体解剖可见BAC处理段结肠肠管狭窄、痉挛,无蠕动,病变近端肠管扩张,肠腔内容物潴留。②术后1、3、5和6周,实验组病理学检查可见肌间及黏膜下神经节细胞逐渐减少,术后7周肌间及黏膜下神经节细胞完全消失,肠壁其他结构完整,未发现瘢痕形成及炎症细胞浸润。对照组和正常组均可见正常的肌间和黏膜下神经节细胞。③术后1、3、5和6周,实验组肌间及黏膜下神经节细胞S-100蛋白和NSE的表达逐渐减少;术后7周S-100蛋白和NSE的表达完全消失,对照组及正常组S-100蛋白和NSE表达均未见明显变化。④术后1、3、5、6和7周,实验组结肠c-Kit的表达逐渐减少,ICC分布均较对照组和正常组明显减少,网络状结构受到破坏,ICC形态出现异常,表现为突起变短、变钝。对照组和正常组c-Kit表达无明显变化。结论本研究成功建立HD乳鼠模型,为进一步研究HD及其相关并发症的病因、病理机制、生理机制及以ICC为靶点治疗HD奠定实验基础。
Objective To establish Hirschsprung′s disease（HD） neonatal rat model similar to human being and evaluate the model.Methods Ninety neonatal SD rats,6-7 days old,were randomly divided into normal group,experimental group and control group.All rats were operated under ether anesthesia.In experimental group,0.1% benzalkonium chloride（BAC） solution was applied to the descending colon for 15 minutes to set up the model,0.9% normal saline was applied in control group.There was no treatmetnt in normal group.At an interval of 1 week,3 weeks,5 weeks,6 weeks and 7 weeks after BAC treatment,general observation was performed.The expression of S-100 protein and neuron-specific enolase（NSE） were detected with immunohistochemistry.The distribution and expression of c-Kit in each group interstitial cells of Cajal（ICC） were detected with immunofluorescence.Results ① 7 weeks after BAC treatment,the rats had abdominal distention,defecation decrease,spirit flagging,emaciation,fecal pellets were significantly larger and drier than the control group and the normal group.Autopsy revealed a narrowed segment at the site of BAC treatment,spasm and no peristalsis,accompanied by distended proximal colon filled with massive feces.② 1 week,3 weeks,5 weeks and 6 weeks after BAC treatment,the results of HE staining showed gradually ganglion cells in the myenteric and submucous plexuses decreased.Seven weeks after BAC treatment,ganglion cells in the myenteric and submucous plexuses completely disappeared without scar formation and inflammatory cell infiltration.The control group and the normal group showed normal ganglion cells in myenteric and submucoous plexuses.③ 1 week,3 weeks,5 weeks and 6 weeks after BAC treatment,the expression of S-100 protein and NSE decreased in ganglion cells of myenteric and submucous plexuses.7 weeks after BAC treatment,ganglion cells of myenteric and submucous plexuses completely disappeared.S-100 protein expressed in cytoplasm and nuclei of ganglion cells and appeared as brown particles,NSE
出处
《中国循证儿科杂志》
CSCD
2010年第3期217-222,共6页
Chinese Journal of Evidence Based Pediatrics
基金
陕西省自然科学基金:2003C2022
