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牛初乳对实验性巨结肠大鼠模型合并小肠结肠炎的预防作用的研究

A Study on the Bovine Colostrum in the Prevention of Enterocolitis in the Rat Model with Experimental Aganglionosis

【作者】 赵萍

【导师】 张钦明;

【作者基本信息】 首都医科大学 , 儿外科学, 2008, 硕士

【摘要】 目的:用化学方法选择性地去除大鼠结肠神经节细胞,成功建立实验性巨结肠大鼠动物模型,从而为本实验第二部分做好准备。方法:8-9周龄Wistar大鼠50只,随机分为两组,麻醉下开腹,实验组用0.1%的苯扎氯铵(BAC)作用于大鼠降结肠远端2cm长的肠管浆膜表面40分钟,温盐水冲洗后关腹,对照组用生理盐水代替BAC。观察两组大鼠的临床症状,并分别于术后1周、2周、3周、4周、5周、6周时每组随机取2只大鼠处死,观察处理段结肠的大体解剖变化,并取作用肠段结肠进行病理检查,行HE染色观察其局部肠神经节细胞消除情况。结果:BAC处理1周后,实验组大鼠略有腹胀,神经节细胞开始减少,部分开始变性;2周后进一步腹胀,处理段结肠开始出现狭窄趋势,神经节细胞略减少;3周后腹胀明显,处理段结肠痉挛狭窄,神经节细胞明显减少;4-5周后腹胀十分明显,处理段结肠明显狭窄,其近端结肠明显扩张,肠内容物滞留,大多数肠神经节细胞消失;6周后极度腹胀,处理段结肠非常狭窄,其近端结肠极度扩张,神经节细胞完全消失。而对照组大鼠降结肠大体及病理组织学均无明显变化。结论:采用0.1%BAC作用于大鼠降结肠浆膜可成功建立实验性无神经节细胞性巨结肠动物模型,为下一步进行牛初乳预防实验性巨结肠合并肠炎的研究奠定了基础。这也为今后进一步研究该病及其并发症的病因、发病机制及治疗等方面提供了适宜的动物模型。目的:探讨服用牛初乳,是否可以预防实验性巨结肠大鼠发生肠炎。方法:按本实验第一部分方法建立的56只实验性巨结肠模型鼠,随机分成两组,每组各28只。实验组从建立模型术后第一天开始口服纯牛初乳粉溶液,每天服用30ml,对照组不服用。两组均正常饮食及饮水。同时观察两组大鼠出现小肠结肠炎的临床症状的情况,并于术后5周处死大鼠,观察肠壁表面情况,取痉挛段及扩张段肠段行病理检查,行HE染色观察肠壁炎症的情况。根据临床症状和组织学检查来进行肠炎的诊断,统计两组各自出现肠炎的发生率,进行统计学分析,从而对两组发生肠炎的情况进行比较。结果:实验组有7只大鼠发生肠炎,发生率为25%,对照组有15只大鼠发生肠炎,发生率为53.6%。两组相比较差异有统计学意义(χ2=4.791, P<0.05)。结论:口服牛初乳粉溶液,可以降低巨结肠大鼠模型发生肠炎的发生率,对巨结肠大鼠发生肠炎有预防作用。

【Abstract】 Objective:To establish a successful rat model of experimental aganglionsis by the chemical ablation of colonic ganglion cells,and make good preparation for the second part of this experiment.Methods: Fifty Wistar rats, 8-9 weeks old, were randomly divided into two groups: experiment group (n=25) and control group(n=25). All animals were operated by laparotomy under anesthesia. In experiment group, 0.1% benzalkonium chloride (BAC) solution was applied to the serosal surface of the two-centimeter-long descending colon for 40 minutes. In control group, 0.9% normal saline was applied with as the same method as in experiment group. The observation of general presentations、morphological changes and histologic examination were performed at an interval of 1 week, 2week, 3week, 4week, 5week and 6week after operation by choosing two animals randomly from each group.Results: 1 week after BAC treatment, the rats showed mild abdominal distention. Histologic examinations appeared the degeneration of partial ganglion cells and the trend of reduction of ganglion cells. 2 weeks after BAC treatment, the rats showed moderate abdominal distention and the ganglion cells decreased further. A mildly narrow colonic segment at the segment of BAC treatment could be found. 3 weeks after BAC treatment, the rats showed obvious abdominal distention and the obvious reduction of ganglion cell. Also, a moderately narrow colonic segment and its distended proximal colon could be found. 4-5 weeks after BAC treatment, the rats showed serious abdominal distention, autopsy revealed a obviously narrow colonic segment accompanied by distended proximal colon filled with massive feces. A majority of ganglion cell disappeared. 6 weeks after BAC treatment, the rats showed more serious abdominal distention, autopsy revealed an extraordinary narrow colonic segment and marked dilation of the proximal segment. The ganglion cell disappeared completely.However, compared to experimental group, the control group showed no visible change in clinical symptom、morphological changes and histologic finding at every period of time.Conclusions: The animal model of experimental aganglionosis can be successfully produced in the rat by applying 0.1% BAC to the serosa of descending colon. This model may provide the appropriate animal model for the further study of this experiment in the prevention of Hirschsprung’s associated enterocolitis. Also,it will provide the good animal model for further studies on the etiology, pathophysiology and treatment of congenital aganglionosis and its complications in the future. Objective:To study the probability of the preventive effect of bovine colostrums on the occurrence of enterocolitis in the rat model with aganglionic megcolon.Methods: Fifty-six rat models with experimental aganglionosis were established according to the method of the first part of this experiment. They were randomly divided into two groups: experimental group (n=28) and control group (n=28). The rats in experimental group were fed to the pure bovine colostrums solution 30ml each day from the first postoperative day in addition to normal food. The control group were not. The clinical manifestations of enterocolitis were observed after operation. All the rats were killed in order to observe the inflammation from both surface of bowel and histological changing of colon (including the contracted and the proximal dilated segment) at the 5th week of post-operation. The diagnosis of enterocolitis was made by combinating clinical manifestations with histological findings. The incidence of enterocolitis of two groups were summarized and statistical analyzed.Results: Seven rats in experimental group had enterocolitis with the incidence of 25%,15 rats in control group had enterocolitis with the incidence of 53.6%. There was significant difference of the incidence of enterocolitis between two groups (χ2=4.791, P<0.05).Conclusions: Oral taking pure bovine colostrums solution may effectively decrease the occurrence of enterocolitis of rat with experimental aganglionosis.

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