【作者】 王宏；

【导师】 唐方；

【作者基本信息】 天津医科大学 ， 中西医结合临床， 2009， 硕士

【摘要】 目的：胃肠的运动功能是消化系统最主要的生理功能之一。临床上将脘腹胀满,恶心,呕吐,暖气,腹泻便秘为主要表现或伴有精神不佳,四肢乏力等症状者归属中医脾胃病范畴。有实验研究表明,缺血再灌注可以导致大鼠精神萎靡、活动欠佳、毛色晦暗、腹部胀满并出现胃肠动力障碍,提示缺血再灌注模型在一定程度上模拟了中医理论中焦壅塞,气机淤滞,脾气下陷,胃气上逆所致的脾胃升降失常的临床表现。本实验分别建立肠系膜上动脉缺血再灌注模型和肢体缺血再灌注动物模型,以不同缺血时间和不同再灌注时间进行分组。通过对大鼠肠黏膜及主要脏器病理形态观察；结肠平滑肌肌条收缩振幅及结肠平滑肌细胞Ca2+-ATP酶活力的检测,比较两种造模方法对结肠动力损伤的动态变化规律,并筛选适合的胃肠动力障碍模型,有助于今后探索临床不同阶段胃肠动力障碍性疾患的最佳给药时间与合理用药,提供实验研究依据。方法：1、建立肠系膜上动脉缺血再灌注动物模型：利用无创动脉夹夹闭大鼠肠系膜上动脉：统一缺血45min,依据不同再灌注时间分为：再灌注6h组、再灌注24h组、再灌注48h组(C45-6、C45-24、C45-48)；统一再灌注时间48h,依据不同缺血时间进行分为：缺血60min组、缺血75min组(C60-48、C75-48)。2、建立肢体缺血再灌注动物模型：采用统一300g拉力橡皮筋束缚大鼠双侧大腿腹股沟处,环绕结扎3周,持续缺血3小时后,迅速解除结扎,进行肢体再灌注,按照再灌注时间不同分为：12h组、18h组、24h组(Z3-12、Z3-18、Z3-24)。3留取小肠、结肠及心脏、肺脏、肾脏,进行HE染色,光镜下观察其病理变化,计数小肠绒毛正常分型的比例,测量小肠黏膜厚度及绒毛高度。4制备大鼠离体结肠平滑肌肌条,通过对大鼠结肠平滑肌肌条收缩波平均振幅的检测,探讨不同时间点结肠平滑肌收缩波振幅动态变化的规律。5制备大鼠结肠平滑肌细胞,运用定磷法测定细胞内Ca2+-ATP酶活力,探讨不同缺血再灌注模型结肠平滑肌细胞Ca2+-ATP酶活力的变化。结果：1、肠系膜上动脉缺血再灌注模型组：统一缺血45min组,在延长再灌注时间后小肠黏膜病理出现形态改变,但正常绒毛分型比例、黏膜厚度绒毛高度与空白对照组均无差异(P>0.05),结肠平滑肌肌条收缩波振幅于再灌注6h、24h呈下降,再灌注48h后上升的趋势,与空白对照组均无统计学差异(P>0.05)。统一再灌注48h组,缺血时间由45min延长至60min后,小肠黏膜正常绒毛分型比例、绒毛高度、黏膜厚度均较空白对照组呈有意下降(P<0.05),结肠平滑肌肌条收缩振幅下降(P<0.05)；延长缺血时间至75min结肠肠壁变薄,肌张力差,不能耐受1g前负荷,无法测量其振幅。模型各组结肠黏膜部分出现轻度病理改变。缺血75min组结肠平滑肌细胞Ca2+-ATP酶活力较空白对照组无统计学差异(P>0.05)。2、肢体缺血再灌注模型各组：小肠粘膜均出现病理形态的改变,正常绒毛分型比例、绒毛高度、绒毛厚度均较空白对照组呈有意下降(P<0.05)。随再灌注时间延长各组小肠黏膜病理损伤呈有意下降(P<0.05),各组心脏、肺脏、肾脏均出现不同程度的病理损伤。肢体缺血再灌注12h、18h组结肠平滑肌肌条收缩波振幅均低于空白对照组(P<0.05,P<0.01)；再灌注24h结肠平滑肌变薄,弹性差,不能耐受1g前负荷拉力,无法测量其振幅。模型各组结肠黏膜部分出现了病理改变。肢体缺血再灌注12h组结肠平滑肌细胞Ca2+-ATP酶活力较空白对照组呈升高趋势,再灌注18、24h后则出现逐渐下降趋势,但均维持在正常水平,较空白对照组均无统计学差异(P>0.05)。结论：两种缺血再灌注模型由于造模方法的不同,存在不同的动态变化规律,均能够造成结肠动力不同程度的损伤,可以作为模拟临床胃肠动力障碍性疾病的动物模型,其中肢体缺血再灌注模型以操作简便、易于统一量化手法为佳。更多还原

【Abstract】 Objective:The gastrointestinal motility function is one of the most significant physiological functions of the digestive system. The clinical symptoms of abdominal distention, nausea, vomiting, diarrhea and constipation as well as depression and debilitation which belong to the category of the spleen and stomach disease of Traditional Chinese medicine. According to the research,IR can lead to depression, less actions, matted fur, abdominal debilitation and also the disorder of colon motion of model animals, which prompt the model of IR can analogue the symptoms of Traditional Chinese medicine:choking of the middle energizer, stasis of the movement of qi, the sag of spleen qi, super inverse of stomach qi.Setting up 2 kinds of animal models: limb ischemia-reperfusion and intestinal ischemia-reperfusion explore the regularity of different reperfusion time injury in rats and find out a better model for functional gastrointestinal disorders and disorders of gastrointestinal motility by observing the mental status and vital signs;recording the average contractile amplitude of colonic muscle strips by a multiscriptor; detection of the Ca2+-ATPase and observing the morphological changes of intestinal mucosa,colon mucosa and capital organs through microscopy.Method:1.Setting up the model of intestinal ischemia-reperfusion:The superior mesenteric arteries in the three groups were occluded for 45 minutes and reperfusion for 6 hours 24 hours and 48 hours. And then different groups were:reperfusion for 48 hours after occluding the superior mesenteric arteries for 60 minutes and 75 minutes.2、Setting up the model of limb ischemia-reperfusion:using rubber band hypodesis the limbs of rats for 3 loops with same tensile force 300g. for 3 hours, and then reperfusion for 12hours,18hours and 24hours3、Observing the morphological changes of intestinal mucosa,colon mucosa and capita organs through microscopy.4、Recording the average contractile amplitude of colonic muscle strips by a multiscriptor. 5、Detection of the Ca2+-ATPase of cells of colonic smooth muscle. Results:1、The model of intestinal ischemia-reperfusion:After extending the time of reperfusion,the mental status and vital signs became worse and there were changes in the small intestine mucosa which was in the ischemia-45min group.But the contractile amplitude of colonic muscle strips ascensused in the C45-48 group.After extending the time of reperfusion from 45 minutes to 60 minutes,both small intestine and colon mucosa had pathological changes, and the mental status and vital signs became worse and worse; average contractile amplitude of colonic muscle strips decreased in the C60-48 group, there were differences between this group and control group;while the colonic muscle strips of group C75-48 became thin,so the amplitude could not be record.There was no differences of activity of Ca2+-ATPase between the group of ischemia 75min and the control group.2、The model of limb ischemia-reperfusion:Extending the time of reperfusion, there were pathological changes on the small intestine mucosa,colon mucosa, heart, lung, and kidney. In comparison with the control group, the average contractile amplitude of colonic muscle strips was decreased in the group of reperfusion 12 hours and the group of reperfusion 18 hours (P<0.05, P<0.01);while the colonic muscle strips of group of reperfusion 24 hours became thin and could not record the amplitude. The activity of Ca2+-ATPase was ascensus in the group of reperfusion 12 hours, while it decreased in the groups of both reperfusion 18 hours and reperfusion 24 hours.Conclusion:The two kinds of models had their own dynamic regulars because of different making methods, but they can both injury the colon motion, so they can be used as models to analogue the functional gastrointestinal disorders and disorders of gastrointestinal motility.The model of limb ischemia-reperfusion is better.更多还原