【作者】 李少群；

【导师】 马遂；

【作者基本信息】 北京协和医学院 ， 临床医学， 2009， 博士

【摘要】 目的：研究急性心理应激对胃肠粘膜形态和屏障功能以及结肠运动的影响。方法：55只雄性wistar大鼠经过3小时限动束缚后随机分入4个组：A组(3小时即时处死组)14只,B组(束缚后休息3小时处死组)14只,C组(束缚后休息9小时处死组)14只,D组(束缚后休息21小时处死组)13只；另外13只大鼠禁食禁水3小时作为对照；造模其间计数大鼠排便数。在各自时间点解剖大鼠采集标本。肉眼评估胃粘膜、回肠粘膜、结肠粘膜形态；光镜下对胃、回肠、结肠粘膜病理形态进行观察；免疫组化法分析回肠、结肠粘膜紧密连接Occludin蛋白的表达；TUNEL法检测回肠上皮细胞凋亡；检测血浆二胺氧化酶(DAO)活性和D-乳酸(D-lac)浓度。结果：束缚成功造成大鼠急性心理应激；急性心理应激导致大鼠胃粘膜广泛糜烂和散在出血；急性心理应激组紧密连接蛋白Occludin蛋白染色变淡,平均光密度略有降低,但与对照组差异不显著；急性心理应激组回肠粘膜上皮细胞凋亡指数明显增高。和对照组比较,急性心理应激各组血浆DAO活性和D-lac浓度未见明显增高；急性心理应激导致大鼠结肠运动增强。结论：1本束缚方法制造心理应激模型是成功的。2本心理应激模型可导致大鼠胃肠粘膜从形态学损害到屏障功能的损害。3本研究中大鼠肠粘膜屏障功能损害可能以细胞凋亡增加为主要途径,细胞间隙紧密连接破坏为次要途径。4急性心理应激中大鼠结肠运动增强。更多还原

【Abstract】 Objective:To observe gastrointestinal membrane barrier function and colonic dynamics in wistar rats under acute psychological stress.Methods:Fifty-five male wistar rats under3hours of restraint stress were randomly divided into4groups, respectively having0hours (14),3hours (14),9hours (14),21hours (13) of recovery. Additional13rats which subjected to3hours of fasting to food and water were treated as control. Feces output were collected and counted after the3hours of stress or fasting. Rats were anesthetized using pentobarbital at respective time point. Blood plasma was collected and stored at-80℃until serving for detection of diamine oxidase(DAO) and D-lactic acid (d-lact). Stomach, ileum and colon were assessed in naked eyes and samples of each were collected for mucosa membrane damage assessments after HE staining. One of the tight junction proteins, occludin, was detected using immunohistochemistry staining in ileum and colon membrane. Apoptosis in ileac membrane were elevated using TUNEL technology.Results:After3hours of restraint stress, gastric membrane showed extensive erosion, which recovered during time. Gastric membrane in HE showed congestion, hemorrhage, epithelial cell necrosis after3hours of restraint stress and recovered during time. Ileac and colonic membrane showed similar reaction but much more moderate. Apoptosis increased after the stress significantly. Anti-occludin staining showed decreased optical density in stress rats, but without statistical significance. There was no significant difference between restraint animals and the control in plasma DAO and d-lact. Feacal output significantly increased in the stressed animals. Conclusions:(1) The restraint method used in the study successfully induced acute psychological stress of wistar rat.(2) Acute psychological stress resulted in damage of the gastrointestinal membrane from morphology to barrier function.(3) Barrier dysfunction of this model might mainly due to epithelial cell apoptosis increase, accompanying with tight junction damage to some extent.(4) Psychological stress promoted colonic dynamics.更多还原