应激状态下大鼠肠屏障功能损伤过程中细胞因子的改变及意义
发布时间：2018-12-17 14:57
【摘要】：目的:探究应激状态下大鼠肠粘膜屏障功能的改变,以及在此过程中某些细胞因子的作用和它们之间的相关性,以期初步了解肠屏障损伤的细胞生物学机制,寻找能够阻断肠屏障功能损伤过程的保护因素,为临床严重应激和危重病患者的治疗和预后探索新的途径。 方法:健康Wistar大白鼠60只,雌雄不限,随机分为实验组、对照组和正常组,每组20只。实验组通过以显微手术用无损伤动脉夹夹闭肠系膜上动脉起始部,1小时后松夹恢复肠系膜上动脉血流制成肠缺血后再灌流模型,对照组只进行同样腹部手术操作但不夹闭肠系膜上动脉。正常组不手术。6小时后处死动物,3组同样进行肠系膜上动脉多聚甲醛灌注,取距回盲部15厘米处肠管1厘米,常规固定、石蜡包埋,组织切片,通过普通HE染色观察肠粘膜组织细胞学形态;用免疫组织化学方法染色显示肠粘膜组织或细胞中肿瘤坏死因子、血管活性肠肽、基质金属蛋白酶-13及层粘连蛋白的表达情况;并用VIDAS图象分析系统进行半定量分析。 结果:普通HE染色结果显示:光镜下,实验组与对照组相比,可见肠壁变薄,上皮细胞水肿明显,部分坏死脱落,小肠绒毛短细,绒毛间距变宽。中性粒细胞及淋巴细胞浸润,个别肠粘膜固有层内腺体出现灶性区域坏死。免疫组织化学染色显示:实验组肠粘膜上皮细胞胞浆及基质肿瘤坏死因子表达较对照组增高,VIDAS图象分析测得光密度值分别为:实验组0.45±0.0183,对照组0.27±0.0196,正常组0.26±0.0187;粘膜组织中基质金属蛋白酶-13表达较对照组增高,VIDAS图象分析测得光密度值分别为:实验组0.29±0.0253,对照组0.16±0.0249,正常组0.15±0.0251;基底膜层粘连蛋白表达较对照组降低,VIDAS图象分析测得光密度值分别为:实验组0.29±0.0672,对照组0.47±0.0685,正常组0.50±0.0679,实验组和对照组之间统计学比较差异有显著性(P0.01),对照组与正常组相比无明显差异
[Abstract]:Objective: to investigate the changes of intestinal barrier function in rats under stress, and the role of some cytokines in the process and the correlation between them, in order to understand the cellular biological mechanism of intestinal barrier injury. To find the protective factors which can block the process of intestinal barrier injury, and to explore a new way for the treatment and prognosis of patients with severe stress and critical disease. Methods: sixty healthy Wistar rats, male and female, were randomly divided into experimental group, control group and normal group with 20 rats in each group. In the experimental group, the initial part of the superior mesenteric artery was clamped by a non-invasive artery clip in microsurgery, and the blood flow of the superior mesenteric artery was restored after 1 hour of operation to make the model of intestinal ischemia and reperfusion. The control group only performed the same abdominal operation without clipping the superior mesenteric artery. In the normal group, the animals were killed after 6 hours. The three groups were also perfused with paraformaldehyde from the superior mesenteric artery. The intestinal canal was taken from the ileocecal area 15 cm, fixed routinely, embedded in paraffin wax, and sectioned by tissue section. The histological morphology of intestinal mucosa was observed by HE staining. The expressions of tumor necrosis factor, vasoactive intestinal peptide, matrix metalloproteinase-13 and laminin in intestinal mucosal tissues or cells were detected by immunohistochemical staining, and semi-quantitative analysis was performed by VIDAS image analysis system. Results: the results of ordinary HE staining showed that compared with the control group, the intestinal wall of the experimental group was thinner, the epithelial cell edema was obvious, some of the intestinal villi were necrotic and shedding, the villi of the small intestine were short and thin, and the villi spacing was wider. Infiltration of neutrophils and lymphocytes and focal necrosis of glands in some lamina propria of intestine. Immunohistochemical staining showed that the expression of cytoplasm and matrix tumor necrosis factor in intestinal epithelial cells in the experimental group was higher than that in the control group. The optical density of the experimental group was 0.45 卤0.0183, and that of the control group was 0.27 卤0.0196, respectively. Normal group 0.26 卤0.0187; The expression of matrix metalloproteinase-13 in mucosal tissue was higher than that in control group. The optical density values measured by VIDAS image analysis were 0.29 卤0.0253 in the experimental group, 0.16 卤0.0249 in the control group and 0.15 卤0.0251 in the normal group. The expression of laminin in the basement membrane was lower than that in the control group. The optical density measured by VIDAS image analysis was 0.29 卤0.0672 in the experimental group, 0.47 卤0.0685 in the control group, 0.50 卤0.0679 in the normal group, respectively. There was significant difference between the experimental group and the control group (P0.01), but there was no significant difference between the control group and the normal group.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R363
本文编号：2384354
[Abstract]:Objective: to investigate the changes of intestinal barrier function in rats under stress, and the role of some cytokines in the process and the correlation between them, in order to understand the cellular biological mechanism of intestinal barrier injury. To find the protective factors which can block the process of intestinal barrier injury, and to explore a new way for the treatment and prognosis of patients with severe stress and critical disease. Methods: sixty healthy Wistar rats, male and female, were randomly divided into experimental group, control group and normal group with 20 rats in each group. In the experimental group, the initial part of the superior mesenteric artery was clamped by a non-invasive artery clip in microsurgery, and the blood flow of the superior mesenteric artery was restored after 1 hour of operation to make the model of intestinal ischemia and reperfusion. The control group only performed the same abdominal operation without clipping the superior mesenteric artery. In the normal group, the animals were killed after 6 hours. The three groups were also perfused with paraformaldehyde from the superior mesenteric artery. The intestinal canal was taken from the ileocecal area 15 cm, fixed routinely, embedded in paraffin wax, and sectioned by tissue section. The histological morphology of intestinal mucosa was observed by HE staining. The expressions of tumor necrosis factor, vasoactive intestinal peptide, matrix metalloproteinase-13 and laminin in intestinal mucosal tissues or cells were detected by immunohistochemical staining, and semi-quantitative analysis was performed by VIDAS image analysis system. Results: the results of ordinary HE staining showed that compared with the control group, the intestinal wall of the experimental group was thinner, the epithelial cell edema was obvious, some of the intestinal villi were necrotic and shedding, the villi of the small intestine were short and thin, and the villi spacing was wider. Infiltration of neutrophils and lymphocytes and focal necrosis of glands in some lamina propria of intestine. Immunohistochemical staining showed that the expression of cytoplasm and matrix tumor necrosis factor in intestinal epithelial cells in the experimental group was higher than that in the control group. The optical density of the experimental group was 0.45 卤0.0183, and that of the control group was 0.27 卤0.0196, respectively. Normal group 0.26 卤0.0187; The expression of matrix metalloproteinase-13 in mucosal tissue was higher than that in control group. The optical density values measured by VIDAS image analysis were 0.29 卤0.0253 in the experimental group, 0.16 卤0.0249 in the control group and 0.15 卤0.0251 in the normal group. The expression of laminin in the basement membrane was lower than that in the control group. The optical density measured by VIDAS image analysis was 0.29 卤0.0672 in the experimental group, 0.47 卤0.0685 in the control group, 0.50 卤0.0679 in the normal group, respectively. There was significant difference between the experimental group and the control group (P0.01), but there was no significant difference between the control group and the normal group.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R363
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