【摘要】：目的:探究應激狀態(tài)下大鼠腸粘膜屏障功能的改變,以及在此過程中某些細胞因子的作用和它們之間的相關(guān)性,以期初步了解腸屏障損傷的細胞生物學機制,尋找能夠阻斷腸屏障功能損傷過程的保護因素,為臨床嚴重應激和危重病患者的治療和預后探索新的途徑。 方法:健康Wistar大白鼠60只,雌雄不限,隨機分為實驗組、對照組和正常組,每組20只。實驗組通過以顯微手術(shù)用無損傷動脈夾夾閉腸系膜上動脈起始部,1小時后松夾恢復腸系膜上動脈血流制成腸缺血后再灌流模型,對照組只進行同樣腹部手術(shù)操作但不夾閉腸系膜上動脈。正常組不手術(shù)。6小時后處死動物,3組同樣進行腸系膜上動脈多聚甲醛灌注,取距回盲部15厘米處腸管1厘米,常規(guī)固定、石蠟包埋,組織切片,通過普通HE染色觀察腸粘膜組織細胞學形態(tài);用免疫組織化學方法染色顯示腸粘膜組織或細胞中腫瘤壞死因子、血管活性腸肽、基質(zhì)金屬蛋白酶-13及層粘連蛋白的表達情況;并用VIDAS圖象分析系統(tǒng)進行半定量分析。 結(jié)果:普通HE染色結(jié)果顯示:光鏡下,實驗組與對照組相比,可見腸壁變薄,上皮細胞水腫明顯,部分壞死脫落,小腸絨毛短細,絨毛間距變寬。中性粒細胞及淋巴細胞浸潤,個別腸粘膜固有層內(nèi)腺體出現(xiàn)灶性區(qū)域壞死。免疫組織化學染色顯示:實驗組腸粘膜上皮細胞胞漿及基質(zhì)腫瘤壞死因子表達較對照組增高,VIDAS圖象分析測得光密度值分別為:實驗組0.45±0.0183,對照組0.27±0.0196,正常組0.26±0.0187;粘膜組織中基質(zhì)金屬蛋白酶-13表達較對照組增高,VIDAS圖象分析測得光密度值分別為:實驗組0.29±0.0253,對照組0.16±0.0249,正常組0.15±0.0251;基底膜層粘連蛋白表達較對照組降低,VIDAS圖象分析測得光密度值分別為:實驗組0.29±0.0672,對照組0.47±0.0685,正常組0.50±0.0679,實驗組和對照組之間統(tǒng)計學比較差異有顯著性(P0.01),對照組與正常組相比無明顯差異
[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 鹵0.0183, and that of the control group was 0.27 鹵0.0196, respectively. Normal group 0.26 鹵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 鹵0.0253 in the experimental group, 0.16 鹵0.0249 in the control group and 0.15 鹵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 鹵0.0672 in the experimental group, 0.47 鹵0.0685 in the control group, 0.50 鹵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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2005
【分類號】：R363

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