AP时肠黏膜通透性的变化及其机制

发布时间：2018-01-04 11:17

本文关键词：AP时肠黏膜通透性的变化及其机制　出处：《山西医科大学》2006年硕士论文　论文类型：学位论文

【摘要】： 目的:在坏死型胰腺炎基础上的继发感染是AP最主要的死因,这与细菌移位密切相关。ANP时伴有IETM已形成共识。细菌移位与IETM的形成均与肠黏膜通透性增强密切相关。因此,观察AP时肠黏膜通透性的改变及其机制就显得十分重要。材料与方法:健康雌性大鼠体重210g-230g共50只,随机分为3组。假手术组(sham operation SO):仅开腹轻翻胰腺,不施加处理因素;急性坏死性胰腺炎模型组(acute necrosis pancreatitis ANP):依据经典的手术模型制备;禁食组(fasting diet FD):胃肠外营养,禁食72h。于72h处死动物,留取血浆、血清与组织标本待测。偶氮基质法测定血浆内毒素(ET)含量,判定肠源性内毒素血症情况;放射免疫法测定血清中TNF-α与IL-4,判定机体免疫功能;采用分光光度法测定血浆与肠组织匀浆中的二胺氧化酶DAO,观察肠组织的损伤情况;碘-淀粉比色法测定血清淀粉酶AMY,判定胰腺炎的严重程度;尾静脉注射Evans蓝检测肠黏膜与血管通透性的变化;采用硫代戊巴比妥钠法测定血清中丙二醛MDA ;采用荧光光度法检测血浆与肠组织匀浆中的组胺含量;肠组织甲苯胺蓝染色观察肠组织肥大细胞;HE染色显示肠与胰腺组织形态学改变;缺口原位末端标记法判定肠上皮细胞与胰腺腺泡细胞凋亡情况。结果:各组之间ET、DAO、TNF-α、IL-4及组胺水平均有显著性差异,其中ANP组各指标水平最高,FD组次之; SO及FD组与ANP组的血清AMY水平有显著性差异;血浆中ET含量与血浆及肠黏膜组胺、血浆DAO、MDA含量有相关性,血浆及肠黏膜组胺与TNF-α、IL-4含量有相关性。甲苯胺蓝染色显示ANP组肥大细胞处于激活状态的数目较多,FD组次之。HE染色与原位细胞凋亡检测结果相一致,以ANP组肠黏膜与胰腺组织损伤最严重、凋亡细胞最多,FD组次之。结论:禁食与ANP时均使肠黏膜与血管通透性增强,肠上皮细胞出现凋亡,肠黏膜受损,导致IETM形成。IETM又通过直接或间接的作用来加重肠黏膜的损害,而后者的损伤加重又促使IETM的形成,形成一个恶性循环,这也是AP死亡率居高不下的主要原因。
[Abstract]:Objective: Based on secondary necrotizing pancreatitis on AP infection is the leading cause of death, and this is closely related to bacterial translocation of.ANP with IETM has formed a consensus. The formation of bacterial translocation and IETM were closely related with the intestinal mucosal permeability increased. Therefore, to observe the AP changes and mechanism of intestinal mucosal permeability is very important.
Materials and methods: weight 210g-230g healthy female rats 50 rats were randomly divided into 3 groups. Sham operation group (sham operation SO): laparotomy only flip the pancreas, without any treatment; acute necrotizing pancreatitis model group (acute necrosis pancreatitis ANP): the basis of the classic model of surgical preparation; the fasting group (fasting diet FD): parenteral nutrition, fasting 72h. 72h were collected in animal plasma, serum and tissue samples were taken for determination of plasma endotoxin azo matrix method (ET) to determine the content of intestinal endotoxemia; Determination of TNF- alpha and IL-4 in serum by radioimmunoassay, determination of immune function was determined by; two monoamine oxidase DAO in plasma and intestinal homogenate in spectrophotometry, to observe the injury of intestinal tissue; iodine starch colorimetric determination of serum amylase AMY, determining the severity of pancreatitis; tail vein injection of Evans detection of intestinal mucosal and vascular blue The change of permeability; Determination of malondialdehyde MDA in serum by pentobarbital sodium thiosulfate method; histamine content in plasma and intestinal tissue were detected by fluorescence spectrophotometry; observation of intestinal tissue mast cells in intestinal tissue toluidine blue staining; HE staining showed that the morphology of intestinal and pancreatic tissue changes; gap TUNEL determination of intestinal epithelial cells and apoptosis of pancreatic acinar cells.
Results: between the groups ET, DAO, TNF- alpha, IL-4 and histamine levels were significantly different in ANP group, the highest level of each index, SO and FD group; FD group and ANP group, the serum AMY levels were significantly different; the content of ET in serum and plasma and intestinal mucosal histamine, plasma DAO, correlation the content of MDA in plasma and intestinal mucosal histamine and TNF- alpha, there is a correlation between the content of IL-4. Toluidine blue staining of mast cells in ANP group is active number, followed by the FD.HE staining and in situ apoptosis detection results consistent with ANP group intestinal and pancreatic tissue injury is the most serious, most of apoptotic cells in FD group..
Conclusion: fasting and ANP were intestinal mucosal and vascular permeability, intestinal epithelial cell apoptosis, intestinal mucosal damage, leading to IETM formation of.IETM through direct or indirect role to aggravate the damage of intestinal mucosa, and damage aggravated form but also to promote the IETM, forming a vicious spiral, which is the main reason of AP the high mortality rate.

【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363

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