【作者】 孙海荣；

【导师】 郭洪志；

【作者基本信息】 山东大学 ， 神经内科， 2005， 硕士

【摘要】 目的 建立大鼠脑出血致多器官功能障碍综合征(MODS)的动物模型,研究大鼠脑出血致MODS后血浆内毒素含量的变化及肺、小肠、肝和肾的组织病理改变,内毒素受体CD14mRNA在肺、小肠、肝、肾等各器官组织的表达,探讨脑出血致MODS的肠道机制。 方法 1.采用立体定向术在大鼠尾状核注入胶原酶加肝素建立脑出血致MODS模型,随机将96只Wister大鼠分为正常对照组(n=6)、假手术组(n=6)、实验1组(0.4 u胶原酶,n=6)和实验2组(0.8u胶原酶,n=6),后两组又分为4h、8h、12h、24h、36h、48h和72h时相点7个亚组。 2.观察术后各时相点症状、体征变化,检测外周血WBC、肝肾功能、心肌酶学改变,采用改良基质显色法鲎试验测定血浆内毒素的含量。光镜下观察脑、肺、小肠、肝和肾组织病理变化,依据盛志勇院士提出的动物全身炎症反应综合征(SIRS)和MODS的诊断标准判断SIRS和MODS的发生率。 3.应用原位杂交实验和CMIA真彩色医学图象分析系统,检测表达CD14mRNA阳性颗粒的面密度和光密度,作为细胞CD14mRNA的相对含量。 4.实验数据用均数±标准差((?)±S)表示,应用SPSS10.0统计软件包进行统计学处理,数据间差异显著性检验采用t检验、方差分析,以P<0.05为差异有显著意义,并进行相关分析。山东大学硕士学位论文 结果 1.实验组在胶原酶注射后4h均有明显血肿形成,24h后血肿基本上不再增大,实验2组形成的血肿大于实验1组,其血肿形态大小相对恒定,其神经功能评分明显高于实验1组(P<0.01)。 2.实验组动物的体温、呼吸、ALT、BLjN、CR、心肌酶与正常对照组、假手术组相比有显著性差异,并且在24h一48h之间变化最大。实验2组与实验1组比较,呼吸、心率、体温及外周血WBC均存在显著性差异(尸<0.05或尸<0.01),尤其以外周血wBC差异更加明显(P<0.01)。实验1组SIRS发生率75.4%,MODS发生率21.4%,实验2组SIRS发生率100%,MODS发生率67.9%。 3.实验组血浆内毒素含量明显升高(P<0.01或P<0.001),且实验2组高于实验1组(P<0.01)。血浆内毒素于术后8h开始升高,24h达高峰,在实验l组72h接近正常;而在实验2组72h仍维持较高水平。 4.实验组各时相点均可见各脏器不同程度的炎性损害,实验1组在24h一36h时相点的脏器病理变化较显著,在48h逐渐消退,72h时相点基本恢复正常;实验2组出血后4h即见肺脏、小肠炎性反应,12h后肺、小肠、肝、肾均见非特异性炎症反应,在24h一48h的脏器病理变化较显著,72h仍见较明显炎症反应。 5.正常对照组各器官组织中有少量CD 14mRNA表达,假手术组仅肺组织CD14mRNA表达较正常对照组显著升高(P<0.01),实验2组肺、小肠、肝组织CD14mRNA表达显著升高(p<0.01),且随时间进展组织CD14mRNA表达迅速升高,24h一36h达高峰,72h仍维持较高水平,且以肺脏变化最显著护<0.001)。 6.相关分析表明,实验2组肺、小肠组织CD 14mRNA表达与血浆内毒素呈正相关。 结论 1.大鼠尾状核注入0.8U胶原酶加肝素法可以成功的建立脑出血致MODS模型。 2.脑出血后存在重要脏器的炎性病理改变,尤其是肠道勃膜的病理改变和肝山东大学硕士学位论文功能障碍,为内毒素血症和肠道内毒素易位的发生提供了条件。 3.脑出血后出现内毒素血症,且各脏器组织内毒素受体CD14的表达上调,提示内毒素可能是刺激组织内毒素受体CD14表达的重要因素之一,内毒素一内毒素受体CD14一炎性介质间相互作用所致SIRS可能是脑出血致MODS的发病机制之一。 关键词脑出血多器官功能障碍综合征CD14内毒素全身炎症反应综合征动物模型更多还原

【Abstract】 Objective To establish a model of cerebral hemorrhage in rats complicated by multiple organ dysfunction syndrome (MODS); to investigate changes in tissue pathology of brain\lung\liver\intestines and kidney; to investigate plasma endotoxin and lipopolysaccharide(LPS) receptor CD14 gene expression in vital organs after cerebral hemorrhage ;to discuss the pathogenesis of cerebral hemorrhage complicated by MODS.Methods1. Cerebral hemorrhage was induced in rats by injecting different dose collagenase into the caudate nuclear. 96 Wister rats were randomly divided into 7 groups: normal control group (n=6) sham-operative group (n=6) and 2 groups of Cerebral hemorrhage(0.4U,0.8U)(n=42)including 4h 8h 12h 24h 36h 48h 72h seven time points.2. Changes of symptoms and signs including temperature heart rate and respiration of each group were recorded, the amount of WBC and function of liver and kidney were assayed, the dose of plasma endotoxin were examined. Pathological changes of brain\lung\liver\intestines and kidney were recorded and pictures were taken through optical microscope.3. The area density and optical density of positive staining expressingCDHmRNA were analyzed for the relative content of CD14mRNA using in situ hybridization and CMIA medical imaging analysis system.4. Results were expressed as x±s. One-way-anova of SPSS 10.0 soft ware was used to evaluate the difference among groups. Correlation analysis of SPSS 10.0 were used,too.Results1. Hematoma was found 4h after collagenase injection in hemorrhage group and the hematoma would not enlarge after 24h. The hematom of group2 was larger than that of group 1 and the hematoma formed was stable. The neurologic score of group 2 was much higher than that of group l(P<0.01).2. The rats of operative group had higher temperature and respiration, and the levels of ALT AST BUN Cr CK and LDH in those group improved. There were significant differences in between operative group and normal control group, between operative group and sham-operative group, between group 1 and group 2. Incidences of systemic inflammatory response syndrome (SIRS) and MODS were respectively 75.4% and 21.4% after cerebral hemorrhage of group 1, Whereas incidences of SIRSand MODS were respectively 100% and 67.9% after cerebral hemorrhage of group2.3. There was endotoxemia administration after acute cerebral hemorrhage. Plasma endotoxin level of experimental group 2 is higher than that of experimental group l(P<0.01). Plasma endotoxin level started to increase at 8 hours after hemorrhage, peaking at 24 hours, then returned to the baseline value at 72 hours in experimental group 1, but it still maintain high level in experimental group 2.4. In experimental group there are various inflammatory lesions on the organ tissue at every time point , in experimental group 1 the organ has significantly pathological changes at 24-36 hour point, the changes gradually extinct in 48 hours, and it almost recovers normal at 72 hour point; in experimental group 2 the inflammatory reaction on lung and small intestine can be seen 4 hours after bleeding, after 12 hours nonspecific inflammatory reaction can be seen on lung\ intestinesMiver and kidney, the pathological changes become significant at 24-48 hour point, and there are still significantly inflammatory reaction at 72 hour.5. In normal matched control group there are a small quantity of CD14mRNAexpression in organ tissue , the lung CD14mRNA expression in sham-operative group is obvious higher than that in normal control group (P<0.01), in experimental group the CD14mRNA expression in various tissue increased significantly (P<0.01), it culminate after 24 hours in experimental group 1 ,and start to decrease after 48 hours; in experimental group 2 the CD14mRNA expression in various tissue increased quickly with the time passed, it culminate after 24 hours, and maintain high level after 72 hours, and the changes in lung is the most significant.6. Relative analysis indicated that there was positive correlation bet更多还原