IL-18在肠缺血—再灌流致肺损伤中作用的研究以及小鼠新型IL-18分子的克

发布时间：2018-05-17 19:48

本文选题：IL-18 + 缺血　；参考：《中国科学院研究生院（上海生命科学研究院）》2005年博士论文

【摘要】：多器官功能障碍综合症(MODS),是临床上危重病人的主要死亡原因,而急性呼吸衰竭综合症(ARDS)是其重要组成部分。肠缺血-再灌流(I/R)不仅造成肠自身的损伤,还是临床上引起MODS和ARDS的主要机制。本文主要研究在小鼠肠缺血-再灌流致肺损伤模型中IL-18表达水平的变化、IL-18的功能及其可能的作用机制。采用随机分组的方法分为3组,用夹闭上端肠系膜动脉30分钟、再灌流3小时作为小鼠I/R模型组;假手术组(sham)除了不进行动脉夹闭,与I/R组一样;以及正常小鼠组(不进行手术操作)。1、为研究外源和内源IL-18的作用,在缺血前15分钟尾静脉分别注射IL-18或抗IL-18中和抗体;2、为研究肺组织血管通透性,于缺血前尾静脉注射伊文斯蓝(EBD);3、为研究诱导性一氧化氮合酶(iNOS)对IL-18作用的影响,iNOS的选择性抑制剂L-NIL于缺血前15分钟尾静脉注射。用Western blot和ELISA的方法检测血清IL-18,用免疫组化的方法显示肺组织IL-18水平的变化,用半定量PCR和ELISA分别检测组织和血清TNF-α的表达,通过检测组织EBD的含量、肺组织MPO活性和组织病理学改变以确定肺部的炎症损伤。与正常小鼠血清IL-18水平相比(174±18pg/mL),肠缺血-再灌流显著促进IL-18水平的迅速升高,在再灌流1小时达到峰值(660±153 pg/mL),然后逐渐降至本底水平(3小时);而假手术并不导致血清IL-18水平的明显变化。与I/R模型组相比,外源注入IL-18(I/R+IL-18),能进一步显著促进肺组织的通透性、肺部MPO活性以及组织损伤,相反,抑制内源IL-18的活性(I/R+IL-18Ab)则明显抑制肺组织的MPO活性、组织通透性以及组织损伤。而且,外源IL-18的注入,能在假手术组小鼠而非正常小鼠中,
[Abstract]:Multiple organ dysfunction syndrome (MODS) is the main cause of death in critically ill patients, and acute respiratory failure syndrome (ARDS) is an important part of it. Ischemia-reperfusion (I / R) not only causes intestinal injury, but also causes the main mechanism of MODS and ARDS. The purpose of this study was to investigate the changes of IL-18 expression level and the possible mechanism of IL-18 expression in a mouse model of intestinal ischemia-reperfusion lung injury. The upper mesenteric artery was clamped for 30 minutes and then perfused for 3 hours as the I / R model group. In order to study the effects of exogenous and endogenous IL-18, IL-18 or anti-IL-18 neutralizing antibody were injected into caudal vein 15 minutes before ischemia to study the vascular permeability of lung tissue in normal mice group. In order to study the effect of inducible nitric oxide synthase (iNOS) on the effect of IL-18, L-NIL, a selective inhibitor of iNOS, was injected into the tail vein 15 minutes before ischemia. Serum IL-18 was detected by Western blot and ELISA, IL-18 level in lung tissue was detected by immunohistochemical method, expression of TNF- 伪 in tissue and serum was detected by semi-quantitative PCR and ELISA, and the content of EBD was detected. Lung tissue MPO activity and histopathological changes to determine lung inflammatory injury. Compared with normal mice, the serum IL-18 level was 174 卤18pg / mL, and intestinal ischemia-reperfusion significantly increased the level of IL-18. After reperfusion, the peak value was 660 卤153pg / mL, and then decreased to the background level for 3 hours, but sham-operation did not result in a significant change in serum IL-18 level. Compared with the I / R model group, exogenous injection of IL-18(I/R IL-18 could further promote the permeability of lung tissue, the activity of MPO and tissue damage. On the contrary, I / R IL-18Ab) significantly inhibited the activity of MPO in lung tissue. Tissue permeability and tissue damage. Moreover, exogenous IL-18 injection could be used in sham-operated mice rather than normal mice.
【学位授予单位】：中国科学院研究生院（上海生命科学研究院）
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R363;Q78

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