郭秉楠;燕宪亮;许铁;

• 1:徐州医科大学卫生应急研究所徐州医科大学附属医院急救中心

摘要(Abstract)：

目的:探讨白细胞介素-33(interleukin-33, IL-33)对鼠巨细胞病毒(murine cytomegalovirus, MCMV)肺炎的影响。方法:通过分子生物学的方法构建IL-33原核表达载体纯化并鉴定纯化产物;通过聚合酶链式反应(polymerase chain reaction, PCR)法鉴定IL-33受体ST2-/-小鼠的基因型;通过鼻腔感染MCMV建立小鼠肺炎模型,并通过病理学评价肺炎模型及与IL-33的相关性;野生型及ST2-/-小鼠尾静脉注射外源性IL-33蛋白后,酶联免疫吸附法(enzymelinked immunoabsorbent assay, ELISA)检测小鼠肺中IL-33蛋白变化情况,病理学比较肺炎模型的严重程度,空斑实验比较肺部病毒载量程度;流式细胞术检测2种小鼠骨髓原性的抑制性细胞(myeloid-derived suppressor cell, MDSC)细胞的ST2表达情况及MDSC细胞在肺中的比例;实时定量聚合酶链式反应(Real-time polymerase chain reaction, Real-time PCR)检测肺中MDSC发挥抑制功能相关的的精氨酸酶-1(Arginase-1)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和Th2型细胞因子。结果:成功构建IL-33原核表达载体并通过SDS-PAGE和Western Blot方法验证;PCR成功鉴定并区分野生型及ST2-/-型小鼠;通过病理学检测成功建立鼻腔感染MCMV肺炎模型,肺炎模型组IL-33 mRNA和蛋白含量显著高于对照组;野生型小鼠尾静脉注射IL-33蛋白后,第7天肺炎病理学表现最严重,肺病毒载量最高;ST2-/-小鼠尾静脉注射IL-33蛋白后,与野生型小鼠相比,肺炎病理学表现严重程度及病毒载量显著降低;外源加入IL-33蛋白后,Arginase-1、iNOS及Th2型细胞因子显著升高。结论:IL-33可以通过活化肺中MDSC细胞促进MCMV肺炎。

关键词(KeyWords)： 巨细胞病毒肺炎;白细胞介素-33;骨髓原性的抑制性细胞;小鼠

Abstract:

Keywords:

基金项目(Foundation): 江苏省高校自然科学基金(18KJB320027);; 江苏省博士后基金(2018K247C)

作者(Authors): 郭秉楠;燕宪亮;许铁;

DOI: 10.16424/j.cnki.cn32-1807/r.2020.06.003

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