NF-κB信号通路和细胞因子在金黄色葡萄球菌PV杀白细胞素相关肺损伤中的作用

发布时间：2017-12-31 10:06

本文关键词：NF-κB信号通路和细胞因子在金黄色葡萄球菌PV杀白细胞素相关肺损伤中的作用　出处：《蚌埠医学院》2011年硕士论文　论文类型：学位论文

【摘要】：目的：研究金黄色葡萄球菌杀白细胞素（Panton-Valentine leukocidin，PVL）所致小鼠急性肺损伤的病变特征，以及核转录因子-κB（NF-κB）信号通路蛋白和细胞因子在肺损伤中的变化及其作用。方法：将60只小鼠随机分为4组，每组15只，采用气管插管法造模，分别注入重组杀白细胞素（A组）、PVL阳性金黄色葡萄球菌培养上清（B组）、PVL阴性金黄色葡萄球菌培养上清（C组），及生理盐水（D组）。造模后，于3h、6h、9h采外周血进行白细胞计数与分类；并于各时间点处死小鼠，取肺组织标本进行病理学检查，收集支气管肺泡灌洗液（BALF）采用流式细胞术（FCM）检测中性粒细胞凋亡率和坏死率；ELISA方法检测肺组织匀浆TNF-α、IL-8、IL-10、MCP-1蛋白含量，RT-PCR方法检测肺组织匀浆NF-κBmRNA的表达，免疫组织化学染色检测肺组织中NF-κB p65蛋白。结果：与D组比较，A组、B组小鼠肺组织病理检查显示有弥漫性炎性细胞浸润、出血、水肿等肺损伤表现；外周血白细胞及中性粒细胞数下降,支气管肺泡灌洗液中中性粒细胞凋亡和坏死率增加；肺组织匀浆中NF-κBmRNA和NF-κB p65蛋白表达量随作用时间延长逐渐增加；TNF-α、IL-8、IL-10含量逐渐升高；C组小鼠无明显的肺损伤表现；外周血白细胞数量以及肺组织中NF-κB信号通路蛋白和细胞因子均与D组小鼠无显著差异。结论：PVL可引起小鼠肺部出血坏死性炎症改变，NF-κB信号通路激活及细胞因子释放在PVL相关肺损伤中发挥重要作用。
[Abstract]:Objective: To study the pathological characteristics of acute lung injury induced by Panton-Valentine leukocidin (PVL) in mice, and the changes and roles of nuclear factor kappa B (NF- kappa B) signaling pathway protein and cytokines in lung injury.
Methods: 60 mice were randomly divided into 4 groups, 15 rats in each group, the model was made by intratracheal intubation, were injected recombinant Leucocidin (A group), PVL positive Staphylococcus aureus culture supernatant (B group), PVL negative Staphylococcus aureus culture supernatant (C group), and physiological salt water (group D). After modeling, 3h, 6h, 9h in peripheral blood counts and classification of white blood cells; and the mice were sacrificed at each time point, lung tissue specimens were taken for pathological examination, collecting bronchoalveolar lavage fluid (BALF) by flow cytometry (FCM) detection of neutrophils the apoptosis rate and necrosis rate; detection of lung tissue homogenate TNF- alpha, ELISA IL-8, IL-10, MCP-1 protein, RT-PCR was used to detect the expression of lung NF- kappa BmRNA, immunohistochemical staining of NF- kappa B p65 protein in lung tissues was detected.
Results: compared with D group, A group, B group of mouse lung tissue pathological examination showed diffuse infiltration of inflammatory cells, hemorrhage, edema and other manifestations of lung injury; peripheral blood leukocyte and neutrophil count decreased, bronchoalveolar lavage neutrophils apoptosis and necrosis rate of increase in the amount of time with the lotion; gradually increased the expression of NF- BmRNA and NF- p65 kappa kappa B protein in lung tissue homogenate; TNF- alpha, IL-8, IL-10 content increased gradually; group C showed no apparent lung injury; the number of peripheral blood cells and lung tissue in NF- B signaling pathway protein and cytokines and D group were no significant the difference.
Conclusion: PVL can induce pulmonary hemorrhage and necrotic inflammation in mice. The activation of NF- kappa B signaling pathway and the release of cytokines play an important role in PVL related lung injury.

【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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