NF-κB信號(hào)通路和細(xì)胞因子在金黃色葡萄球菌PV殺白細(xì)胞素相關(guān)肺損傷中的作用
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本文關(guān)鍵詞:NF-κB信號(hào)通路和細(xì)胞因子在金黃色葡萄球菌PV殺白細(xì)胞素相關(guān)肺損傷中的作用 出處:《蚌埠醫(yī)學(xué)院》2011年碩士論文 論文類型:學(xué)位論文
更多相關(guān)文章: 金黃色葡萄球菌 殺白細(xì)胞素 壞死性肺炎 細(xì)胞因子 核轉(zhuǎn)錄因子-κB 信號(hào)轉(zhuǎn)導(dǎo)途徑
【摘要】:目的:研究金黃色葡萄球菌殺白細(xì)胞素(Panton-Valentine leukocidin,PVL)所致小鼠急性肺損傷的病變特征,以及核轉(zhuǎn)錄因子-κB(NF-κB)信號(hào)通路蛋白和細(xì)胞因子在肺損傷中的變化及其作用。 方法:將60只小鼠隨機(jī)分為4組,每組15只,采用氣管插管法造模,分別注入重組殺白細(xì)胞素(A組)、PVL陽(yáng)性金黃色葡萄球菌培養(yǎng)上清(B組)、PVL陰性金黃色葡萄球菌培養(yǎng)上清(C組),及生理鹽水(D組)。造模后,于3h、6h、9h采外周血進(jìn)行白細(xì)胞計(jì)數(shù)與分類;并于各時(shí)間點(diǎn)處死小鼠,取肺組織標(biāo)本進(jìn)行病理學(xué)檢查,收集支氣管肺泡灌洗液(BALF)采用流式細(xì)胞術(shù)(FCM)檢測(cè)中性粒細(xì)胞凋亡率和壞死率;ELISA方法檢測(cè)肺組織勻漿TNF-α、IL-8、IL-10、MCP-1蛋白含量,RT-PCR方法檢測(cè)肺組織勻漿NF-κBmRNA的表達(dá),免疫組織化學(xué)染色檢測(cè)肺組織中NF-κB p65蛋白。 結(jié)果:與D組比較,A組、B組小鼠肺組織病理檢查顯示有彌漫性炎性細(xì)胞浸潤(rùn)、出血、水腫等肺損傷表現(xiàn);外周血白細(xì)胞及中性粒細(xì)胞數(shù)下降,支氣管肺泡灌洗液中中性粒細(xì)胞凋亡和壞死率增加;肺組織勻漿中NF-κBmRNA和NF-κB p65蛋白表達(dá)量隨作用時(shí)間延長(zhǎng)逐漸增加;TNF-α、IL-8、IL-10含量逐漸升高;C組小鼠無(wú)明顯的肺損傷表現(xiàn);外周血白細(xì)胞數(shù)量以及肺組織中NF-κB信號(hào)通路蛋白和細(xì)胞因子均與D組小鼠無(wú)顯著差異。 結(jié)論:PVL可引起小鼠肺部出血壞死性炎癥改變,NF-κB信號(hào)通路激活及細(xì)胞因子釋放在PVL相關(guān)肺損傷中發(fā)揮重要作用。
[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.
【學(xué)位授予單位】:蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2011
【分類號(hào)】:R363
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 王鳳玲;劉靜;楊青;;金黃色葡萄球菌耐藥基因及致病毒素基因的研究[J];中國(guó)感染與化療雜志;2009年04期
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