目的:探討不同濃度香煙煙霧提取物濃度在體外對(duì)外周血中性粒細(xì)胞生成中性粒細(xì)胞外誘捕網(wǎng)（NETs）的影響,吸煙相關(guān)慢性阻塞性肺疾病患者較正常人對(duì)煙草煙霧刺激外周血中性粒細(xì)胞產(chǎn)生NETs敏感性的異同及紅霉素的干預(yù)作用。方法:招募平均年齡在64歲的穩(wěn)定期COPD患者及健康自愿者各10名,分離純化其外周血中性粒細(xì)胞后進(jìn)行刺激分組:（1）分別使用0.1%、0.3%、0.5%、1%及2%濃度的香煙煙霧提取物（CSE）對(duì)正常人中性粒細(xì)胞進(jìn)行刺激,利用熒光DNA定量法觀察各濃度CSE刺激下中性粒細(xì)胞發(fā)生NETosis的情況;（2）使用PMA、CSE分別對(duì)COPD組及正常人組其進(jìn)行刺激,應(yīng)用免疫熒光染色法觀察各組NETs的生成情況及CSE誘導(dǎo)生成NETs的形態(tài)特征及組成結(jié)構(gòu);應(yīng)用熒光DNA定量法對(duì)各組中性粒細(xì)胞的NETs生成情況進(jìn)行定量;（3）對(duì)正常人外周血中性粒細(xì)胞在固定使用0.1%CSE進(jìn)行刺激的前提下,分別用濃度為1μg/mL、5μg/mL及10μg/mL的紅霉素對(duì)其進(jìn)行干預(yù),利用熒光DNA定量法定量各濃度紅霉素干預(yù)下對(duì)煙草煙霧誘導(dǎo)中性粒細(xì)胞發(fā)生NETosis化的抑制情況;（4）在固定使用0.1%C... 

【文章來源】：廣西醫(yī)科大學(xué)廣西壯族自治區(qū)

【文章頁數(shù)】：73 頁

【學(xué)位級(jí)別】：碩士

【部分圖文】：

各濃度香煙煙物提取物對(duì)人外周血中性粒細(xì)胞生成NETs情況的比較

性粒細(xì)胞較正常人的敏感性增高，但究竟是何種原因?qū)е麓朔N敏感性增高目前還尚未得知，有待進(jìn)一步的研究考證。圖2-1 正常人組及COPD病人組人外周血中性粒細(xì)胞在不同刺激物影響下發(fā)生NETosis的比較Figure 2 Comparision between NETosis of helthy people and NETosis of COPD patients, of peripheralblood neutrophils under different stimuli. Compare the control group,CSE and PMA stimulation for 4 hoursbetween peripheral blood of normal people(A) or from stable period COPD patients(B)neutrophils separatedand purified from peripheral blood.To compare the difference of NETs between healthy people and COPDpatients, we quantified NETs from both groups. After collecting samples of supernatants from both the healthyand the COPD patients, Diluted Quant-iT PicoGreen Reagent was added,then samples were placed in themultimode reader for fluorescent quantification(D).Data are expressed as means±SD.The comparisons weredetermined by one-way ANOVA

圖2-2煙草煙霧誘導(dǎo)的NETs形態(tài)學(xué)特征及蛋白和酶類構(gòu)成Figure 2-2.Characteristics of NETs induced by CSE.Neutrophils isolated from Chronic Obstructive PulmonaDisease were stimulated for NETs foramtion with CSE as described and then stained with（A）citrullinatehistoe H3（CitH3,green）or（B）neytrophil elastase（NE,green）,and DNA were stained with Propidium iod（PI）。NETs were then examined by fluorescence microscope.Original magnification,X40

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