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烟草烟雾暴露对气道上皮细胞转分化改变的初探

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烟草烟雾暴露对气道上皮细胞转分化改变的初探

作者:李雯曦

来源:《中国医学创新》2017年第23期

【摘要】目的:观察烟草烟雾暴露人气道上皮细胞(16HBE)后,发生间充质转分化(EMT)样改变。方法:(1)以不同浓度(5~50 μg/mL)CSC刺激16HBE 7 d,用CCKit-8检测细胞总体活性,筛选最佳浓度。(2)用CSC最大活性影响浓度刺激16HBE 7 d,观察细胞形态改变,经Western-blot法和细胞免疫荧光技术检测上皮细胞标志物E钙黏蛋白(E-cad)及EMT标记物1型胶原(COL1)、波形蛋白(Vimentin)和基质金属蛋白酶9(MMP-9)的表达水平。结果:(1)不同浓度CSC刺激后,16HBE在

10 μg/mL时OD值最高,为(2.562±0.045)。各浓度间OD值比较差异均有统计学意义(P

【关键词】烟草烟雾凝集物;气道上皮细胞;上皮细胞间充质转分化

【Abstract】 Objective:To observe the epithelial-mesenchymal transition in human bronchial epithelial cells(16HBE) exposed to cigarette smoke.Method:Cells were stimulated by cigarrette smoke condensate(CSC) with different concentrations(5-50 μg/mL) for 7 days,then chosen the optimal concentration by cell counting kit-8.Western blot and Immunofluorescence method were used to estimate the expression levels of E-cad,type 1 collagen,Vimentin and MMP-9 in 16HBE induced with CSC (10 μg/mL) for 7 days.Result:16HBE manifested a morphological characteristic of loss of cell-cell contact and elongated shape,after 7 days induced with CSC.The level of E-cad downregulated in the experimental group[Western blot(0.301±0.041) vs

(0.591±0.068),P

【Key words】 Cigarette smoke condensate; Bronchial epithelial cell; Epithelial-mesenchymal transition

First-author’s address:Guangzhou First People’s Hospital,Guangzhou 510180,China

doi:10.3969/j.issn.1674-4985.2017.23.008

气道重塑是慢性阻塞性肺疾病(Chronic Obstructive Pulmonary Disease,COPD)的主要病理特征和引起不可逆性气流受限的主要原因之一[1],其中上皮间充质转分化(Epithelial-Mesenchymal Transition,EMT)与气道重构密切相关[2],EMT通过使气道壁中的成纤维细胞增加,从而参与气道重构。吸烟是公认的导致COPD发病和不可逆气流受限进展的最为重要的