香烟提取物对气道平滑肌细胞增殖作用的研究

目的 探讨香烟提取物(CSE)对人气道平滑肌细胞(ASMCs)增殖以及钙网织蛋白(CRT)、CCAAT增强子结合蛋白α(CEBPα)表达的影响及机制。方法 (1)通过收集经不同浓度CSE处理24 h的ASMCs,分为对照组、2.5%CSE组、5%CSE组、10%CSE组。以MTT法分析各组细胞增殖情况,采用逆转录聚合酶链反应(RT-PCR)检测各组细胞CEBPα的m RNA水平;免疫印迹法检测4组细胞CRT、CEBPα的蛋白水平。(2)在10%CSE组,分别在ASMCs中转染阴性对照si RNA、CRT的si RNA,比较2组ASMCs的CEBPα、CRT表达及细胞增殖情况。结果 (1)对照组...

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Published in天津医药 Vol. 43; no. 9; pp. 978 - 981
Main Author 管频 于化鹏 吴智勇 李伟 吴洁
Format Journal Article
LanguageChinese
Published 广东广州南方医科大学 邮编510000 2015
海口,海南省人民医院医疗保健四区%广东广州南方医科大学 邮编510000%海口,海南省人民医院医疗保健四区%海口,海南医学院
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ISSN0253-9896
DOI10.11958/j.issn.0253-9896.2015.09.005

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Summary:目的 探讨香烟提取物(CSE)对人气道平滑肌细胞(ASMCs)增殖以及钙网织蛋白(CRT)、CCAAT增强子结合蛋白α(CEBPα)表达的影响及机制。方法 (1)通过收集经不同浓度CSE处理24 h的ASMCs,分为对照组、2.5%CSE组、5%CSE组、10%CSE组。以MTT法分析各组细胞增殖情况,采用逆转录聚合酶链反应(RT-PCR)检测各组细胞CEBPα的m RNA水平;免疫印迹法检测4组细胞CRT、CEBPα的蛋白水平。(2)在10%CSE组,分别在ASMCs中转染阴性对照si RNA、CRT的si RNA,比较2组ASMCs的CEBPα、CRT表达及细胞增殖情况。结果 (1)对照组、2.5%CSE组、5%CSE组、10%CSE组ASMCs的CEBPα蛋白表达依次递减,CRT和细胞增殖程度呈依次增高(P〈0.05);而CEBPαm RNA表达差异无统计学意义(P〉0.05)。(2)在10%CSE作用下,CRTsi RNA组中ASMCs的CEBPα表达较阴性对照si RNA组增强(P〈0.05),而CRT和细胞增殖程度均较相应阴性对照si RNA组减弱(P〈0.05)。结论 CSE可使ASMCs中CRT的表达增强,从而抑制CEBPαm RNA的翻译,使CEBPα的表达减少,最终促进ASMCs的增殖。
Bibliography:myocytes,smooth muscle;cell proliferation;pulmonary disease,chronic obstructive;in vitro;cigarette smoke extract;calreticulin;CCAAT enhancer-binding protein
Objective To explore the effects and mechanism of cigarette smoke extract (CSE) on the proliferation of airway smooth muscle cells (ASMCs) and the expression of CCAAT/enhancer-binding protein (CEBPα) and calreticulin. Methods (1) The ASMCs were stimulated with different concentrations of CSE for twenty-four hours. According to the concentrations of CSE,the cells were divided into control group, 2.5%CSE group, 5%CSE group and 10%CSE group. The proliferation of ASMCs was measured by MTT colrimetric method. The CEBPαmRNA was analyzed by RT-PCR. Western bloting assay was performed to detect the levels of CRT and CEBPαprotein. (2) In 10%CSE group, transfection of the siRNA respectively for negative control or calreticulin was performed in accordance with instructions. The cell proliferation and the expression of calreticulin and CEBPαwere compared in negative c
ISSN:0253-9896
DOI:10.11958/j.issn.0253-9896.2015.09.005