军事医学
軍事醫學
군사의학
BULLETIN OF THE ACADEMY OF MILITARY MEDICAL SCIENCES
2015年
5期
325-328
,共4页
刘莎莎%敖登其木格%王红丽%胡永亮%黎卫平%宋伦
劉莎莎%敖登其木格%王紅麗%鬍永亮%黎衛平%宋倫
류사사%오등기목격%왕홍려%호영량%려위평%송륜
PM2.5%NF-κB%支气管%上皮细胞%血管内皮生长因子类
PM2.5%NF-κB%支氣管%上皮細胞%血管內皮生長因子類
PM2.5%NF-κB%지기관%상피세포%혈관내피생장인자류
PM2.5%NF-κB%bronchi%epithelial cells%vascular endothelial growth factors
目的：探讨细颗粒物（particulate matter 2．5，PM2．5）诱导支气管上皮细胞（bronchial epithelial cell，Beas－2B）表达血管内皮生长因子（vascular endothelial growth factor，VEGF）的分子机制。方法 PM2．5粉末溶解于DMEM培养基中，倍比稀释成不同浓度（0、12．5、25、50、100μg／ml）的溶液，超声30 min后加入血清及双抗（链霉素和青霉素），使终浓度为2％血清的体系，刺激人支气管上皮细胞；双荧光素酶报告基因分析系统检测核因子－κB （nuclear factor－kappa B，NF－κB）活化水平和vegf基因启动子的转录活化水平；Western印迹检测NF－κB组成亚基p65磷酸化水平、阻遏蛋白IκBα及VEGF蛋白表达水平。结果 PM2．5呈剂量依赖性诱导Beas－2B细胞VEGF表达；PM2．5诱导Beas－2B细胞中NF－κB活化，在浓度为100μg／ml活化强度最高，阻遏蛋白IκBα降解和p65亚基磷酸化水平升高；抑制NF－κB p65亚基表达后Beas－2B细胞VEGF蛋白表达水平下调，vegf基因启动子的转录活化水平下降。结论 PM2．5通过活化NF－κB途径诱导支气管上皮细胞中VEGF表达。
目的：探討細顆粒物（particulate matter 2．5，PM2．5）誘導支氣管上皮細胞（bronchial epithelial cell，Beas－2B）錶達血管內皮生長因子（vascular endothelial growth factor，VEGF）的分子機製。方法 PM2．5粉末溶解于DMEM培養基中，倍比稀釋成不同濃度（0、12．5、25、50、100μg／ml）的溶液，超聲30 min後加入血清及雙抗（鏈黴素和青黴素），使終濃度為2％血清的體繫，刺激人支氣管上皮細胞；雙熒光素酶報告基因分析繫統檢測覈因子－κB （nuclear factor－kappa B，NF－κB）活化水平和vegf基因啟動子的轉錄活化水平；Western印跡檢測NF－κB組成亞基p65燐痠化水平、阻遏蛋白IκBα及VEGF蛋白錶達水平。結果 PM2．5呈劑量依賴性誘導Beas－2B細胞VEGF錶達；PM2．5誘導Beas－2B細胞中NF－κB活化，在濃度為100μg／ml活化彊度最高，阻遏蛋白IκBα降解和p65亞基燐痠化水平升高；抑製NF－κB p65亞基錶達後Beas－2B細胞VEGF蛋白錶達水平下調，vegf基因啟動子的轉錄活化水平下降。結論 PM2．5通過活化NF－κB途徑誘導支氣管上皮細胞中VEGF錶達。
목적：탐토세과립물（particulate matter 2．5，PM2．5）유도지기관상피세포（bronchial epithelial cell，Beas－2B）표체혈관내피생장인자（vascular endothelial growth factor，VEGF）적분자궤제。방법 PM2．5분말용해우DMEM배양기중，배비희석성불동농도（0、12．5、25、50、100μg／ml）적용액，초성30 min후가입혈청급쌍항（련매소화청매소），사종농도위2％혈청적체계，자격인지기관상피세포；쌍형광소매보고기인분석계통검측핵인자－κB （nuclear factor－kappa B，NF－κB）활화수평화vegf기인계동자적전록활화수평；Western인적검측NF－κB조성아기p65린산화수평、조알단백IκBα급VEGF단백표체수평。결과 PM2．5정제량의뢰성유도Beas－2B세포VEGF표체；PM2．5유도Beas－2B세포중NF－κB활화，재농도위100μg／ml활화강도최고，조알단백IκBα강해화p65아기린산화수평승고；억제NF－κB p65아기표체후Beas－2B세포VEGF단백표체수평하조，vegf기인계동자적전록활화수평하강。결론 PM2．5통과활화NF－κB도경유도지기관상피세포중VEGF표체。
Objective To investigate the molecular mechanism of vascular endothelial growth factor ( VEGF) expression in bronchial epithelial cells (Beas-2B)induced by particulate matter 2.5(PM2.5).Methods PM2.5 powder was dis-solved in DMEM medium and diluted into five concentrations , 0,12.5,25,50 and 100μg/ml, respectively.The double an-tibiotics ( streptomycin and penicillin ) and FBS were added into the solution to a 2% final concentration of serum system after being treated by ultrasound for 30 minutes.The cultured Beas-2B cells were then treated with different doses of PM2.5.Subsequently, nuclear factor-kappa B(NF-κB) transactivity and the transcriptional activation of vegf gene promot-er were tested by dual-luciferase reporter gene analysis system while phosphorylation of p 65 , expression levels of IκBαand VEGF were detected by Western blotting .Results PM2.5 induced up-regulation of VEGF expression in Beas-2B cells in a dose-dependent manner , accompanied by NF-κB transactivation at the highest level under 100 μg/ml of PM2.5 treatment. Moreover, PM2.5 induced degradation of the repressor protein IκBαand increase in the phosphorylation level of p 65 sub-unit in Beas-2B cells.Knockdown of NF-κB p65 expression significantly inhibited vegf gene promoter transcriptional activa-tion as well as VEGF protein expression in Beas-2B cells induced by PM2.5.Conclusion PM2.5 induces VEGF expres-sion via activation of NF-κB pathway in bronchial epithelial cells .
