摘要
为了更好地理解苯并芘(Benzo(a)pyrene,B(a)P)在气道细胞诱导的毒理作用机制.应用二维凝胶电泳(two-dimensional electrophoresis,2-DE)分离B(a)P处理组(B(a)P-A549)、对照组(DMSO-A549)的总蛋白质,图像分析识别差异表达的蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定差异蛋白质点,Western blot验证差异蛋白热激蛋白27(heat shock protein 27,HSP 27)以及锰超氧化物歧化酶(Mn superoxide dismutase,Mn SOD)的表达,使用0.1,1,10μmol/L B(a)P处理A549细胞,应用Western blot及RT-PCR检测各组细胞中Mn SOD的表达,检测各组细胞中总抗氧化活性(total antioxidant capacity)、总SOD活性(total activity of SOD)、过氧化氢酶活性(catalase activity,CAT)、谷胱甘肽还原酶活性(glutathione reductase,Gr).本研究首先建立了B(a)P处理组、对照组的2-DE图谱,质谱鉴定了23个差异蛋白质,Western blot证实HSP 27及Mn SOD的表达水平在B(a)P处理组中较对照组明显下调;使用0.1,1,10μmol/L B(a)P处理A549细胞后,发现随着B(a)P的浓度升高,虽然Mn SOD的表达及SOD活性出现先诱导后抑制的现象,但CAT及Gr活性的增高依然提高了机体的总抗氧化活性.研究结果提示:HSP 27及Mn SOD与B(a)P在气道细胞诱导的毒理作用相关,CAT及Gr在对抗B(a)P带来的氧化损伤方面可能具有十分重要的作用.
In order to better understand the molecular mechanisms of Benzo（a）pyrene mediated toxicity on airway epithelial cells,comparative proteomics involving two-dimensional gel electrophoresis（2-DE） and MALDI-TOF Mass Spectrometry（MS） was performed on total proteins extracts from B（a）P-A549 and its control cell line DMSO-A549.Then the expression of heat shock protein 27（HSP 27） and Mn superoxide dismutase（Mn SOD）,two of the identified differential expressed protein,was analyzed using western blot.After 0.1,1,10 μmol/L B（a）P treatment,the Mn SOD expression levels were detected by reverse transcription polymerase chain reaction（RT-PCR） and western blot analysis,the total antioxidant capacity,total activity of SOD,catalase activity（CAT） and glutathione reductase activity（Gr） were deter-mined also.In this study,23 differentially expressed proteins was identified.Western blot confirm that B（a）P decreased heat shock protein 27 and Mn SOD expression levels in A549 cells and the expression lev-els of this two genes were increased to the control level after detoxification.After 0.1,1,10 μmol/L B（a）P treatment on A549 cells,the expression level of Mn SOD and SOD activity were significantly induced after 0.1 μmol/L B（a）P exposure and then reduced after 1 μmol/L and 10 μmol/L B（a）P exposure.With the in-creased concentration of B（a）P,the catalase activity,the total antioxidant capacity and the glutathione re-ductase activity increased.Our results indicate that HSP 27 and Mn SOD may be related to the B（a）P medi-ated toxicity in airway epithelial cells.CAT and Gr may play an important role to antioxidant injury induced by B（a）P.
出处
《化学学报》
SCIE
CAS
CSCD
北大核心
2011年第21期2609-2617,共9页
Acta Chimica Sinica
基金
国家自然科学基金(No.30770647)
湖南省自然科学基金(No.07JJ3055)
湖南长沙市环境保护局专项资金(No.2010)资助项目
