Acta Chimica Sinica ›› 2011, Vol. 69 ›› Issue (21): 2609-2617.DOI: 10.6023/A1012161J Previous Articles     Next Articles

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应用毒理蛋白质组学技术研究多环芳烃类物质诱导气道上皮细胞毒理作用的机制

闵凌峰1,何淑雅2,陈琼*,1,谢明萱1,彭红兵1,曹兰玉1,王怀石1,朱小燕1   

  1. (1中南大学湘雅医院老年医学科、呼吸内科 长沙 410008)
    (2南华大学生物化学与分子生物学系 衡阳 421001)
  • 投稿日期:2010-12-16 修回日期:2011-06-18 发布日期:2011-07-02
  • 通讯作者: 陈琼 E-mail:qiongch@yahoo.com.cn
  • 基金资助:

    耐辐射奇球菌基因文库构建及其修改蛋白质相互作用网络研究;蛋白质组学技术筛选验证吸烟相关性肺癌预警标志物

Using Proteomic Analysis to Screen for Differentially Expressed Proteins in Airway Epithelial Cells Toxicity Mediated by Polycyclic Aromatic Hydrocarbons

Min Lingfeng1 He Shuya2 Chen Qiong*,1 Xie Mingxuan1 Peng Hongbing1 Cao Lanyu1 Wang Huaishi1 Zhu Xiaoyan1   

  1. (1 Department of Respiratory, Department of Geriatric Medicine, Xiangya Hospital of Central South University, Changsha 410008)
    (2 Department of Biochemistry and Molecular Biology, University of South China, Hengyang 421001)
  • Received:2010-12-16 Revised:2011-06-18 Published:2011-07-02

In order to better understand the molecular mechanisms of Benzo(a)pyrene [B(a)P] 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 determined 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 levels 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 increased concentration of B(a)P, the catalase activity, the total antioxidant capacity and the glutathione reductase activity increased. Our results indicate that HSP 27 and Mn SOD may be related to the B(a)P mediated toxicity in airway epithelial cells. CAT and Gr may play an important role to antioxidant injury induced by B(a)P.

Key words: PAHs, airway epithelial cells, proteomics, HSP 27, Mn SOD

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