摘要
[目的]了解氢醌(HQ)对人骨髓间充质干细胞蛋白表达谱的影响,筛选HQ骨髓毒性作用的靶蛋白。[方法]体外分离并培养人的骨髓间充质干细胞至第3代,随即将细胞分为实验组和对照组。实验组用5×10-5mol/L的HQ染毒24h;对照组加入相同浓度的磷酸缓冲液处理24h。染毒完毕后即刻抽提两组细胞的总蛋白,然后利用双向凝胶电泳技术和LC-Qq-TOF质谱技术进行蛋白表达谱分析,找到可能的HQ骨髓毒性靶蛋白。[结果]在3次重复实验中,对照组与HQ处理组相比较,均有约20个差异蛋白点。其中3次中有11个重复的差异蛋白点,质谱分析鉴定出膜联蛋白-Ⅱ、甲酰-谷胱苷肽水解酶、尿苷二磷酸葡萄糖脱氢酶和醛缩酶-A4个蛋白点。[结论]HQ可诱导膜联蛋白-Ⅱ、醛缩酶-A和尿苷二磷酸葡萄糖脱氢酶表达下调及甲酰-谷胱苷肽水解酶表达的上调。
[Objective] To investigate the change of proteomics expression in human bone marrow mesenchymal stem cells induced by hydroquinone and to screen for the target protein.[ Methods] The human bone marrow mesenchymal stem cells were isolated and cultivated till the third generation,then were divided randomly into two groups：the experiment group and the control group.In the experiment group,the stem cells were exposed to hydroquinone at 5×10^-5 mol/L for 24 hours while in the control group,the cells were treated with the same concentration of phosphate buffer solution.The total protein was isolated and purified after exposure.The two dimensional gel electrophoresis and mass spectrum were used for detecting the changes in proteomics expression to find the target protein which hydroquinone affected.[ Results] Among 3 repeated experiments,each had about 20 different proteins between the control group and the hydroquinone treated group.But there were only 11 well repeated protein spots,in which 4 proteins were identified by LC-Qq-TOF MS/MS,i.e.Annexin-Ⅱ,Formyl-glutathione hydrolase,UDP-glucose dehydrogenase and Aldolase A.[ Conclusion] Hydroquinone exposure can induce a decrease of Annexin-Ⅱ,Aldolase A, UDP-glucose dehydrogenase,and an increase of Formyl-glutathione hydrolase protein.
出处
《环境与职业医学》
CAS
北大核心
2009年第5期464-467,共4页
Journal of Environmental and Occupational Medicine
关键词
氢醌
骨髓间充质干细胞
双向凝胶电泳
质谱
蛋白质组学
hydroquinone
bone marrow mesenchymal stem cells
two dimensional gel electrophoresis
mass spectrum
proteomics
