醋酸铅对大鼠胰岛瘤细胞INS-1自噬与凋亡的影响Effects of Lead Acetate on the Autophagy and Apoptosis of the Rat Insuloma Cells INS 1
付荣;孙丽;王晓丽;胡士涛;季虹;荣海钦;
摘要(Abstract):
为了研究环境中常见重金属污染物铅对大鼠胰岛瘤细胞INS-1自噬与凋亡的影响及其可能的分子机制,使用不同浓度醋酸铅暴露大鼠胰岛瘤细胞INS-1,利用磺酰罗丹明B(sulforhodamine B,SRB)法检测细胞存活率;Western blot法检测不同浓度醋酸铅对细胞中自噬与凋亡标志蛋白及哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路标志分子表达的影响;利用活性氧(reactive oxygen species,ROS)试剂盒检测细胞内ROS水平;用ROS清除剂N-乙酰-L-半胱氨酸(N-acetylcysteine,NAC)预处理细胞,检测醋酸铅引起的细胞自噬与凋亡是否受到影响。使用统计学软件SPSS 17.0对数据进行统计学分析后,与对照组相比,醋酸铅导致INS-1细胞的存活率下降(p<0.05),且呈时间-剂量依赖性;随醋酸铅浓度增大,自噬和凋亡标志蛋白表达增加,mTOR信号途径标志分子没有明显变化;NAC预处理后醋酸铅引起的细胞自噬和凋亡减少。实验结果表明,醋酸铅可通过mTOR-非依赖途径诱导INS-1细胞的自噬与凋亡,其可能机制为刺激细胞内ROS的产生。
关键词(KeyWords): 醋酸铅;INS-1细胞;自噬;凋亡;活性氧(ROS)
基金项目(Foundation): 中国科学院生态环境研究中心开放基金(KF2011-14)
作者(Author): 付荣;孙丽;王晓丽;胡士涛;季虹;荣海钦;
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参考文献(References):
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