癌变·畸变·突变 ›› 2006, Vol. 18 ›› Issue (2): 88-092.doi: 10.3969/j.issn.1004-616x.2006.02.003

• 论著 • 上一篇    下一篇

三种马兜铃酸类化合物对HK-2细胞的毒性比较

郭永超;林哲绚;李 慧;罗文鸿   

  1. 汕头大学医学院中心实验室,广东 汕头 515041
  • 收稿日期:2005-10-24 修回日期:2005-12-01 出版日期:2006-03-30 发布日期:2006-03-30
  • 通讯作者: 罗文鸿

The Toxic Effects of Three Aristolochia Compounds on HK-2 Cell

GUO Yong-chao;LIN Zhe-xuan; LI Hui;LUO Wen-hong   

  1. The Central Laboratory, Shantou University Medical College, Shantou 515041, Guangdong,China
  • Received:2005-10-24 Revised:2005-12-01 Online:2006-03-30 Published:2006-03-30
  • Contact: LUO Wen-hong

摘要: 背景与目的: 研究Aristolochic acid I(AAI)、Aristolochic acid II(AAII) 和 Aristolochic acid Ia (AAIa) 3种马兜铃酸类化合物在体外对肾小管上皮细胞株(HK-2)的毒性作用的差异。 材料与方法: 倒置显微镜下观察药物作用后细胞形态学改变;设计药物作用组、阴性对照组和空白调零组,采用四甲基偶氮唑蓝[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT]法检测药物作用下细胞活力抑制率;免疫组化SABC法观察α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达;流式细胞术分析细胞周期及细胞凋亡。 结果: 形态学观察表明,在30 μg/ml剂量浓度下作用24 h,AAI、AAIa药物作用组的细胞收缩变圆、脱壁;免疫组化结果表明,20 μg/ml 的剂量浓度下作用72 h,3种化合物分别作用下的细胞均出现α-SMA的表达;MTT结果显示3种药物和细胞生长抑制率之间存在着浓度和时间依赖性关系,根据半数抑制浓度IC50值,抑制作用AAI>AAIa>AAII;3种药物都能影响细胞周期甚至诱导细胞发生不同程度的凋亡。 结论: 3种药物对HK-2的毒性作用存在差异,其引起毒性效应的作用机制也可能有所不同。

关键词: 马兜铃酸, HK-2 细胞, 细胞毒性

Abstract: BACKGROUND & AIM: To investigate the toxic effects of aristolochic acid I,aristolochic acid II and aristolochic acid Ia on renal proximal tubular epithelial cell (cell line HK-2) in vitro. MATERIAL AND METHODS: The morphological changes of HK-2 cell were examined with contrast microscopy and the growth inhibition of HK-2 after treatment with the three aristolochia compounds was studied by[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT]assay. The drug treatment group, negative control group and blank control group were set up. The expression of α-smooth muscle actin was defected by immunohistochemistry and cell cycle and apoptosis rate were defermined by flow cytometry. RESULTS: Under the light microscope, HK-2 cells became contracted and rounded in AAI and AAIa groups.The expression of α-smooth muscle actin increased in the AAI,AAII and AAIa groups compared with control; the three compounds all induced the inhibition the growth of HK-2 cells in concentration-and time-dependent manner.The median inhibitory concentration(IC50) revealed that inhibition intensities were AAI>AAIa>AAII. All these compounds could affect cell cycle and even induce cell apoptosis. CONCLUSION: AAI,AAII and AAIa were cytotoxic to various extents which might have resulted from different mechanisms.

Key words: aristolochic acid, HK-2 cell, cytotoxicity

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