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-di methylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide,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30μg/ml剂量浓度下作用24h,AAI、AAIa药物作用组的细胞收缩变圆、脱壁;免疫组化结果表明,20μg/ml的剂量浓度下作用72h,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。
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 MLTHODS： 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-diphenyltetmzolium 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 ＇immunnhistochemistry and cell cycle and apoptosis rate were defermined by flow cytometry. RESULTS： Under the light microscope, HK-2 cells became contracted and rounded in AM and AAⅠa groups.The expression of α-smooth muscle actin increased in the AAⅠ, AⅡ 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, AⅡ and AAⅠa were cytotoxic to various extents which might have resulted from different mechanisms.
出处
《癌变．畸变．突变》
CAS
CSCD
2006年第2期88-92,共5页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
国家自然科学基金(No.30271139)
广东省自然科学基金(No.022124)
2003年度教育部"优秀青年教师资助计划"项目(教人司[2003]355号)
