中国中药杂志

2008, (07) 793-797

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马兜铃内酰胺-Ⅰ进入人肾小管上皮细胞及细胞内分布和蓄积的观察
Observation of penetration,distribution and accumulation in human renal proximal tubular epithelial cells by aristololactam-Ⅰ

商朴;王璇;李晓玫;唐嘉薇;蔡少青;
SHANG Pu1,WANG Xuan1,LI Xiao-mei2,TANG Jia-wei2,CAI Shao-qing1(1.School of Pharmaceutical Sciences,Peking University,Beijing 100083,China;2.Renal Division,Department of Medicine,First Hospital,Peking University,and Key Lab of Renal Diseases,Public Health Ministry of China,Beijing 100034,China)

摘要(Abstract):

目的:观察马兜铃内酰胺-Ⅰ(AL-Ⅰ)能否进入肾小管上皮细胞及其在细胞内的分布、蓄积情况。方法:以体外培养的人类近端肾小管上皮细胞系(HK-2)为研究对象,用不同质量浓度的AL-Ⅰ(5~20μg.mL-1)处理细胞,采用荧光倒置显微镜观察细胞内由AL-Ⅰ产生荧光的有无及其分布,并在洗去含AL-Ⅰ的培养液后继续培养HK-2细胞48h,观察细胞内AL-Ⅰ荧光的持续情况以判断其蓄积性。结果:不同质量浓度的AL-Ⅰ(5~20μg.mL-1)处理细胞0.5h内,均能在肾小管上皮细胞内观察到蓝绿色的AL-Ⅰ荧光,并且荧光仅分布于细胞浆中,而在细胞核内未观察到。在洗去含AL-Ⅰ培养液后继续培养时AL-Ⅰ的荧光可在细胞内持续存在48h以上,其持续存在部位仍为细胞浆。结论:AL-Ⅰ可以在短时间内迅速进入肾小管上皮细胞,其分布部位在细胞浆,不进入细胞核;AL-Ⅰ在细胞浆内的蓄积可能与其致肾损伤的毒性作用机制有关,即通过进入肾细胞并蓄积于细胞内在肾病过程中持续发挥毒性作用。
Objective:To study whether aristololactam Ⅰ(AL-Ⅰ)can enter renal proximal tubular epithelial cells and the situation of intracellular distribution and accumulation.Method:Cultured human renal proximal tubular epithelial cell line(HK-2)was used as the subject.Intracellular fluorescence from AL-Ⅰ and its distribution are examined by fluorescence microscopy after a treatment with different concentration of AL-Ⅰ,the intracellular accumulation of AL-Ⅰ was also investigated by incubated cells in AL-Ⅰ-free medium for 48 h after washing-out the media containing AL-Ⅰ.Result:After treatment of AL-Ⅰ(concentration from 5 μg·mL-1 to 20 μg·mL-1),glaucous fluorescence could be observed inside renal proximal tubular epithelial cells at 0.5 h,and the fluorescence distributed only in cytoplasm while not be observed in nuclei.Moreover,the fluorescence of AL-Ⅰ could be kept in cytoplasm for more than 48 h after washing out the media containing AL-Ⅰ.Conclusion:AL-Ⅰ is able to enter renal proximal tubular epithelial cells in short time and accumulate in cytoplasm,but not enter nuclei.This property may contribute to the cytotoxic mechanism of renal injury induced by AL-Ⅰ,which may partially explain the persistent renal toxicity of AAs and its metabolites in the development of aristolochic acid nephropathy.

关键词(KeyWords): 马兜铃内酰胺-Ⅰ;肾小管上皮细胞;荧光显微镜观察;分布;蓄积
aristololactam Ⅰ;renal tubular epithelial cell;fluorescence microscopy;distribution;accumulation

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(30371748);; 国家科技支撑计划项目(2006BAI14B01);; 国家教育部985项目

作者(Author): 商朴;王璇;李晓玫;唐嘉薇;蔡少青;
SHANG Pu1,WANG Xuan1,LI Xiao-mei2,TANG Jia-wei2,CAI Shao-qing1(1.School of Pharmaceutical Sciences,Peking University,Beijing 100083,China;2.Renal Division,Department of Medicine,First Hospital,Peking University,and Key Lab of Renal Diseases,Public Health Ministry of China,Beijing 100034,China)

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