中国医学科学院学报

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中国医学科学院学报

中国医学科学院学报 ›› 2009, Vol. 31 ›› Issue (4): 476-480.doi: 10.3881/j.issn.1000-503X.2009.04.019

• 论著 • 上一篇    下一篇

有机阴离子转运蛋白介导马兜铃酸进入肾小管上皮细胞

王艳艳1,2;谌贻璞1,2;芮宏亮2   

  1. 1中国医学科学院北京协和医学院研究生院,北京100730
    2卫生部中日友好医院肾病中心,北京100029
  • 收稿日期:2008-09-04 修回日期:1900-01-01 出版日期:2009-08-30 发布日期:2009-08-30
  • 通讯作者: 谌贻璞

Organic Anion Transporter System Mediates the Transport of Aristolochic Acid in Tubular Epithelial Cells

WANG Yan-yan1,2;CHEN Yi-pu1,2;RUI Hong-liang2   

  1. 1Graduate School, CAMS and PUMC, Beijing 100730, China
    2Center of Nephrology, China-Japan Friendship Hospital, Beijing 100029, China
  • Received:2008-09-04 Revised:1900-01-01 Online:2009-08-30 Published:2009-08-30
  • Contact: CHEN Yi-pu

摘要: 摘要:目的 探讨马兜铃酸是否能进入人近端肾小管上皮细胞(HKC)及其进入HKC的可能途径。方法 液相色谱-串联质谱法检测经马兜铃酸钠作用后HKC破碎液中马兜铃酸的含量。乳酸脱氢酶释放实验检测马兜铃酸钠对HKC的毒性作用,并比较有机阴离子转运蛋白抑制剂丙磺舒与有机阳离子转运蛋白抑制剂四乙基铵对马兜铃酸钠导致的细胞毒作用的影响。逆转录实时聚合酶链反应和免疫印迹法检测马兜铃酸钠对HKC表达结缔组织生长因子的影响。结果 马兜铃酸钠孵育后的细胞破碎液中可检测到马兜铃酸。60 mg/L马兜铃酸钠可使HKC的乳酸脱氢酶释放率显著升高(P<0.01),丙磺舒可显著抑制上调的乳酸脱氢酶释放(P<0.05),而四乙基铵却无此作用。丙磺舒可抑制马兜铃酸钠诱导的结缔组织生长因子高表达(P<0.05)。结论 马兜铃酸钠能通过HKC的细胞膜有机阴离子转运蛋白进入细胞发挥生物学效应和毒性作用。

关键词: 马兜铃酸, 肾小管上皮细胞, 液相色谱-串联质谱, 有机阴离子转运蛋白, 结缔组织生长因子

Abstract: ABSTRACT:Objective To investigate whether aristolochic acid can be transported into human kidney proximal tubular cell (HKC) and its potential mechanism. Methods Intracellular aristolochic acid was measured by liquid chromatography-tandem mass spectrometry. The release of lactate dehydrogenase (LDH) induced by aristolochic acid in the presence of organic anion transporter inhibitor (probenecid) or organic cation transporter inhibitor (tetraethylammonium) was evaluated. The effects of probenecid on aristolochic acid induced connective tissue growth factor (CTGF) mRNA and protein expression were also examined by real time polymerase chain reaction and Western blot, respectively. Results Aristolochic acid was detected in the suspension of the denatured HKC after incubation with aristolochic acid sodium salt. The release of LDH from HKC, which was induced by 60 mg/L aristolochic acid sodium salt, was significantly inhibited by 1 mmol/L probenecid (P<0.01), but not by 1 mmol/L tetraethylammonium. The increased CTGF mRNA and protein expression in HKC stimulated by 40 mg/L aristolochic acid sodium salt was significantly down-regulated by 1 mmol/L probenecid (P<0.05), with an inhibition rate of 16% and 21%, respectively. Conclusion Aristolochic acid can be transported into HKC by organic anion transport system, and then exerts its biological effects.

Key words: aristolocdic acid, renal tubular epitdelial cells, liquid cdromatocrapdy-tandem mass spectrometry, orcanic anion transporter, connective tissue crowtd factor