基于肝细胞毒价检测的雷公藤质量评价方法研究  

Establishment of hepatotoxic bioassay for quality control of Tripterygium wilfordii

作　　者：赵庆国[1,2] 王艳辉[3] 马致洁[4] 张萍[3] 赵艳玲[3] 赵奎君[4] 格小光[5] 郝庆秀[5] 黄璐琦[5] 王伽伯[3] 肖小河[3] 

机构地区：[1]解放军军事医学科学院毒物药物研究所,北京100850 [2]解放军第三〇二医院药学部,北京100039 [3]解放军第三〇二医院全军中医药研究所,北京100039 [4]首都医科大学附属北京友谊医院,北京100050 [5]中国中医科学院中药资源中心,北京100070

出　　处：《中草药》2015年第3期378-383,共6页Chinese Traditional and Herbal Drugs

基　　金：国家科技支撑计划课题(2012BAI29B02);国家公益性行业科研专项(201207002);中央本级课题(20603020101);北京市中医局科技发展基金项目(QN2014-18);首都医科大学附属北京友谊医院院启动项目(yyqdkt2014-20)

摘　　要：目的建立基于肝细胞毒价检测的雷公藤药材质量评价方法。方法以正常人肝细胞（L02细胞系）为研究载体,以细胞抑制率为检测指标,优化建立肝细胞毒价检测方法 ,评价不同产地雷公藤及其地方习用品昆明山海棠药材的质量。结果以对乙酰氨基酚（毒价定义为400 U/g）为阳性对照品,50%乙醇超声提取雷公藤后真空干燥为干膏,用培养基配制生药量3 mg/m L的供试液,稀释比1∶0.65,量反应平行线法检测,18份雷公藤样品肝细胞毒价17.78~4 131.4 U/g（相差超过200倍）,5份昆明山海棠样品肝细胞毒价209.42~7 422.2 U/g（相差超过30倍）,不同产地雷公藤及昆明山海棠药材的肝细胞毒价差异显著;并且所检测样品鲜品较市售干品毒性大。结论初步建立了基于肝细胞毒价检测的雷公藤质量评价方法 ,直接关联其临床肝脏毒副反应,对从质量控制角度提高雷公藤临床用药安全具有参考价值,也为雷公藤肝毒性成分筛选提供准确定量的方法 。Objective To establish a method suitable to the quality evaluation of the roots in Tripterygium wilfordii based on hepatotoxic potency. Methods The inhibitory rates of T. wilfordii and its regional substitute--T, hypoglaucum, with normal human hepatocytes （L02 cell line） as carrier, were established by optimizing a series of factors, such as test samples prepared with conditions. Results Taking p-acetaminophenol as positive control （toxic potency was 400 U/g）, The 50% ethanol extract of T. wilfordii was dried under ultrasonic vacuum to obtaine the dry paste. The test solution with crude drug of 3 mg/mL was prepared by medium. The dilution ratio was l： 0.65. The hepatotoxic potencies of 18 batches of T. wilfordii and 5 batches of T. hypoglaucum were 17.78-4 131.4 and 209.42-7 422.2 U/g, respectively, detected by the reaction parallel line method, and the differences were over 200 and 30 times. There was the significant hepatotoxic potency of T. wilfordii and T. hypoglaucum from the various origins. In addition, the fresh collected samples had the larger hepatotoxic potency than that in the dried samples collected in the market. Conclusion The preliminarily established hepatotoxic potency bioassay is useful to evaluate the quality of T. wilfordii which directs the correlation with the drug＇s hepatotoxic adverse effect in clinic. The method can provide the reference for the clinical safety use of T. wilfordii based on the quality control and be as the quantitative method used to screen the toxic components from T. wilfordii

关 键 词：雷公藤 昆明山海棠 肝毒性 毒价 质量控制 

分 类 号：R285.5[医药卫生—中药学]