仙灵骨葆同方提取物的肝细胞毒性研究

Study on hepatotoxicity of Xianling Gubao extract and its unilateral extracts

目的:考察仙灵骨葆同方提取物的肝细胞毒性。方法:不同浓度的仙灵骨葆同方、缺方及各单方提取物作用于人肝癌Hep3B细胞后,用CCK-8试剂盒检测肝细胞毒性。以阳性药物FCCP验证该细胞模型并获得量效参考值,进而筛选出不同提取物的半数抑制浓度(IC50),比较各提取物的肝细胞毒性的差异。结果:全方、缺方和各单方提取物的IC50分别为:全方提取物153.2μg/ml,缺淫羊藿提取物312.2μg/ml,缺续断提取物217.1μg/ml,缺丹参提取物256.5μg/ml,缺知母提取物229.6μg/ml,缺补骨脂提取物246.9μg/ml,缺地黄提取物259.4μg/ml;淫羊藿提取物91.77μg/ml,续断提取物179.0μg/ml,丹参提取物410.6μg/ml,知母提取物307.9μg/ml,补骨脂提取物73.22μg/ml,地黄提取物180.2μg/ml。其中全方提取物、淫羊藿提取物和补骨脂提取物的IC50值明显低于其他提取物,故初步判断仙灵骨葆处方中造成肝毒性的主要是淫羊藿和补骨脂。结论:本研究采用CCK-8法,筛选出仙灵骨葆处方中的肝毒性成分主要为淫羊藿和补骨脂提取物,为深入研究仙灵骨葆的肝毒性打下了基础。

Objective:To study the hepatotoxicity of Xianling Gubao extract.Methods:With different concentrations of Xianling Gubao extract and its unilateral extracts acting on human hepatocyte Hep3B,hepatotoxicity was detected by using CCK-8 kit.The cell model was validated by positive drug FCCP,and the dose effect reference value was thus obtained.IC 50 of different extracts was screened out,and the differences in hepatotoxicity were compared between various extracts.Results:The IC 50 values of various extracts were as follows:Xianling Gubao extract was 153.2μg/ml,Epimedium brecicornu omission extract was 312.2μg/ml,Dipsacus asper omission extract was 217.1μg/ml,Salvia miltiorrhiza omission extract was 256.5μg/ml,Anemarrhena asphodeloides omission extract was 229.6μg/ml,Psoralea corylifolia omission extract was 246.9μg/ml,Rehmannia glutinosa omission extract was 259.4μg/ml,Epimedium brevicornu extract was 91.77μg/ml,Dipsacus asper extract was 179.0μg/ml,Salvia miltiorrhiza extract was 410.6μg/ml,Anemarrhena asphodeloides extract was 307.9μg/ml,Psoralea corylifolia extract was 73.22μg/ml and Rehmannia glutinosa extract was 180.2μg/ml.The IC 50 values of Xianling Gubao,Epimedium brecicornu and Psoralea corylifolia were significantly lower than those of other extracts.Therefore,it could be estimated that the substances that caused hepatotoxicity were mainly Epimedium brevicornu and Psoralea corylifolia.Conclusion:With CCK-8,the main hepatotoxic ingredients screened out in the formula of Xianling Gubao were chiefly Epimedium brecicornu and Psoralea corylifolia extracts.The research results could provide solid evidence for further study of the hepatotoxicity of Xianling Gubao.