目的 筛选体外心肌毒性代谢组学研究中模型药物盐酸多柔比星(DOX)的受试浓度及受试时间。方法 采用MTS法测定DOX作用24、48、96 h后对原代心肌(PC)细胞活力的影响,确定最佳的受试总时长;并在最佳受试时间内,测定不同浓度DOX对PC细胞活力的影响。通过显微镜下观察细胞形态改变,综合分析确定DOX的低、高浓度及不同的受试时间点。结果 DOX的最佳受试总时长为48 h,作用48 h的IC₅₀值为(1.03±0.40) μmol/L。综合分析确定DOX的受试低、高浓度分别为0.09和3.44 μmol/L,分别于药物暴露0、6、24和48 h时收获并测定样品。结论 建立了一种确定体外心肌毒性代谢组学研究中受试药物浓度及受试时间的方法,为此类体外代谢组学研究提供参考依据。

Objective To select the test concentrations and test times of model drug doxorubicin (DOX) for metabonomic study of the in vitro cardiotoxicity. Methods The cell viability of DOX on cardiomyocytes for 24, 48 and 96 h were determined by MTS assay, respectively, so as to define the optimum total test time. Afterwards, the cell viability of DOX at different concentrations on primary cardiomyocytes were determined, then the low and high concentration of test drug DOX and different test time points for metabonomic study of the in vitro cardiotoxicity were defined through observe the morphological changes of cells under microscope. Results The optimum total test time was 48 h, during which the concentration of DOX resulting in 50% viability of primary cardiomyocytes (IC₅₀) was (1.03±0.40) μmol/L. The test low and high concentrations for metabonomic experiment of the in vitro cardiotoxicity were defined as 0.09 and 3.44 μmol/L, respectively. The cardiomyocytes samples for metabonomics were harvested and determined at 0, 6, 24 and 48 h after drug exposure, respectively. Conclusions A method of selection of test drug concentrations and test times for metabonomic study of the in vitro cardiotoxicity was established, which provide references for such in vitro metabonomic studies.

重大新药创制科技重大专项(2012ZX09505001-001)