目的 篩選體外心肌毒性代謝組學(xué)研究中模型藥物鹽酸多柔比星(DOX)的受試濃度及受試時(shí)間。方法 采用MTS法測(cè)定DOX作用24、48、96 h后對(duì)原代心肌(PC)細(xì)胞活力的影響,確定最佳的受試總時(shí)長(zhǎng);并在最佳受試時(shí)間內(nèi),測(cè)定不同濃度DOX對(duì)PC細(xì)胞活力的影響。通過(guò)顯微鏡下觀察細(xì)胞形態(tài)改變,綜合分析確定DOX的低、高濃度及不同的受試時(shí)間點(diǎn)。結(jié)果 DOX的最佳受試總時(shí)長(zhǎng)為48 h,作用48 h的IC₅₀值為(1.03±0.40) μmol/L。綜合分析確定DOX的受試低、高濃度分別為0.09和3.44 μmol/L,分別于藥物暴露0、6、24和48 h時(shí)收獲并測(cè)定樣品。結(jié)論 建立了一種確定體外心肌毒性代謝組學(xué)研究中受試藥物濃度及受試時(shí)間的方法,為此類體外代謝組學(xué)研究提供參考依據(jù)。

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.

重大新藥創(chuàng)制科技重大專項(xiàng)(2012ZX09505001-001)