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目的改良心肌细胞培养方法,评价抗肿瘤药物多柔比星(doxorubicin,DOX)、顺铂(cisplatinum,DDP)和氯法拉滨(clofarabine,CLO)的心肌细胞毒性。方法采用双酶依次消化法充分分离心脏组织中的心肌细胞,优化差速贴壁法进行细胞纯化的时长为20 min,台盼蓝染色法检测细胞成活率,免疫组织化学法鉴定心肌细胞纯度,MTS法分别测定多柔比星、顺铂和氯法拉滨对心肌细胞的毒性。结果较低浓度的多柔比星(0.17μmol·L-1)、顺铂(17.50μmol·L-1)和氯法拉滨(0.33μmol·L-1)即能引起与对照组细胞存在显著性差异(P<0.05),3种药物对心肌细胞的半数抑制浓度IC50值分别为(1.17±0.28)、(7.90±2.05)和(34.07±14.43)μmol·L-1。结论与各自对照组细胞相比,3种抗肿瘤药物的心肌细胞毒性呈浓度依赖性增加,且心肌细胞在药物暴露早期对其敏感性较高,提示临床需警惕药物引起的心脏毒性。
Objective To improve the cardiomyocyte culture method and evaluate the cardiomyocyte cytotoxicity of antitumor drug doxorubicin (DOX), cisplatin (DDP) and clofarabine (CLO). Methods The cardiomyocytes were isolated by digestion and digestion method. The optimal cell culture time was 20 min by differential adherence method, the survival rate of cells was detected by trypan blue staining, the purity of cardiomyocytes was identified by immunohistochemistry, MTS assay of doxorubicin, cisplatin and clofarabine on myocardial cell toxicity. Results Low concentration of doxorubicin (0.17μmol·L-1), cisplatin (17.50μmol·L-1) and clofarabine (0.33μmol·L-1) (P <0.05). The IC50 values of the three drugs on cardiomyocytes were (1.17 ± 0.28), (7.90 ± 2.05) and (34.07 ± 14.43) μmol·L-1, respectively. Conclusion Compared with the control cells, the cardiomyocyte cytotoxicity of three kinds of antitumor drugs increased in a concentration-dependent manner, and the sensitivity of cardiomyocytes in the early stage of drug exposure was high, suggesting that the cardiotoxicity induced by drugs should be vigilant in clinic.