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目的利用聚酰胺-胺(polyamidoamine,PAMAM)树枝状大分子包裹抗癌药物阿霉素,建立细胞内测定阿霉素浓度的HPLC检测方法,研究PAMAM对阿霉素在HepG 2细胞内的摄取行为的影响。方法将盐酸阿霉素脱去盐酸后利用疏水作用进入PAMAM内腔制备载药胶束,以柔红霉素为内标,粉碎细胞后提取细胞内的阿霉素进行浓度测定。测定细胞摄取、外排期间的药物浓度,绘制时间-浓度曲线,并计算动力学参数。结果 PAMAM包裹脱盐酸阿霉素后得到阿霉素的质量浓度为1.034g·L-1的溶液,成功建立胞内阿霉素的含量测定方法。细胞摄取动力学结果显示PAMAM使得阿霉素胞内达峰时间为1h,胞内浓度明显增高;消除动力学结果显示,k减小为原来的0.347倍,t1/2延长为原来的2.878倍,MRT增加为原来的2倍。结论经PAMAM包裹后阿霉素能够在HepG 2细胞内快速达峰,胞内浓度增加,消除速率减慢,滞留时间延长,有利于药物药效的发挥。
OBJECTIVE: To establish an HPLC assay for the determination of doxorubicin in cells by polyamidoamine (PAMAM) dendrimer coated with doxorubicin, and investigate the effect of PAMAM on uptake of doxorubicin in HepG2 cells Impact. Methods After the doxorubicin hydrochloride was removed by hydrochloric acid, the drug-loaded micelles were prepared by hydrophobic interaction into the PAMAM cavity. The daunorubicin was used as an internal standard, and the intracellular doxorubicin was extracted after being smashed to determine the concentration. The concentration of drug during cell uptake and efflux was determined. The time-concentration curve was plotted and the kinetic parameters were calculated. Results After PAMAM parched with acidified doxorubicin, a doxorubicin solution with a concentration of 1.034 g · L -1 was obtained. The intracellular doxorubicin assay was successfully established. The kinetics of cellular uptake showed that intracellular PAMAM concentration reached the intracellular peak of doxorubicin for 1 h and the intracellular concentration was significantly increased. The elimination kinetics showed that k decreased to 0.347 times and t1 / 2 to 2.878 times, MRT increased 2 times the original. CONCLUSION: Adriamycin can rapidly reach the peak in HepG 2 cells after PAMAM encapsulation, and the intracellular concentration increases, the elimination rate slows down and the residence time prolongs, which is beneficial to the drug efficacy.