摘要
目的:观察非细胞毒性浓度(抑制率≤5%)半夏醇提取液对耐阿霉素的人白血病细胞系K562/A02多药耐药性的逆转作用,并探讨其逆转机制。方法:将细胞悬液接种于72孔培养板中,细胞数为2×108个/L,半夏醇提取液(2.5,5,10,20,40μg.L-1)处理K562/A02细胞72 h后,采用MTT法测定半夏醇提取液的细胞毒性及其对K562/A02细胞敏感性的影响,用流式细胞仪检测非细胞毒性浓度的半夏醇提取液处理后K562/A02细胞膜表面糖蛋白p170表达的变化与细胞内柔红霉素的浓度,应用SPSS 16.0软件包对实验结果进行统计学处理。结果:半夏醇提取液对K562/A02细胞有一定的细胞毒作用,其非细胞毒性质量浓度为6.5μg.L-1,非细胞毒性浓度半夏醇提取液可显著降低阿霉素对K562/A02细胞的半数抑制浓度(IC50),使K562/A02细胞的IC50由原来的(30.9±0.11)μg.L-1降低至(10.1±0.21)μg.L-1,其逆转倍数为3.06倍;半夏醇提取液作用于K562/A02细胞后,细胞膜糖蛋白p170的表达从91.21%下调至45.12%(P<0.01);柔红霉素外渗试验显示,半夏醇提取液作用于K562/A02细胞后,可使细胞内化疗药物的浓度明显增加。结论:半夏醇提取液可部分逆转多药耐药细胞系K562/A02细胞对阿霉素的耐药性,半夏醇提取液逆转白血病细胞耐药性是通过下调了白血病细胞膜糖蛋白p170的表达,使其将化疗药物泵出细胞外的功能受到了抑制,从而使化疗药物在白血病细胞内能达到有效浓度,从而可以杀灭耐药的白血病细胞系。
Objective：To investigate the effect and mechanism of alcoholic extract of Pinellia ternate at non-cytotoxic concentration（inhibit rate≤5%）reversing the multidrug resistant（MDR） leukemia cell lines K562/A02.Method：Cells were seeded into 72-well plates at 2×108/L with varying concentrations（2.5,5,10,20,40 μg · L-1） of P.ternate for 72 h,MTT colorimetry was used to determine the cytotoxic effect of alcoholic extract of P.ternate and the sensibility to adriamycin of K562/A02 cell line.Flow cytometry was used to determine the expressive and functional changes of glycoprotein-170（p170）on the cell membrane dealed with alcoholic extract of P.ternate at non-cytotoxic concentration.And daunorubicin concentration in the cells was detected by flow cytometry.The statistical treatment with SPSS 16.0 was applied to deal with all the experimental data.Result：Alcoholic extract of P.ternate had definite cytotoxic effect on K562/A02,the non-cytotoxic dose of oxymatrine was 6.5 μg · L-1.Non-cytotoxic dose of alcoholic extract of P.ternate can significantly decrease the medium inhibition concentration（IC50） value of K562/ADM cell line to adriamycin（ADM）.The IC50was obviously reduced from（30.9±0.11） μg · L-1 to（10.1±0.21） μg · L-1,alcoholic extract of P.ternate at non-cytotoxic concentration could partly reverse MDR of K562/A02（3.06times）,and could descend the expression of p170 from 91.21% to 45.12%（P〈0.01）.The exosmosis test of daunorubicin showed that intracellular concentration of chemotherapeutics concentration increased evidently.Conclusion：Alcoholic extract of P.ternate is able to partly reverse MDR of K562/A02 cell line could descend the expression of P170,and can restrain the function of pumping chemotherapeutics out of the cel1 and enhance the efficient intracellular concentration of chemotherapeutics which can kill leukemia cells of multidrug resistance,and partly reversed MDR of K562/A02.
出处
《中国实验方剂学杂志》
CAS
北大核心
2012年第18期157-160,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(30771156)
山东省自然科学基金项目(2009ZRB01762)
山东省高校科技计划项目(J12LM56)
关键词
半夏醇提取液
K562/A02细胞系
多药耐药
alcoholic extract of Pinellia ternate； K562/A02 cell line； multidrug resistance
