五味子甲素对K562/ADR、HL60/ADR、MCF-7/ADR多药耐药逆转机制的研究  

Reversing mechanism of schizandrin A on multi-drug resistance of K562/ADR,HL60/ADR,MCF-7/ADR cell lines

作　　者：秦小清 梁宇光[1] 高洪志[1] 曲恒燕[1] 董瑞华[1] 吴行伟[1] 李海燕[1] 刘泽源[1] 

机构地区：[1]军事医学科学院附属医院临床药理室,北京100071

出　　处：《中国药理学通报》2011年第3期329-334,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30973561/H3105)

摘　　要：目的探讨五味子甲素(schizandrin A or deoxyschizan-drin,schA)对白血病细胞K562/ADR、HL60/ADR、乳腺癌细胞MCF-7/ADR多药耐药的逆转作用,并初步探讨其逆转机制。方法 MTT法检测schA对耐药细胞的逆转作用;流式细胞仪检测schA对细胞内柔红霉素、罗丹明-123含量和细胞表面P-gp表达水平的变化;用Real-time PCR方法检测schA对细胞内mdr1 mRNA和mrp1 mRNA表达;生化检测法检测schA对细胞内GSH含量的变化。结果耐药逆转实验显示:不同浓度的schA对作用机制不同的化疗药物耐药产生不同的逆转效果;蓄积实验表明schA可增加柔红霉素、罗丹明123在耐药细胞内的蓄积,并且有良好的剂量依赖关系;schA处理K562/ADR、HL60/ADR细胞24 h后,能降低P-gp蛋白和mdr1、mrp1基因的表达;schA处理K562/ADR、HL60/ADR细胞4 h后,可降低细胞内谷胱甘肽含量。结论 schA对耐药机制不同的细胞株K562/ADR、HL60/ADR均有耐药逆转作用,推测可能是与抑制细胞表面的P-gp蛋白功能和表达,降低mdr1、mrp1耐药基因的表达和降低细胞内谷胱甘肽含量有关,schA通过影响上述机制,进而增加细胞内的药物浓度,达到有效杀灭肿瘤细胞的作用。Aim To study the reversal effect of schA on the K562/ADR,HL60/ADR,MCF-7/ADR,and explore its reversal mechanism.Methods schA′s reversal effect was evaluated by MTT assay;accumulation of daunorubicin（DNR）and rhodamine-123（Rh123）and the expression of P-glycoprotein and multidrug resistance associated protein 1（MRP1）were evaluated by flow cytometry（FCM）;the expression of intracellular mdr1 mRNA and mrp1 mRNA was detected with Real-time PCR;the changes of GSH content were detected by biochemical tests.Results The result of reversion of multidrug resistance showed schA had different reversal effects on different mechanisms of chemotherapeutic agents;the experiments of accumulation showed that the schA could significantly increase daunorubicin,rhodamine 123 contents in resistant cells,which had a good dose dependent effect;the treatment of schA could reduce remarkably the expressions of the P-gp protein and mdr1,mrp1 gene after 24 h treatment;and it could reduce GSH content after 4 h treatment in K562/ADR,HL60/ADR.Conclusions schA has the reversal effect of drug resistance in different mechanisms of the two cell lines of K562/ADR and HL60/ADR.It increases the concentration of the drug resistant cells mainly by inhibiting the function and expression of P-gp,MRP1 protein and reducing mdr1,mrp1 gene expression and GSH content,and then it enhances the sensitivity and reversal effects of resistant cell lines.

关 键 词：五味子甲素 MDR1 MRP1 K562/ADR、HL60/ADR、MCF-7/ADR细胞株 多药耐药 逆转效应 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]