熊果酸对K562/ADR细胞多药耐药基因1 mRNA和P-糖蛋白表达影响及机制研究  

Influence of ursolic acid on multidrug resistance-1 mRNA/P-glycoprotein in K562/ADR cells

作　　者：胡锦芳[1,2] 冯慧玲[1,3] 孙文雄 丁薇[1] 朱丹丹 刘名义 张红[1] 黄世博[1] 熊玉卿[1] HU Jin-fang;FENG Hui-ling;SUN Wen-xiong;DING Wei;ZHU Dan-dan;LIU Ming-yi;ZHANG Hong;HUANG Shi-bo;XIONG Yu-qing(Institute of Clinical Pharmacology,Nanchang University,Nanchang 330006,China;Department of Pharmacy,First Affiliated Hospital of Nanchang University,Nanchang 330006, China;Department of Pharmacy,People's Hospital of Ganzhou City, Ganzhou 341000, Jiangxi Province, China;Department of Good Clinical Practice,Second Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学临床药理研究所,南昌330006 [2]南昌大学第一附属医院药学部,南昌330006 [3]赣州市人民医院药剂科,江西赣州341000 [4]南昌大学第二附属医院药物临床试验机构办公室,南昌330006

出　　处：《中国临床药理学杂志》2018年第23期2739-2742,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81673506);江西省科技厅研究基金资助项目(20161BBG70182)

摘　　要：目的研究熊果酸对多药耐药基因1 (MDR1) mRNA和P-糖蛋白(P-gp)表达的影响及机制。方法空白组给予0. 5%二甲基亚砜(DMSO),核因子-κB(NF-κB)激动剂组给予5μmol·L^(-1)阿霉素,NF-κB抑制剂组给予25μmol·L^(-1)吡咯烷二硫代氨基甲酸盐(PDTC),实验组单独或联合激动剂或抑制剂给予5,10,25,50μmol·L^(-1)熊果酸。培养48 h后,用逆转录聚合酶链式反应(RT-PCR)检测MDR1 mRNA水平,用Western blot法测定P-糖蛋白和核蛋白P65的表达水平。结果与空白组比较,5,10,25,50μmol·L^(-1)熊果酸分别使MDR1 mRNA的表达下调了7. 41%,15. 58%,28. 75%和44. 21%(IC50=66. 04μmol·L^(-1)),P-糖蛋白的表达下调了23. 36%,37. 23%,43. 07%和64. 23%(IC50=26. 27μmol·L^(-1))。与空白组相比,5,10,25,50μmol·L^(-1)熊果酸分别使P65的表达下调了13. 73%,28. 92%,44. 07%和56. 77%(IC50=34. 07μmol·L^(-1)),且熊果酸和PDTC在5～50μmol·L^(-1)均能明显抑制阿霉素诱导的P65的表达,差异均有统计学意义(均P <0. 05)。结论熊果酸可显著抑制K562/ADR细胞MDR1 mRNA和P-糖蛋白的表达,且对抗阿霉素的诱导作用;其机制可能是通过抑制MDR1 mRNA和P-糖蛋白的上游调控信号通路NF-κB位点核蛋白P65的表达;减少NF-κB的核转位,从而抑制NF-κB活性并干扰MDR1基因转录,最终导致MDR1 mRNA及P-糖蛋白表达量的下调。Objective To study the influence of ursolic acid(UA) on the expression of multidrug resistance-1(MDR1) mRNA/P-glycoprotein( P-gp) and the molecular biological mechanisms.Methods Blank group was given 0.5% dimethyl sulfoxide (DMSO) ,The nuclear factor-κB (NF-κB) agonist group was given 5 μmol·L^-1 doxorubicin, and the NF-κB inhibitor group was given 25 μmol·L^-1 pyrrolidine dithiocarbamate (PDTC) .The test groups were administered 5,10, 25,50 μmol·L^-1 ursolic acid alone or in combination with an agonist or inhibitor,After 48 h,the expression of MDR1 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) ,the P-gp and nucleoprotein p65 in K562 /ADR cells were detected by Western blot.Results Compared with blank group, 5, 10, 25 and 50 μmol·L^-1 UA could down-regulated the expression of MDR1 mRNA by 7.41%,15.58%,28.75%,44.21% (IC50 = 66.04 μmol·L^-1) ,and the expression of P-gp decreased 23.36%,37.23%,43.07%,64.23% (IC50 = 26.27 μmol·L^-1) ,and the expression of p65 decreased 13.73% ,28.92% ,44.07% ,56.77% (IC50 = 34.07 μmol·L^-1) .Meanwhile, UA and 5-50 μmol·L^-1 PDTC could significantly inhibit the expression of P65 induced by adriamycin,all had significant differences(all P<0.05) .Conclusion UA could inhibit the expressions of MDR1 mRNA and P-gp in K562 /ADR cells and antagonize the induction of adriamycin.The mechanisms may be related to the inhibition of the expression of NF-κB nuclear protein P65 in upstream of MDR1 mRNA and P-gp in K562 /ADR cells and thedecrease of NF-κB nuclear translocation,thus inhibiting the NF-κB activation and interfering with the transcription of MDR1,eventually leading to the down-regulation of MDR1 mRNA and the expression of P-gp.

关 键 词：熊果酸 P-糖蛋白 核蛋白P65 

分 类 号：R28[医药卫生—中药学]