益气养阴方对白血病干细胞MDR1 mRNA和P-gp表达的影响  

Effect of Yiqi Yangyin Recipe on the mRNA Expression of MDR1 and P-glycoprotein of Leukemic Stem Cells

作　　者：闫理想 史哲新[1] 杨向东[1] 李德冠[2] 刘富春 王秀婷 高宏[1] 姚芳[1] 王兴丽[1] 

机构地区：[1]天津中医药大学第一附属医院血液科,天津300381 [2]中国医学科学院放射医学研究所仪器中心,天津300192 [3]天津中医药大学研究生院,天津300193

出　　处：《中国中西医结合杂志》2017年第12期1491-1495,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：天津市应用基础及前沿技术研究计划重点资助项目(No.12JCZDJC25400)

摘　　要：目的探讨益气养阴方对白血病干细胞多药耐药基因1(multidrug resistance gene 1,MDR1)mRNA和P-糖蛋白(P-glycoprotein,P-gp)表达的影响。方法从KG1a细胞株中分选出CD34+CD38-KG1a细胞,采用CCK-8法检测益气养阴方联合柔红霉素(daunorubicin,DNR)对KG1a干细胞的抑制率及IC50值;RT-PCR法检测益气养阴方干预KG1a干细胞后MDR1mRNA的表达量;流式细胞术检测益气养阴方干预KG1a干细胞后各组P-gp表达。结果经过免疫磁珠法分选CD34+CD38-KG1a细胞比例为(97.2±2.5)%,IC50(48h)值为(0.50±0.07)μg/mL。与DNR组和对照兔血清+DNR组比较,20、40μL含药兔血清+DNR组24、48、72h的抑制率均增高(P<0.05)。与空白组比较,DNR组MDR1mRNA表达增高(P<0.05);与对照兔血清组比较,对照兔血清+DNR组MDR1 mRNA表达增高(P<0.05);与含药兔血清组比较,含药兔血清+DNR组MDR1mRNA表达降低(P<0.05)。与空白组比较,DNR组P-gp表达量增高,但差异无统计学意义(P>0.05);分别与DNR组、含药兔血清组比较,含药兔血清+DNR组P-gp表达量均明显降低(P<0.05)。结论益气养阴方可逆转KG1a干细胞对DNR的耐药性,其机制可能与其协同DNR降低多药耐药基因编码的P-gp蛋白表达,从而提高对DNR的敏感性。Objective To observe the effect of Yiqi Yangyin Recipe （YYR） on the expressions of multidrug resistance gene 1 （MDR1） mRNA and P-glycoprotein （P-gp） of leukemic stem cells. Methods CD34 ＋ CD38 - KG1a cells were isolated from KG1 a cell lines. CCK-8 method was used to detect the inhibi- tory rate and IC50 value of daunorubicin （DNR） on KGla stem cells. The mRNA expression of MDR1 in KG1 a stem cell after intervened by YYR was detected by RT-PCR. P-gp expression in each group was de- tected by flow cytometry after KGla stem cells were intervened by YYR. Results The proportion of CD34 ＋ CD38- KG1 a cells after isolated by immunomagnetic beads was 97.2% ±2.5%, IC50 value at 48 h was （0.50 ±0.07） μg/mL. Compared with the DNR group and the control rabbit serum ＋DNR group, the inhibition rate increased in 20, 40 μL drug containing rabbit serum ＋ DNR groups at 24, 48, and 72 h （P 〈0.05）. Compared with the blank group, mRNA expression of MDR1 increased in the DNR group （P 〈0.05）. Compared with the control rabbit serum group, the expression of MDR1 mRNA in the controlrabbit serum ＋ DNR group increased （P 〈0.05）. Compared with the drug containing rabbit serum group, the expression of MDR1 mRNA decreased in drug containing rabbit serum ＋DNR group （P 〈0.05）. Com- pared with the blank group, the expression of P-gp was higher in the DNR group, but with no statistical significance （P 〉0.05）. Compared with the DNR group and the drug containing rabbit serum group, the expression of P-gp was significantly decreased in the drug containing rabbit serum ＋ DNR group （P 〈 0.05）. Conclusions YYR could reverse drug resistance of KGla cells to DNR. Its possible mechanism might lower P-gp protein expression encoded in MDR1 by coordinating DNR, thus improving its sensitivity to DNR.

关 键 词：益气养阴方 白血病干细胞 KG1a细胞 多药耐药基因1 P-糖蛋白 

分 类 号：R285[医药卫生—中药学]