穿心莲内酯抑制铜绿假单胞菌外排泵MexAB-OprM的作用  

Studies of andrographolide on inhibiting MexAB-OprM efflux pump of Pseudomonas aeruginosa

作　　者：郭威[1] 周莹 叶露[1] 王丽丽[1] 吴春明[3] 覃慧敏[4] 李洪涛[1] 宋建新[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院感染科,湖北武汉430030 [2]湖北省中山医院肝病科,湖北武汉430013 [3]温州市中医院肝病科,浙江温州325000 [4]三峡大学第一临床医学院宜昌市中心人民医院,湖北宜昌443003

出　　处：《中国医院药学杂志》2010年第16期1343-1347,共5页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金资助项目(编号:30571647);湖北省卫生厅科研基金项目(编号:NX200506);国家重点基础研究发展(973计划)(编号:2007CB512900)

摘　　要：目的:研究穿心莲内酯对铜绿假单胞菌外排泵MexAB-OprM的作用及其机制。方法:采用实时荧光定量PCR分别检测PAO1和MexAB-OprM过度表达株Mex1、Mex2、Mex3培养3h及加入穿心莲内酯0,100,200mg·L-1再培养6h的MexB基因表达量。采用Mueller-Hinton倍比稀释法,检测头孢他啶、头孢匹罗、氯霉素、左氧氟沙星、卡那霉素、依米配能、美罗培南对上述菌株的最小抑菌浓度(MIC)。结果:培养9h与3h相比,PAO1、Mex1、Mex2、Mex3的MexB表达量显著升高,P值分别为0.031,0.027,0.030,0.012。穿心莲内酯100,200mg·L-1组与对照组相比,头孢他啶、头孢匹罗、左氧氟沙星、美罗培南对PAO1、Mex1、Mex2、Mex3株菌的MIC分别降低50%,75%,75%和50%～75%,而对PAO1株QS信号分子C4-HSL的分泌差异无显著性。结论:外排泵MexAB-OprM主要在稳定生长期表达,穿心莲内酯对铜绿假单胞菌外排泵MexAB-OprM转录表达的抑制作用使其可恢复部分抗菌药物的敏感性,而这种作用并非通过抑制C4-HSL的分泌而实现。OBJECTIVE To investigate the effect and mechanism of andrographolide on inhibiting MexAB-OprM efflux pump of Pseudomonas aeruginosa. METHODS The MexB mRNA expressions of PAO1 and MexAB-OprM overexpressing strain （Mexl, Mex2, Mex3） were measured by using real time PCR at 3 h and 9 h after inoculation. In presence of 0,100,200 mg·L^-1 andrographolide, the minimal inhibitory concentration（MIC） of CAZ, CPR, CP, L-OFLX, KM, IPM, MER to those strains was detected by using Mueller-Hinton broth dilution. The effect of andrographolide on quorum sensing signal C4- HSL in PAO1 strain was also determined by a sensor strain. RESULTS The relative MexB mRNA amount of PAO1 and Mex1 ,Mex2,Mex3 cultured for 9 h was significantly higher than those strains cultured for 3 h and P value was 0. 031,0. 027,0. 030,0. 012 sepa rarely. In contrast, the MIC of CAZ,CPR,L-OFLX, MER to those strains in the concentration of 100,200 mg·L^-1 andrographolide also decreased 50%, 75%,75% and 50%-75%, respectively, compared with the control. In addition, no significant difference was observed in the level of C4-HSL secretion in PAO1 strain treated with andrographoiide.. CONCLUSION Mex AB-OprM efflux pump was mainly transcribed in stationary growth phases. The effect of andrograpbolide on inhibiting the transcriptional expression of MexAP-OprM might contribute to recover several antibiotics susceptibility in Pseudomonas aeruginosa. However, this effect should not depend on inhibition of C4-HSL secretion.

关 键 词：穿心莲内酯 铜绿假单胞菌 外排泵 密度感应 

分 类 号：R285.6[医药卫生—中药学]