硝苯地平调控铜绿假单胞菌群体感应系统的初步研究  

Preliminary Study on the Regulation of Nifedipine on Quorum Sensing System in Pseudomonas Aeruginosa

作　　者：晏子俊[1] 吴贤丽 颜晓燕[3] 代军[4] 袁晓燕[1] 张景勍[5] 张良明[1] Yan Zijun;Wu Xianli;Yan Xiaoyan;Dai Jun;Yuan Xiaoyan;Zhang Jingqing;Zhang Liangming(Department of Pharmacy,Panzhihua Central Hospital,Sichuan Panzhihua 617067,China;Department of Laboratory,Panzhihua Central Hospital;School of Pharmacy,Chengdu Medical College;Department of Oncology,Panzhihua Central Hospital;Chongqing Research Center for Pharmaceutical Engineering,College of Pharmacy,Chongqing Medical University)

机构地区：[1]攀枝花市中心医院药学部,四川攀枝花617067 [2]攀枝花市中心医院检验科 [3]成都医学院药学院 [4]攀枝花市中心医院肿瘤科 [5]重庆医科大学药学院重庆药物高校工程研究中心

出　　处：《中国药师》2020年第10期2045-2048,共4页China Pharmacist

基　　金：四川省科技厅面上项目(编号:18YYJC1856);四川省医学会高血压疾病(施慧达)专项科研课题(编号:2018SHD2-7);攀枝花市指导性科技计划项目(编号:2019ZD-S-8);攀枝花市中心医院院内科研课题(编号:201813)。

摘　　要：目的:探讨硝苯地平调控铜绿假单胞菌(PA)群体感应(QS)系统抑制毒力因子的表达。方法:应用倍比稀释法测定硝苯地平对PA的体外最低抑菌浓度(MIC);设置空白对照组和实验组,体外构建适宜环境,培养菌株24 h,测定600 nm处硝苯地平对PA增殖的影响,并绘制生长曲线;蛋白水解酶实验测定硝苯地平对PA蛋白水解酶的抑制作用;弹性蛋白酶实验测定硝苯地平对PA弹性蛋白酶的抑制作用;绿脓毒素实验测定硝苯地平对PA绿脓毒素的抑制作用。结果:硝苯地平在体外对PA表现出良好的抗菌活性:硝苯地平抗PA的MIC为256μg·ml^-1;生长曲线显示,加样4 h时后,硝苯地平浓度为256,128,64,32μg·ml^-1时可明显抑制PA生长(P<0.05或P<0.01);毒力因子表达实验中,硝苯地平浓度为512,256,128,64,32,16,8μg·ml^-1时可明显抑制蛋白水解酶和弹性蛋白酶的活性(P<0.05或P<0.01),硝苯地平浓度为512,256,128,64,32,16μg·ml^-1时可明显抑制绿脓毒素的表达(P<0.05或P<0.01);且硝苯地平对PA毒力因子表达的抑制作用表现出浓度依赖性。结论:硝苯地平可调控QS系统抑制PA毒力因子表达的作用,在体外可有效抑制PA的生长,有望成为一种新型抗菌增效药物。Objective:To evaluate the inhibitory effect of nifedipine on the quorum sensing system in pseudomonas aeruginosa(PA).Methods:The minimum inhibitory concentration(MIC)of nifedipine to PA was determined by disc-diffusion method.After PA were cultured with suitable medium for 24 hours,the absorbency at 600 nm was detected and the growth curve was drawn.The production of proteolytic enzymes was determined by protease assay.The production of elastase was determined by elastase assay.The production of pyocyanin was determined by pyocyanin assay.Results:Nifedipine showed good antibacterial activity against PA in vitro.The MIC of nifedipine against PA was 256μg·ml^-1.The growth of PA was inhibited by nifedipine when the concentration of nifedipine was 256,128,64 and 32μg·ml^-1(P<0.05 or P<0.01).In the bactrical virulence factors experiment,the concentration of nifedipine was 512,256,128,64,32,16 and 8μg·ml^-1,the activities of proteolytic enzymes and elastase were significantly inhibited(P<0.05 or P<0.01).The concentration of nifedipine at 512,256,128,64,32 and 16μg·ml^-1 significantly inhibited the expression of pyotoxin(P<0.05 or P<0.01).The inhibitory effect of nifedipine on the expression of PA virulence factors showed a concentration-dependent manner.Conclusion:The quorum sensing can be inhibited by nifedipine in PA.Nifedipine effectively inhibit the growth of PA in vitro,which may be developed into a new type of antibacterial drug.

关 键 词：硝苯地平 调控 铜绿假单胞菌 群体感应系统 

分 类 号：R965.1[医药卫生—药理学]