亚苄基环戊酮介导的光动力对多重耐药铜绿假单胞菌的体外杀伤效应  

In Vitro Activities of Benzylidend Cyclopentanone Photosensitizers Mediated Photodynamic Therapy Against Multidrug-Resistant Pseudomonas Aeruginosa

作　　者：邢丽梅[1] 王颖[1] 邱海霞[1] 曾晶[1] 周少娜[1] 赵榆霞[2] 赵洪友[1] 张佳莹[1] 顾瑛[1] XING Li-mei;WANG Ying;QIU Hai-xia;ZENG Jing;ZHOU Shao-na;ZHAO Yu-xia;ZHAO Hong-you;ZHANG Jia-ying;GU Ying(Department of Laser Medicine,The First Medical Center of The General Hospital of Chinese PLA,Beijing 100853,China;Technical institute of Physics and Chemistry CAS)

机构地区：[1]解放军总医院第一医学中心激光医学科,北京市100853 [2]中国科学院物理化学技术研究所

出　　处：《中国激光医学杂志》2019年第5期241-245,296,共6页Chinese Journal of Laser Medicine & Surgery

基　　金：国家自然基金资助（61575208)

摘　　要：目的探讨新型光敏剂亚苄基环戊酮化合物P3介导的光动力对多重耐药铜绿假单胞菌的体外杀伤效应。方法实验对象为铜绿假单胞标准菌(ATCC27853)1株和临床多重耐药菌(PA1、PA2、PA3)3株。(1)检测实验菌株与光敏剂P3的结合特性:以荧光光谱检测法检测孵育时间和孵育浓度对实验菌株与光敏剂P3结合的影响,先将4株铜绿假单胞菌与10μM光敏剂分别孵育不同时间(5、15、30、60、120和150 min),根据前期的检测结果再选择不同浓度(2. 5、5、10、25和50μM)的光敏剂P3孵育30 min。(2)观察光敏剂P3介导的PDT对铜绿假单胞菌的体外光动力抗菌效应,即PDT组(B组),按照不同浓度的光敏剂P3分为4组分别为2. 5μM(B1组)、5μM(B2组)、10μM(B3组)和25μM(B4组),药物与4种菌株的孵育时间30 min后,进行PDT处理,激光波长532 nm,功率密度40 mW/cm2,照射时间600 s,同时设立3个对照组(A组):空白对照组(A1组)、单纯照光组(A2组)和单纯光敏剂组(A3组),用稀释平板法培养24 h进行菌落计数。结果孵育时间5~30 min时,四株铜绿假单胞菌与光敏剂P3的结合量随孵育时间延长而逐渐增加;30 min后趋于饱和。浓度梯度实验结果显示,四株铜绿假单胞菌与光敏剂P3的结合量呈孵育浓度剂量依赖性增加。在相同孵育浓度和相同孵育时间条件下,四株铜绿假单胞菌与光敏剂P3的结合量未见显著差异。光敏剂P3对四株铜绿假单胞菌的PDT杀伤作用随着光敏剂P3浓度增高逐渐增强,当光敏剂P3浓度为25μM时,PDT对4株铜绿假单胞菌株均达到有效杀伤,即活性下降均>4Log;光敏剂P3对铜绿假单胞标准菌和临床耐药菌的PDT杀伤效应比较,差异无统计学意义(P>0. 05);单纯照光和单纯光敏剂对细菌的存活无影响。结论光敏剂P3介导的PDT对铜绿假单胞菌有良好的体外杀伤作用,其作用不受细菌耐药性的影响。Objective To assess the killing efficacy of P3 mediated photodynamic therapy against multidrug-resistant pseudomonas aeruginosa in vitro.Methods ATCC27853 strain and 3 clinical drug-resistance strains(PA1,PA2 and PA3)of pseudomonas aeruginosa were applied inthis study. Firstly,4 tested strains were incubated with 10 μM P3 for different lengths of time(5,15,30,60,120 and 150 min)orwith P3 of different concentrations(2. 5,5,10 and 25 μM)for 30 min. The amount of the P3 combined with strains was measuredwith fluorescence spectrometry. Additionally,these strains were incubated with P3 of different concentrations(2. 5,5,10,25 μM)for 30 min in the dark. Then,these strains were treated with the same dose of PDT(40 m W/cm2×600 s,532 nm). The control group,sole irradiation group and sole PS group were set. The dilution plate method was used to count colonies after 24 h.Results The amount of the P3 combined with strains increased gradually for the first 5-30 min period,which then reached to asaturation point. The results from the other experiment showed that the amount of P3 combined with strains grew with theconcentration. Given the same incubation concentration and incubation time,the amount of the four P3 combined with strains keptstable. In the PDT group,the efficacy of PDT against strains was enhanced with the increase of P3 concentrations. At a concentrationof 25 μM,the activity of the 4 tested strains decreased by more than 4 Log,indicating the loss of activity of 99. 99% bacteria. Theviability of the 4 tested strains were not affected in the sole irradiation group or the sole PS group.Conclusion P3 mediated PDT is rather effective to kill pseudomonas aeruginosa,which is not affected by the drug resistance ofbacterial.

关 键 词：亚苄基环戊酮光敏剂 抗菌光动力疗法 多重耐药 铜绿假单胞菌 

分 类 号：R454.2[医药卫生—治疗学] R826.23[医药卫生—临床医学]