[1]庞家胤,和亚雄,郑梦雪,等.光动力抗菌疗法对多重耐药铜绿假单胞菌体外杀伤作用的研究[J].陆军军医大学学报(原第三军医大学学报),2021,43(07):599-605.
 PNAG Jiayin,HE Yaxiong,ZHENG Mengxue,et al.Efficacy of antimicrobial photodynamic therapy for multidrug resistant Pseudomonas aeruginosa in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2021,43(07):599-605.
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光动力抗菌疗法对多重耐药铜绿假单胞菌体外杀伤作用的研究(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
43卷
期数:
2021年第07期
页码:
599-605
栏目:
基础医学
出版日期:
2021-04-15

文章信息/Info

Title:
Efficacy of antimicrobial photodynamic therapy for multidrug resistant Pseudomonas aeruginosa in vitro
作者:
庞家胤和亚雄郑梦雪尹锐
陆军军医大学(第三军医大学)第一附属医院皮肤科
Author(s):
PNAG Jiayin HE Yaxiong ZHENG Mengxue YIN Rui

Department of Dermatology, First Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400038, China

关键词:
多重耐药菌铜绿假单胞菌光动力抗菌疗法甲苯胺蓝O外排泵基因
Keywords:
 
分类号:
R378.991; R454.2; R915
文献标志码:
A
摘要:

目的研究甲苯胺蓝O(toluidine blue O,TBO)介导的光动力抗菌疗法(antimicrobial photodynamic therapy,APDT)对多重耐药铜绿假单胞菌的杀伤作用。方法以烧伤患者创面分离的多重耐药铜绿假单胞菌和标准铜绿假单胞菌为研究对象。实验组采用不同浓度TBO(2.5、5.0、7.5、10.0  μmol/L)和不同剂量(10、20、30、40 J/cm2)光照处理[红光,波长为(635±5)nm];设单纯TBO组、单纯光照组和空白(不处理)3个对照组。采用菌落计数法计算细菌存活率。采用激光共聚焦电子显微镜、透射电镜、扫描电镜观察细菌形态学改变。采用Vitek-2 Compact型全自动药敏分析系统检测亚致死量APDT(TBO浓度2.5  μmol/L、光照剂量10 J/cm2)处理前后细菌对哌拉西林/他唑巴坦、头孢他啶、头孢吡肟、亚胺培南、美罗培南等抗生素最低抑菌浓度的变化。采用RT-qPCR测定多重耐药铜绿假单胞菌APDT处理前后耐药基因表达。结果随TBO浓度和光照剂量的增加,APDT对细菌的杀伤效应随之增加(P<0.05)。亚致死量APDT处理后,扫描电镜观察发现细菌出现皱缩、凹陷、穿孔等改变,透射电镜观察发现菌毛消失、细胞壁连续性中断、胞质电子密度降低、拟核区紊乱等变化。亚致死量APDT治疗后,多重耐药铜绿假单胞菌对上述抗生素敏感度升高,外排泵耐药基因表达下调(P<0.05)。结论TBO介导的APDT在体外对多重耐药铜绿假单胞菌有良好的杀伤作用,并且通过对拟核DNA作用降低耐药基因表达,提升了耐药菌对抗生素的敏感性。

Abstract:

ObjectiveTo investigate the bactericidal effect of toluidine blue O (TBO)-mediated photodynamic antimicrobial therapy (APDT) on multi-drug resistant Pseudomonas aeruginosa (P. aeruginosa). MethodsMulti-drug resistant P. aeruginosa isolated from 1 patient with burn wounds was used as study subject, and so was the standard strain from ATCC (No. 27853). The study bacteria were treated with different concentrations of TBO (2.5, 5.0, 7.5 and 10.0 μmol/L) combined with different doses of light (10, 20, 30 and 40 J/cm2, Red light, wavelength of 635±5 nm). While the control bacteria were divided into blank control group, TBO group and light group. The bacterial survival rate was determined by colony counting. Morphological alterations of the bacteria were observed with laser confocal electron microscopy (CLSM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The effect of sub-lethal dose of APDT (2.5 μmol/L TBO, 10 J/cm2 lighting)  on minimum inhibitory concentrations (MICs) of several antibiotics were detected, including piperacillin/tazobactam, ceftazidime, cefepime, imipenem and meropenem. The expression levels of drug-resistant genes related to the multi-drug resistant P. aeruginosa were detected using RT-qPCR before and after APDT. ResultsThe antibacterial efficacy of APDT was improved with the increases of TBO concentrations and light dose (P<0.05). After sub-lethal dose of APDT, the bacteria showed shrinkage, depression and perforation of bacterial cell wall by SEM, and fimbriae disappearance, cell wall discontinuity and decreased cytoplasmic electron density and nucleus mimics disorder by TEM. And the sub-lethal dose treatment resulted in enhanced sensitivity of the multi-drug resistant strain to above antibiotics, and down-regulation of efflux pump genes (P<0.05). ConclusionTBO-mediated APDT exerts a bactericidal efficacy on multidrug-resistant P. aeruginosa. APDT could alter the expression of drug-resistant genes by attacking nucleoid DNA, and thereby improve the sensitivity of drug-resistant bacteria to antibiotics.

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更新日期/Last Update: 2021-04-02