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

庞家胤; 和亚雄; 郑梦雪; 尹锐

doi:10.16016/j.1000-5404.202012135

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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

基础医学 | 更新时间：2026-01-09

- 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
- Efficacy of antimicrobial photodynamic therapy for multidrug resistant Pseudomonas aeruginosa in vitro
- 第三军医大学学报 2021年43卷第7期页码：599-605
- 作者机构：
  
  400038 重庆，陆军军医大学（第三军医大学）第一附属医院皮肤科
- 作者简介：
  
  [尹锐，E-mail：swyinrui@163.com YIN Rui, E-mail: swyinrui@163.com
- 基金信息：
  
  国家自然科学基金面上项目(81571902)
- DOI：10.16016/j.1000-5404.202012135
  中图分类号： R378.991;R454.2;R915
- 收稿：2020-12-16，
  
  修回：2020-12-27，
  
  纸质出版：2021-04-15
- 稿件说明：
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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]. 第三军医大学学报, 2021,43(7):599-605.

Jiayin PNAG, Yaxiong HE, Mengxue ZHENG, et al. Efficacy of antimicrobial photodynamic therapy for multidrug resistant Pseudomonas aeruginosa in vitro[J]. Journal of Third Military Medical University, 2021, 43(7): 599-605.
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]. 第三军医大学学报, 2021,43(7):599-605. DOI： 10.16016/j.1000-5404.202012135.

Jiayin PNAG, Yaxiong HE, Mengxue ZHENG, et al. Efficacy of antimicrobial photodynamic therapy for multidrug resistant Pseudomonas aeruginosa in vitro[J]. Journal of Third Military Medical University, 2021, 43(7): 599-605. DOI： 10.16016/j.1000-5404.202012135.

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/cm

）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/cm

）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（

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（

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

Objective

To investigate the bactericidal effect of toluidine blue O (TBO)-mediated photodynamic antimicrobial therapy (APDT) on multi-drug resistant

Pseudomonas aeruginosa

(

P. aeruginosa

Methods

Multi-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

5 and 10.0 μmol/L) combined with different doses of light (10

30 and 40 J/cm

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/cm

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.

Results

The antibacterial efficacy of APDT was improved with the increases of TBO concentrations and light dose (

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 (

0.05).

Conclusion

TBO-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.

关键词

Keywords

references

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