光敏疗法对慢性鼻-鼻窦炎患者耐药葡萄球菌体外杀伤效果的研究  

In-vitro study of photodynamic therapy of antibiotic-resistant staphylococcus from patients with chronic rhinosinusitis

作　　者：刘春红[2] 杨晨[1] 丁国强[1] 马英[3] 陈小英[4] 吴旸[5] 郑春泉[1] 赵可庆[1] 

机构地区：[1] 复旦大学附属眼耳鼻喉科医院耳鼻咽喉科, 上海200031 [2] 复旦大学附属眼耳鼻喉科医院检验科, 上海200031 [3] 复旦大学附属华山医院皮肤科, 上海200040 [4]上海交通大学医学院附属瑞金医院皮肤科, 200025 [5]复旦大学基础医学院病原生物学系,医学分子病毒学教育部/卫生部重点实验室, 上海200032

出　　处：《中华耳鼻咽喉头颈外科杂志》2016年第3期164-168,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

摘　　要：目的 观察光敏疗法（photodynamic therapy,PDT）对慢性鼻-鼻窦炎（CRS）患者鼻腔多重耐药金黄色葡萄球菌（金葡菌）和表皮葡萄球菌（表葡菌）的杀伤作用.方法 选取保守治疗无效的CRS患者45例,术中使用拭子自中鼻道处取鼻腔分泌物,分离金葡菌和表葡菌并进行药敏鉴定,筛选能够形成生物膜的菌株.使用发光二极管（light emitting diode,LED）光动力治疗仪、5-氨基酮戊酸（5-aminolevulinic acid,ALA）光敏剂进行PDT参数测定及细菌杀伤实验.将获得的存活菌落形成单位（colony forming unit,CFU）进行以10为底的对数转换后,以Graph Pad Prism 5软件对数据进行统计分析.结果 45例患者中共收集金葡菌13株、表葡菌16株,皆为多重耐药菌,其中能够形成生物膜的菌株金葡菌4株,表葡菌5株.实验结果以lg（CFU）显示,游离金葡菌对照组为8.32±0.31,实验组为6.47±0.67,差异有统计学意义（=9.01,P＜0.01）;表葡菌对照组为8.34±0.20,实验组为6.97±0.59,差异有统计学意义（t=8.84,P＜0.01）.生物膜金葡菌对照组为8.68±0.05,实验组为6.90±0.96,差异有统计学意义（t=3.68,P＜0.05）;表葡菌对照组为8.67±0.05,实验组为7.29±0.61,差异有统计学意义（t=5.07,P＜0.01）.结论 利用ALA作为光敏剂,LED作为光源的PDT能于体外有效杀伤CRS患者鼻腔来源的多重耐药金葡菌和表葡菌,为后续的安全性和体内实验及该治疗手段的临床应用奠定了基础.Objective To evaluate the photodynamic therapy （PDT） against multi-antibioticresistant Staphylococcus aureus （S.aureus） and Staphylococcus epidermidis （S.epidermidis） obtained from patients with chronic rhinosinusitis（CRS）.Methods Forty-five CRS patients who had been given medical treatment but still needed endoscopic surgery were included in this study.The mucus from middle meatus was collected from these patients during surgery, followed by separation of S.aureus and S.epidermidis and drug sensitive test.The strains which could form biofilm were selected.Light emitting diode （LED） array with a major wavelength of （633 ± 10）nm was used as light source and 5-Aminolevulinic acid （ALA） was used as photosensitizer in this PDT experiment.The safe range of LED dose and ALA concentration which were not toxic to bacteria by themselves were confirmed, and then did PDT experiment on S.aureus and S.epidermidis.The data of bacterial colony forming unit were transformed to lgCFU before statistical analysis.The Graph Pad Prism 5 software was used to analyzed the data.Results Thirteen S.aureus and 16 S.epidermidis were included in this experiment （from 45 patients）, all of them were multi-antibioticresistant bacteria, and four of S.aureus and five of S.epidermidis could form biofilm in each group.In planktonic S.aureus experiment, the mean lgCFU was 8.32 ± 0.31 in control group whereas the experiment group was 6.47 ± 0.67 （t =9.01, P 〈 0.01）, and in planktonic S.epidermidis experiment the final data was 8.34 ± 0.20 （control group） and 6.97 ± 0.59 （experiment group） （t =8.84, P 〈 0.01）.In biofilm S.aureus experiment, the mean lgCFU was 8.68 ± 0.05 （control group） ,6.90 ± 0.96 （experiment group） （t =3.68 ,P 〈0.05）;and in biofilm S.epidermidis experiment the data was 8.67 ± 0.05 （control group）, 7.29 ±0.61 （experiment group, t =5.07, P 〈 0.01）.Conclusion Our results demonstrated that ALA-mediated PDT on multi-antibiotic-resistant S.aureus and S.e

关 键 词：光化学疗法 鼻窦炎 葡萄球菌 金黄色 葡萄球菌 表皮 抗药性 细菌 

分 类 号：R765.4[医药卫生—耳鼻咽喉科]