吩噻嗪类光敏剂介导的光动力抗菌疗法对体外白色念珠菌杀伤作用的研究  

STUDY ON PHENOTHIAZINE DYES-MEDIATED PHOTODYNAMIC INACTIVATION OF CANDIDA ALBICANS

作　　者：李庆妮[1] 甄秀梅[1] 黄力毅[1] 全昌园 周丽君[1] 吴清茂[1] 宣伟军[2] 

机构地区：[1]广西医科大学第一附属医院感染性疾病科,南宁530021 [2]广西中医药大学第一临床医学院耳鼻咽喉头颈外科,南宁530001

出　　处：《广西医科大学学报》2016年第2期200-204,共5页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81260239;No.81472002);广西科技攻关资助项目(No.2013BC26041)

摘　　要：目的：比较不同吩噻嗪类光敏剂介导的光动力抗菌疗法（APDT）对白色念珠菌的体外杀伤作用。方法：以白色念珠菌标准株DAY286为实验对象,将其制备成浓度为107个/mL的细胞悬液,实验分为对照组、APDT组、单纯光照组和单纯光敏剂组。（1）对照组：不加入光敏剂,不予以光照射。（2）APDT组：将白色念珠菌细胞悬液分别与不同浓度的吩噻嗪类光敏剂亚甲蓝（MB）、甲苯胺蓝（TBO）、新亚甲蓝（NMB）及二甲基蓝（DMMB）避光混合孵育30 min,再予以同等能量不同波长光源照射。（3）单纯光照组：不加入光敏剂,仅予以同APDT组相同的光源照射。（4）单纯光敏剂组：仅加入浓度相同的MB、TBO、NMB及DMMB,不予以任何光照。用菌落形成单位（CFU）测定各组处理后白色念珠菌的生存率。结果：当光敏剂浓度为15μmol/L时,不同吩噻嗪类光敏剂（MB、TBO、NMB、DMMB）介导的APDT作用下,白色念珠菌生存率分别为78.38%、68.47%、62.37%、0.000 1%（P〈0.05）。在浓度为0.1～15μmol/L时,DMMB浓度越高,APDT对白色念珠菌的杀伤作用越强（P〈0.05）。在浓度为5～100μmol/L时,TBO和MB的杀伤作用也随着光敏剂浓度的增高而增强（P〈0.05）。单纯光敏剂组、单纯光照组与对照组的比较差异均无统计学意义（均P〉0.05）。结论：吩噻嗪类光敏剂中DMMB对白色念珠菌的体外杀伤作用最强,且在一定浓度范围内,DMMB、TBO和MB的抗白色念珠菌作用随着光敏剂浓度的增高而增强。Objective：To evaluate the efficacy of antimicrobial photodynamic therapy（APDT）by phenothiazine dyes on Candida albicans in vitro.Methods：Candida albicans standard strain DAY286 was used to prepare fungus suspension at a concentration of 107colony-forming units（CFU）/mL.Four groups were designed as the following：（1）The blank control group,the fungus suspension without photosensitizers（PSs）and illumination;（2）The APDT group,the fungus suspension was mixed with different concentrations of phenothiazine photosensitizers such as toluidine blue O（TBO）,methylene blue（MB）,new methylene blue（NMB）and 1,9-dimethyl methylene blue（DMMB）,and incubated in the dark at room temperature for 30 min.Then the mixed suspension was exposed to the illuminants at different wavelength but with same energy according to the experimental design;（3）The light group,the fungus suspension was exposed to the illuminant only,which was same as the APDT group;（4）The photosensitizer group,the fungus suspension was incubated with different PSs（MB,TBO,NMB and DMMB）for 30 min without illumination.The survival rate of fungus was calculated by counting CFU after above treatments.Results：Different PSs（MB,TBO,NMB and DMMB）showed various APDT effects against Candida albicans,the survival rate was 78.38%,68.47%,62.37% and 0.0001%,respectively,at the concentration of 15μmol/L（P 〈0.05）.The DMMB-mediated photodynamic inactivation of Candida albicans was increased with the increment of the concentration from 0.1μmol/L to 15μmol/L（P 〈0.05）.At the concentrations from 5μmol/L to 100μmol/L,the APDT effects of TBO and MB showed dose-dependent killing effect（P 〈0.05）.There was no statistically significant difference among the blank control group,the light group and the photosensitizer group（P〉 0.05）.Conclusion：DMMB is the most effective phenothiazinium dye against Candida albicans in vitro.In a certain range of concentration,the APDT effects of DMMB,TBO and MB showed dose-dependen

关 键 词：光动力抗菌疗法 光敏剂 吩噻嗪类 白色念珠菌 体外实验 

分 类 号：R-519.3[医药卫生]