氨基酮戊酸光动力对痤疮丙酸杆菌生物膜的作用  

Effect of 5-aminolevulinic Acid Photodynamics Therapy on Biofilm of Propionibacterium Acnes

作　　者：刘宇甄 曾荣[1] 段志敏[1] 徐浩翔[1] 吴秋菊[1] 陈青[2] 林彤[3] 李岷[1] LIU Yuzhen;ZENG Rong;DUAN Zhimin;XU Haoxiang;WU Qiuju;CHEN Qing;LIN Tong;LI Min(Department of Laser Surgery,Institute of Dermatology,CAMS and PUMC,Nanjing 210042,China;Jiangsu Province Blood Center,Nanjing 210042,China;Jiangsu Key Laboratory of Molecular Biology for Skin Diseases&STDs,Institute of Dermatology,CAMS and PUMC,Nanjing 210042,China)

机构地区：[1]中国医学科学院,北京协和医学院,皮肤病研究所激光科,南京210042 [2]江苏省血液中心,南京210042 [3]中国医学科学院,北京协和医学院,皮肤病研究所江苏省皮肤性病学分子生物学重点实验室,南京210042

出　　处：《中国医学科学院学报》2020年第3期283-288,共6页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(81502739和81371750);江苏省自然科学基金(BK20150068);中国医学科学院医学与健康科技创新工程项目(CIFMS-2017-I2M-1-017和CIFMS-2018-I2-AI-018)。

摘　　要：目的探讨氨基酮戊酸光动力治疗(ALA-PDT)对痤疮丙酸杆菌生物膜的作用。方法在细胞爬片上培养构建痤疮丙酸杆菌生物膜,然后进行ALA-PDT处理。将实验分为不同光照剂量(50、100、200 J/cm^2)的ALA-PDT组(ALA-PDT1、ALA-PDT2、ALA-PDT3组)和单纯ALA作用组(ALA组)、单纯LED光照射组(LED组)及阴性对照组(ALA-PDT-组),采用激光共聚焦显微镜(CLSM)观察生物膜结构及死菌/活菌比值,四甲基氮盐法(XTT)检测生物膜活力。结果CLSM观察结果显示,与ALA-PDT-组相比,ALA组和LED组的生物膜结构无明显差异;ALA-PDT1组、ALA-PDT2组和ALA-PDT3组的生物膜结构则明显被破坏,且光照强度越强,生物膜结构破坏越严重。ALA-PDT-组、ALA组、LED组、ALA-PDT1组、ALA-PDT2组和ALA-PDT3组的痤疮丙酸杆菌生物膜死菌/活菌比值分别为0.350±0.033、0.305±0.046、0.330±0.032、1.525±0.439、2.293±0.148和3.092±0.189,其中,ALA组(md=0.003,P=1.000)和LED组(md=-0.025,P=1.000)与ALA-PDT-组差异无统计学意义;ALA-PDT1组(md=-0.162,P<0.001)、ALA-PDT2组(md=-0.254,P<0.001)和ALA-PDT3组(md=-0.352,P<0.001)明显高于ALA-PDT-组。XTT检测结果显示,ALA-PDT-组、ALA组、LED组、ALA-PDT1组、ALA-PDT2组和ALA-PDT3组的痤疮丙酸杆菌生物膜活力分别为0.462±0.028、0.465±0.044、0.437±0.047、0.301±0.040、0.207±0.001和0.110±0.007,其中,ALA组(md=-0.044,P=1.000)和LED组(md=-0.020,P=1.000)与ALA-PDT-组相比差异没有统计学意义;ALA-PDT1(md=1.175,P<0.001)、ALA-PDT2(md=1.942,P<0.001)和ALA-PDT3组(md=2.742,P<0.001)明显低于ALA-PDT-组。结论ALA-PDT对痤疮丙酸杆菌生物膜有抑制作用,不仅能够破坏生物膜的结构,还能够抑制生物膜的活力。Objective To investigate the effect of 5-aminolevulinic acid photodynamic therapy(ALA-PDT)on Propionibacterium acnes(P.acnes)biofilm.Methods P.acnes biofilms were constructed on a cell slide and treated with ALA-PDT.According to different light doses,the biofilms were divided into six groups:ALA-PDT group[ALA-PDT1(50 J/cm^2),ALA-PDT2 group(100 J/cm^2),ALA-PDT3 group(200 J/cm^2)],ALA-only group(ALA group),light-only group(LED),and a negative control group(ALA-PDT-group).The biofilm structure and the ratio of the dead bacteria/live bacteria were observed using a laser confocal microscope(CLSM).Biofilm viability was measured using the XTT assay.Results CLSM showed that the biofilm structures of ALA group and LED group were not significantly different from that of ALA-PDT-group,whereas the biofilm structure was more seriously damaged in ALA-PDT1 group,ALA-PDT2 group,and ALA-PDT3 group than in the ALA-PDT-group.The ratios of the dead/live bacteria in ALA-PDT-group,ALA group,LED group,ALA-PDT1 group,ALA-PDT2 group,and ALA-PDT3 group were 0.350±0.033,0.305±0.046,0.330±0.032,1.525±0.439,2.293±0.148 and 3.092±0.189,respectively.ALA group(md=0.003,P=1.000)and LED group(md=-0.025,P=1.000)did not significantly differ from the ALA-PDT-group.However,the ratio of dead/live bacteria in ALA-PDT-group was significantly lower than those in ALA-PDT1 group(md=-0.162,P<0.001),ALA-PDT2 group(md=-0.254,P<0.001),and ALA-PDT3 group(md=-0.352,P<0.001).The values of the XTT assay were were 0.462±0.028,0.465±0.044,0.437±0.047,0.301±0.040,0.207±0.001,and 0.110±0.007,respectively,in ALA-PDT-group,ALA group,LED group,ALA-PDT1 group,ALA-PDT2 group,and ALA-PDT3 group.Although the values of XTT assay in ALA(md=-0.044,P=1.000)and LED groups(md=-0.020,P=1.000)did not significantly differ from that in ALA-PDT-group,it was significantly higher in ALA-PDT-group than in ALA-PDT1 group(md=1.175,P<0.001),ALA-PDT2 group(md=1.942,P<0.001),and ALA-PDT3 group(md=-0.352,md=2.742,P<0.001).Conclusions ALA-PDT has an inhibitory effect on P.acnes biofi

关 键 词：痤疮丙酸杆菌 生物膜 氨基酮戊酸光动力 

分 类 号：R378.8[医药卫生—病原生物学]