DNaseⅠ对痤疮丙酸杆菌生物膜的抑制效应研究  

Inhibitory effect of deoxyribonucleaseⅠagainst Cutibacterium acnes biofilms

作　　者：周蒙 郑娜娜 曾荣 徐浩翔 段志敏 刘宇甄 李岷 Zhou Meng;Zheng Nana;Zeng Rong;Xu Haoxiang;Duan Zhimin;Liu Yuzhen;Li Min(Hospital of Dermatology,Chinese Academy of Medical Sciences and Peking Union Medical College,Nanjing 210042,China;Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs,Nanjing 210042,China;Department of Dermatology,The Affiliated Jiangning Hospital of Nanjing Medical University,Nanjing 211100,China)

机构地区：[1]中国医学科学院北京协和医学院皮肤病医院,南京210042 [2]江苏省皮肤病与性病分子生物学重点实验室,南京210042 [3]南京医科大学附属江宁医院,南京211100

出　　处：《中华皮肤科杂志》2023年第10期920-924,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金(82173432、82103749);江苏省“双创博士”项目(JSSCBS20211610)。

摘　　要：目的探讨DNaseⅠ对痤疮丙酸杆菌生物膜的抑制效应。方法构建痤疮丙酸杆菌生物膜并给予DNaseⅠ处理,分为不同DNaseⅠ浓度组(0、5、10、20 U/ml)。采用四甲基氮盐法检测生物膜活力,结晶紫半定量法检测生物膜含量,激光共聚焦显微镜观察生物膜结构及活菌/死菌比值。组间差异比较采用单因素方差分析。结果5、10、20 U/ml的DNaseⅠ处理后痤疮丙酸杆菌生物膜活力值分别为1.882±0.421、1.653±0.287、1.473±0.154,与阴性对照组(2.668±0.245)相比存在显著抑制,且DNaseⅠ浓度越高抑制作用越显著(F=9.68,P=0.005)。5、10、20 U/ml DNaseⅠ组生物膜含量值分别为1.039±0.003、0.489±0.079、0.147±0.034,与阴性对照组(1.359±0.071)相比显著降低,且DNaseⅠ浓度越高生物膜含量越低(F=174.40,P<0.001)。激光共聚焦显微镜观察结果显示,与阴性对照组相比,5、10、20 U/ml DNaseⅠ组生物膜结构被破坏,且DNaseⅠ浓度越高生物膜结构破坏越严重。5、10、20 U/ml DNaseⅠ组痤疮丙酸杆菌生物膜活菌/死菌比值分别为2.303±0.457、1.534±0.526、1.263±0.354,比阴性对照组(4.475±0.146)显著降低,且浓度越高生物膜活菌/死菌比值越低(F=56.75,P<0.0001)。结论DNaseⅠ对痤疮丙酸杆菌生物膜的结构有破坏作用,对其活力有抑制作用。Objective To investigate the inhibitory effect of deoxyribonucleaseⅠ(DNaseⅠ)on Cutibacterium acnes biofilms.Methods Cutibacterium acnes biofilms were constructed,and then were divided into 4 groups(negative control group,5,10 and 20 U/ml DNaseⅠgroups)to be treated with DNaseⅠat different concentrations of 0,5,10 and 20 U/ml respectively.The biofilm viability was evaluated by tetrazolium salt colorimetric assay,the biofilm content was determined by crystal violet staining-based semi-quantitative analysis,the biofilm structure was observed by confocal laser scanning microscopy,and the live/dead bacteria ratio was calculated.One-way analysis of variance was used to analyze differences between groups.Results After the treatment with DNaseⅠ,the biofilm viability was significantly inhibited in the 5,10 and 20 U/ml DNaseⅠgroups(1.882±0.421,1.653±0.287,1.473±0.154,respectively)compared with the negative control group(2.668±0.245),and the inhibitory effect was gradually enhanced with the increase in concentrations of DNaseⅠ(F=9.68,P=0.005).Crystal violet semi-quantitative analysis showed that the biofilm content was also significantly lower in the 5,10 and 20 U/ml DNaseⅠgroups(1.039±0.003,0.489±0.079,0.147±0.034,respectively)than in the negative control group(1.359±0.071),and the higher the DNaseⅠconcentration,the lower the biofilm content(F=174.40,P<0.001).Confocal laser scanning microscopy showed that the biofilm structure was destroyed in the 5,10 and 20 U/ml DNaseⅠgroups compared with the negative control group,and the higher the DNaseⅠconcentration,the more severe the destruction of biofilm structure.Additionally,the live/dead bacteria ratio was significantly lower in the 5,10 and 20 U/ml DNaseⅠgroups(2.303±0.457,1.534±0.526,1.263±0.354,respectively)than in the negative control group(4.475±0.146),and the ratio decreased with the increase in concentrations of DNaseⅠ(F=56.75,P<0.0001).Conclusion DNaseⅠhad a destructive effect on the structure of Cutibacterium acnes biofilms,and c

关 键 词：痤疮丙酸杆菌 生物膜 脱氧核糖核酸酶Ⅰ 

分 类 号：R758.733[医药卫生—皮肤病学与性病学]