中华皮肤科杂志 ›› 2023, Vol. 56 ›› Issue (10): 920-924.doi: 10.35541/cjd.20230196

• 论著 • 上一篇    下一篇

DNaseⅠ对痤疮丙酸杆菌生物膜的抑制效应研究

周蒙1，2    郑娜娜1    曾荣1    徐浩翔1    段志敏1    刘宇甄3    李岷1，2   

  1. 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
  • 收稿日期:2023-04-10 修回日期:2023-05-25 发布日期:2023-10-08
  • 通讯作者: 刘宇甄；李岷 E-mail:liuyuzhen409@126.com; limin@pumcderm.cams.cn
  • 基金资助:
    国家自然科学基金（82173432、82103749）；江苏省“双创博士”项目（JSSCBS20211610）

Inhibitory effect of deoxyribonucleaseⅠ against Cutibacterium acnes biofilms

Zhou Meng1,2, Zheng Nana1, Zeng Rong1, Xu Haoxiang1, Duan Zhimin1, Liu Yuzhen3, Li Min1,2   

  1. 1Hospital of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042, China; 2Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Nanjing 210042, China; 3Department of Dermatology, The Affiliated Jiangning Hospital of Nanjing Medical University, Nanjing 211100, China
  • Received:2023-04-10 Revised:2023-05-25 Published:2023-10-08
  • Contact: Liu Yuzhen; Li Min E-mail:liuyuzhen409@126.com; limin@pumcderm.cams.cn
  • Supported by:
    National Nature Science Foundation of China （82173432, 82103749）; Jiangsu Provincial Double Innovation Doctor Project （JSSCBS20211610）

PDF

3

摘要： 【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 < 0.000 1）。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。

关键词: 痤疮丙酸杆菌, 生物膜, 脱氧核糖核酸酶Ⅰ

Abstract: 【Abstract】 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 DNaseⅠ groups） to be treated with DNaseⅠ at 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 DNaseⅠ groups （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 DNaseⅠ groups （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 DNaseⅠ concentration, 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 DNaseⅠ groups compared with the negative control group, and the higher the DNaseⅠ concentration, 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 DNaseⅠ groups （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 < 0.000 1）. Conclusion DNaseⅠ had a destructive effect on the structure of Cutibacterium acnes biofilms, and could inhibit their viability.

Key words: Cutibacterium acnes, Biofilms, DeoxyribonucleaseⅠ

引用本文

周蒙, 郑娜娜 曾荣 徐浩翔 段志敏 刘宇甄 李岷, . DNaseⅠ对痤疮丙酸杆菌生物膜的抑制效应研究[J]. 中华皮肤科杂志, 2023,56(10):920-924. doi:10.35541/cjd.20230196

Zhou Meng, Zheng Nana, Zeng Rong, Xu Haoxiang, Duan Zhimin, Liu Yuzhen, Li Min, . Inhibitory effect of deoxyribonucleaseⅠ against Cutibacterium acnes biofilms[J]. Chinese Journal of Dermatology, 2023, 56(10): 920-924.doi:10.35541/cjd.20230196