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原花青素对铜绿假单胞菌生物膜形成的影响

Effect of proanthocyanidins on biofilm formation by Pseudomonas aeruginosa
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摘要 目的观察原花青素对铜绿假单胞菌群体感应系统和生物膜形成的影响。方法采用微孔板法检测原花青素对铜绿假单胞菌生物膜形成的影响;采用运动平板法检测原花青素对铜绿假单胞菌涌动的抑制作用;通过RNA测序分析原花青素对铜绿假单胞菌基因表达的影响。结果原花青素在不影响铜绿假单胞菌生长的情况下显著促进铜绿假单胞菌生物膜的形成且均呈剂量依赖性;24 h组在PAC浓度为1.95μg/ml时,对PAO1生物膜的形成有促进作用(t=4.903,P<0.01),而12h组在PAC浓度低至0.98μg/ml时便能促进PAO1生物膜的形成(t=4.289,P<0.05);原花青素在浓度100μg/ml时可抑制PA的涌动距离及降低PA涌动复杂性,同时也能促进青脓素分泌。原花青素还可引起铜绿假单胞菌142个基因差异性表达,其中上调差异基因78个,下调差异基因56个;与环境中微生物代谢通路有关的上调基因10个,下调基因8个;Pathway分类可知与群体感应系统相关的有9个上调基因如PqsB、PqsC、PqsD等,6个基因下调;与烯二炔类抗生素生物合成、硝基甲苯降解和二甲苯降解有关通路的基因有1个;GO功能分析显示参与生物过程的基因居多,为284个,其中参与细菌代谢的基因75个;行使分子功能的基因146个,与细胞成分有关的基因100个。结论原花青素可促进铜绿假单胞菌生物膜的形成和青脓素的产生并抑制铜绿假单胞菌的涌动。 Objective To study the effects of proanthocyanidins on the quorum sensing system of and biofilm formation by Pseudomonas aeruginosa.Methods The effect of proanthocyanidins on biofilm formation by P.aeruginosa was determined using the microplate method.Inhibition of the swarming motility of P.aeruginosa by proanthocyanidins was determined using a moving plate.The effect of procyanidins on gene expression by P.aeruginosa was analyzed using RNA sequencing.Results Proanthocyanidins significantly promoted biofilm formation by P.aeruginosa without affecting its growth.After 24 h,the concentration of PACs was 1.95 μg/ml(P<0.01,t=4.903),while after 12 h the concentration of PACs was lower at 0.98 μg/ml(P<0.05,t=4.289),promoting biofilm formation by PAO1.At 100 μg/ml,procyanidins reduced the swarming distance and complexity of PAO1,but they also promoted the production of pyocyanin.In addition,procyanidins induced the differential expression of 142 genes in P.aeruginosa,78 of which were up-regulated and 56 of which were down-regulated.There were 10 up-regulated genes and 8 down-regulated genes related to the microbial metabolic pathways in the environment.Pathway classification revealed that there were 9 up-regulated genes associated with the quorum sensing system,such as PqsB,PqsC,and PqsD,and 6 genes were down-regulated.There was only one gene linked to the pathway of biosynthesis,degradation of nitrotoluene,and degradation of xylene.GO function analysis indicated that most of the 284 genes involved in biological processes included 75 genes involved in bacterial metabolism,146 genes associated with molecular functions,and 100 genes in connection with cell composition.Conclusion Proanthocyanidins promoted biofilm formation and production of pyocyanin by P.aeruginosa,and they also inhibited its swarming motility.
作者 刘艺清 佘鹏飞 王妍乐 谭芳 周林颖 陈丽华 罗振 伍勇 LIU Yi-qing;SHE Peng-fei;WANG Yan-le;TAN Fang;ZHOU Lin-ying;CHEN Li hua;LUO Zhen;WU Yong(Clinical Laboratory The Third Xianya Hospital Affiliated xvith Central South University,Changsha,Hunan,China 110013)
出处 《中国病原生物学杂志》 CSCD 北大核心 2019年第3期264-269,共6页 Journal of Pathogen Biology
关键词 铜绿假单胞菌 群体感应系统 生物膜 青脓素 原花青素 Pseudomonas aeruginosa quorum sensing system biofilm pyocyanin proanthocyanidins
作者简介 刘艺清(1994-),女,贵州人,硕士研究生。研究方向:临床检验诊断学。E-mail:liuyiqing824@163.com;通讯作者:伍勇,E-mail:wuyong_xy@126.com.
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