血链球菌cpnp培养上清液抑制铜绿假单胞菌生长和生物膜的作用
Inhibition of growth and biofilm of Pseudomonas aeruginosa by Streptococcus sanguinis cpnp culture supernatant
高毅敏 1李娜 1李新鸣 1金慧 2张家鑫 2耿大升 3孙贺 4刘光焱 2马明月2
作者信息
- 1. 沈阳医学院基础医学院,沈阳 110034;辽宁省环境污染与微生态重点实验室
- 2. 辽宁省环境污染与微生态重点实验室
- 3. 沈阳医学院基础医学院,沈阳 110034;沈阳医学院附属中心医院检验科
- 4. 沈阳医学院基础医学院,沈阳 110034;沈阳医学院附属第二医院检验科
- 折叠
摘要
目的 研究血链球菌cpnp培养上清液抑制铜绿假单胞菌生长和生物膜形成、破坏成熟生物膜的作用,以及其联合抗生素的抑菌作用,为利用呼吸道共生菌制剂干预铜绿假单胞菌呼吸道定植,减少呼吸道感染提供依据.方法 采用牛津杯法测定血链球菌cpnp对铜绿假单胞菌生长的抑制作用.使用96孔板法测定血链球菌cpnp培养上清液联合头孢他啶对铜绿假单胞菌生长的抑制作用.体外建立铜绿假单胞菌生物膜模型,利用结晶紫半定量法、刚果红染色法、RT-qPCR法测定血链球菌cpnp培养上清液对生物膜形成的抑制作用;利用结晶紫半定量法、MTT法测定血链球菌cpnp培养上清液对成熟生物膜的破坏作用;采用光镜和电镜观察生物膜的形态.结果 血链球菌cpnp培养上清液对铜绿假单胞菌生长有抑制作用,与头孢他啶联用时表现出协同作用(P＜0.001).血链球菌cpnp培养上清液可减少铜绿假单胞菌生物膜形成量(P＜0.001)、胞外多糖产量(P＜0.001)及降低胞外多糖基因表达水平(P＜0.050).血链球菌cpnp培养上清液可减少成熟生物膜总量(P＜0.001)和降低生物膜代谢活性(P＜0.001).结论 血链球菌cpnp上清液和其联用抗生素能拮抗铜绿假单胞菌生长,并且上清液对其生物膜有良好的抑制和破坏作用.
Abstract
Objective To explore the role of Streptococcus sanguinis cpnp(hereinafter the cpnp)culture supernatant(CS)in inhibiting the growth and biofilm formation of Pseudomonas aeruginosa(P.aeruginosa)and destroying the ma-ture biofilm,as well as its inhibitory effect in combination with antibiotics,in order to provide a basis for the use of respir-atory commensal bacterial agents to intervene in the colonization of P.aeruginosa in respiratory tract and reduce the respir-atory tract infections.Methods The inhibitory effect of the cpnp on the growth of P.aeruginosa was determined using Oxford cup method.96-well plate method was used to determine the inhibitory effect of the cpnp CS combined with ceftazidime on the growth of P.aeruginosa.The biofilm model of P.aeruginosa was established in vitro,and the inhibit-ory effect of the cpnp CS on the formation of its biofilm was determined with crystal violet semiquantitative method,congo red staining method,and RT-qPCR method;the destructive effect of the cpnp CS on mature biofilm was measured by us-ing crystal violet semiquantitative method and MTT method;and the morphology of the biofilm wasobserved using light microscopy and electron microscopy.Results The cpnp CS inhibited the growth of P.aeruginosa,showed synergistic ef-fect when combined with ceftazidime(P＜0.001),and reduced the amount of P.aeruginosa biofilm formed(P＜0.001),the production of extracellular polysaccharides(P＜0.001),and the level of expression of extracellular polysaccharide genes(P＜0.050).The cpnp CS reduced the total amount of mature biofilm(P＜0.001)and decreased the metabolic activ-ity of biofilm(P＜0.001).Conclusion The cpnp CS and combined antibiotics antagonized the growth of P.aeruginosa,and the supernatant had a good inhibitory and destructive effect on its biofilm.
关键词
血链球菌/铜绿假单胞菌/生物膜/协同作用Key words
Streptococcus sanguinis/Pseudomonas aeruginosa/Biofilm/Synergistic effect引用本文复制引用
基金项目
辽宁省创新能力提升联合基金(2022-NLTS-14-05)
沈阳医学院研究生科技创新基金(Y20220510)
出版年
2024
NETL
NSTL
万方数据