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小檗碱对铜绿假单胞菌密度感应缺陷株及其分泌的致病因子的作用研究 预览 被引量:1

Inhibitory Effect of Berberine on the Growth and Virulence of Clinical Isolates of Quorum Sensing-deficient Pseudomonas aeruginosa
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摘要 目的 观察小檗碱对铜绿假单胞菌(Pseudomonas aeruginosa,PA)密度感应缺陷株的生长及其分泌的致病因子的影响,初步探索小檗碱对铜绿假单胞菌密度感应系统及调控致病因子表达的作用机制。方法 不同浓度小檗碱分别与铜绿假单胞菌野生株PAO1、密度感应系统(quorum sensing system,QS)基因缺陷株PAO JP2、临床分离QS缺陷株共培养,运用倍比稀释法测定最小抑菌浓度,紫外分光光度法测定绿脓菌素分泌量,苔黑酚法测定鼠李糖脂水平,结晶紫法测定生物被膜起始量,荧光定量PCR检测各株QS基因表达。结果 小檗碱对铜绿假单胞菌野生株PAO1 MIC值为12.50mg/mL,临床分离株C84 MIC值为1.56mg/mL,PAO JP2株与C39株MIC值均为3.12mg/mL,C104和C117株MIC值均为6.25mg/mL。在低于MIC浓度下,小檗碱对PAO1株、C104株、C117株绿脓菌素水平有显著抑制,且对各QS缺陷株鼠李糖脂有不同程度抑制作用。此外,PAO JP2株、C39株、C84株生物被膜可被小檗碱显著抑制。各株残留的LasR和rhI转录均受到小檗碱显著抑制。结论 小檗碱对临床分离QS缺陷株有良好的抑制作用,可能通过调节QS基因多种不同细菌毒力因子进行调控。 Objective To investigate the effects of berberine on the growth and virulence of clinical isolates of Pseudomonas aeruginosa(P, aeruginosa)with quorum sensing(QS) deficiency and the underlying mechanism. Methods P. aeruginosa PAO1, QS-deficient PAO JP2 and clinically isolated QS-deficient strains were cultured with berberine at difference concentrations. The minimum inhibitory concentrations (MICs)of berberine against different isolates were determined by double-dilution analy- sis. The secretion of.pyocyanin was measured by ultraviolet spectroscopy,the amount of rhamnolipid by oricinol assay, the biofilm formation by crystal violet staining,and the transcription levels of QS gene by real-time PCR. Results The MICs of berberine against P. aeruginosa were 12.50 mg/mL in the PAO1 strain,1.56 mg/mL in the C84 strain,3.12 mg/mL in both JP2 and C39 strains,and 6.25 mg/mL in both C104 and Cl17 stains. Berberine at sub-MIC could significantly decrease the pyocyanin levels in strains PAO1, C104, and Cl17, and suppress the rhamnolipid levels in QS-deficient stains to varying degrees. Additionally,the biofilm formation was profoundly inhibited by berberine in strains JP2, C39, and C84. The transcrip- tion of remaining LasR and rhI was significantly down-regulated by berberine at sub-MIC. Conclusion Berberine can signifi- cantly inhibit the growth of clinically isolated QS-deficient P. aeruginosa stains and down-regulate the QS-dependent virulence by reducing the transcription levels of QS gene.
作者 夏飞 肖红兵 杜鸣 廖亚玲 张明伟 王平 Xia Fei1 ,Xiao Hongbing2 ,Du Minglet al( 1 Department of Pharmacy, The Third People's Hospital of Hubei Province ,Wuhan 430033 ,China 2Health Center of Caidian Street of Huangpi District,Wuhan 430323, China)
出处 《华中科技大学学报:医学版》 CSCD 北大核心 2017年第2期155-159,174共6页 Journal of Huazhong University of Science and Technology(Health Sciences)
基金 湖北省自然科学基金资助项目(No.2011CDB302)
关键词 铜绿假单胞菌 小檗碱 密度感应缺陷 致病因子 Pseudomonas aeruginosa berberine quorum-sensing deficiency pathogenic factors
作者简介 夏飞,男,1978年生,助理研究员,E-mail:53042940@qq.com 通讯作者,Corresponding author,E-mail:WP0166@139.com
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