Quercetin inhibits Pseudomonas aeruginosa biofilm formation via the vfr-mediated lasIR system

Pseudomonas aeruginosa is one of the most common opportunistic pathogens that cause biofilm-associated infections. Biofilm formation is partially regulated by the quorum sensing (QS) system, and quercetin can inhibit QS, biofilm formation and virulence factors. We therefore speculated that quercetin would inhibit the formation of P. aeruginosa biofilm via the QS system. In this study, we successfully constructed lasI, rhlI and lasI/rhlI geneknockout strains. The knockout of the lasI and lasI/rhlI genes resulted in decreases in adhesion, biofilm formation, swarming motility and the expression of biofilm-associated genes, whereas deletion of the rhlI gene had no obvious influence on these biofilm-related indicators with the exception of the swarming motility. After treatment with quercetin, the lasI- and lasI/rhlI-mutant strains exhibited increased adhesion, biofilm formation, swarming motility and biofilm-associated gene expression compared with the control group. However, quercetin still exerted an inhibitory effect on these physiological factors and the biofilm-associated gene expression in the rhlI-mutant strain. The knockout of QS genes reduced the production of pyocyanin and protease activity, but after the virulence factors of the QS-mutant strains treated with quercetin showed almost no differences compared with those of the control group. In addition, quercetin could significantly inhibit vfr gene expression regardless of the presence of QS genes. The results indicated that quercetin might inhibit the lasIR system through the vfr gene and ultimately the formation of P. aeruginosa biofilms.