Pseudomonas aeruginosa PAO1’de QS ilişkili genlerin ekspresyon seviyeleri üzerine Lactobacillus sp. metabolitlerinin anti-quorum sensing etkilerinin belirlenmesi

Öz

Amaç: Pseudomonas aeruginosa hastane enfeksiyonları ile ilişkili önemli bir patojen olup patojenitesi çoğunlukla quorum sensing QS sistemi ile ilişkilidir. Bu çalışmanın amacı, vajinal Lactobacillus izolatlarının metabolitlerinin anti-QS aktivitelerinin değerlendirilmesi ve metabolitlerin P. aeruginosa PAO1’in QS ilişkili genlerinin transkripsiyonel regülasyonu üzerindeki etkilerinin araştırılmasıdır.Yöntem: Çalışmamızda daha önce 16S rRNA gen dizi analizi ile tanımlanmış olan 13 adet Lactobacillus izolatı kullanılmıştır. Bu izolatların metabolitlerinin, Chromobacterium violaceum CV12472 suşu kullanılarak anti-QS aktiviteleri değerlendirilmiştir. Metabolitlerin QS ile ilişkili genlerin ekspresyonları üzerindeki etkisi kantitatif revers transkriptaz polimeraz zincir reaksiyonu RT-qPZR ile araştırılmıştır. Bulgular: Test edilen tüm metabolitler C. violaceum’un şeffaf zon bölgesi görünümü ile karakterize edilen anti-QS aktivite göstermişlerdir. Metabolitlerle temas sonrasında P. aeruginosa PAO1’de, test edilen tüm quorum sensing ilişkili genler lasI, lasR, rhlR ve mvfR anlamlı bir down-regülasyon göstermişlerdir. Sonuç: Antibakteriyel direncin artması nedeniyle farklı etki mekanizmasına sahip doğal kaynaklardan köken alan, enfeksiyonlara karşı yeni ajanlar gerekmektedir. Çalışmamız, in vitro Pseudomonas biyofilmlerinin kontrolünde Lactobacillus metabolitlerinin olası anti-QS ajanı olarak kullanılabileceklerini vurgulamaktadır

Anahtar Kelimeler

• anti-quorum sensing aktivitesi, Lactobacillus sp., metabolitler, RT-qPCR, Pseudomonas aeruginosa

Assessment of the anti-quorum sensing effect of Lactobacillus sp. metabolites on expression levels of QS-related genes in Pseudomonas aeruginosa PAO1

Abstract

Objective: Pseudomonas aeruginosa is an important pathogen associated with nosocomial infections and its pathogenicity is mostly linked with the quorum sensing QS system. The aim of this study was to evaluate the anti-QS activity of the metabolites of vaginal Lactobacillus isolates and to investigate the effect of these metabolites on transcriptional regulation of QS related genes in P. aeruginosa PAO1.Methods: In this study, 13 Lactobacillus isolates that were previously identified by 16S rRNA gene sequence analysis were used. Metabolites of these isolates were assessed for the anti-QS activity by using Chromobacterium violaceum CV12472. The influence of metabolites on the expression of QS related genes was also examined by reverse transcription-quantitative polymerase chain reaction RT-qPCR .Results: All tested metabolites exhibited anti-QS activity with the appearance of a non-pigmented zone of C. violaceum. All tested quorum sensing-related genes lasI, lasR, rhlR and mvfR in P. aeruginosa PAO1 showed significant down-regulation after treating with the metabolites Conclusion: New anti-infection agents from natural resources with a different mode of action are necessary due to the increasing occurrence of antibacterial resistance. Our study highlights the possible usage of Lactobacillus metabolites as an anti-QS agent against P. aeruginosa biofilm cells in vitro.

Keywords

• anti-quorum sensing activity, Lactobacillus sp., metabolites, RT-qPCR. Pseudomonas aeruginosa

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