Synergistic Activity of Berberine with Azithromycin against Pseudomonas Aeruginosa Isolated from Patients with Cystic Fibrosis of Lung In Vitro and In Vivo

Abstract

Background/Aims: Pseudomonas aeruginosa (PA) is one of the major opportunistic pathogens which can cause chronic lung infection of cystic fibrosis (CF). The formation of PA biofilm promotes CF development and restricts the antimicrobial efficacies of current antibiotics. Methods: The antimicrobial effects of azithromycin (AZM) and berberine (BER) alone and in combination were evaluated using microdilution method, checkerboard assay, time-kill test, qRT-PCR analysis and absorption method. The treatments of AZM and/or BER were further evaluated in an animal lung infection model via observing survival rate, bacterial burden and histopathology of lung, the levels of pro-/anti-inflammatory cytokines. Results: AZM-BER were demonstrated to be synergistic against ten clinical PA isolates as well as the standard reference PA ATCC27853, in which PA03 was the most susceptible isolate to AZM-BER with FICI of 0.13 and chosen for subsequent experiments. The synergism of AZM-BER was further confirmed against PA03 in time-kill test and scanning electron microscope (SEM) at their concentrations showing synergism. In PA03, we found that AZM-BER could significantly attenuate productions of a series of virulence factors including alginate, LasA protease, LasB protease, pyoverdin, pyocyanin, chitinase as well as extracellular DNA, and remarkably inhibit the levels of quorum sensing (QS) molecules and the expressions of lasI, lasR, rhlI, rhlR at 1/2×MIC, 1×MIC and 2×MIC. In the infection model, the mice survival were increased markedly, the inflammations of infected lungs were improved greatly along with reduced IL-6, IL-8 and ascended IL-10 at 0.8 mg/kg of AZM combined with 3.2 mg/kg of BER. Conclusion: BER might be a promising synergist to enhance the antimicrobial activity of AZM in vitro and in vivo.

The Interaction of N-Acetylcysteine and Serum Transferrin Promotes Bacterial Biofilm Formation

“…In hospitals, biofilms occur on implanted medical devices such as urinary or intravenous catheters, prosthetic cardiac valves, orthopedic implants, and contact lenses [1]. They can also be found on soft tissues, with notable examples including the Pseudomonas aeruginosa biofilms in cystic fibrosis (CF) patients and the bacterial biofilms that develop on chronic wound surfaces such as diabetic foot ulcers and burns [2-5]. Because biofilms can effectively shield pathogens from host immune responses and antimicrobial treatment, and may lead to refractory infections or biofilm-associated diseases, they present a great health risk [6, 7].…”

Metabolomic Analysis by Nuclear Magnetic Resonance Spectroscopy as a New Approach to Understanding Inflammation and Monitoring of Pharmacological Therapy in Children and Young Adults With Cystic Fibrosis

“…However, it is unknown whether these clinical effects of azithromycin were due to its potential anti-inflammatory effects as inflammatory markers were not measured in this study. Antibiofilm activities against P. aeruginosa at therapeutic concentrations ( Li et al, 2017 ) and effects on bacterial communication (quorum sensing) ( Skindersoe et al, 2008 ) might explain, at least partially, the clinical efficacy of azithromycin reported in this study ( Saiman et al, 2003 ). However, the antibacterial effect of azithromycin is unlikely to explain pulmonary function improvement as P. aerugino sa is considered naturally resistant to macrolides ( Girard et al, 1987 ) and no differences in the airway colonization by P. aeruginosa were observed during the treatment with azithromycin for more than 12 months in a retrospective study in CF patients ( Samson et al, 2016 ).…”

<p>Quorum Quenching: A Potential Target for Antipseudomonal Therapy</p>

“…Moreover, the above-mentioned compounds, alone and in combination, can considerably decrease motility phenotypes and biofilm production of P. aeruginosa PAO1. 132 Li et al 133 also found that azithromycin and berberine could significantly reduce the production of several pathogenic factors such as biofilm production, as well as secretion of pyocyanin and elastase, and remarkably inhibition of the QS system and the expressions of the genes regulated by QS. In their study, it was also demonstrated that LasA activity was drastically decreased after the administration of azithromycin and berberine, separately and in combination.…”

“…LasR and RhlR receptors [143] Ajoene, a Sulfur-Rich Molecule from Garlic Pathogenic factors production [144] Berberine Production of biofilm, secretion of protease, pyoverdin, pyocyanin and the expressions of lasI, lasR, rhlI, rhlR genes [133] Trans-cinnamaldehyde Secretion of protease, elastase, pyocyanin and production of biofilm and the expressions of lasI, lasR, rhlI, rhlR genes [145] Salicylic acid Secretion of protease, elastase, pyocyanin and production of biofilm and the expressions of lasI, lasR, rhlI, rhlR genes [145] Phillyrin Pyocyanin, rhamnolipid, elastase, swimming and twitching motility and biofilm formation [146] Trans-anethole Swarming motility, secretion of protease, elastase, pyocyanin and the expressions of lasB gene [147] Cinnamic acid Secretion of pyocyanin, proteases, elastase and production biofilm [148] Hordenine Swarming motility, secretion of pyocyanin, elastase, rhamnolipid, production of biofilm and the expressions of lasI, lasR, rhlI, rhlR genes [149] Eugenol from clove extract Pathogenic factors and production of biofilm [150] Ocimum sanctum Secretion of pyocyanin, protease, elastase and production of biofilm [151] Musa paradisiaca Pyocyanin, protease, elastase and biofilm formation [151] Caffeine Motility phenotypes [151] Methanolic extract of Phyllanthus amarus Motility phenotypes, pyocyanin secretion [152] Zingerone Motility phenotypes, production of biofilm and pathogenic factors production [153] Combretum albiflorum Elastase secretion and production of biofilm [154] Allium sativum (garlic) extract Production of biofilm, elastase secretion [155,156] FL fraction of Psidium guajava L Virulence factors production and production of biofilm [157] Clove oil Protease, chitinase and pyocyanin secretion, swimming motility and production of biofilm [158] The methanol extract of fenugreek Protease, LasB elastase, pyocyanin and chitinase production, swarming motility and production of biofilm [159] (Continued) a wide spectrum, so it can be considered as an alternative for the treatment of P. aeruginosa infections. 105 Other attractive compounds are Engineered nanoparticles (ENPs), which are used as QSIs.…”