title: Ellagic acid derivatives from Terminalia chebula Retz. downregulate the expression of quorum sensing genes to attenuate Pseudomonas aeruginosa PAO1 virulence.
abstract: BACKGROUND
  Burgeoning antibiotic resistance in Pseudomonas aeruginosa has necessitated the development of anti pathogenic agents that can quench acylhomoserine lactone (AHL) mediated QS with least risk of resistance. This study explores the anti quorum sensing potential of T. chebula Retz. and identification of probable compounds(s) showing anti QS activity and the mechanism of attenuation of P. aeruginosa PAO1 virulence factors.
  
  
  RESULTS
  Methanol extract of T. chebula Retz. fruit showed anti QS activity using Agrobacterium tumefaciens A136. Bioactive fraction (F7), obtained by fractionation of methanol extract using Sephadex LH20, showed significant reduction (p<0.001) in QS regulated production of extracellular virulence factors in P. aeruginosa PAO1. Biofilm formation and alginate were significantly (p<0.05) reduced with enhanced (20%) susceptibility to tobramycin. Real Time PCR of F7 treated P. aeruginosa showed down regulation of autoinducer synthase (lasI and rhlI) and their cognate receptor (lasR and rhlR) genes by 89, 90, 90 and 93%, respectively. Electrospray Ionization Mass Spectrometry also showed 90 and 64% reduction in the production of 3-oxo-C(12)HSL and C(4)HSL after treatment. Decrease in AHLs as one of the mechanisms of quorum quenching by F7 was supported by the reversal of inhibited swarming motility in F7-treated P. aeruginosa PAO1 on addition of C(4)HSL. F7 also showed antagonistic activity against 3-oxo-C(12)HSL-dependent QS in E. coli bioreporter. C. elegans fed on F7-treated P. aeruginosa showed enhanced survival with LT50 increasing from 24 to 72 h. LC-ESI-MS of F7 revealed the presence of ellagic acid derivatives responsible for anti QS activity in T. chebula extract.
  
  
  CONCLUSIONS
  This is the first report on anti QS activity of T. chebula fruit linked to EADs which down regulate the expression of lasIR and rhlIR genes with concomitant decrease in AHLs in P. aeruginosa PAO1 causing attenuation of its virulence factors and enhanced sensitivity of its biofilm towards tobramycin.
authors: Department of Microbiology, Panjab University, Chandigarh, India.
authors: Sajal
authors: S
authors: Sarabhai
authors: Prince
authors: P
authors: Sharma
authors: Neena
authors: N
authors: Capalash
mesh_terms: D000818:Animals; D018441:Biofilms / Q000187:drug effects / Q000254:growth & development; D017173:Caenorhabditis elegans / Q000382:microbiology; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology*; D004610:Ellagic Acid / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000494:pharmacology; D005798:Genes, Bacterial / Q000187:drug effects*; D008025:Ligases / Q000235:genetics; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics / Q000472:pathogenicity*; D053038:Quorum Sensing / Q000187:drug effects / Q000235:genetics*; D029766:Terminalia / Q000737:chemistry / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics; D014774:Virulence / Q000187:drug effects / Q000235:genetics*
chemical_list: D010936:Plant Extracts; D014157:Transcription Factors; D004610:Ellagic Acid; D008025:Ligases; C096955:RHLI protein, Pseudomonas aeruginosa