title: Membrane-damaging potential of natural L-(-)-usnic acid in Staphylococcus aureus.
abstract: The purpose of this investigation was to try to understand the antibacterial mechanism of L-(-)-usnic acid isolated for the first time from fruticose lichen Usnea subfloridana using clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentration (MIC) of L-(-)-usnic acid against the clinical isolates of MRSA and reference strain S. aureus MTCC-96 (SA-96) was in the range 25-50 μg/ml. Treatment of both reference and clinical strains (MRSA-ST 2071) with four-fold MIC concentrations (100-200 μg/ml) of L-(-)-usnic acid reduced the viability of cells without damaging the cell wall. However, the loss of 260 nm absorbing material and increase in propidium iodide uptake was observed in both of the strains. Similarly, a combined effect of L-(-)-usnic acid (25-50 μg/ml) and 7.5 % NaCl resulted in a reduced number of viable cells within 24 h in comparison to the control. These observations clearly indicate that L-(-)-usnic acid exerts its action by disruption of the bacterial membrane. Further, in vivo efficacy showed that L-(-)-usnic acid significantly (p < 0.001) lowered the microbial load of spleen at doses ranging from 1 to 5 mg/kg. Further, toxicity studies in infected mice at doses 20 times higher than the efficacious dose indicated L-(-)usnic acid to be safe. Paradoxically, L-(-)usnic acid exhibited changes in serum triglycerides, alkaline phosphatase (ALKP) and liver organ weight in the healthy mice administered with only 25 mg/kg body weight. The results obtained in this study showed that natural L-(-)-usnic acid exerts its antibacterial activity against MRSA by disruption of the cell membrane. Further, the natural L-(-)-usnic acid was found to be safe up to 100 mg/kg body weight, thereby, making it a probable candidate for treating S. aureus infections.
authors.affiliation: CSIR-Central Institute of Medicinal and Aromatic Plants, P.O. CIMAP, Lucknow, 226 015, India.
authors.forename: V K
authors.initials: VK
authors.lastname: Gupta
authors.forename: S
authors.initials: S
authors.lastname: Verma
authors.forename: S
authors.initials: S
authors.lastname: Gupta
authors.forename: A
authors.initials: A
authors.lastname: Singh
authors.forename: A
authors.initials: A
authors.lastname: Pal
authors.forename: S K
authors.initials: SK
authors.lastname: Srivastava
authors.forename: P K
authors.initials: PK
authors.lastname: Srivastava
authors.forename: S C
authors.initials: SC
authors.lastname: Singh
authors.forename: M P
authors.initials: MP
authors.lastname: Darokar
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000009:adverse effects / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D001572:Benzofurans / Q000009:adverse effects / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D002462:Cell Membrane / Q000187:drug effects*; D015169:Colony Count, Microbial; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects* / Q000302:isolation & purification; D051379:Mice; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects; D013154:Spleen / Q000382:microbiology; D013203:Staphylococcal Infections / Q000188:drug therapy / Q000382:microbiology; D020949:Usnea / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D001572:Benzofurans; C073339:usnic acid