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KMID : 0861020150300050015
Korea Journal of Herbology
2015 Volume.30 No. 5 p.15 ~ p.21
Antibacterial activity of Tonghyeonipal-dan against Methicillin-resistant Staphylococcus aureus
Kim In-Won

Kang Ok-Hwa
Kong Ryong
Kwon Dong-Yeul
Abstract
Objectives: Methicillin-resistant Staphylococcus aureus (MRSA) is a human pathogen. New antibacterial agents are needed to treat MRSA-related infections. This study investigated the antibacterial activity of EtOH 70% extracts of Tonghyeonipal-dan (THD) which prescription is composed of oriental medicine against MRSA.

Methods: The antibacterial activity of THD was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method, Checkerboard dilution test, and Time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors were used.

Results: The minimum inhibitory concentration (MIC) of THD is 1,000~2,000 ¥ìg/mL against MRSA. In the checkerboard dilution test, fractional inhibitory concentration index (FICI) of THD in combination with antibiotics indicated synergy or partial synergism against S. aureus. Furthermore, a time-kill assay showed that the growth of the tasted bacteria was considerably inhibited after 24 h of treatment with the combination of THD with selected antibiotics. For measurement of cell membrane permeability, THD 500 ¥ìg/mL along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (TRIS) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide (NaN3) were used as an inhibitor of ATPase. TX, TRIS, DCCD and NaN3 cooperation against S. aureus showed synergistic action.

Conclusions: Accordingly, antimicrobial activity of THD was affected by cell membrane and inhibitor of ATPase were assessed. These results suggest that THD has antibacterial activity, and that THD extract offers great potential as a natural antibiotic against MRSA.
KEYWORD
Tonghyeonipal-dan, MRSA, Synergy effect, Time-kill assay, Membrane-permeability, ATPase inhibition
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