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Antibacterial Effect of Haedokgeumhwa-san against Methicillin-Resistant Staphylococcus aureus

해독금화산(解毒金花散)의 Methicillin-Resistant Staphylococcus aureus에 대한 항균효과

  • Lee, Ha-Il (Department of Rehabilitation Medicine of Korean Medicine, College of Korean Medicine, Won-Kwang University) ;
  • Lee, Su-Kyung (Department of Rehabilitation Medicine of Korean Medicine, College of Korean Medicine, Won-Kwang University) ;
  • Kwon, Young-Mi (Department of Radiology, College of Korean Medicine, Won-Kwang University) ;
  • Song, Yung-Sun (Department of Rehabilitation Medicine of Korean Medicine, College of Korean Medicine, Won-Kwang University)
  • 이하일 (원광대학교 한의과대학 한방재활의학교실) ;
  • 이수경 (원광대학교 한의과대학 한방재활의학교실) ;
  • 권영미 (원광대학교 한의과대학 영상의학과) ;
  • 송용선 (원광대학교 한의과대학 한방재활의학교실)
  • Received : 2015.03.16
  • Accepted : 2015.03.26
  • Published : 2015.04.30

Abstract

Objectives Methicillin-Resistant Staphylococcus aureus (MRSA) is a human pathogen and a major cause of hospital-acquired infections. New antibacterial agents that have not been compromised by bacterial resistance are needed to treat MRSA-related infections. In this study, we investigated the antimicrobial activity ofethanol extract of Haedokgeumhwa-san (HGH) which prescription is composed of korean medicine against MRSA. Methods The antibacterial activity of HGH extract was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method (minimal inhibitory concentration; MIC), checkerboard dilution test, and time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors. The checkerboard dilution test was used to examined synergistic effect of ampicillin, oxacillin, ciprofloxacin, vancomycin, gentamicin and norfloxacin in combination with HGH ethanol extract. A time-kill assay was performed a survival curve which was obtained by plotting viable colony counts depending on time on bacterial growth. Results The minimum inhibitory concentration (MIC) of ethanol extract (HGH) ranged from 1,000 to $2,000{\mu}g/mL$ against all the tested bacterial strains, respectively. We are able to confirm that HGH extract has potentially strong antibacterial activity. In the checkerboard dilution test, fractional inhibitory concentration index of HGH in combination with antibiotics indicated synergy or partial synergism against S. aureus. A time-kill study showed that the growth of the tested bacteria was considerably inhibited after 8 hr of treatment with the combination of HGH with selected antibiotics. For measurement of cell membrane permeability, HGH $250{\sim}1,000{\mu}g/mL$ 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 ($NaN_3$) was used as an inhibitor of ATPase. TX, Tris, DCCD and $NaN_3$ cooperation against S. aureus showed synergistic action. Accordingly, antimicrobial activity of HGH was affected by cell membrane and inhibitor of ATPase. Conclusions These results suggest that Haedokgeumhwa-san extract has antibacterial activity, and that HGH extract offers a potential as a natural antibiotic against MRSA.

Keywords

Haedokgeumhwa-san;MRSA;Antibacterial effect

Acknowledgement

Supported by : 원광대학교

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