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만응환(萬應丸) 에탄올 추출물의 메티실린 내성 포도상구균에 대한 항균활성 및 내성억제 효과

Antibacterial Activity and Inhibition of Resistance in Methicillin-resistant Staphylococcus aureus by Maneung-hwan Ethanol Extract

한방재활의학과학회지 2020년 30권 1호 p.31 ~ 45
나용수 ( Na Yong-Su ) - 원광대학교 한의과전문대학원 제3의학과

김종규 ( Kim Jong-Gyu ) - 원광대학교 한의과대학 한방재활의학교실
송용선 ( Song Yung-Sun ) - 원광대학교 한의학전문대학원 제3의학과

Abstract


Objectives : In this study, we investigated the antimicrobial activity of a 70% ethanol extract of Maneung-hwan (MEH), which is prescribed by practitioners of oriental medicine for use against methicillin-resistant Staphylococcus aureus (MRSA).

Methods : The antibacterial activity of MEH against MRSA strains was evaluated using the disc diffusion method, broth microdilution method (minimal inhibitory concentration, MIC), checkerboard dilution test, and time-kill test. The mechanism of action of MEH was investigated by bacteriolysis using detergents or ATPase inhibitors Additionally, mRNA and protein expression were investigated by quantitative reverse transcription-polymerase chain reaction and western blot assay, respectively.

Results : The MIC of MEH was 25~1,600 μg/mL against all the tested bacterial strains. We showed that MEH extract exerts strong antibacterial activity. In the checkerboard dilution test, the fractional inhibitory concentration index of MEH in combination with antibiotics indicated synergism or partial synergism against S. aureus. The time-kill study indicated that the growth of the tested bacteria was considerably inhibited after a 24-h treatment with MEH and selected antibiotics. To measure the cell membrane permeability, MEH (3.9 μg/mL) was combined with Triton X-100 (TX) at various concentrations N,N-dicyclohexylcarbodimide (DCCD) was also tested as an ATPase inhibitor. TX and DCCD cooperation against S. aureus exhibited synergistic action. Accordingly, the antimicrobial activity of MEH in the context of cell membrane rupture and ATPase inhibition was assessed. Additionally, the expression of genes and proteins associated with resistance was reduced after exposing MRSA to MEH.

Conclusions : These results suggest that MEH possesses antibacterial activity and acts as a potential natural antibiotic against MRSA.

키워드

Methicillin-resistant Staphylococcus aureus; Maneung-hwan; Antibacterial activity; Resistance inhibition
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