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현토단(玄兎丹)의 RAW 264.7 대식 세포에서의 항염증 효과에 관한 연구
김마룡 ( Ma-ryong Kim ),강옥화 ( Ok-hua Kang ),공룡 ( Ryong Kong ),서윤수 ( Yun-soo Seo ),주전 ( Tian Zhou ),김상아 ( Sang-a Kim ),김은수 ( Eun-su Kim ),신민아 ( Min-a Sin ),이영섭 ( Young-seob Lee ),권동렬 ( Dong-yeul Kwon ) 대한본초학회 2017 大韓本草學會誌 Vol.32 No.2
Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan (HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. PGE2 were measured using EIA kit. Proinflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. NF-κB/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, TNF-α, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of NF-κB in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of NF-κB delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced PGE2,NO, IL-6 and TNF-α
황형칠 ( Hyeong-chil Hwang ),공룡 ( Ryong Kong ),강옥화 ( Ok-hwa Kang ),권동렬 ( Dong-yeul Kwon ) 대한본초학회 2018 大韓本草學會誌 Vol.33 No.5
Objectives : The aim of this study is to investigate the preventive effect of Puerariae Flos ethanol extract (PE) on methionin and choline deficient (MCD)-diet-induced Nonalcoholic fatty liver disease (NAFLD) in C57BL/6J mice. Methods : In the in vivo experiments, C57BL/6J mice were divided into 4 groups; Normal group, Control group, MCD+PE 100 group, and MCD+PE 300 group. After 4 weeks, body weight, liver weight, biochemical parameters for liver function test, histological changes, reverse transcription polymerase chain reaction (RT-PCR), and Western blot were assessed. Results : Mice lost body weight with the MCD-diet and the MCD+PE 100 group and MCD+PE 300 groups lost less than the control group, though showed no statistical significance. Liver weights were decreased by the MCD diet, but MCD+PE 300 groups were increased significantly. In the liver function test, all the values were decreased with the MCD-diet, MCD+PE 100 group and MCD+PE 300 groups were increased significance. In histological findings of the livers, MCD-diet induced severe fatty accumulation in the livers, but this fatty change was reduced in the MCD+PE 100 group and MCD+PE 300 groups was inhibited respectively. In lipid accumulation factors (such as SREBP-1c, C/EBPα, PPAR-γ), MCD+PE 100 group and MCD+PE 300 groups showed inhibitory effect on liver lipogenesis by reducing associated gene expressions caused by MCD diet. Conclusions : We were able to know that Puerariae Flos ethanol extract (PE) shown hepatoprotective effects via a decrease on the hepatic lipogenesis factors in the experimental NAFLD Models.
대황고(大黃膏)의 메티실린 내성 황색 포도상구균에 대한 항균활성
이순애 ( Sun-ae Lee ),공룡 ( Ryong Kong ),강옥화 ( Ok-hua Kang ),서윤수 ( Yun-soo Seo ),주전 ( Tian Zhou ),김상아 ( Sang-a Kim ),송옥희 ( Ok-hee Song ),김민철 ( Min-chul Kim ),한형선 ( Hyoung-sun Han ),최지나 ( Ji-na Choi ),이영섭 대한본초학회 2017 大韓本草學會誌 Vol.32 No.2
Objectives : Infectious diseases by Methicillin-Resistant Staphylococcus aureus (MRSA) are a growing problem worldwide. Characteristic of MRSA is endlessly mutation to resist antibiotics. Daehwanggo (DHG) is one of the oriental medicine prescriptions contained in Principles and Practice of Eastern Medicine. Daehwanggo was mainly used for external preparation from old times. The purpose of this study is to confirm possibility as supplementary drug of DHG about antibiotics through observation of synergy effect between DHG and commercial antibiotics and to observe restriction on growth of MRSA on any pathway through observation of mechanism. Methods : The minimum inhibitory concentration (MIC) of DHG against MRSA is 500 ~ 2000 ㎍/㎖ by broth dilution method. In the checkerboard method, the combinations of DHG with antibiotics has partial synergistic effect or synergy effect and DHG markedly reduced the MICs of the antibiotics oxacillin (OX), gentamicin (GT) against MRSA. In the inhibition of resistance mechanism of DHG against MRSA, the expression of resistance gene and protein about β-lactam antibiotic was reduced. Also, we observed the effect of DHG about cell membrane permeability against MRSA, and confirmed that DHG suppressed growth of strains by increasing cell membrane permeability. Results : Basis on the result, we speculate that DHG increase antibacterial activity of antibiotics against MRSA by changing the structure of cell wall of MRSA. Conclusions : These data suggest that Daehwanggo possesses possibility as supplementary drug about antibiotics against MRSA.
산사의 NAFLD 보호 효과 및 그 작용기전에 관한 연구
김민철 ( Min-chul Kim ),공룡 ( Ryong Kong ),한형선 ( Hyoung-sun Han ),강담희 ( Dam-hee Kang ),이승진 ( Seung-jin Lee ),이천천 ( Cheon-cheon Lee ),왕서 ( Seo Wang ),권동렬 ( Dong-yeul Kwon ),강옥화 ( Ok-hwa Kang ) 대한본초학회 2018 大韓本草學會誌 Vol.33 No.6
Objectives : Non-alcoholic fatty liver disease (NAFLD) is characterized by the accumulation of hepatic triglycerides (TG) that leads to inflammation and fibrosis. Crataegi Fructus ethanol extract (CE) is a korean traditional herb that used for digestive diseases. It has been investigated that CE has the effect that prevent hepatotoxicity caused by CCl4 or GaIN and regulate the inflammatory in several organs. However, a hypolipidemic effect of CF has not been reported. Methods : The purpose of this study is that examine the lipid accumulation inhibitory effect of CE on NAFLD. We checked the body and liver weight change of MCD-diet induced mice with/without administration of CE. The blood lipid levels of C57BL/6J mice were checked by biochemistry. Also we observed the liver histology of MCD-diet induced mice and investigate the molecular mechanisms in MCD-diet-induced NAFLD in C57BL/6J mice. Results : CE improved MCD-diet-induced lipid accumulation and TG and TC levels. Also, CE decreased hepatic lipogenesis such as SREBP-1, C/EBPα, PPARγ, ACC and FAS. Besides, we also found out that CE increased AMPK phosphorylation. These results indicated that CE has the same ability to activate AMPK and then reduce SREBP-1, and FAS expression, finally leading to inhibit hepatic lipogenesis and hepatic antioxidative ability. Conclusions : In this report, we found CE exerted a regulatory effect on lipid accumulation by decreasing lipogenesis in MCD-diet induced NAFLD model. Therefore, CE extract may be active in the prevention of fatty liver.
황흑산(黃黑散)의 메티실린 내성 황색포도상구균에 대한 항균 활성과 옥사실린과의 시너지 효과
최성훈 ( Sung-hoon Choi ),강옥화 ( Ok-hua Kang ),주전 ( Tian Zhou ),공룡 ( Ryong Kong ),이승진 ( Seung-jin Lee ),강담희 ( Dam-hee Kang ),정혜인 ( Hye-in Jung ),이영섭 ( Young-seob Lee ),권동렬 ( Dong-yeul Kwon ) 대한본초학회 2016 大韓本草學會誌 Vol.31 No.5
Objectives : Hwangheuk-san is a complex prescription composed of oriental traditional medicine and has been reported for antioxidant, antimicrobial and anticancer effects in the recent study. Methicillin-resistant Staphylococcus aureus (MRSA) is one of important causes of fatal infectious diseases such as septicemia, endocarditis, toxic shock syndrome, pneumonia, skin and soft tissue infections (SSTIs). S. aureus is reported as being for a variety of human diseases and its epidemiological relevance is mainly due to their ability of becoming highly resistant to common antimicrobials such as tetracycline, penicillin, cphalosporin and aminoglycoside. The objective of this study is to determine the antimicrobial effect of Hwangheuk-san ethanol extracts (HHS) and synergistic effects with antibiotics oxacillin against MRSA. Methods : The antimicrobial activity of HHS was measured by the disc diffusion method, broth microdilution method and the checkerboard dilution test, time-kill curve assay was performed to investigate synergistic effects with antibiotics oxacillin against MRSA. Results : HHS showed antimicrobial activity against MRSA with a MIC value of 125 ㎍/㎖. In the checkerboard test, the interaction of HHS with antibiotics oxacillin produced almost synergy or partial synergy against MRSA. This study showed that HHS reduced the MICs of oxacillin tested, and a remarkable antibacterial effect of HHS, with membrane permeability enhancers. Conclusions : These results suggest that HHS has the antimicrobial effect and synergistic effects with antibiotics oxacillin against MRSA. This study thus can be a valuable source for the development of a new drug with low MRSA resistance.
통현이팔단 에탄올 추출물의 Methicillin Resistant Staphylococcus aureus에 대한 항균활성
김인원 ( In Won Kim ),강옥화 ( Ok Hwa Kang ),공룡 ( Ryong Kong ),권동렬 ( Dong Yeul Kwon ) 대한본초학회 2015 大韓本草學會誌 Vol.30 No.5
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 μg/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 μg/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.