해독금화산 물추출물이 LPS로 유도된 대식세포의 염증반응에 미치는 영향

임재수 ( Jae Soo Lim ),강옥화 ( Ok Hwa Kang ),서윤수 ( Yun Soo Seo ),권동렬 ( Dong Yeul Kwon ) 대한본초학회 2015 大韓本草學會誌 Vol.30 No.5

Objectives: The Haedokgumhwa-san water extract (HDKHS) is used in Korea, Japan and China as a traditional therapeutic agent to cure an infectious disease. But its study is not enough. Therefore, the present study focused on the elucidation of HDKHS to investigate the anti-inflammatory effects and to established the possible mechanisms involved in its action on LPS-stimulated immune response in murine macrophages. Methods: Inflammatory status was induced by LPS and measured by increasement of inflammatory mediators. LPS induced secretions of NO and PGE2 in RAW 264.7 cells were measured using griess reagent and enzyme-linked immunosorbent assay (ELISA) kit respectively. production of IL-6 was examined using ELISA kit and expression of IL-6 mRNA was measured by RT-PCR method. To investigate the effects of HDKHS on inflammatory mediators, such as iNOS, COX-2 and MAPKs, western blot and RT-PCR were performed. Results: HDKHS significantly reduced production of NO and PGE2 which were induced by LPS. Also, activation of IL-6 was reduced both protein and mRNA levels. The expressions of inflammatory mediator include iNOS and COX-2 were decreased by pretreatment with HDKHS. futhermore The result showed HDKHS down-regulate the LPS induced phosphorylation of ERK 1/2, one of the MAPK family, which is considered as a main regulator of transmission from pathogens to nucleus of immune cells. Conclusions: Our results suggest that the anti-inflammatory properties of HDKHS may stem from the inhibition of pro-inflammatory mediators via suppression of initiation of inflammatory response by inhibiting MAPKs signaling pathways.

속단(續斷) 추출물의 C57BL/6 마우스를 이용한 2주 경구투여 독성시험

허혜윤 ( Hye Yun Heo ),신동호 ( Dong-ho Shin ),이지혜 ( Ji Hye Lee ),서윤수 ( Yun-soo Seo ),김용범 ( Yong-bum Kim ),신인식 ( Insik Shin ),강소희 ( Sohi Kang ),손미경 ( Mee-kyung Son ),김중선 ( Joong-sun Kim ) 대한본초학회 2021 大韓本草學會誌 Vol.36 No.5

Objectives : A root of Dipsacus asperoides C. Y. Cheng et T. M. Ai (D. asperoides) has been traditionally used as a medicinal resource in several Asian countries, including Korean and traditional Chinese medicine that has been traditionally used for treating several medical conditions including pain, arthritis, and bone fractures in Korea. In the present study, we investigated potential subacute toxicities of D. asperoides extract. Methods : C57BL/6 mice (male, 7weeks) were randomly divided into 4 groups of 5 mice. Except for the control group, the mice were orally administrated D. asperoides extract at doses of 50, 150, or 450 mg/kg/day for 2 weeks. At the end of the treatment period, all mice were euthanized, and the following parameters were examined: mortality, body weight, clinical signs, gross findings, hematology, serum biochemistry, organ weight, and histopathology. Results : There were no abnormalities in mortality, clinical signs, body weight, gross findings, or organ weight after repeated administration of D. asperoides extract for 2 weeks, compared with the control group. In addition, there were no significant changes in hematological, serum biochemical, and histopathological parameters between the control group and D. asperoides extract administrated groups with doses of up to 450 mg/kg/day. Conclusion : In this study, D. asperoides extract showed no significant toxicities at a dose of up to 450 mg/kg/day in mice. Although we could not confirm the toxic dose of D. asperoides extract, it can be considered safe for further pharmacological use.

Human mast cell에서 升麻葛根湯(승마갈근탕)의 항염증 효과에 대한 연구

금준호 ( Joon Ho Keum ),서윤수 ( Yun Soo Seo ),강옥화 ( Ok Hwa Kang ),최장기 ( Jang Gi Choi ),권동렬 ( Dong Yeul Kwon ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.5

Objectives : Seungmagalgeun-tang (SMGGT) is traditional medicine widely used for inflammatory disease and flu. But SMGGT exhibits potent anti-inflammatory activity with an unknown mechanism. To elucidate the molecular mechanisms of SMGGT water extract on pharmacological and biochemical actions in inflammation, we examined the effect of SMGGT on pro-inflammatory mediators in Phorbol-12-myristate-13-acetate (PMA)+A23187 -stimulated mast cells. Methods : In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to measure the activation of MAPKs. Cells were treated with SMGGT 1 h prior to the addition of 50 nM of PMA and 1 μM of A23187. Cell viability was measured by MTS assay. The investigation focused on whether SMGGT inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8) and mitogen-activated protein kinases (MAPKs) in PMA+A23187 -stimulated mast cells. Results : SMGGT has no cytotoxicity at examined concentration (100, 250, and 500 μg/ml). Also, gene expression of IL-6 and IL-8 in HMC-1 cells stimulated by PMA+A23187 was down regulated by SMGGT. Furthermore, SMGGT suppressed the PMA+A23187-induced phosphorylation of extracellular signal-regulated kinase (ERK) and c-jun N-terminal Kinase(JNK). But, SMGGT could not regulate phosphorylation of p38 MAPK. Conclusions : These results suggest that SMGGT has inhibitory effects on PMA+A23187-induced IL-6 and IL-8 production. These inhibitory effects occur through blockades on the phosphorylation of ERK and JNK.

백강잠 추출물의 C57BL/6 마우스를 이용한 2주경구투여 독성시험

남현화 ( Hyeon-hwa Nam ),서윤수 ( Yun-soo Seo ),문병철 ( Byeong-cheol Moon ),이지혜 ( Ji-hye Lee ),김용범 ( Yong-bum Kim ),김중선 ( Joong-sun Kim ) 대한본초학회 2020 大韓本草學會誌 Vol.35 No.2

Objectives : Bombyx batryticatus L. is the dried larval form of the silkworm (Bombyx mori L.) infected by Beauveria bassiania (Bals.) Vuill. It is used as a food and medicinal resource to treat asthma, headaches, epilepsy, and convulsions in traditional Korean and Chinese medicines. However, the research of the toxicity about B. batryticatus is not enough yet. Here, we investigate the effects of potential subacute toxicity following the repeated oral administration of B. batryticatus water extract to C57BL/6 mice, at various doses of 0, 50, 150, and 450 ㎎/㎏/day during a two-week period. Methods : The following parameters were examined during the study period: body weight, gross findings, clinical signs, organ weight, hematology, serum biochemistry, histopathology, and mortality. At the end of the treatment period, all the mice were euthanized. Results : No changes were observed in the body weights, gross findings, clinical signs, organ weights, and mortality after two weeks of administration of the B. batryticatus extract. In addition, compared with the normal control group, no noticeable treatment-related changes were observed in the hematological, serum biochemical, and histopathological parameters in the treated group following treatment with doses of up to 450 ㎎/㎏/day. Conclusion : Based on these findings, we conclude that the treatment of mice with the water extract of B. batryticatus did not cause considerable C57BL/6 toxicity, and therefore, it could be considered safe for further pharmacological studies.1)

한의학 기반 반려동물용 치료기술 개발 관련 특허 분석

이지혜,김성호,이수은,남현화,박준홍,천진미,서윤수,김보혜,이정민,문창종,김중선,문병철,Lee, Ji Hye,Kim, Sung Ho,Lee, Sueun,Nam, Hyeon-Hwa,Park, Jun Hong,Chun, Jin Mi,Seo, Yun-Soo,Kim, Bohye,Lee, Jeongmin,Moon, Changjong,Kim, Joong Sun,Moon 대한본초학회 2022 大韓本草學會誌 Vol.37 No.6

Objective : In this study, we conducted to analysis the patents about Korean medicine based treatment technology for pet. Methods : We collected the Korean, Japan, USA, China, EU and PCT patent for the Korean medicine-based treatment technology for pet using WIPSON data base registered by October 4, 2020 were selected. Results : Patents for the development of pet Korean medicine-based treatment technology have been constantly applied since the mid-2000s, starting in 1990, with a total of 84 applications. The number of patent applications filed in Korea was the largest, after Japan and China. The detailed composition of the applications reported until now is pharmaceutical compositions for the treatment of pet were the most applied with 39%(33 cases), followed by feed used as stocks for pet at 36%(30 cases). According to the patent application rate by disease, most of applications rate composed to be associated with immunity in 2009 to 2013 with 40% of applications. In 2014 to 2018 43% of patent applications are associated with gastrointestinal/digestive system. In this regard, multiple applications were filed in the order of immunity, digestive system, obesity and urinary system. Conclusion : This study analyzed the trend of 84 patents about Korean medicine-based treatment technology for pet to date to establish the basis for future research and development. There were many applications related to immunity from 2009 to 2013, and since then, related to digestive disease from 2014 to 2018.

뇌전증 동물 모델에 대한 백출 추출물의 보호 효과

강소희 ( Sohi Kang ),이수은 ( Su Eun Lee ),이아영 ( Ayeong Lee ),서윤수 ( Yun-soo Seo ),문창종 ( Changjong Moon ),김성호 ( Sung Ho Kim ),이지혜 ( Jihye Lee ),김중선 ( Joong Sun Kim ) 대한본초학회 2021 大韓本草學會誌 Vol.36 No.6

Objectives : Atractylodis rhizoma Alba has been traditionally used as a medicinal resource that is used for enhancing Qi (氣) in traditional medicine in Korea, China, and Japan. This study investigated the protective effects of Atractylodis rhizoma Alba extract (ARE) against trimethyltin (TMT), a neurotoxin that causes selective hippocampal injury, using both in vitro and in vivo models. Methods : We investigated the effects of ARE on TMT- (5mM) induced cytotoxicity in primary cultures of mouse hippocampal cells (7 days in vitro ) and on hippocampal injury in C57BL/6 mice injected with TMT (2.6 mg/kg). Results : We observed that ARE treatment (0 - 50 μg/mL) significantly reduced TMT-induced cytotoxicity in cultured hippocampal neurons in a dose-dependent manner, based on results of lactate dehydrogenase and 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assays. Additionally, this study showed that orally administered ARE (5 mg/kg; between -6 and 0 days before TMT injection) significantly attenuated seizures in adult mice. Furthermore, quantitative analysis of allograft inflammatory factor-1 (Iba-1)- and glial fibrillary acidic protein (GFAP)- positive cells showed significantly reduced levels of Iba-1- and GFAP-positive cell bodies in the dentate gyrus of mice treated with ARE prior to TMT injection. These findings indicate the significant protective effects of ARE against the TMT-induced massive activation of microglia and astrocytes in the hippocampus. Conclusions : We conclude that ARE minimizes the detrimental effects of TMT-induced hippocampal neurotoxicity, both in vitro and in vivo . Our findings may serve as useful guidelines to support ARE administration as a promising pharmacotherapeutic approach to hippocampal degeneration.

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

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.

Single Dose Oral Toxicity Study of Cicadidae Periostracum Extracts in Sprague-Dawley Rats

전병석 ( Byung-suk Jeon ),정희영 ( Huiyeong Jeong ),이수은 ( Sueun Lee ),서윤수 ( Yun-soo Seo ),김중선 ( Joong-sun Kim ),남현화 ( Hyeon Hwa Nam ),이지혜 ( Ji Hye Lee ) 대한본초학회 2024 大韓本草學會誌 Vol.39 No.3

Objective : Cicadae Periostracum (CP), which is the discarded shell of the Cryptotympana atrata (Fabricius, 1775), is a recognized component of oriental medicine for treatment sore throat, itching, shock, sedation, edema. However, the safety and toxicity of CP have not yet been established. It has been reported that symptoms of addiction or side effects may occur in patients who take high doses of CP or who are hypersensitive to it. Therefore, we investigated the acute toxicity of an CP extracts in Sprague-Dawley (SD) rats. Methods : To study acute toxicity, five SD rats of each sex per group were treated with CP extracts at single doses of 0, 500, 1000, or 2000 mg/kg administrated by oral gavage, and body weight, clinical signs, and mortality were observed after dosing. At the end of 14-day observation period, all animals were sacrificed and complete hematological and macroscopic examinations were performed. Results : There were no dead animal and test article-related effects on body weight change or the gross finding. No toxicologically significant results were observed between control and treated groups in hematology. Although salivation related to stress at the highest dose was observed in clinical signs immediately after administration, it is considered to have no toxicological significance. Conclusion : As the results, we did not find any adverse effect at the dose levels of 500, 1000, or 2000 mg/kg in rats. The minimal lethal dose was considered to be over 2000 mg/kg body weight in rats.

인간 비만세포에서 PMA와 A23187에 의해 유도된 전염증 매개체에 대한 신효월도산 추출물의 항염증 효과

위경 ( Gyeong Wi ),양다운 ( Da Wun Yang ),강옥화 ( Ok Hwa Kang ),김성배 ( Sung Bae Kim ),문수현 ( Su Hyun Mun ),서윤수 ( Yun Soo Seo ),강다혜 ( Da Hye Kang ),임재수 ( Jae Soo Lim ),김마룡 ( Ma Ryong Kim ),곽남원 ( Nam Won Kwak ) 대한본초학회 2014 大韓本草學會誌 Vol.29 No.6

Objectives : Sinhyowoldo-san (SHWDS) is said to be a traditional medicine used for shigellosis, abdominal pain, diarrhea. But mechanism of SHWDS mediated-modulation of immune function is not sufficiently understood. To ascertain the molecular mechanisms of SHWDS 70% EtOH extract on pharmacological and biochemical actions in inflammation, we researched the effect of pro-inflammatory mediators in phorbol-12-myristate-13-acetate (PMA)+ A23187-activated human mast cell line (HMC-1). Methods : In the present research, cell viability was measured by MTS assay. pro-inflammatory cytokine production was measured by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to analyze the activation of mitogen-activated protein kinases (MAPKs), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-кB). The investigation focused on whether SHWDS inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8), MAPKs and NF-кB in PMA+A23187-activated HMC-1 cells. Results : SHWDS has no cytotoxicity at measured concentration (50, 100, and 250 μg/ml). SHWDS (250 μg/ml) inhibits pro-inflammatory cytokine expression in PMA+ A23187-activated HMC-1 cells. Moreover, SHWDS inhibited cyclooxygenase (COX)-2 expression. In activated HMC-1 cells, SHWDS suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2) and c-jun N-terminal Kinase (JNK 1/2). Then, SHWDS suppressed activation of nuclear factor NF-кB in nuclear, degradation of IkB α in cytoplasm. Conclusions : We propose that SHWDS has an anti-inflammatory therapeutic potential, which may result from inhibition of ERK 1/2, JNK 1/2 phosphorylation and NF-кB activation, thereby decreasing the expression of pro-inflammatory genes.