당귀 물추출물의 면역증강 효과

조성기,이성태,유영법,박혜란 경산대학교 생명자원개발연구소 1999 생명자원과 산업 Vol.3 No.-

천연물로부터 생체의 방어력을 증강시킬 수 있는 면역조절물질을 발굴하기 위한 목적의 일환으로, 한방에서 보혈에 관련하여 널리 사용되고 있는 당귀가 면역계에 미치는 영향을 검토하였다. 당귀추출물은 실험관 내에서 lipopoly-saccharides(LPS), phytohemagglutinin(PHA)와 같은 수준의 lymphcoyte 증식 효과를 보였으며, 그 효과는 LPS보다 오랫동안 지속되었다. LPS가 강력한 B cell mitogen이지만 생체내 독성이 심하여 투여가 어려운 반면, 당귀추출물에 의한 생체내 독성은 거의 없으므로 앞으로 생체내 투여 가능성이 있을 것으로 사료된다. Mouse에 투여하였을 때에도 당귀추출물은 SRBC에 대한 항체를 생성하는 lymphocytes의 수를 증가시켰으며, GVH 반응도 증가시켜 주었다. 따라서 당귀의 면역계에 대한 효과는 B cell의 증식과 그에 따른 항체 생성의 증가 뿐만아니라, T cell 매개 면역반응의 증가에 의한 것으로 생각되며, 앞으로 인체의 면역계에 미치는 효과 및 그 기작에 관한 많은 연구를 통하여 당귀가 면역증강제로 개발될 가능성이 있을 것으로 사료된다. The effects of Angelica gigas Nakai, which has been used as a blood-building decoction for recovery from weakness in the Chinese medicine, were examined for the purpose of screening a new nontoxic immunomodulator. The water extract of Angelica gigas Nakai showed a strong mitogenic effect on splenocytes in vitro to the same level of lipopolysaccharide(LPS) and phytohemagglutinin(PHA). Splenocytes stimulated with the extract in vitro, survived longer than that with lipopolysaccharide(LPS). LPS is a powerful B cell stimulant. However, it is now entirely restricted to only experimental purposes because of the highly toxic nature. Compared with this, the extract of Angelica gigas Nakai did not show any acute toxicity in mice injected intraperitoneally at a dosage of 1g/kg-body weight. It was tested in a hemolytic plaque assay whether in vivo treatment of the extract of Angelica gigas Nakai could influence the secretion of antibodies against sheep red blood cells(SRBC). It turned out that intraperitoneal(i.p.) injection of the extract of Angelica gigas Nakai for 4 days increased the number of antibody-secreting cells remarkably. Splenomegaly, due to graft versus host reaction, was also increased after 7 days i.p. treatment of the extract of Angelica gigas Nakai. These results indicated that the extract of Angelica gigas Nakai might be a useful immunomodulator, especially since it is a relatively nontoxic natural product.

참당귀의 흰쥐 4-Vessel Occlusion 전뇌허혈에 대한 신경방어효과

김영옥,하니나,부영민,박선영,박주영,유영법,신준식,안덕균,김호철 대한본초학회 2002 大韓本草學會誌 Vol.17 No.2

Objectives : Angelicae Radix, the root of Angelica gigas N_AKAI (Umbelliferae) , which has been used to nourish the blood(補血), regulate menstruation(調經), relieve pain and promote bowel movements(崩漏) and activate blood circulation for the treatment of fatigue, headache, constipation and blood troubles in traditional Korean medicine. It's flavour and channel tropism is sweet(甘), pungent(辛), warm(溫), acting on the lung, heart and spleen(肺, 心, 脾). The purpose of study we report here was to determine the neuroprotective effect of Angelica gigas on global ischemia induced by 4-vessel occlusion in Wistar rats. Methods : Angelica gigas extracts was lyphilized after extraction with 85% methanol. We induced 4-VO for 10 minutes and reperfused again. The number of CA1 pyramidal neurons were counted after 7 days of reperfusion under the cresyl violet staining. Results : The results showed that Angelica gigas had significantly neuroprotective effects (100, 250 and 500mg/kg of AR extracts, p<0.001) compared with control group. Each neuroprotective ratio was about 66.1, 79.3 and 78.6% respectively. Immunohistochemical data partially explained the protective effects of Angelica gigas via attenuation of COX-2 induction in hippocampus. Conclusion : Angelica gigas gas neuroprotective effects on the global ischemia induced by 4-vessel occlusion in Wistar rats and the attenuation of COX-2 induction is thought to be its possible mechanism.

Phenolic Compounds from the Rachis of Cedrela sinensis

Yu, Young-Beob,Lee, Jong-Ho,Choi, Jae-Sue,Ok, Kwang-Dae,Park, Jong-Cheol The Korean Society of Pharmacognosy 1996 생약학회지 Vol.27 No.3

From the rachis of Cedrela sinensis A. Juss., methyl gallate, quercitrin, bis-(p-hydroxyphenyl)ether, adenosine, isoquercitrin, rutin, (+)-catechin and (-)-epicatechin were isolated and characterized by spectral data.

The Effective Preparation of Flavonoids from Scutellaria baicalensis G_EORGI by Diaion HP-20 Resin

Yu, Young-Beob The Plant Resources Society of Korea 2014 한국자원식물학회지 Vol.27 No.6

Scutellaria baicalensis $G_{EORGI}$ (Scutellariae Radix) has been used to clear heat and to dry dampness in the stomach or intestines, which manifests as diarrhea or dysenteric disorder. In this study, we investigated the effective preparation of active components in Scutellariae Radix using the methods of solvent extraction and absorption fractionation for the development of new functional food or pharmaceuticals. The marker substances, baicalin, baicalein, wogonoside, and wogonin were directly isolated from the Scutellariae Radix. There chemical structures were elucidated by spectroscopic analysis. The Scutellariae Radix was extracted with hot water. To enhance yield of flavonoids in Scutellariae Radix, the hot water extract was dissolved in ethanol with concentration dependent manner. The precipitates were separated using centrifugal techniques at 10,000 rpm. Supernatant liquid was applied to the HPLC for quantification of major compounds. Separately, the hot water extract was absorbed on Diaion HP-20 resin. And then, the absorbed fraction was eluted with methanol for HPLC. The contents of baicalin, baicalein, wogonoside and wogonin in different treatment methods were analyzed by HPLC. Total amount of four major components were 16.9% in 50% ethanol extract, 21.7% in 70% ethanol extract, 20.5% in 90% ethanol extract, and 39.3% in absorbed fraction of Diaion HP-20 resin. In these results, we found that resin absorption method is suitable for the extraction of enriched flavonoids from Scutellariae Radix.

Human Immunodeficiency Virus Type Ⅰ에 대한 음나무 추출물의 억제활성

Yu Young Beob,Shim Bum Sang,Ahn Kyoo Seok,Choi Seung Hoon,Park Jong Cheol,Miyashiro H.,Hattori M. 대한동의생리학회 2004 동의생리병리학회지 Vol.18 No.4

For the purpose of developing new anti-HIV agents from natural sources, the extracts of Kalopanax pictus were tested for their inhibitory effects on HIV-1 replication and its essential enzymes as the reverse transcriptase (RT). protease and α-glucosidase. In the assay of HIV-1-infected human T-cell line, water extracts of stem and leafstalk inhibited the HIV-1-induced cytopathic effects with Ie (inhibitory concentration) of 25 and 50㎍/㎖, respectively. Moreover water extracts (100㎍/㎖) of stem and leafstalk showed strong activity of 80% and 90% on anti-HIV-1 RT using Enzyme Linked Oligonucleotide Sorbent Assay (ELOSA) method. In the HIV-1 protease inhibition assay, aqueous stem extract inhibited the activity of the enzyme to cleave an oligopeptide, resembling one of the cleavage sites in the viral polyprotein which can only be processed by HIV-1 protease with 58%, but no glucosidase inhibitory activities. We found out this result, for these samples it is possible that the inhibition of the viral replication in vitro is due to the inhibition at least one of RT and protease. It would be of great interest to identify the compounds which are responsible for this inhibition, since all therapeutically useful agent up to date are RT, PR and α-glucosidase inhibitors.

Effects of Triterpenoids and Flavonoids Isolated from Alnus firma on HIV-1 Viral Enzymes

Yu, Young-Beob,Miyashiro, Hirotsugu,Nakamura, Norio,Hattori, Masao,Park, Jong-Cheol 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.7

Triterpenoids and flavonoids isolated from Alnus firma S. Z. were found to inhibit HIV-1 virus replication and controlled its essential enzymes. In this study, the inhibition of HIV-1 viral replication and its essential enzymes, such as reverse transcriptase, protease and ${\alpha}-glucosidase$, were observed using 18 Korean plant extracts. Among the extracts, the methanol extract of Alnus firma leaves showed potent inhibition against the HIV-1 induced cytopathic effect (CPE) in MT-4 cells on microscopic observation (the minimum concentration for complete inhibition of HIV-1 induced CPE, IC=50 ${\mu}g/mL$). Thus, 14 compounds were isolated and identified from the methanol extract of Alnus firma leaves. Of these compounds, the alnustic acid methyl ester exhibited inhibition against HIV-1 pretense, with an $IC_{50}$ of 15.8 ${\mu}M$, and quercetin, quercitrin and myricetin $3-O-{\beta}-D-galactopyranoside$ displayed inhibition against H IV-1 reverse transcriptase, all with $IC_{50}$ values of 60 ${\mu}M$. Based on these results, the viral replication inhibition of the methanol extract of Alnus firma leaves was adjudged to be acutely related to the protease inhibition activation of the alnustic acid methyl ester as well as the reverse transcriptase inhibition activation of flavonoids.

Analysis of Qing-xin Niu-huang Wan by HPLC-ESI-MS and Its Neuro-protective Effect

Young Beob Yu,Yoo Sik Yoon,Bum Sang Shim,Sung Hoon Kim,Seung Hoon Choi,Kyoo Seok Ahn 한의병리학회 2005 대한동의병리학회 학술대회논문집 Vol.2005 No.8

다래나무 추출물의 HIV-1 효소억제활성과 구조활성상관(QSAR)예측

유영법 ( Young Beob Yu ) 대한본초학회 2006 大韓本草學會誌 Vol.21 No.4

일본에서의 본초자원 재배 관리현황 : 사물탕(당귀, 천궁, 작약, 지황)구성약재를 중심으로

유영법 ( Young Beob Yu ),신현규 ( Hyun Kyu Shin ) 대한본초학회 2006 大韓本草學會誌 Vol.21 No.3

감마선 조사 당귀(Angelica gigas Nakai)의 유효성분 안정성 및 유전독성학적 안전성 연구

유영법(Young-Beob Yu),조성기(Sung-Kee Jo) 한국식품영양과학회 2000 한국식품영양과학회지 Vol.29 No.2

건조 생약재 당귀의 감마선 조사 위생화의 가능성을 검토하기 위하여 감마선 조사후 유효성분 안정성 및 유전독성학적 안전성을 평가하고자 하였다. 감마선 조사 당귀의 유효성분 안정성을 검증하기 위해 당귀의 유효성분인 decursin과 decursinol angelate를 분리, 정제하고 이를 표준품으로 하여 HPLC법으로 정량분석을 실시하였다. 구조 이성체인 decursin과 decursinol angelate는 normal phase column에서 좋은 분리능을 보여주었으며, 역상 column에서는 column의 길이와 내경이 분리능에 영향을 미침을 확인할 수 있었다. 감마선 조사(10 kGy) 당귀와 비조사 당귀(0 kGy)의 유효성분 정량결과, 조사 당귀에서 decursin의 함량이 0.27% 정도 증가하였으나 유의성은 관찰되지 않았으며, decursinol angelate는 조사 및 비조사당귀의 비교 결과 함량변화가 인정되지 않았다. Chromatogram의 pattern에서도 감마선 조사에 따른 변화가 관찰되지 않았다. 이는 방사선 조사 건조 생약재의 유효성분의 안정성을 확보하는데 중요한 결과로 생각된다. 또한 10 kGy 감마선 조사 당귀의 열수 추출물은 Salmonella thyphimurium 균주를 이용한 복귀돌연변이 실험에서 복귀돌연변이 수를 증가시키지 않았으며, CHO 세포를 이용한 소핵유발성 검증에서도 소핵빈도를 증가시키지 않았다. 이로써 감마선 조사된 당귀의 추출물이 돌연변이원으로 작용하지 않으며, 세포 핵분열중에 독성을 나타내지 않음을 알 수 있었다. 이상의 결과로 보아 생약재 당귀를 감마선으로 조사한 후에도 당귀내 유효성분이 그대로 보전되는 것으로 사료되며, 감마선 조사 당귀 추출물의 독성학적 안전성은 앞으로 생체내 유전독성시험, 만성독성 및 생식독성시험 등을 통하여 확인될 수 있을 것으로 사료된다. In the present studies, we assessed the stability of active components and toxicological safety of irradiated Angelica gigas Nakai (Danggui). In order to confirm the stability of active components in the γ-irradiated roots of Danggui, the quantitative analysis of decursin and decursinol angelate of γ-irradiated sample was carried out by high performance liquid chromatographic (HPLC) methods using reverse phase columns and normal phase columns. From the root of Danggui, decursin and decursinol angelate were isolated by a silica gel column chromatography (toluene : ether (1 : 1), Hexane : EtOAc (15 : 1)). And then the struc- tures were confirmed in the ¹H and ¹³C-NMR analysis. The HPLC chromatograms of decursin and decursinol angelate in γ-irradiated Danggui were similar with those of non-irradiated sample. In the examination of in vitro genotoxicity of the water extract from γ-irradiated Danggui using Salmonella reversion assay (Ames test) and micronucleus test in Chinese hamster ovary (CHO) cells, mutagenicity was not exhibited in the two assays with or without metabolic activation. These results suggest that active components in the γ-irradiated Danggui should be stable and that the safety of γ-irradiated Danggui could be revealed in further tests in vivo.