• 한국미생물·생명공학회지
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• 제26권2호
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• pp.130-136
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• 1998

참당귀 세포(Angelica gigas H4)를 현탁배양하여 면역 증강성 다당을 생산하기 위한 최적배지 설계 및 생산조건 최적화 연구를 수행하였다. 우선 세포의 성장에 가장 좋은 배지로서 Schenk and Hildebrandt(SH) 배지를 선택하여 배양한 결과 $25^{\circ}C$, 암배양조건에서 각각 15.8g DCW/l와 0.85 g/l의 세포 및 다당을 얻을 수 있었다. 한편 다당 생산용 배지의 개발을 위하여 여러 가지 식물세포배양용 배지 중에서 다당 생산능이 가장 높은 배지로서 Gamborg B5 배지를 선정하였으며 B5 배지를 변형하여 면역증강성 다당생산을 위한 최적배지(PPM)를 개발하였다. 그리고 최적배지를 이용한 당귀 세포증식과 다당 생산을 위한 최적 pH는 6.0-6.5, 배양온도는 $20^{\circ}C$, 그리 고 광조건보다는 암조건이 좋은 것으로 밝혀졌다. 상기 최적조건에서의 최대 세포증식과 다당생산은 각각 14.8 g/l, 1.5 g/l 이었다.

• BMB Reports
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• 제40권5호
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• pp.636-643
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• 2007

This study was designed to identify and characterize the mechanism of macrophage activation by AAP, an acidic polysaccharide fraction isolated from the roots of Angelica sinensis (Oliv.) Diels. As a result, AAP significantly enhanced nitric oxide (NO) production and cellular lysosomal enzyme activity in murine peritoneal macrophages in vitro and in vivo. Furthermore, L-NAME, a specific inhibitor of inducible nitric oxide synthase (iNOS), effectively suppressed AAP-induced NO generation in macrophages, indicating that AAP stimulated macrophages to produce NO through the induction of iNOS gene expression and the result was further confirmed by the experiment of the increase of AAP-induced iNOS transcription in a dose-dependent manner. To further investigate, AAP was shown to strongly augment toll-like receptor 4 (TLR4) mRNA expression and the pretreatment of macrophages with anti-TLR4 antibody significantly blocked AAP-induced NO release and the increase of iNOS activity, and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) secretion.

• KSBB Journal
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• 제21권5호
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• pp.331-335
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• 2006

참당귀(Angelica gigas Nakai) 현탁세포의 면역 증강성 다당의 생산을 증진시키기 위하여 초기 당농도와 생장조절제인 2,4-D의 농도를 변화시켰다. 당의 농도를 높일수록 세포의 생장과 ECP의 생산은 각각 1.6배, 1.1배 증가하여 효과적이었다. 2,4-D는 첨가하지 않은 것이 세포의 생장이 대조군에 비해 1.2배 높았지만, ECP의 생산의 경우 2mg/L 2,4-D를 첨가한 경우 대조군에 비해 10% 이상 더 생산되었다. 또한 배양 중 여러 가지의 elicitor를 첨가함으로써 ECP의 생산을 증진시키기 위하여 fungal elicitor들을 처리한 결과 Verticillium dahliae 90 mg/L을 배양 4일째에 처리한 경우 세포의 생장을 다소 저해시킨 반면, ECP 생산량이 1.7배나 증가함을 확인하였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.158-161
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• 2003

Root cultures of Angelica gigas Nakai were found to be sensitive to elicitation by poly-saccharide elicitors, such as methyl-$\beta$-cyclodextrin, glucan, carboxymethyl-$\beta$-chitin, chitosan, yeast extract and pectin. For the production of decursinol angelate, ca rboxymethyl-$\beta$-chitin and glucan were found to be the most efficient elicitors. The e nhanced accumulation of decursinol angelate was proportional to the increase of the phenylalanine ammonialyase (PAL) activity after the treatment with most of the elicitors. However, carboxymethyl-$\beta$-chitin treatment did not stimulate the PAL activity, despite the 1.6-fold increase in the decursinol angelate production.

• KSBB Journal
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• 제21권5호
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• pp.336-340
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• 2006

본 연구에서는 참당귀 현탁세포배양에 의한 면역 증강성 ECP의 생산을 증진시키기 위하여 배지교환식 배양을 수행하였으며, ECP의 분비를 촉진하는 초음파 처리와 Pluronic F-68의 영향을 조사하였다. 참당귀의 현탁세포배양시, 최대 세포생장은 6일째에 16.8 g DCW/L였고, ECP의 생산은 세포생장과 함께 증가하다가 8일째에 최고 0.9 g/L가 생산되었다. 식물세포 고농도배양법인 배지교환식 배양을 적용해, 초기 당 농토를 높이고 접종 후 5일부터 연속적으로 배지교환을 해주어 23.8 gDCW/L의 고농도의 현탁세포를 얻을 수 있었다. 초음파 처리 및 Pluronic F-68의 첨가는 참당귀 세포의 세포막의 투과성을 증진시켜 ECP의 생산성을 높임과 동시에 세포 내의 다당의 배지로의 배출을 유도한 것으로 판단된다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3715-3721
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• 2015

Leukemia is a clonal disorder with blocked normal differentiation and cell death of hematopoietic progenitor cells. Traditional modalities with most used radiation and chemotherapy are nonspecific and toxic which cause adverse effects on normal cells. Differentiation inducing therapy forcing malignant cells to undergo terminal differentiation has been proven to be a promising strategy. However, there is still scarce of potent differentiation inducing agents. We show here that Angelica sinensis polysaccharide (ASP), a major active component in Dong quai (Chinese Angelica sinensis), has potential differentiation inducing activity in human chronic erythro-megakaryoblastic leukemia K562 cells. MTT assays and flow cytometric analysis demonstrated that ASP inhibited K562 cell proliferation and arrested the cell cycle at the G0/G1 phase. ASP also triggered K562 cells to undergo erythroid differentiaton as revealed by morphological changes, intensive benzidine staining and hemoglobin colorimetric reaction, as well as increased expression of glycophorin A (GPA) protein. ASP induced redistribution of STAT5 protein from the cytoplasm to the nucleus. Western blotting analysis further identified that ASP markedly sensitized K562 cells to exogenous erythropoietin (EPO) by activating EPO-induced JAK2/STAT5 tyrosine phosphorylation, thus augmenting the EPO-mediated JAK2/STAT5 signaling pathway. On the basis of these findings, we propose that ASP might be developed as a potential candidate for chronic myelogenous leukemia inducing differentiation treatment.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6549-6556
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• 2013

Leukemia stem cells (LSCs) play important roles in leukemia initiation, progression and relapse, and thus represent a critical target for therapeutic intervention. Hence, it is extremely urgent to explore new therapeutic strategies directly targeting LSCs for acute myelogenous leukemia (AML) therapy. We show here that Angelica sinensis polysaccharide (ASP), a major active component in Dong quai (Chinese Angelica sinensis), effectively inhibited human AML $CD34^+CD38^-$ cell proliferation in vitro culture in a dose-dependent manner while sparing normal hematopoietic stem and progenitor cells at physiologically achievable concentrations. Furthermore, ASP exerted cytotoxic effects on AML K562 cells, especially LSC-enriched $CD34^+CD38^-$ cells. Colony formation assays further showed that ASP significantly suppressed the formation of colonies derived from AML $CD34^+CD38^-$ cells but not those from normal $CD34^+CD38^-$ cells. Examination of the underlying mechanisms revealed that ASP induced $CD34^+CD38^-$ cell senescence, which was strongly associated with a series of characteristic events, including up-regulation of p53, p16, p21, and Rb genes and changes of related cell cycle regulation proteins P16, P21, cyclin E and CDK4, telomere end attrition as well as repression of telomerase activity. On the basis of these findings, we propose that ASP represents a potentially important agent for leukemia stem cell-targeted therapy.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.319-319
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• 2005

• 한국약용작물학회지
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• 제10권4호
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• pp.237-242
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• 2002

It was found that the purified extract from A. gigas Nakai (polysaccharide, M.W., 25 kD) controled differentiating human ES cells. Its optimal supplementation concentration was decided as 0.8 $({\mu}g/ml)$ to efficiently control the differentiation. It also enhanced the cell growth, compared to the control. However, most widely used and commercially available differentiating agent, Leukemia Inhibitory Factor (LIF) negatively affected on the cell growth even though it controls the differentiation of ES cells, down to 40-50 % based on morphological observation and telomerase activity. It was presumed that the extract first affected on cell membrane and resulted in controlling signal system, then amplify gene expression of telomere, which enhanced the telomerase activity up to three times compared to the control. LIF only increased the enzyme activity up to two times. It was confirmed that the extract from A. gigas Nakai could be used for substituting currently used differentiation controlling agent, LIF from animal resources as a cheap plant resource and not affecting the cell growth. It can broaden the application of the plants not only to functional foods and their substitutes but also to fine chemicals and most cutting-edge biopharmaceutical medicine.

• 한국식품과학회지
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• 제51권4호
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• pp.336-342
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• 2019

참당귀 유래 면역활성 다당의 활성과 구조의 상관관계를 규명하고자 조다당 AGE-0로부터 두 차례의 연속적인 컬럼 크로마토그래피를 실행하여 단일 정제 다당 AGE-2c-I을 얻었다. AGE-2c-I은 농도의존적으로 우수한 항보체 활성을 보여주었으며, 일반화학적 특성을 분석한 결과, 분자량 약 140 kDa의 고분자 다당으로 4종의 구성당과 13종의 당쇄 결합양식으로 구성되어 있음을 확인할 수 있었다. 이 다당은 ${\beta}$-glucosyl Yariv reagent와의 높은 반응성을 보임으로써 arabino-3,6-galactan의 구조를 가진 rhamnogalacturonan-I 구조임을 추정할 수 있었다. 한편, AGE-2c-I의 미세구조의 해명과 항보체 활성에 관여하는 다당 중의 활성부위 검토를 위해 ${\alpha}$-L-arabinofuranosidase와 endo-1,4-${\beta}$-galactanase를 이용한 연속적 가수분해를 행하고 얻은 단편획분들을 이용, 구성당 및 당쇄결합 양식 분석, ${\beta}$-glucosyl Yariv reagent와의 반응성 검토 및 항보체 활성 결과를 분석한 결과, 참당귀 유래 항보체활성 다당 AGE-2c-I은 rhamnogalacturonan-I과 유사한 구조를 소유하고 있음이 확인되었으며, AGE-2c-I의 측쇄 구조인 arabino-${\beta}$-3,6-galactan 사슬이 항보체 활성 발현에 주요 역할을 수행하며, 5-linked Araf와 3,5-branched Araf로 구성된 ${\alpha}$-arabinan 측쇄가 활성에 부분적으로 관여하고 있음을 최종 확인할 수 있었다.