• 분석과학
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• 제13권5호
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• pp.624-629
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• 2000

Mistletoe lectin I(ML I)에 대한 간단하고 빠른 용액상 효소결합 분석법을 렉틴의 당 특이성을 이용하여 개발하였다. ML I에 특이성을 가지고 있는 D-galactose를 사용하였으며, 용액상 분석법의 효소로는 malate dehydrogenase(MDH)를 사용하였다. 분석신호물질로 사용되는 MDH-galactose 접합체는 isothiocyanate 방법을 통해 합성하였으며, 이 접합제는 thiourea 결합을 하고 있다. ML I의 존재하에, ML I은 D-galactole와의 특이 인식결합을 통해 MDH-galartose 접합체의 활동도를 억제한다. 그러므로, 존재하는 ML I의 농도는 MDH-galactose 접합제의 촉매활동도의 억제도에 비례하게 된다. 따라서, 본 용액상 효소결합 분석법을 통하여 ${\mu}g/mL$ 수준의 ML I의 측정이 분석 시간 10분 이내에 가능하였다.

• Archives of Pharmacal Research
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• 제20권4호
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• pp.306-312
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• 1997

We attempted to isolate and characterize the lectins from stem and leaves of Korean mistletoe (Viscum album var. coloratum) by affinity chromatography. Lectin I was isolated only from stem. Lectin II was not isolated from Korean mistletoe, whereas lectin III was isolated from the stem and leaves. The hemagglutinating activity of lectin I was 16HU and inhibited by D-galactose, lactose, and N-acetyl-D-galactosamine. The lectin I has molecular weight of 60, 000D being composed of two basic subunits with molecular weights of 32, 000D and 28, 000D which are linked by a disufide bond. The lectin III from stem has molecular weight of 66, 000D being two basic subunits which have molecular weights of 34, 000D and 29, 000D and are linked by a disufide bond. The activity of lectin I was stable at the pH range of 4.00-8.50 and at a wide range of temperature (0-42.deg. C). The lectin I showed more potent mitogenic activity to murine lymphocytes than concanavalin A.

• Bulletin of the Korean Chemical Society
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• 제25권1호
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• pp.115-118
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• 2004

We have examined the feasibility of using the specific interaction between mistletoe lectin I (ML I) and ${\beta}$-Dgalactose instead of the anti-ML I antibody in developing a homogeneous type competitive binding assay for ML I. We also have examined the feasibility of adapting the biotin/avidin mediated homogeneous assay for this system. Alkaline phosphatase (AKP) was employed as a single substrate enzyme label. The dose-response curve shows a detection range of 1-25 ${\mu}$g/mL and a linear response with a correlation coefficient of 0.99. To demonstrate the analytical utility of this method, 10 ${\mu}$g/mL of ML I was spiked into distilled water. The results show that the mean recovery was 10.03 ${\mu}$g/mL with an SD of 0.18. The difference between the spiked value and the mean recovery was 0.03 ${\mu}$g/mL, with a relative error of 0.3 and 1.6 % of RSD.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.213.2-214
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• 2003

Three mistletoe lectins (ML -I, ML -IIU, ML -IIL) have been identified in Europe based on sugar specificities for galactose(Gal) and N-acetyl galactosamine(GalNAc). Korean mistletoe lectins have been known as mainly ML -II type. In previous results, we suggested that there are two lectins, 64 KDa and 60 KDa, in Korean mistletoe lectin (KML -C). (omitted)

• Archives of Pharmacal Research
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• 제26권10호
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• pp.861-867
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• 2003

Inhibitory effect of the lectins (KML-C) isolated from Korean mistletoe (KM; Viscum album coloratum) on tumor metastases produced by murine tumor cells (B16-BL6 melanoma, colon 26M3.1 carcinoma and L5178Y-ML25 lymphoma cells) was investigated in syngeneic mice. An intravenous (i.v.) administration of KML-C (20-50 ng/mouse) 2 days before tumor inoculation significantly inhibited lung metastases of both B16-BL6 and colon 26-M3.1 cells. The prophylactic effect of 50 ng/mouse of KML-C on lung metastasis was almost the same with that of 100 $\mu$ g/mouse of KM. Treatment with KML-C 1 day after tumor inoculation induced a significant inhibition of not only the experimental lung metastasis induced by B16-BL6 and colon 26M3.1 cells but also the liver and spleen metastasis of L5178Y-ML25 cells. Furthermore, multiple administration of KML-C given at 3 day-intervals after tumor inoculation led to a significant reduction of lung metastasis and suppression of the growth of B16-BL6 melanoma cells in a spontaneous metastasis model. In an assay for natural killer (NK) cell activity. i.v. administration of KML-C (50 ng/mouse) significantly augmented NK cytotoxicity against Yac-1 tumor cells 2 days after KML-C treatment. In addition, treatment with KML-C (50 ng/mouse) induced tumoricidal activity of peritoneal macrophages against B16-BL6 and 3LL cells. These results suggest that KML-C has an immunomodulating activity to enhance the host defense system against tumors, and that its prophylactic and therapeutic effect on tumor metastasis is associated with the activation of NK cells and macrophages.

• 대한한방내과학회지
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• 제34권1호
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• pp.31-45
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• 2013

Objectives : Gokgisaeng (Korean mistletoe) is used for the treatment of inflammatory and cancer diseases in traditional Korean medicine and its major component lectins have been reported to induce nitric oxide (NO) in RAW 264.7 macrophages, and also induce apoptosis of various types of cancer cells, although its modulatory effects on cancer cell migration and macrophage activation is poorly understood. The aim of this study is to clarify molecular mechanisms of action responsible for the anti-inflammatory and antitumor migration potentials of Korean mistletoe extract (KME). Methods : We investigated the anti-inflammatory activity of KME on NO production and inducible nitric oxide synthase (iNOS) expression by lipopolysaccharide (LPS) in both RAW 264.7 macrophages and rat C6 glioma cells, and also evaluated inhibitory efficacy on glioma cell growth and migration. For assessment, XTT assay, nitrite assay, RT-PCR, scratch-wound and Boyden chamber assay, and western blot analysis were performed. Results : Previously reported, unlike the efficacy of Gokgisaeng lectin, KME inhibited NO production and iNOS expression, and suppressed pro-inflammatory mediators including IL-$1{\beta}$, IL-6, COX-2, iNOS in LPS-stimulated RAW 264.7 cells. Furthermore, KME suppressed tumor cell growth and migration, and it also inhibited LPS-induced NO release and iNOS activation by down-regulating expression of protein kinase C (PKC) and phosphorylation of ERK in C6 glioma cells. Conclusions : Our research findings provide evidence that KME can play a significant role in blocking pro-inflammatory reaction and malignant progression of tumors through the suppression of NO/iNOS by down-regulating of inflammatory signaling pathways, PKC/ERK.

• 약학회지
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• 제45권2호
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• pp.180-189
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• 2001

We have reported that water-extracted Korean mistletoe (KM-110) had various biological activities such as antitumor and immunomodulatory activity, and the pectin fraction (KML-C) of the extract was one of major factors related to its biological functions. In this paper, we produced murine monoclonal antibody (mAb) against KML-C. The cAbs obtained were largely classified into two groups according to specificity to KML-C and ML-I, a pectin from European mistletoe. One group mAbs (9H7-D10 and 3C2-lH4) strongly reacted with KML-C, but not ML-I. In contrast, another group cAbs (8Bll-2C5, BE12-3E9 and 5E10-Fl) reacted with both KML-C and ML-1. The subisotypes of these mobs were shown to be IgGl (9H7-lD10, 3C2-lH4 and 8Bll-2C5) or IgM (8E12-3E9 and 5E10-Fl). To develop an assay system for determination of the amount of KML-C, we established the sandwich ELISA (enzyme-linked immunosorbent assay) method using these mAbs and horse radish peroxidase (HRP)-labelled cAbs. In various combinations of the cAbs for coated antibody and detection antibody, the sandwich ELISA quantitatively detected KML-C, showing the detection limit ranging from 7-5,000 ng/ml. Especially reproducibility (C.V) of the sandwich ELISA, in which 8E12-3E9 was used for coating antibody and 8Bll-2C5-HRP for detection antibody, was 4.59-5.83 in intra assay, and 3.9-9.4 in inter assay.

• 한국식품과학회지
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• 제32권2호
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• pp.461-468
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• 2000

본 연구에서는 Comet assay를 이용해서 차 및 전통 약재로 사용하고 있는 한국산 겨우살이의 항유전독성을 측정하고자 하였다. 겨우살이 조추출물을 분자량에 따라 14,000 이상 분획(I), $8,000{\sim}14,000$의 분획(II), $3,500{\sim}8,000$ 분획(III) 그리고 3,500 이하 분획(IV)으로 분리하였다. 이중 분획 IV만이 비종양성 3T3 세포에서 농도에 따른 MNNG에 의한 DNA 손상을 감소시켰으며 겨우살이 lectin과 viscotoxin이 포함된 분획 I, III은 활성을 전혀 나타내지 않았다. 열처리에 의해서도 분획 IV는 시간이 지남에 따라서 활성이 약간 감소됨을 나타내었으나 여전히 유의적인 효과를 지니고 있었다. 겨우살이 분획 IV로부터 얻어진 유기용매 분획의 항유전독성 평가결과, 분획 IV의 항유전독성은 주로 수용성 물질에 의해 나타나는 것임을 알 수 있었다. Sephadex G-25 gel filtration chromatography에서, 겨우살이 분획 IV의 항유전독성에 기여하는 성분은 분자량 1,000 미만의 수용성 물질이었으며 이 저분자량 물질을 분리해내어 암 예방을 위한 기능성 식품소재로써 이용할 수 있는 가능성을 확인할 수 있었다.