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마우스 Macrophage의 IL-1 및 TNF-${\alpha}$의 분비유도에 있어서 한국산 겨우살이 추출물이 미치는 영향
윤택준,유영춘,홍은경,조영호,이석원,유보림,김종배,Yoon, Taek-Joon,Yoo, Yung-Choon,Hong, Eun-Kyung,Cho, Young-Ho,Lee, Suk-Won,Azuma, I.,Yoo, Bo-Im,Kim, Jong-Bae 한국생약학회 1994 생약학회지 Vol.25 No.2
To investigate the effect of Korean mistletoe on stimulation of macrophage, the activity to induce interleukine-1(IL-1) and tumor necrosis factors-${\alpha}(TNF-{\alpha})$ from murine peritoneal macrophage by its extracts originated from oak was examined. From in vitro analysis of the cytokines using the culture supernatants of macrophages stimulated with its extracts for 1hr, it was found that Korean mistletoe induces IL-1 and $TNF-{\alpha}$ from murine macrophage. Furthermore, both extracts of Korean mistletoes that were extracted with distilled water and 2% acetic acid exhibited a significant activity to induce two cytokines. In the stimulation for 30 min, Korean mistletoe at concentration of $1{\sim}100\;\mu/ml$ showed a significant induction of IL-1 from macrophage until 24 hrs after stimulation, showing maximal activity on $5{\sim}10\;hrs\;at\;10{\sim}100\;\mu/ml$. On the other hand, $TNF-{\alpha}$ was induced on the early period, 2 hrs, after stimulation at a wide range of concentration, $1{\sim}500\;\mu/ml$. In addition, the fraction of Korean mistletoe from 80% saturated ammonium sulphate precipitation showed a significant activity to induce both cytokines from macrophage. The present study demonstrates that Korean mistletoe contains immunoregulatory factors responsible for stimulating murine macrophage to secrete IL-1 and $TNF-{\alpha}$ which play an important role in immune responses, and suggests that the activity of Korean mistletoe to induce two cytokines is functioned by a possible independent stimulation manner.
한국산 겨우살이 유산균 발효 추출물의 독성 및 면역증강 효과
윤택준(Taek Joon Yoon),양웅석(Woong Suk Yang),박성민(Sung Min Park),정회윤(Hoe Yune Jung),이안나(An Na Lee),정진혁(Jin Hyuk Jung),강태봉(Tae Bong Kang),유영춘(Yung Choon Yoo),김종배(Jong Bae Kim) 한국식품과학회 2009 한국식품과학회지 Vol.41 No.5
Lactobacillus casei KCTC 2180 및 Lact. plantarum KCTC 3103으로 발효된 한국산 겨우살이 추출물인 FKM-110의 렉틴 성분의 함량 변화, 독성 및 면역증강 효과를 검토하여 다음과 같은 결과를 얻었다. 두 가지 유산균을 이용하여 KM-110을 1-3일간 발효시킨 결과, 렉틴 성분은 발효 전에 비하여 53-71% 정도의 함량을 나타냄으로서 감소되는 경향을 보였다. 마우스에 대한 시료의 독성시험을 위하여 각 시료를 피하 주사하고 LDsub{50} 값을 측정한 결과, KM-110은 50-100 mg/kg인 반면 유산균으로 발효된 겨우살이 추출물인 FKM-110은 150-200 mg/kg인 결과를 보임으로 발효에 의하여 in vivo 독성효과는 감소하는 경향을 보였다. 각 시료를 macrphage에 직접 자극하여 염증성 cytokine인 IL-1β 및 TNF-α의 생산능력을 조사한 결과, FKM-110이 KM-110에 비하여 낮은 cytokine의 생산양식을 나타냈다. 그러나 FKM-110의 항원 KLH에 대한 면역증가 효과를 항체 생산능으로 조사한 결과, KM-110과 차이 없이 항원 단독 투여군에 비하여 높은 역가의 항체가 생산되었다. 따라서 겨우살이 추출물을 유산균으로 발효시킨 결과, in vivo 독성은 감소하였으나 면역증강 활성은 KM-110과 동일하게 유지되는 결과를 얻었다. 따라서 겨우살이의 면역증강 활성에 관여하는 성분으로 렉틴 이외에 물질이 관여할 수 있음을 강하게 시사하였다. In this study, Korean mistletoe extract (KM-110) was fermented with two strains of Lactobacillus (FKM-110) and then toxicity, lectin content, and immune activities were investigated. The lectin content of FKM-110 was about 53- 71% lower than that of KM-110. When mice were subcutaneously administered with KM-110 and FKM-110, the LDsub{50} obtained for KM-110 treatment was 50-100 mg/kg as compared to 150-200 mg/mL for FKM-110. Each preparation stimulated macrophages directly and enhanced productivity of inflammatory cytokines such as TNF-α and IL-1β. FKM-110 treatment resulted in lower cytokine production compared to KM-110. When mice were immunized with Keyhol limpet hemocyanin (KLH) antigen along with KM-110 or FKM-110 administration, higher antibody titers to KLH were observed in the KM-110 or FKM-110 groups compared to mice immunized with KLH alone, thereby showing no difference between KM-110 and FKM-110. Therefore, fermentation of Korean mistletoe extract with these Lactobacillus strains decreased toxicity in vivo while the enhancement of immune activity by KM-110 and FKM-110 was similar. These data suggest that KM-110 fermentation tended to decrease lectin content and in vivo toxicity. In addition, other components in the fermented mistletoe extract appear to stimulate immuno-adjuvant activity instead of lectin.
윤택준 ( Taek Joon Yoon ),조선영 ( Seon Yoong Jo ) 한국식품영양학회 2010 韓國食品營養學會誌 Vol.23 No.4
This study was conducted to investigate the effects of Acanthopanax senticosus extracts(ASE) on alcohol administered mice. The administration of Acanthopanax senticosus extracts(60㎎/㎏) had beneficial actions toward alcohol degradation in acute alcohol treated mice. In the acute alcohol degradation experiment, serum alcohol concentration were lower 3 and 6 hours after taking ethanol(5g/㎏) in ASE treated mice. The oral administration of ASE showed decreased gastric mucous membrane damage produced in ethanol treated mice. In addition, intraperitoneal(i.p.) administration of ASE showed anti-inflammatory effects in inhibition tests of vascular permeability produced by acetic acid. ASE also reduced concentrations of nitric oxide(NO), tumor necrosis alpha(TNF)-α and interleukin(IL)-6 in macrophages that were activated by LPS. These results demonstrate that Acanthopanax senticosus extracts possesses the potential to stimulate alcohol degradation and inhibit inflammatory effects in mice.
민들레 뿌리 물 추출물의 마우스 선천 및 획득 면역계에 미치는 효과
윤택준 ( Taek Joon Yoon ) 한국식품영양학회 2008 韓國食品營養學會誌 Vol.21 No.3
Hot-water (100℃) and cold-water (4℃) extracts of Taraxacum officinale root were assessed for the effects of innate and adaptive immune responses in mice. Hot water extracts (TO-100) and cold water extracts (TO-4) did not affect the viability of macrophages at concentrations below to 18 mg/㎖ and 8 mg/㎖, respectively. The thioglycollate-induced macrophages cultured with TO-100 and TO-4 produced a significantly higher quantity of various cytokines, such as IL-6 and IL-12, than those treated with medium. This shows that the extracts potently stimulated the innate immune response. When mice were subcutaneously immunized(sc) with OVA+FIA (Freund`s incomplete adjuvant)-emulsified TO-100, TO-100 did not affect the production of IgE, but enhanced the production of IgG I, IgG2a and IgG2b. The culture supernatant obtained from the splenocytes of mice treated with OVA+FIA-emulsified TO-100 also evidenced elevated levels of both OVA-specific Th1-type (IFN-γ) and Th2-type cytokines (IL-4, IL-6 and IL-10). These results suggested that TO-100 can modulate the immune responses to allergens in mice.
생약 조성물, DTS20의 알코올 분해 및 항염증 효과
윤택준(Taek Joon Yoon),조선영(Sun Young Jo),이수정(Sue Jung Lee),김은영(Eun Young Kim),신광순(Kwang-Soon Shin),서형주(Hyung Joo Suh) 한국생물공학회 2011 KSBB Journal Vol.26 No.5
These studies were conducted to investigate the alcohol degradation effects of the extract of herbal composition (DTS20) containing Viscum album L., Lycium chinense L., Inonotus obliquus and Acanthopanax senticosus H., on the alcohol administered mice. To investigate anti-hangover effect, alcohol and alcohol dehydrogensae (ADH) concentration of blood were measured after oral administration of ethanol. The administration of DTS20 (200-500 mg/kg) had beneficial actions toward alcohol degradation in acute alcohol treated mice model. The oral administration of DTS20 showed decreased gastric mucous membrane damage produced in ethanol treated mice. In addition, intraperitoneal administration of DTS20 showed anti-inflammatory effects in inhibition tests of vascular permeability produced by acetic acid. DTS20 also reduced the concentration of nitric oxide (NO), tumor necrosis factor (TNF)-α in macrophages that were activated by LPS. These results demonstrate that DTS20 possesses potential to stimulate the alcohol degradation and inhibit the inflammatory effects in mice.
윤택준 ( Taek Joon Yoon ),문원국 ( Won Kook Moon ),이광호 ( Kwang Ho Lee ) 한국식품영양학회 2007 韓國食品營養學會誌 Vol.20 No.3
In experimental lung metastasis of colon26-M3.1 carcinoma cells, we found that the prophylactic or therapeutic administration of Inonotus obliquus extracts significantly inhibited lung metastasis. In an in vitro cytotoxicity analysis, the extracts did not affect colon26-M3.1 cell growth at concentrations up to 1000 ㎍/㎖. Peritoneal macrophages that were stimulated with the extracts produced TNF-α. These data suggest that Inonotus obliquus extract has antitumor activity to inhibit tumor metastasis, and its antitumor effects are partially associated with macrophages activation.
한국산 겨우살이 숙주별 렉틴 함량과 지표물질로서의 특성 조사
김인보,윤택준,박춘호,이우경,이소희,김종배,Kim, Inbo,Yoon, Taek Joon,Park, Choon Ho,Lee, Woo Kyoung,Lee, So Hee,Kim, Jong Bae 한국식품영양학회 2015 韓國食品營養學會誌 Vol.28 No.6
겨우살이는 전통적으로 항암활성이 있는 약용식물의 하나로 알려져 왔고, 렉틴은 세포독성 및 면역자극 자극 활성을 가지는 대표성분으로 인정되고 있다. 한국산 겨우살이에 함유되는 렉틴은 유럽산의 그것과는 달리 galactose와 N-acetylgalactosamine(GalNAc) 특이성을 동시에 가지는 렉틴 성분인 KML인 것으로 나타났다. Sandwich ELISA법을 이용하여 각기 다른 종류의 숙주나무에서 유래된 5종의 겨우살이로부터 렉틴 함량을 비교한 결과, 숙주나무별 차이가 인정되어 밤나무 겨우살이는 참나무 겨우살이에 비하여 약 10배 많은 렉틴을 함유하고 있었다. L5178Y-ML25 lymphoma 세포에 약 90%의 세포독성을 나타내는 농도의 KML과 한국산 참나무 유래 겨우살이 추출물인 KM-100에 두 종류의 단일클론 항체(9H7-10 and 8B11-2C5)를 동시처리한 후 세포독성 중화효과를 조사한 결과, KML의 경우 약 10%, KM-110의 경우 약 30%의 세포독성을 보였다. 이러한 결과는 겨우살이에서 렉틴 외에도 세포독성을 가지는 다른 성분이 존재할 것으로 사료되었다. RAW 264.7 대식 세포주에 KM-110과 KM-110으로부터 렉틴이 제거된 분획인 LFKM-110을 자극시킨 결과, LFKM-110에서 $TNF-{\alpha}$ 및 IL-6와 같은 cytokine의 생산을 증진시키는 결과를 보였다. 따라서 KM-110에서 면역 세포를 자극하는 다른 성분의 존재하고 있음을 강하게 제시되었다. Traditionally, mistletoe is known as an effective anti-cancer medicinal plant, and lectin is recognized as a major component with cytotoxic and immuno-stimulant activity in mistletoe. A Korean mistletoe lectin (KML) has specificity to galactose and galactosamine and is distinguish from European mistletoe lectin (EML). When we examined the concentration of lectin in mistletoe originated from five different types of host trees, the result indicate that the lectin concentration is variable depending on the host tree. Noticeably, mistletoe from chestnut tree contains ten folds higher lectins than that of an oak tree. We also tested the concentration of KML and crude extract (KM-110) of Korean mistletoe that shows 90% cytotoxicity in L5178Y-ML25 lymphoma cell. In addition, the cells show 90% and 70% viability by the treatment of two neutralizing antibodies of KML, 9H7-D10 and 8B11-2C5 neutralization effect with two monoclonal antibodies of KML, 9H7-D10 and 8B11-2C5. Therefore, the result expected that the mistletoe contain some other cytotoxic components except lectin. Finally, the production of $TNF-{\alpha}$ and IL-6 by RAW 264.7 cells stimulated with lectin free-crude extract (LFKM-110) following neutralization by 9H7-D10 monoclonal antibody shows higher than that of lectin containing-crude extract (KM-110). These results suggest that the Korean mistletoe lectin ha a great potential to be developed as therapeutic agent of cancer.