• 한국식품영양학회지
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• 제32권1호
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• pp.1-10
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• 2019

In order to examine the functionality of broccoli sprout (Brassica oleracea, BS), solvent extracts were prepared and their anti-oxidative and immunomodulating activities were compared with those of broccoli (B). EtOH extracts (E) were potently higher than hot-water extracts (HW) in the antioxidant contents and radical scavenging activity. In particular, the total polyphenolic contents in addition to ABTS and DPPH radical scavenging activity were significantly higher in EtOH extract of broccoli sprout (BS-E; 9.15 mg GAE/g, 4.52 mg AEAC/g, and 1.14 mg AEAC/g) compared with that of broccoli (B-E; 7.83 mg GAE/g, 3.63 mg AEAC/g, and 0.97 mg/AEAC/g). Whereas, total flavonoid content was significantly higher in B-E (1.60 mg QE/g) than BS-E (1.43 mg QE/g). Anti-inflammatory activity was investigated using LPS-induced cell line model at a concentration of $10{\sim}100{\mu}g/mL$, in which all solvent extracts of both broccoli sprouts and broccoli were not toxic to RAW 264.7 cell lines. In anti-inflammatory activity of broccoli sprouts, EtOH extracts also showed significantly more potent activity than hot-water extracts in all sample concentrations tested. In addition, BS-E ($100{\mu}g/mL$) inhibited nitric oxide (NO) and IL-6 production to 60.9% and 68.9% compared with the LPS inflammation group (without extracts), whereas B-E inhibited 49.6% and 54.9%. On the other hand, in immunostimulating activity by splenocytes and macrophages, hot-water extract showed significantly higher activity than EtOH extract. Especially, BS-HW stimulated the splenocyte proliferation (1.2-fold against saline group) and IFN-${\gamma}$ production (264.39 pg/mL) at $100{\mu}g/mL$, and the production of IL-6 (1.33-fold), IL-12 (1.09-fold) and TNF-${\alpha}$ (1.49-fold) from macrophages was also significantly enhanced over broccoli. In conclusion, broccoli sprouts showed more potent anti-oxidative, anti-inflammatory and immunostimulating activity than broccoli, suggesting the possibility of using broccoli sprouts as functional food materials.

• Preventive Nutrition and Food Science
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• 제21권3호
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• pp.245-254
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• 2016

Water-soluble polysaccharides isolated from the rhizome of Polygonatum sibiricum and fractionated using ionexchange chromatography were investigated to determine their structure and immunostimulating activity. Crude and fractions ($F_1$ and $F_2$) consisted of carbohydrates (85.1~88.3%) with proteins (4.51~11.9%) and uronic acid (1.79~7.47%), and included different levels of mannose (62.3~76.3%), glucose (15.2~20.3%), galactose (4.35~15.3%), and arabinose (4.00~7.65%). The crude contained two peaks with molecular weights (Mw) of $151{\times}10^3$ and $31.8{\times}10^3$, but $F_1$ and $F_2$ exhibited one major peak with Mw of $103{\times}10^3$ and $628{\times}10^3$, respectively. Little immunostimulatory activity was observed by the crude; however, $F_1$ and $F_2$ significantly activated RAW264.7 cells to release nitric oxide and various cytokines, suggesting they were potent immunostimulators. The backbone of the most immunostimulating fraction ($F_1$) was ($1{\rightarrow}4$)-manno- and ($1{\rightarrow}4$)-gluco-pyranosyl residues with galactose and glucose attached to O-6 of manno-pyranoside.

• 대한수의학회지
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• 제46권3호
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• pp.215-224
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• 2006

In order to search the availability of the lectin extracted from Korean mistletoe(Viscum album coloratum) as an adjuvant for the avian vaccines, attempts were made to determine toxicity of the lectin in chicks and its immunostimulating activity on the inactivated vaccines against Newcastle disease virus(NDV). For the determination of toxicity, the lectin was injected into the thigh muscle of SPF chicks(Charles River) of 1-week-old and observed hematologically and pathologically. For the determination of immunostimulating effects, lectin-adjuvanted, inactivated NDV vaccines were injected into the thigh muscle of SPF chicks in the same age group. Sera of the chicks were examined for the hemagglutination-inhibition(HI) antibodies induced, their HI titers and reaction to the NDV antigens. The data were further compared with those from aluminum hydroxide [$Al(OH)_3$]-adjuvanted vaccines and vaccines without adjuvant, and the results are as follows. There were no significant changes observed in the values of RBC, WBC, Hb, PCV, MCV, MCH, MCHC, AST, ALT, BUN, creatinine and total proteins in the chicks administered with lectin of 1.1, 2.2 and $22.2{\mu}g/kg$ body weight, which means the lectin has no effects on blood values and functions of liver and kidney. In histopathologic observation, no lesions were observed in the brain, heart, liver, lung, spleen, kidney, thymus and bursa of Fabricius of the chicks administered with lectin of 1.1, 2.2 and $22.2{\mu}g/kg$ body weight. There were inflammatory lesions, such as congestion, hemorrhage, edema, infiltration of macrophages and coagulation necrosis observed in the thigh muscle of chicks administered with lectin of $22.2{\mu}g/kg$ body weight, whereas no changes were observed in 1.1 and $22.2{\mu}g/kg$ lectin administered chicks. In chicks immunized with lectin($4.4{\mu}g/kg$ of body weight)-adjuvanted B1, LaSota and Ulster 2C vaccines, HI titers in reciprocal values for $log_2$ were 1.8-2.2 at 1 week after vaccination, which was similar with those of 1.5-2.9 by $Al(OH)_3$-adjuvanted vaccines. The HI titers by the lectin-adjuvanted vaccines reached to 3.9-5.3 at 4 weeks, whereas those by the $Al(OH)_3$-adjuvanted vaccines were more high as 7.3-9.3. Meanwhile, the immunostimulating effects of the lectin were recognized while compared to the HI titers with 2.4-3.7 in chicks immunized with vaccines without adjuvants at 4 weeks after vaccination. The chicks immunized with lectin-adjuvanted vaccines were enough to resist challenges by Kyojeongwon strain, a very virulent NDV at 4 weeks after vaccination as well as chicks immunized with $Al(OH)_3$-adjuvanted vaccines. The HI titers by the lectin-adjuvanted vaccines reached to high level as 8.7-10.3 as those with 8.2-9.6 by the $Al(OH)_3$-adjuvanted vaccines at 6 weeks after vaccination, which may be the booster effects by the challenge virus. Antibodies specific to the HN and F antigens of NDV were observed in the sera of both chicks immunized with lectin-adjuvanted vaccines and $Al(OH)_3$-adjuvanted vaccines.

• 한국임상수의학회지
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• 제17권1호
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• pp.21-27
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• 2000

고양이 말초혈액 다형핵백혈구 (PMN)의 유주활성에 있어서 계난백유래물질 (EWD)의 면역증강 효과를 검토하였다. PMN의 유주활성은 EWD 그 자체 및 EWD로 배양한 PMN의 배양상층액에서는 활성이 존재하지 않았다. 그러나 EWD로 배양한 MNC 배양상층액에서는 PMN에 대한 유주활성이 증가되었다. 본 활성은 checkerboard assay를 통해 진성 유주활성으로 밝혀졌다. 이러한 고양이 PMN 유주활성은 human recombinant interleukin-8(hr IL-8)에 의해서도 증가되었다. IL-8의 또 다른 특성인 세포 형태변형 실험에서도 EWD로 배양한 MNC의 배양상층액과 hr IL-8 모두에서 높은 수준의 세포 형태변형효과를 나타내었다. 이와 같이 EWD로 배양한 MNC의 배양상층액과 hr IL-8 모두에서 증가된 PMN의 유주활성 및 세포변형효과는 anti-human IL-8mAb에 의해 농도의존적으로 억제되었다. 이상의 결과로부터 EWD는 고양이 PMN의 유주활성에 있어서 면역증강 효과가 있으며, 이것은 EWD에의해 활성화된 MNC에서 분비되는 IL-8양 유주성 인자에 의해 PMN의 유주활성 증가 및 세포 형태 변형이 일어나는 것으로 사료되었다.

• Food Science and Biotechnology
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• 제15권6호
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• pp.877-883
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• 2006

The immunostimulatory activities of Lactobacillus plantarum, the major microorganism in kimchi fermentations were investigated. Five strains of L. plantarum exhibited weak immunopotentiating activity, but L. plantarum PS-21 showed as strong a mitogenic activity as Bifidobacterium adolescentis M101-4, a known positive strain. It is of interest that, L. plantarum PS-21 stimulated proliferation of Peyer's patch cells, one of the most important tissues in the gut-associated lymphoreticular system. Cell' wall fractions from L. plantarum PS-21 also showed strong mitogenic activity compared with the soluble cytoplasmic fraction. A peptidoglycan fraction (PG) extracted from the cell wall of L. plantarum PS-21 was identified as an active mitogenic component when used in murine lymph node and spleen cell test systems. PG showed dose-dependent mitogenic activity and significantly enhanced antibody production in lymph node cells when studied in vitro. The lysosomal enzyme activity of murine peritoneal macrophages was increased when analyzed following injection of PG to the host animal. Furthermore, PG enhanced the production of cytokines such ($TNF-{\alpha}$ and IL-6) in the in vitro culture of RAW 264.7 macrophage cells.

• 한국식품과학회지
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• 제48권3호
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• pp.289-295
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• 2016

국내 시판중인 인스턴트 커피에서 다당류를 추출하여 새로운 면역 활성 물질을 확인하고자 하였다. 시판중인 인스턴트 커피를 구입하여 냉수추출 후, 에탄올 침전과 투석을 통하여 약 24%의 높은 수율로 다당류(ICP-0)를 얻을 수 있었다. ICP-0은 중성당 89.9%, 산성당 10.1%로 이루어진 다당체로써, 구성 성분으로는 갈락토스 55.5%, 만노스 25.7%와 소량의 아라비노스, 람노스 등으로 이루어진 것을 확인하였으며, 갈락토만난(galactomannan)이나 팩틴 다당류(pectic polysaccharide)가 혼합된 구조일 것이라고 추정되었다. 인스턴트 커피 유래 ICP-0에 대하여 대식세포 자극에 대한 세포증식능을 확인한 결과, NC 대비 유의적으로 높은 증식활성을 보였으며, IL-6, IL-12 및 TNF-${\alpha}$의 경우, $10{\mu}g/mL$에서도 NC 대비 우수한 사이토카인 분비 유도능을 보임이 확인되었다. 또한 NO, ROS를 분석한 결과에서도 NO의 경우 $10{\mu}g/mL$ 이상에서 NC 대비 양호한 활성을 보여 주었다. ICP-0에 대하여 2차 면역기관 중 하나인 비장세포상의 증식능을 확인한 결과, NC 대비 115% 정도 증식하는 것이 확인되어 면역세포를 직접 증진시키는 마이토겐 활성이 있음을 나타내었다. 인스턴트 커피 유래 ICP-0가 장관 내 분포되어 있는 Peyer's patch를 경유하여 장관면역에 공헌할 수 있는지 여부를 확인한 결과, NC 대비 유의적으로 골수세포의 증식능을 확인하였으며 GM-CSF와 같은 관련 사이토카인의 분비 유도가 확인되었다. 이상의 결과는 시판 인스턴트 커피로부터 분리한 다당류가 대식세포의 기능과 장관면역계를 증진시킬 수 있는 가능성을 제시한다.

• 한국식품영양과학회지
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• 제41권10호
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• pp.1378-1387
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• 2012

추출조건에 따른 생리활성 영향을 검토하기 위하여 차가버섯(Inonotus obliquus)을 물($50^{\circ}C$ reflux, $90^{\circ}C$ 이상 decoction, $12^{\circ}C$ 가압추출)과 에탄올(50, 70과 $90^{\circ}C$ reflux)로 추출한 후 물 추출물을 조다당획분(IO-CP)으로 분획하였을 때 IO-CP(4.8~16.8%)의 수율은 에탄올추출물(IO-E, 1.9~2.7%)보다 높았으며 추출온도 증가에 따라 그 차이는 더 컸다. 가압추출로 조제된 조다당획분(IO-CP-121)은 가장 높은 총 폴리페놀 및 플라보노이드 함량(35.10 mg TAE/g와 18.48 mg QE/g)과 DPPH와 ABTS를 이용한 자유 라디칼소거능(26.08과 27.99 mg AEAC/100 mg)을 나타내었다. IO-CP-D(decoction)와 IO-CP-50(reflux)은 IO-CP-121보다 유의적으로 높은 마이토젠(saline 대조군의 2.10과 1.95배, 100 ${\mu}g/mL$)과 장관면역 활성(6.30과 5.74배)을 보였으나 에탄올추출물 활성은 확인되지 않았다. 또한, 모든 IO-CP는 0.1 mg/mL 농도에서 RAW 264.7 세포주에 대하여 독성을 나타내지 않았으나, IO-CP-121이 LPS-자극 RAW 264.7 세포주에 대하여 가장 유의적으로 염증성 인자인 TNF-${\alpha}$와 nitric oxide(NO) 생성을 억제하였다(29.2와 63.5%). 에탄올추출물도 0.1 mg/mL 농도에서 독성을 보이지 않았으나 TNF-${\alpha}$와 NO의 생성 억제능은 IO-CP-121보다 현저히 낮음을 알수 있었다. 한편, 활성 다당획분을 분리하기 위하여 면역활성 획분인 IO-CP-D를 DEAE-Sepharose CL-6B column에서 비흡착 획분(IO-CP-I)과 7개의 흡착 획분(IO-CP-II~VIII)으로 분획한 결과, IO-CP-II가 가장 우수한 마이토젠과 마크로파지 활성(4.51과 1.64배)을 나타내었다. 이와 같이 분리된 면역활성 획분인 IO-CP-II는 주로 중성당(61.86%)과 함께 소량의 산성당(2.96%)을 포함하고 있는 다당류임이 밝혀졌으며, 주요 구성당으로서 Glc, Gal와 Man(molar ratio of 1.00:0.55:0.31)를 포함하고 있음을 확인할 수 있었다. 이러한 결과로부터 추출조건은 차가버섯 생리활성에 중요하게 작용하고 있으며, decoction 추출방법으로 제조하여 분획한 면역활성 획분으로부터 차가버섯 면역활성은 적어도 중성 다당류가 관여하고 있음을 알 수 있었다.

• Biomolecules & Therapeutics
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• 제8권2호
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• pp.199-205
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• 2000

On the inter-order protoplast fusants of Lentinus edodes and Ganoderma lucidum was the antitumor activity test performed and the fusant P22 was selected. The hot water extract of the cultured mycelia of P22 were purified by DEAE-cellulose chromatographya and the resulting purified fraction was designated as P22A. It was found to be a proteoglycan whose molecular weight was 47 kDa. When examined for immunopotentiation activity, P22A increased the number of colony forming unit in the bone marrow stem cells to 3-folds. It also potentiated the secretion of nitric oxide in activated macrophages to 2-folds. In humoral immune response, it increased the activities of the alkaline phosphatase in differentiated B cells to 1.6-folds and the number of plaque forming cells to 1.8-folds. In cellular immune response, it restored the depressed response of delayed type hypersensitivity in tumor bearing mice to normal level. These results suggest that P22A have potential to restore the decreased immune activity of the tumor bearing mice to normal level.

• 한국식품과학회지
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• 제51권3호
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• pp.278-285
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• 2019

Tomato (Lycopersicon esculentum) is widely known for its beneficial effects on human health. To investigate the beneficial effects of polysaccharides from cherry tomato, cherry tomato polysaccharides (CTP) were prepared, the component sugars were analyzed, and the immunomodulatory activities in RAW 264.7 macrophages were assessed. CTP mainly contained arabinose (Ara) and galactose (Gal), suggesting that CTP might be enriched with an arabinogalactan (AG) moiety. The Ara and Gal present in CTP are likely components of AG-II (35.4%), namely $arabino-{\beta}-(3,6)-galactan$. To investigate the immunomodulatory activity of CTP, cytokine levels and iNOS2, COX-2, and $NF-{\kappa}B$ protein levels were analyzed, and $NF-{\kappa}B$ nuclear translocation and phagocytosis were observed by immunofluorescence. CTP significantly increased the levels of $TNF-{\alpha}$, MCP-1, and IL-6. CTP also increased iNOS2 and COX-2 expression as well as $NF-{\kappa}B$ nuclear translocation in RAW 264.7 cells. CTP significantly stimulated phagocytosis activity. These results showed that CTP stimulates macrophage activity, which can boost the innate immune response. CTP with high AG-II content could be used as a prebiotic to strengthen immunity.

• 한국미생물·생명공학회지
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• 제30권1호
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• pp.39-45
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• 2002

실크 피브로인의 가수분해를 통해 생산된 펩타이드 성분에 의해 murine macrophage RAW264.7 세포에 의한 nitric oxide 생성이 촉진됨을 발견하였다. 산 및 효소적 가수분해물의 가수분해도를 비교한 결과 실크 피브로인 단백질은 산 가수분해에 의해 가장 효과적으로 분해되었으며 효소적 가수분해의 경우에는 pepsin, trypsin, Alcalase의 순으로 가수분해도가 높았다. 산 가수분해물을 단독으로 macrophage에 처리하였을 때 처리농도에 따라 NO 생성촉진활성이 높아졌으나 이 활성은 가수분해물 내의 펩타이드 성분들과 오염되어 혼재하는 LPS 성분의 상호작용에 의한 것임이 확인되었다. 함유된 LPS 성분들을 한외여과에 의해 제거한 효소적 가수분해물들의 NO 생성촉진활성은 peptic hydrolysate가 가장 높았고 tryptic-, Alcalase hydrolysate 순이었다. 이러한 활성의 차이는 가수분해물 내의 고분자량 펩타이드 분포가 많을수록 활성이 높다는 관계에 기인하였으나 산 가수분해물의 경우에는 예외적으로 나타났다. 각 가수분해물의 아미노산 조성을 분석한 결과 arginine, lysine의 함량이 높을수록 활성이 높으며 alanine의 glycine에 대한 비율이 커질수록 활성이 높아졌다. 산 가수분해물의 경우에는 낮은 분자량의 펩타이드들이 많이 분포하지만 arginine 및 alanine의 함량이 높아 비교적 높은 NO 생성 촉진활성을 나타내는 것으로 확인되었다.