• 한국식품과학회지
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• 제47권3호
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• pp.286-292
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• 2015

다당의 생물활성은 다당의 구조적인 특징과 분자량 분포에 의해 중요한 영향을 미치기 때문에, 특정 다당의 정제는 다당 연구를 위해 필수적이다. 따라서 본 연구에서는 서로 다른 특성을 소유한 다당의 분획을 위한 신속 분리 방법을 개발하고 대표 화합물로 한국산 귤피로부터 조제한 다당 혼합물을 이용, 본 분리법을 최적화 하였다. 귤피는 펙티나아제 처리 후 에탄올 침전법을 통해 조다당 획분인 CPE로 조제되었으며, CPE는 재차 농도별로 연속 희석된 에탄올 용액(EtOH:DIW=8:1, 4:1, 3:1, 2:1, 1.5:1, 1:1, 및 0.5:1)을 이용하여 7가지 획분(CPE8-CPE0.5)으로 분획되었다. CPE8-CPE1획분은 구성당 분석 결과 람노갈락투로난-I과 람노갈락투로난-II 다당의 지표 구성당인 총 11종의 서로 다른 당으로 구성되어 있었으며, arabino-${\beta}$-3,6-galactan 잔기를 함유하고 있는 것으로 확인되었다. 그러나 CPE0.5 획분에서는 람노갈락투로난-II 및 arabino-${\beta}$-3,6-galactan 잔기를 함유하고 있지 않았다. 한편, CPE8-CPE1 획분을 처리한 mouse 복강 대식세포에서는 농도의존적으로 IL-6의 생산 증가가 관찰된 반면, CPE1 및 CPE0.5 획분에서는 활성이 급격히 감소됨을 확인 할 수 있었다. 따라서 이상의 결과로부터 분리방법이 다양한 특성을 갖는 다당의 혼합물로부터 생물활성을 갖는 람노갈락투로난류를 신속히 분리하는데 매우 유용한 것으로 판단되었다.

• 한국식품과학회지
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• 제47권4호
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• pp.511-516
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• 2015

발아 전후 보리 당단백질 추출물에 대한 영양성분 변화와 RAW 264.7 세포에 처리 후 생성하는 산화질소 II와 사이토카인(인터류킨-6, 종양괴사인자-${\alpha}$) 양을 측정하여 면역 증진 효능을 예측하였다. 보리의 발아가 진행됨에 따라 당단백질 추출물의 단백질과 지방질 함량은 감소하고 총당 함량은 증가하는 추세를 확인하였다. 주요 중성당은 아라비노스, 포도당, 자일로스이며 발아함에 따라 6탄당인 포도당은 감소한 반면, 5탄당인 아라비노스와 자일로스는 증가하였다. 아미노산 함량은 GEB에 비해 24-GEGB와 48-GEGB에서 각각 1.03, 1.24배 증가하였다. 면역 활성을 측정하기 위하여 RAW 264.7 세포에 24-GEGB와 48-GEGB를 처리한 결과 GEB에 비해 더 높은 산화질소 II와 사이토카인(인터류킨-6, 종양괴사인자-${\alpha}$)을 생성하였다. 결론적으로, 보리 종자가 발아하면 종자 내 단백질, 지방질 및 당질 등의 영양성분 조성이 변하고, 그 결과 발아보리 당단백질 추출물은 외부 침입에 대해 1차적으로 방어하는 대식세포를 자극하여 세포에 독성을 미치지 않는 범위의 면역매개물질 생성하여 면역 증진 효과를 기대할 수 있는 것으로 판단된다.

• Journal of Periodontal and Implant Science
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• 제31권4호
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• pp.787-801
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• 2001

The molecular mechanisms control the function of PDL(periodonta1 ligament) cells and/or fibroblasts remain unclear. PDLsl7, PDL-specific gene, had previousely　identified the cDNA for a novel protein from cultured　PDL fibroblasts using subtraction hybridization between gingival fibroblasts and　PDL fibroblasts. The purpose　of this study was to determine the regulation by growth factors and cytokines on　PDLsl7 gene expression in　cultured human periodontal ligament cells and observe the immunohistochemical　localization of PDLsl7 protein in various tissues of mouse. Primary PDL fibroblasts isolated by scraping the root of the extracted human mandibular third molars. The cells were incubated with various concentration of human recombinant $IL-1{\beta}$, PDGF-BB and TGF\;${\beta}$ for 48h nd 2 weeks. At each time point total RNA was extracted and the levels of transcription ere assessed by reverse transcription-polymerase chain reaction (RT-PCR assay). polyclonal antiserum raised against PDLsl7 peptides, CLSVSYNRSYQINE and SEAVHETDLHDGC, were made, and stained the tooth, periodontium, developing bone, bone marrow and mid-palatal suture of the mouse. The results were as follows. 1. PDLsl7 mRNA levels were increased in response to PDGF (10ng/ml) and $TGF\;{\beta}$(20ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF{\beta}$for 48 h. 2. PDLsl7 was up-regulated only by $TGF{\beta}$(20 ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF\;{\beta}$ for 2 weeks and unchanged by the other stimulants. 3. PDLsl7 was a novel protein coding the 142 amino acid peptides in the ORF and the nucleotide sequences of the obtained cDNA from RT-PCR was exactly same as the nucleotides of the database. 4. Immunohistochemical analysis showed that PDLsl7 is preferentially expressed in the PDL, differentiating osteoblast-like cells and stromal cells of the bone marrow in the adult mouse. 5. The expression of PDLsl7 protein was barely detectable in gingival fibroblasts, hematopoetic cells of the bone marrow and mature osteocytes of the alveolar bone. These results suggest that PDLsl7 might upregulated by PDGF-BB or $TGF{\beta}$ and acts at the initial stage of differentiation when the undifferentiated mesenchymal cells in the bone marrow and PDL differentiate into multiple cell types. However, more research needs to be performed to gain a better understanding of the exact function of PDLsl7 during the differentiation of bone marrow mesenchymal and PDL cells.

• Journal of Periodontal and Implant Science
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• 제50권3호
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• pp.159-170
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• 2020

Purpose: Immunization with Porphyromonas gingivalis heat shock protein 60 (PgHSP60) may have an immunoregulatory effect on atherogenesis. The aim of this study was to determine whether nasal immunization with a PgHSP60 peptide could reduce atherosclerotic plaque formation in apolipoprotein E knockout (ApoE KO) mice. Methods: Seven-week-old male ApoE KO mice were assigned to receive a normal diet, a Western diet, a Western diet and challenge with PgHSP60-derived peptide 14 (Pep14) or peptide 19 (Pep19), or a Western diet and immunization with Pep14 or Pep19 before challenge with Pep14 or Pep19. Results: Atherosclerotic plaques were significantly smaller in mice that received a Western diet with Pep14 nasal immunization than in mice that received a Western diet and no Pep14 immunization with or without Pep14 challenge. An immunoblot profile failed to detect serum reactivity to Pep14 in any of the study groups. Stimulation by either Pep14 or Pep19 strongly promoted the induction of CD4⁺CD25⁺ forkhead box P3 (FoxP3)⁺ human regulatory T cells (Tregs) in vitro. However, the expression of mouse splenic CD4⁺CD25⁺FoxP3⁺ Tregs was lower in the Pep14-immunized mice than in the Pep14-challenged or Pep19-immunized mice. Levels of serum interferon gamma (IFN-γ) and transforming growth factor beta were higher and levels of interleukin (IL) 10 were lower in the Pep14-immunized mice than in the other groups. Induction of CD25^- IL-17⁺ T helper 17 (Th17) cells was attenuated in the Pep14-immunized mice. Conclusions: Nasal immunization with Pep14 may be a mechanism for attenuating atherogenesis by promoting the secretion of IFN-γ and/or suppressing Th17-mediated immunity.

• 대한본초학회지
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• 제27권5호
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• pp.21-25
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• 2012

Objective : This study was performed to estimate the effects of OMC-2010 extract on cytokine production in mouse spleen cells. Methods : Mouse spleen cells were pre-treated with ethanol and water extract of OMC-2010 for 1 h, then stimulated with lipopolysaccharide (LPS, 1 ${\mu}g/ml$) for 48 h. Then the cells were harvested for real-time reverse transcription polymerase chain reaction to detect cytokines. Results : OMC-2010 ethanol extract significantly inhibited the LPS-induced interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha and IL-5 mRNA expressions, but not shown such changes in IL-6, IL-4, IL-13. OMC-2010 water extract significantly inhibited the LPS-induced TNF-alpha, and IL-5 mRNA expressions, but not shown such changes in IL-1beta, IL-6, IL-4, IL-13. Conclusions : Theses results could suggest that both ethanol and water OMC-2010 extract could inhibit the TNF-alpha and IL-5 mRNA expression.

• 한방비만학회지
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• 제19권2호
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• pp.106-112
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• 2019

Objectives: The aim of this study was to observe the reduction of lipid accumulation by treatment with Akkermansia muciniphila extract on 3T3-L1 adipocytes. Methods: After treating pasteurized Akk. muciniphila strains in HT-29 colorectal cancer cell, the relative expression of interleukin (IL)-8, tumor necrosis factor-α, IL-6, and IL-1β mRNA was analyzed by real time polymerase chain reaction, respectively. 27 strains of Akk. muciniphila which have anti-inflammatory effects were selected. 3T3-L1 pre-adipocytes were treated with Akk. muciniphila for 24 hr and then measured the toxicity using water soluble tetrazolium salt assay. The cells were incubated for 4 days and then differentiated into adipocytes using the medium including adipogenic reagents for 10 days. The Akk. muciniphila was treated when the medium was exchanged for differentiation medium at 4^th day and insulin medium at 6^th day. To observe the lipid accumulation, the cells were stained with Oil red O dye and were measured using a spectrophotometer. Results: In the cytotoxicity test, the cell viability of 3T3-L1 pre-adipocytes was significantly increased compared to the control group which untreated with Akk. muciniphila, and there was no cytotoxicity of Akk. muciniphila at 1×10⁷ CFU/mL. The results on Oil red O staining and absorbance measurements were showed a significant decrease in lipid accumulation in the group which was treated with Akk. muciniphila compared to the control group. Conclusions: In our results, Akk. muciniphila has the inhibitory effect of lipid accumulation in 3T3-L1 adipocytes. This suggests that Akk. muciniphila could be help to improve obesity.

• 생명과학회지
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• 제23권4호
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• pp.569-577
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• 2013

간장은 한국의 전통 발효식품으로써 최근 항암 효과, 항균작용, 항산화능, 항혈전 효과 등의 효능이 보고되고 있다. 한국의 간장은 콩과 밀을 주요 원료로 사용한다. 콩의 세포벽으로부터 유래되는 다당류는 효소적 가수분해에 저항성이 있으므로, 발효가 끝난 후에도 간장에 잔존하게 되며 이것을 간장 유래 다당류(Kanjang polysaccharides, KPS)라 부른다. 본 연구에서는 산분해법과 양조법으로 제조된 시판 간장인 A~T의 20종의 투석물로부터 다당류를 제조하고, 비만세포에서 염증성 cytokine의 방출과 mRNA의 발현에 대한 KPS의 효과를 실험함으로써 항염증 효과를 조사하였다. RBL-2H3 세포에서 KPS의 처리는 histamine과 ${\beta}$-hexosaminidase의 방출을 현저히 억제시켰다. 자극된 HMC-1 세포에서 KPS의 처리는 염증성 cytokine인 IL-6, IL-8, TNF-${\alpha}$의 방출 및 mRNA의 발현을 감소시켰다. 특히, 양조간장으로부터 유래된 KPS는 산분해 간장보다 비만세포에서 우수한 항염증 효과를 나타내었다. 따라서, KPS는 알레르기성 염증반응을 억제시키는데 효과적일 것이라 사료된다.

• 한국식품영양학회지
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• 제29권3호
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• pp.431-437
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• 2016

본 연구는 전통약재로 주로 사용되는 구절초에서 분리한 다당류(CZPS)가 선천 및 적응면역에서 중추적인 역할을 수행하는 대식세포의 활성화에 미치는 영향에 관하여 관찰하였다. 구절초에서 분리한 다당류를 마우스 유래 대식세포주인 RAW 264.7 cell에 처리하였을 때, iNOS의 발현을 조절하여 NO의 생성이 증가되었으며, 또한 면역활성 물질인 TNF-${\alpha}$, IL-$1{\beta}$ 및 IL-6 등의 cytokine의 분비능이 증가되었으며, 이러한 면역활성 매개자인 NO 및 cytokine의 증가의 원인에 관한 정확한 면역세포내 신호전달에 관하여 알아본 결과, CZPS 처리는 MAPKs(ERK, p38)의 인산화를 촉진시키며, 이로 인한 전사인자인 NF-${\kappa}B$의 인산화를 증가시켜, 대식세포의 활성화를 유도시키는 것으로 관찰되었다.

• Journal of Periodontal and Implant Science
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• 제42권6호
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• pp.185-195
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• 2012

Purpose: (-)-epigallocatechin-3-gallate (EGCG) has been reported to exert anti-inflammatory and antibacterial effects in periodontitis. However, its exact mechanism of action has yet to be determined. The present in vitro study evaluated the anti-in-flammatory effects of EGCG on human periodontal ligament fibroblasts (hPDLFs) and human periodontal ligament stem cells (hPDLSCs) affected by bacterial lipopolysaccharide (LPS) extracted from Porphyromonas gingivalis. Methods: hPDLFs and hPDLSCs were extracted from healthy young adults and were treated with EGCG and/or P. gingivalis LPS. After 1, 3, 5, and 7 days from treatment, cytotoxic and proliferative effects were evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and bromodeoxyuridine assay, respectively. And then, the gene expressions of hPDLFs and hPDLSCs were observed for interleukin (IL)-$1{\beta}$, IL-6, tumor necrosis factor (TNF)-${\alpha}$, osteoprotegerin (OPG), receptor activator of nuclear factor kappa-B ligand (RANKL), and RANKL/OPG using real-time polymerase chain reaction (PCR) at 0, 6, 24, and 48 hours after treatment. The experiments were performed with the following groups for hPDLFs and hPDLSCs; 1) No treat, 2) EGCG alone, 3) P. gingivalis LPS alone, 4) EGCG+P. gingivalis LPS. Results: The 20 ${\mu}M$ of EGCG and 20 ${\mu}g/mL$ of P. gingivalis LPS had the lowest cytotoxic effects, so those concentrations were used for further experiments. The proliferations of hPDLFs and hPDLSCs increased in all groups, though the 'EGCG alone' showed less increase. In real-time PCR, the hPDLFs and hPDLSCs of 'EGCG alone' showed similar gene expressions to those cells of 'no treat'. The gene expressions of 'P. gingivalis LPS alone' in both hPDLFs and hPDLSCs were highly increased at 6 hours for IL-$1{\beta}$, IL-6, TNF-${\alpha}$, RANKL, and RANKL/OPG, except the RANKL/OPG in hPDLSCs. However, those increased gene expressions were down-regulated in 'EGCG+P. gingivalis LPS' by the additional treatment of EGCG. Conclusions: Our results demonstrate that EGCG could exert an anti-inflammatory effect in hPDLFs and hPDLSCs against a major pathogen of periodontitis, P. gingivalis LPS.

• 한국식품저장유통학회지
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• 제24권8호
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• pp.1122-1128
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• 2017

애호박의 부위 중 덩굴손 유래의 열수 추출물에 대한 세포 독성 및 염증제어활성 대하여 조사하였다. 애호박 덩굴손의 열수에 대한 세포의 독성에 미치는 영향을 조사한 결과 0.313-2.5 mg/mL의 농도일 때 물 추출물은 세포의 생장에 영향을 미치지 않아 세포에 대한 독성이 없다고 판단되었다. BMDMs를 배양한 세포에 호박덩굴손 열수 추출물과 LPS와 함께 처리한 결과 염증성 사이토카인인 IL-6 및 TNF-${\alpha}$의 농도 의존적으로 감소함을 확인하였다. 호박 덩굴손 열수 추출물로부터 단리 정제된 rutin에 의한 염증성 사이토카인의 분비가 억제되었다. 국내에서 재배되고 있는 호박의 부위별로 조사해본 결과 호박부위 중 유일하게 덩굴손부분에서 염증억제 활성을 보였다. 쥬키니 호박, 애호박 및 늙은 호박의 덩굴손에서 유효성분인 rutin을 함유하고 있었고 향후 염증 억제 소재로서의 활용가능성을 시사하였다.