• 제목/요약/키워드: enzyme assay

검색결과 2,164건 처리시간 0.033초

청미래덩굴 잎 물추출물이 처리된 HepG2 세포에서의 포도당흡수기전 연구 (Study of the mechanisms underlying increased glucose absorption in Smilax china L. leaf extract-treated HepG2 cells)

  • 강윤환;김대중;김경곤;이성미;최면
    • Journal of Nutrition and Health
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    • 제47권3호
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    • pp.167-175
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    • 2014
  • 본 연구에는 SCLE를 이용하여 시도된 바가 없는 glucose uptake 유도 실험을 수행하여 HepG2 세포에서 포도당흡수가 증가함을 확인하였다. 또한 이런 포도당의 흡수는 HNF-$1{\alpha}$라는 transcription factor의 활성화를 통해 GLUT-2의 발현을 증가시키기 때문인 것을 실험적으로 증명하였다. 뿐만 아니라 Bacillus stearothermophilus 유래의 GK를 이용하여 활성을 측정한 결과 SCLE가 직접적으로 GK를 활성화하여 포도당의 인산화에 영향을 주는 것을 확인할 수 있었으며 그 실험결과들은 Fig. 8에서 도식화 하였다. 본 연구를 통해 SCLE가 ${\alpha}$-glucosidase inhibition 활성에 의한 혈당의 개선과 당뇨예방 효과뿐만 아니라 다양한 세포내 기전을 통해 혈당 및 당뇨의 개선을 유도할 수 있음을 확인하였고, 이는 SCLE가 neutraceuticals 소재로서의 개발가치가 높음을 시사한다.

Effects of 17β-Estradiol on Colonic Permeability and Inflammation in an Azoxymethane/Dextran Sulfate Sodium-Induced Colitis Mouse Model

  • Song, Chin-Hee;Kim, Nayoung;Sohn, Sung Hwa;Lee, Sun Min;Nam, Ryoung Hee;Na, Hee Young;Lee, Dong Ho;Surh, Young-Joon
    • Gut and Liver
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    • 제12권6호
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    • pp.682-693
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    • 2018
  • Background/Aims: Intestinal barrier dysfunction is a hallmark of inflammatory bowel diseases (IBDs) such as ulcerative colitis. This dysfunction is caused by increased permeability and the loss of tight junctions in intestinal epithelial cells. The aim of this study was to investigate whether estradiol treatment reduces colonic permeability, tight junction disruption, and inflammation in an azoxymethane (AOM)/dextran sodium sulfate (DSS) colon cancer mouse model. Methods: The effects of $17{\beta}$-estradiol (E2) were evaluated in ICR male mice 4 weeks after AOM/DSS treatment. Histological damage was scored by hematoxylin and eosin staining and the levels of the colonic mucosal cytokine myeloperoxidase (MPO) were assessed by enzyme-linked immunosorbent assay (ELISA). To evaluate the effects of E2 on intestinal permeability, tight junctions, and inflammation, we performed quantitative real-time polymerase chain reaction and Western blot analysis. Furthermore, the expression levels of mucin 2 (MUC2) and mucin 4 (MUC4) were measured as target genes for intestinal permeability, whereas zonula occludens 1 (ZO-1), occludin (OCLN), and claudin 4 (CLDN4) served as target genes for the tight junctions. Results: The colitis-mediated induced damage score and MPO activity were reduced by E2 treatment (p<0.05). In addition, the mRNA expression levels of intestinal barrier-related molecules (i.e., MUC2, ZO-1, OCLN, and CLDN4) were decreased by AOM/DSS-treatment; furthermore, this inhibition was rescued by E2 supplementation. The mRNA and protein expression of inflammation-related genes (i.e., KLF4, NF-${\kappa}B$, iNOS, and COX-2) was increased by AOM/DSS-treatment and ameliorated by E2. Conclusions: E2 acts through the estrogen receptor ${\beta}$ signaling pathway to elicit anti-inflammatory effects on intestinal barrier by inducing the expression of MUC2 and tight junction molecules and inhibiting pro-inflammatory cytokines.

황정(黃精)으로부터 유래한 조다당류의 선천면역 활성에 의한 유방암 세포주 전이 억제 효과 (Antimetastatic Effects of Crude Polysaccharide Isolated from Polygonati Rhizoma on 4T1 Breast Cancer Cells by Activation of Innate Immune System)

  • 지해리;황덕상;이창훈;장준복;이진무
    • 대한한방부인과학회지
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    • 제32권4호
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    • pp.1-13
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    • 2019
  • Objective: This study is aimed to investigate the anti-tumor metastasis by innate immunomodulating effects of crude polysaccharide isolated from Polygonati Rhizoma (CP-PR) on 4T1 breast cancer cells. Methods: CP-PR was isolated from Polygonati Rhizoma. Antimetastatic experiments were conducted in vivo mouse model by using 4T1 breast cancer cells. The cell viability of CP-PR was tested with normal spleen and 4T1 breast cancer cells. To observe the activation of macrophages with/without 4T1 breast cancer cells, production of tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin-6 (IL-6), IL-10 and IL-12 were measured with enzyme-linked immunosorbent assay (ELISA), respectively. In addition, the lysis of YAC-1 cells and the production of granzymes were measured to observe the activation of natural killer (NK) cell. Results: Intravenous administration of CP-PR significantly inhibited metastasis of 4T1 breast cancer cells. In an in vitro cytotoxicity analysis, CP-PR affected the growth of normal spleen and 4T1 breast cancer cells above specific concentration. The production of $TNF-{\alpha}$, IL-6, IL-10 and IL-12 were significantly increased in macrophages with CP-PR. As compared with control, CP-PR showed significantly higher production of $TNF-{\alpha}$, IL-10 and IL-12 in macrophages co-cultured with 4T1 breast cancer cells. The lysis of YAC-1 cells and the production of granzymes were significantly up regulated by CP-PR. Conclusion: CP-PR appears to have considerable activity on the anti-metastasis by activation of innate immune system.

고온고압 처리된 파극천 추출물의 MMP-1 발현 억제 효능 증진 (Effects of Autoclaved Morinda officinalis Root Extract on the Suppressive Efficacy of MMP-1 Enzyme)

  • 강정욱;오정영;배준태;김진화;이근수;표형배
    • 대한화장품학회지
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    • 제41권1호
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    • pp.35-43
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    • 2015
  • 파극천은 전통적으로 피부염증 치료에 사용되어온 약초이다. 본 연구에서는 파극천 뿌리를 고온($132^{\circ}C$) 및 고압($1.2kgf/cm^2$)에서 15분 동안 처리한 후 추출물을 분리하여 이들의 MMP-1 효소 억제 효능을 확인하였다. 고온고압 처리된 파극천 추출물의 성분변화를 확인한 결과, 고온고압 처리가 파극천 추출물의 페놀 및 플라보노이드 함량을 약 1.5배 이상 증가시키는 것으로 나타났다. DPPH 및 superoxide 라디칼 소거능을 확인해 본 결과 $500{\mu}g/mL$에서 각각 79.25%, 94.5%의 라디칼 소거 효과가 나타났으며, 기존 추출물 대비 농도 의존적으로 활성 증가를 확인하였다. 또한, 항염 효과를 5-LOX 및 COX-2 저해능을 통해 확인한 결과 고온고압 처리된 파극천 추출물에서 약 45% 증가된 저해능이 나타났다. 이에 자외선에 대한 고온고압 처리된 파극천 추출물의 피부 콜라겐 단백질 분해효소 발현 저해 양상도 농도 의존적으로 발현이 억제되었으며 약 16% 정도 저해 효과가 유의적으로 증가되었다. UVB의 세포보호 효과도 고온고압 처리된 파극천 추출물을 함께 처리하였을 때 세포생존율이 증가됨을 확인할 수 있었다. 결론적으로 파극천 추출물에 고온고압 처리를 통해 기존 추출물보다 뛰어난 항산화, 항염 및 MMP-1 발현 억제 효과의 증가를 확인하였으며, 앞으로 기능성 소재로서 화장품에 응용될 수 있을 것으로 사료된다.

Utilization of Multiple Carbon Sources by Plant Cells

  • Lee, Taek-Kyun;Suh, Jung-Bin;Kim, Se-Hee;Lee, Sun-Min;Lee, Woo-Sung
    • 한국동물학회:학술대회논문집
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    • 한국동물학회 1999년도 한국생물과학협회 학술발표대회
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    • pp.11-11
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    • 1999
  • ;It has been reported that suspension-cultured rice cells grown on mixed carbon sources of glucose (GIc) and acetate exhibited diauxic growth in which acetate was the preferred carbon source (Lee and Lee, 1996). Carrot (Daucus carota L.) suspension cells, showing a diauxic growth very similar to that of rice cells, were used to delineate the mechanisms underlying this preferential use of acetate over GIc. Uptakes of both GIc and 3-0-methylglucose (3-0MG), a non-metabolizable GIc analogue, were similarly inhibited when acetate or butylate, weak acids which are capable of transporting protons into the cytosol, were present in the uptake assay mixture containing cells harvested during the GIc-utilizing second growth phase. Inhibition of GIc uptake by these weak acids was similar when equivalent experiments were carried out with isolated plasma membranes. It was further shown that Glc uptake, which requires a proper proton gradient across the plasma membranes, was inhibited during the first growth phase by acetate-mediated alkalization of growth medium and/or simultaneous acidification of cytosol. This study strongly suggests that Glc utilization in plant cells is inhibited by co-presenting carbon source(s) which can alter the proton gradient across the plasma membrane. We further examined diauxic growth in culture containing GIc and malate. Unlike the case in the culture with GIc and acetate, carrot cells used GIc first. Malate was utilized only after Glc is depleted from medium. These results indicate that GIc can be a preferred or less-preferred carbon source depending on the competing carbon source. It was noted that malate was not directly taken up by cells. Instead it was converted extracellularly into fumarate which was subsequently transported into cells. During the malate-growth phase malate uptake was negligible, and fumarate uptake was active and pH-sensitive. It was shown that fumarase released into medium was responsible for the extracellular conversion of malate into fumarate. An immunoblot experiments showed that fumarase antibody raised against Arabidopsis fumarase provided positive signals only in medium in malate culture, not in fumarate or GIc cultures. This study demonstrates the first example in that fumarase, a mitochondria marker enzyme, can be present in places other than mitochondria.ndria.

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Dietary glucosinolates inhibit splenic inflammation in high fat/cholesterol diet-fed C57BL/6 mice

  • Gu, HyunJi;Gwon, Min-Hee;Kim, Sang-Min;Yun, Jung-Mi
    • Nutrition Research and Practice
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    • 제15권6호
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    • pp.798-806
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    • 2021
  • BACKGROUND/OBJECTIVES: Obesity is associated with chronic inflammation. The spleen is the largest organ of the lymphatic system and has an important role in immunity. Obesity-induced inflammatory responses are triggered by Toll-like receptor (TLR)-myeloid differentiation primary response 88 (MyD88) pathway signaling. Phenethyl isothiocyanate (PEITC) and 3,3'-diindolylmethane (DIM), major dietary glucosinolates present in cruciferous vegetables, have been reported to produce anti-inflammatory effects on various diseases. However, the effects of PEITC and DIM on the obesity-induced inflammatory response in the spleen are unclear. The purpose of this study was to examine the anti-inflammatory effects of PEITC and DIM on the spleen and their mechanism in high fat/cholesterol diet (HFCD)-fed C57BL/6 mice. MATERIALS/METHODS: We established an animal model of HFCD-induced obesity using C57BL/6 mice. The mice were divided into six groups: normal diet with AIN-93G diet (CON), high fat diet (60% calories from fat) with 1% cholesterol (HFCD), HFCD with PEITC 30 mg/kg/day or 75 mg/kg/day (HFCD+P30, HFCD+P75), and HFCD with DIM 1.5 mg/kg/day or 7.5 mg/kg/day (HFCD+D1.5, HFCD+D7.5). Enzyme-linked immunosorbent assay was used to evaluate pro-inflammatory cytokine secretion. Western blot and quantitative polymerase chain reaction were used to analyze protein and mRNA levels of nuclear factor kappa B (NF-κB) p65, interleukin 6 (IL-6), cyclooxygenase 2 (COX-2), TLR2, TLR4, and MyD88 in spleen tissue. RESULTS: Serum IL-6 levels were significantly higher in the HFCD group than in groups fed a HFCD with PEITC or DIM. Levels of NF-κB p65 protein and TLR2/4, MyD88, NF-κB p65, IL-6, and COX-2 mRNA were significantly higher in the HFCD group than in the CON group and were reduced by the PEITC and DIM supplements. CONCLUSIONS: PEITC- and DIM-supplemented diets improved splenic inflammation by modulating the TLR2/4-MyD88 pathway in HFCD-fed mice. We suggest that dietary glucosinolates may at least partially improve obesity-induced inflammation of the spleen.

한국인에서 뎅기바이러스 항체의 혈청 유병률 연구 (Seroprevalence of Dengue Virus Antibody in Korea)

  • 이지현;김한울;김경효
    • Pediatric Infection and Vaccine
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    • 제25권3호
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    • pp.132-140
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    • 2018
  • 목적: 뎅기바이러스 감염 환자 발생은 해외 여행이 증가함에 따라 증가하고 있다. 최근 감염 경험이 없는 사람에게 백신을 접종하면 이후 야생 뎅기바이러스 감염 시 중증 뎅기열 증상을 보일 수 있다고 연구된 바 있다. 따라서, 본 연구는 한국의 다양한 연령군에서 뎅기바이러스에 대한 혈청역학을 연구하였다. 방법: 건강한 영아 98명(2개월-1세), 청소년 152명(13-19세), 성인 90명(20-50세) 및 노인 106명(65세 이상)에서 수집한 혈청 총 446명을 대상으로 하였다. 각 연령군의 뎅기바이러스 immunoglobulin G (IgG) 항체 검사를 ELISA을 통해 측정하였다. 또한 뎅기바이러스 IgG 항체 검사에서 양성 또는 equivocal을 보이는 혈청에 한하여 일본뇌염 바이러스의 IgG 항체를 검사하였다. 결과: 총 446명 검체 중, 청소년군에서 1명(0.2%)만 뎅기바이러스 항체 검사에서 양성으로 나왔다. Equivocal은 14명(3.1%)으로, 청소년군 10명과 노인군 4명이 해당하였다. 뎅기바이러스 IgG 양성이 나온 1명에서 일본뇌염 바이러스 또한 IgG 양성으로 나왔다. 뎅기바이러스 IgG equivocal이 나온 14명에서는 일본뇌염 바이러스 IgG 양성이 6명, equivocal이 3명이었고 음성은 5명이었다. 결론: 한국인에서 뎅기바이러스에 대한 항체 보유율은 매우 낮았다. 본 연구는 향후 뎅기열 예방을 위한 보건 정책 수립에 중요한 자료가 될 수 있을 것이며 향후 지속적인 혈청면역 평가도 필요할 것이다.

포제 및 발효 가공에 따른 오미자와 구기자 물 추출물의 항염증 및 숙취해소 효과 (Anti-inflammation and hangover relief effects of Schisandra chinensis (SC) and Lycium chinense (LC) water extracts depending on drug processing and fermentation)

  • 김하림;김상준;김솔;김홍준;정승일;유강열;김선영
    • 대한한의학방제학회지
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    • 제26권4호
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    • pp.295-306
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    • 2018
  • Schisandra chinensis (SC) and Lycium chinense (LC) were widely distributed in Asia and the fruit has been used traditionally for medicinal herbs. The processing method was solid-state fermentation using Aspergillus oryzae for 48 h after stir-frying treatment at $220^{\circ}C$ for 12 min. In this study, in vitro the anti-inflammatory effect and in vivo hangover reduction were compared to unprocessed SC and LC water extract. Anti-inflammatory effects have been evaluated in pro-inflammatory mediators which were secreted by lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Nitric oxide (NO) was determined using Griess reaction. Proinflammatory cytokines such as tumor necrosis factor $(TNF)-{\alpha}$ and interleukin $(IL)-1{\beta}$ were measured by enzyme-linked immunosorbent assays (ELISA). Alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities were compared to processed SC or LC and mixtures thereof (1:1). In vivo study was compared to hangover relief in alcohol-fed mice. After administering a mixture of SC and LC (300 mg/kg) water extract (1:1), mice were fed 3 g/kg of ethanol. Serum was collected at 1, 3, and 5 h intervals to analyze ethanol and acetaldehyde levels using a colorimetric assay kit. The processed SC and LC water extracts compared to raw materials significantly inhibited LPS-induced NO and inflammatory cytokine production in RAW 264.7 cells. The results of the hangover mouse model are also consistent with anti-inflammatory effects. These results suggest that processed SC and LC extracts may be functional materials for the treatment of inflammation and hangover.

사과 왜성대목 무독묘 생산을 위한 바이러스 제거 방법 비교 (Comparison of Virus Elimination Methods for Disease-free Seedlings of the Apple Dwarfing Rootstock)

  • 권영희;이정관;김희규;김경옥;박재성;허윤선;윤여중
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2019년도 춘계학술대회
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    • pp.54-54
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    • 2019
  • Apple (Malus domestica) is one of the most economically important fruits in Korea. But virus infection has decreased sustainable production of apple and caused the serious problems such as yield loss and poor fruit quality. Virus or viroid infection including Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple mosaic virus (ApMV) and Apple scar skin viroid (ASSVd) has been also reported in Korea. In many cases, apple is infected with virus and viroid with no specific symptoms, the damage caused by the virus are unaware significantly. In our research, we tried to eliminate viruses in the rootstock for the disease-free seedlings of the apple dwarfing rootstock M.9 and M.26. The method of virus elimination was meristem culture, heat($37^{\circ}C$, 6weeks) treatment and chemistry($Ribavirin^{(R)}$) treatment. The analytical methods commonly used for the detection of virus is Enzyme-linked Immuno-Sorbent Assay(ELlSA) and Reverse Transcription-polymerase Chain Reaction(RT-PCR). RT-PCR method was more 30% sensitive than ELISA method. Efficiency of method eliminate virus appeared meristem method > heat treatment > chemistry treatment. The higher acquisition rate of disease-free seedlings is 30~40% on meristem treatment. In meristem treatment, the apple dwarfing rootstock M.9 gained infection ratio of ACLSV, ASPV and ASGV were 45%, 60% and 50% respectively. In the apple dwarfing rootstock M.26, infection ratio of ACLSV, ASPV and ASGV were 40%, 55%, 55%, respectively. Based on our results, it was found that most effective method of disease-free seedlings apple dwarfing rootstocks was by meristem treatment than heat method and chemistry treatment.

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가시오가피 첨가가 영지버섯의 라카아제 및 셀룰라아제 효소 활성에 미치는 영향 (Effect of Eleutherococcus senticosus supplementation on laccase and cellulase activity of Ganoderma lucidum)

  • 김홍일;정용운;이창수
    • 한국버섯학회지
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    • 제16권4호
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    • pp.347-351
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    • 2018
  • 가시오가피의 첨가는 영지버섯 균사체를 이용한 분비 단백질 라카아제 및 셀룰라아제의 생산량을 효율적으로 증가시켰다. 영지버섯 균사체의 생장률 측정에서 가시오가피의 첨가는 영지버섯 균사의 생장률이 톱밥에 비해 절반 수준에 밖에 못미치는 결과를 나타내었다. 반면에 영지버섯 균사의 분비 단백질인 라카아제와 셀룰라아제의 양을 증가시키는데 톱밥에 비해 큰 효과가 있음을 확인할 수 있었다. 가시오가피가 첨가된 배양액에서의 라카아제 활성은 톱밥이 첨가된 배양액에서보다 영지 1호, 영지 2호 및 녹각 영지에서 0.61~2.37배 증가하였고, 이 중에서도 영지 2호는 0.947 U/min으로 라카아제 활성이 가장 높게 평가되었다. 셀룰라아제 활성 평가에서는 가시오가피가 첨가된 배양액에서의 셀룰라아제 활성은 톱밥이 첨가된 배양액에서보다 영지 1호, 영지 2호 및 녹각 영지에서 1.77~2.81배 증가하였고, 이 중에서도 영지 1호 및 영지 2호에 비해 녹각 영지에서 0.172 U/min으로 셀룰라아제 활성이 가장 높게 평가되었다. 따라서 가시오가피의 첨가가 영지버섯 균사의 분비 단백질 생산을 증가시키는데 유용한 소재로 사료된다.