• 제목/요약/키워드: Assay system

검색결과 1,683건 처리시간 0.03초

Expression of Fas/FasL in CD8+ T and CD3+ Foxp3+ Treg Cells - Relationship with Apoptosis of Circulating CD8+ T Cells in Hepatocellular Carcinoma Patients

  • Guo, Cun-Li;Yang, Xiu-Hua;Cheng, Wen;Xu, Yi;Li, Jie-Bing;Sun, Yi-Xin;Bi, Yu-Mei;Zhang, Lei;Wang, Qiu-Cheng
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권6호
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    • pp.2613-2618
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    • 2014
  • Aims: Dysfunction of the host immune system in cancer patients can be due to a number of factors, including lymphocyte apoptosis. Several studies showed that $Foxp3^+T$ cells take part in inducing this process by expressing FasL in tumor patients. However, the relationship between apoptosis, $CD8^+T$ cells and $Foxp3^+T$ cells in HCC patients is still unclear. The present study was designed to investigate the correlation between apoptosis levels and Fas/FasL expression in $CD8^+T$ lymphocytes and $Foxp3^+T$ cells in patients with HCC. Methods: $CD8^+T$ cells and $CD3^+Foxp3^+T$ cells were tested from peripheral blood of HCC patients and normal controls and subjected to multicolor flow cytometry. The expression of an apoptosis marker (annexin V) and the death receptor Fas in $CD8^+T$ cells and FasL in $CD3^+Foxp3^+T$ cells were evaluated. Serum TGF-${\beta}1$ levels in patients with HCC were measured by enzyme-linked immunosorbent assay. The relationship between apoptosis and Fas expression, as well as FasL expression in $CD3^+Foxp3^+T$ cells was then evaluated. Results: The frequency of $CD8^+T$ cells binding annexin V and Fas expression in $CD8^+T$ cells, were all higher in HCC patients than normal controls and the proportion of apoptotic $CD8^+T$ cells correlated with their Fas expression. Serum TGF-${\beta}1$ levels correlated inversely with $CD3^+Foxp3^+T$ cells. Conclusions: Fas/FasL interactions might lead to excessive turnover of $CD8^+T$ cells and reduce anti-tumor immune responses in patients with HCC. Further investigations of apoptosis induction in $Fas^+CD8^+T$ cells in vitro are required.

군리탕가감방(君理湯加減方)이 항종양(抗腫瘍) 면역반응(免疫反應)과 항암제로 유발(誘發)한 부작용(副作用)에 미치는 영향(影響) (Effects of Gunleetang Gagambang Extract on Antitumoral Immunological Response and the Side Effect Induced by Antitumoral Agents)

  • 유경태;문석재;문구;원진희
    • 대한한방종양학회지
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    • 제4권1호
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    • pp.71-87
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    • 1998
  • Even though appropriate immune response is necessary for the survival of the individual, excessive or insufficient immune Response might cause autoimmune or allergic disease. So the immune response must be controlled to the degree that is beneficial for the well being of the individual. This study was undertaken to know the effects of Gunleetang Gagambang on the immune system of the mouse. Gunleetang Gagambang has been used for cure of tumor as a traditional medicine without any experimental evidence to support the rational basis for its clinical use. This study was carried out to evaluate the possible therapeutic or antitumoral effects of Gunleetang Gagambang extract against tumor, and to carry out some mechanisms responsible for its effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene(MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(S180 cells). Treatment of the Gunleetang Gagambang on water-extract(dailly 1mg/mouse, i. p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 20 days. Against squamous cell carcinoma induced by MCA, Gunleetang Gagambang decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice(TBM). Gunleetang Gagambang on also significantly suppressed the development of 3LL cell and S180 cell-implanted tumors in occurrence-frequency and their size. and some developed tumors were regressed by the continuous treatment of Gunleetang Gagambang extract into TBM. In vitro, treatment of Gunleetang Gagambang extract had no effect on the growth of some kinds of cell line such as FsaII, A431 strain but significantly inhibited the proliferation of 3LL, S180 cells and augmented the DNA synthesis of mitogen-activated lymphocytes. Gunleetang Gagambang also stimulated the migrative ability of leukocyte, the MIF and IL-2 production of T lymphocytes, but not IL 6 production of B cells. Gunleetang Gagambang administration to mice enhanced NK cells activities. These results demonstrated that Gunleetang Gagambang extract exhibited a significant prophylactic benefits against tumors and its antitumor activity was manifested depending on the type of tumor cells. And these results also suggested that effect of Gunleetang Gagambang might be chiefly due to nonspecitie enhancement of NK cell activities and cell-mediated immune responses.

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마가목 가지 추출물의 항산화 및 항노화에 관한 연구 (Antioxidative and Antiaging Effects of Sorbus commixta Twig Extracts)

  • 임규남;박민아;박수남
    • 한국응용과학기술학회지
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    • 제28권4호
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    • pp.482-490
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    • 2011
  • In this study, the antioxidative effects and inhibitory effects on tyrosinase and elastase of Sorbus commixta (S. commixta) twig extracts were investigated. The aglycone fraction of S. commixta twig extract showed the prominent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity($FSC_{50}$, $13{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities of S. commixta twig extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated by the luminol-dependent chemiluminescence assay. The 50 % ethanol extract among extracts showed the most prominent ROS scavenging activity ($OSC_{50}$, $0.189{\mu}g/mL$). The cellular protective effects of extract/fractions of S. commixta twig on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The 50 % ethanol extract and ethyl acetate fraction showed the cellular protective effects against ROS in a concentration dependent manner ($5{\sim}50{\mu}g/mL$). The inhibitory effect of S. commixta twig extract on tyrosinase was investigated to assess the whitening efficacy. The ethyl acetate ($IC_{50}$, $113.2{\mu}g/mL$) and aglycone fraction($IC_{50}$, $105.3{\mu}g/mL$) on tyrosinase showed more remarkable inhibitory effect than arbutin($IC_{50}$, $226.88{\mu}g/mL$), known as the whitening agent. The inhibitory effect of aglycone fraction ($IC_{50}$, $6.9{\mu}g/mL$) on elastase was simillar to quercetin($IC_{50}$, $6.1{\mu}g/mL$), flavonoid known as reference compound. These results indicate that extract/fractions of S. commixta twig can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. S. commixta twig extracts can be applicable to new functional cosmetics for anti-aging products.

Endlicheria anomala (Nees) Mez 추출물의 항산화, 항염증 및 미백 활성 (Anti-Oxidative, Anti-Inflammatory, and Anti-Melanogenic Activities of Endlicheria Anomala Extract)

  • 진경숙;이지영;권현주;김병우
    • 한국미생물·생명공학회지
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    • 제41권4호
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    • pp.433-441
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    • 2013
  • 본 연구에서는 Endlicheria anomala (Nees) Mez 메탄올 추출물(EAME)의 항산화, 항염증 및 미백 생리활성을 in vitro assay 및 cell culture model system을 이용하여 분석하였다. EAME의 항산화능을 분석한 결과 DPPH, $H_2O_2$로 유도한 ROS, LPS로 유도한 NO 등 다양한 산화적 스트레스원을 효과적으로 소거하였다. 대표적인 항산화 효소들로 천연물에 의한 항산화능 활성에 의해 주로 발현이 유도되는 세효소인 HO-1, TrxR1, NQO1 및 그 전사 인자인 Nrf2의 단백질 발현에 미치는 영향을 분석한 결과 시료 처리 농도의 증가에 따라 세 효소 및 Nrf2의 발현이 유의적으로 증가됨을 보였다. 또한 EAME는 in vitro DOPA oxidation을 강하게 저해하여 tyrosinase inhibitor로서 작용할 가능성을 시사하였고 이에 B16F10 melanocyte를 이용하여 미백 효능을 분석한 결과 유의적인 melanin 생성억제능 및 tyrosinase 효소 활성 억제능을 보였으며 이는 tyrosinase, TRP-1, TRP-2 등 melanin 생성의 핵심 작용 효소들의 단백질 발현 저해를 통해 일어나는 것으로 나타났다. 이러한 결과를 통해 EAME가 높은 항산화능과 항염증 활성, 그리고 미백 활성을 보유함을 처음으로 밝혔으며 향후 기능성 식품 및 피부 미용 소재로서유용하게 활용될 수 있을 것으로 판단된다.

Mutant p53-Notch1 Signaling Axis Is Involved in Curcumin-Induced Apoptosis of Breast Cancer Cells

  • Bae, Yun-Hee;Ryu, Jong Hyo;Park, Hyun-Joo;Kim, Kwang Rok;Wee, Hee-Jun;Lee, Ok-Hee;Jang, Hye-Ock;Bae, Moon-Kyoung;Kim, Kyu-Won;Bae, Soo-Kyung
    • The Korean Journal of Physiology and Pharmacology
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    • 제17권4호
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    • pp.291-297
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    • 2013
  • Notch1 has been reported to be highly expressed in triple-negative and other subtypes of breast cancer. Mutant p53 (R280K) is overexpressed in MDA-MB-231 triple-negative human breast cancer cells. The present study aimed to determine whether the mutant p53 can be a potent transcriptional activator of the Notch1 in MDA-MB-231 cells, and explore the role of this mutant p53-Notch1 axis in curcumin-induced apoptosis. We found that curcumin treatment resulted in an induction of apoptosis in MDA-MB-231 cells, together with downregulation of Notch1 and its downstream target, Hes1. This reduction in Notch1 expression was determined to be due to the decreased activity of endogenous mutant p53. We confirmed the suppressive effect of curcumin on Notch1 transcription by performing a Notch1 promoter-driven reporter assay and identified a putative p53-binding site in the Notch1 promoter by EMSA and chromatin immunoprecipitation analysis. Overexpression of mutant p53 increased Notch1 promoter activity, whereas knockdown of mutant p53 by small interfering RNA suppressed Notch1 expression, leading to the induction of cellular apoptosis. Moreover, curcumin-induced apoptosis was further enhanced by the knockdown of Notch1 or mutant p53, but it was decreased by the overexpression of active Notch1. Taken together, our results demonstrate, for the first time, that Notch1 is a transcriptional target of mutant p53 in breast cancer cells and suggest that the targeting of mutant p53 and/or Notch1 may be combined with a chemotherapeutic strategy to improve the response of breast cancer cells to curcumin.

라울리아 신초 추출물의 항산화 및 항염 효능에 관한 연구 (Study on Anti-oxidant and Anti-inflammatory Activity of the Tissue-cultured Shoot Clumps from Raoulia australis)

  • 박창민;한나경;정민석;백기엽;최종완
    • 대한화장품학회지
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    • 제40권4호
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    • pp.349-357
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    • 2014
  • 본 연구에서는 생물반응장치를 이용하여 조직 배양된 라울리아 신초에 대하여 화장품 성분으로써 응용가치를 평가하였다. 조직 배양한 라울리아 신초에 대한 항산화 및 항염 활성 효과를 연구하였다. 라울리아는 뉴질랜드나 호주에서 자생하는 국화과의 야생초본식물이다. 이미 몇몇 보고 된 논문에서 라울리아는 기관지염, 수막염 그리고 호흡기 질병 등을 유발하는 바이러스에 대한 증식 억제 활성이 있다고 보고되었다. 실험 결과 조직배양된 라울리아 신초 추출물은 자연 상태의 라울리아 추출물과 비교하여 항산화 활성 및 항염 활성 효과가 우수하였다. 조직 배양된 라울리아 신초 추출물은 자연에서 자란 라울리아 추출물보다 $50{\mu}L/mL$ 농도에서 10~25% 항산화 활성을 증가시켰다. 또한 조직 배양된 라울리아 신초 추출물은 LPS로 유도된 대식세포에서 iNOS와 COX-2의 단백질 발현이 자연에서 자란 라울리아 추출물보다 억제되었다. 본 연구의 결과들로, 조직배양 한 라울리아 신초 추출물은 피부 보호를 위한 천연 화장품 성분으로써 우수한 가능성을 제공할 수 있을 것으로 사료된다.

조망거리에 따른 농촌경관의 농촌다움 평가에 관한 연구 (A Study on Evaluation of the Rurality by the Distance of View)

  • 신민지;신지훈
    • 농촌계획
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    • 제21권2호
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    • pp.63-77
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    • 2015
  • Improve the rural views of the Government and the municipalities with the intention of rural landscape planning. However, rural scenery evaluation of landscape types and element categories, landscape scenes, such as the valuation of resources selected, form a coherent system and ecological point of view is not only a Visual perspective of evaluation, or evaluation is made. This study evaluates the rural landscape through the rural landscape for rurality is implemented can explain the image. Rural amenity 100 represented the rural landscape in rurality with photos of photos, and scenes, photos, location-based rural Vista rurality landscapes drawn components. Selected landscape photography scenic landscapes in the u.s. survey and image vocabularies through surveys is representative of the landscape in rurality of each Vista location-specific photos and images began extracted factors. As a result, the Distant view is a town and village in the rolling hills of the surrounding environment is an image behind the Vista Skyline, Farmland Rural village forests, Behind the hills, Individual housing roof, Housing arrangement, The number of household includes landscape components. Factor analysis 'openness', 'intimacy', 'activity', 'complexity', 'safety' was down to five factors, such as a description of the overall factors that force was 66.45 percent. Middle distance view is close to being out of town houses and village environment, an image in the skyline hills Vista, farmlands, village forests, rolling hills, behind the scenes of the housing component of the neck, the sperm will honor an individual factor analysis results are 'intimacy', 'safety', 'openness', 'specificity', 'complexity' five factors the whole description of the capacity factors compared to 67.24 percent. Close-range view is mainly in Vista village embraced the individual elements and an image of a harmonious location in the surroundings of the sperm, individual houses, fences, gates, courtyard, Garden, garage, agricultural facilities, including, but not limited to, factors assay but an 'intimacy', 'complexity', 'safety', 'activity', 'openness' five factors, such as a description of the overall factors that force was 65.29%. This research seeks to determine the extent to which rurality of rural landscape Vista by location attributes and elements and representative officer in photo selection and rurality to extract the image. To date, followed by a comprehensive study, which was presented through the readjustment could not overlook the Visual elements. The future rural development officer for the evaluation of rural landscape classification and quantitative and objective assessment will have to come up with ways to do.

활성산소종의 조절을 통한 음료 '별의별간'의 급성간독성 보호효과 (Protective effect of STAR of STAR series on CCl4 induced acute hepatotoxicity by regulation of reactive oxygen species)

  • 장보윤;오준석;한지혜;김다은;홍재희;김성연
    • 한국식품저장유통학회지
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    • 제23권2호
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    • pp.275-282
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    • 2016
  • 본 연구는 4종의 별의별간(SS) 음료의 간보호 소재로서의 이용 가능성을 조사하고자 항산화능 평가, t-BHP 와 $CCl_4$로 산화적 손상 및 급성 간독성 유도한 in vitro, in vivo 모델을 활용하여 간보호능을 평가하였다. 실험결과, 별의별간 01~04는 $50{\mu}M$ vitamin C 와 유사한 항산화 효과를 나타내었다. HepG2 세포에 t-BHP로 산화 스트레스를 유도한 뒤 나타나는 세포독성에 대해 별의별간 01 및 04에서 농도 의존적인 세포 보호효과를 보였으며, ROS 생성 억제에서 별의별간 01, 03, 04에서 농도의존적인 억제를 나타내었다. 미나리가 혼합된 별의별간 04에 대한 급성 간손상 in vivo 모델을 활용하여 간보호능 검증 결과, 별의별간 04는 $CCl_4$로 증가된 혈중 ALT, AST의 유의적 감소, 간 조직중 증가된 MDA 함량 감소 및 감소된 GSH의 유의적 증가를 나타내었다. 또한, 혈청 및 간 조직에서 증가된 중성지방과 콜레스테롤을 유의적으로 감소시켰다. 이러한 결과를 종합하며, 별의별간 04는 in vitro 및 in vivo 모델에서 산화적 손상에 대해 간보호 효과를 나타내었다.

연(蓮) 잎과 뿌리의 항산화 및 항암활성 (Antioxidant and Anticancer Activities of Lotus (Nelumbo nucifera) Leaf and Root)

  • 정창호;손기봉;김진희;강선경;박은영;서권일;심기환
    • 한국식품저장유통학회지
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    • 제17권1호
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    • pp.131-138
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    • 2010
  • 연을 새로운 기능성 식품의 재료로 활용하기 위하여 잎과 뿌리로 구분한 후 항산화 및 항암활성을 측정한 결과는 다음과 같다. 총 플라보노이드와 총 페놀릭 화합물은 백련 잎에서 각각 12.84 mg/g 및 24.33 mg/g으로 높게 나타났다. 연의 부위별 DPPH radical 소거활성을 측정한 결과 분획물의 농도가 증가함에 따라 DPPH radical 소거활성이 증가하는 경향을 보였으며, 특히 백련잎의 부탄올 분획물에서 가장 높은 DPPH radical 소거활성을 보였다. 환원력도 분획물의 농도가 증가함에 따라 환원력이 증가하는 경향을 보였으며, 환원력 또한 백련잎의 부탄올 분획물에서 가장 높게 나타났다. Linoleic acid를 이용한 자동산화 억제활성을 실험한 결과 다른 분획물에 비하여 잎의 부탄올 분획물에서 가장 높은 과산화 억제활성을 나타내었고, tyrosinase 저해 활성 역시 잎의 부탄올 분획물에서 가장 높은 활성을 보였다. 연의 부위별 아질산염 소거활성을 측정한 결과 분획물의 농도가 증가함에 따라 아질산염 소거활성 역시 증가하는 경향을 보였으며, 특히 백련과 홍련잎 부탄올 분획물을 1mg/mL의 농도로 첨가하였을 때 각각 95.61%와 92.15%의 높은 아질산염 소거활성을 보였다. 폐암세포와 결장암세포에 대한 생육억제활성을 측정한 결과 백련과 홍련잎의 부탄올 분획물에서 가장 높은 생육억제활성을 보였다.

Expression of $interferon$ $regulatory$ factor-1 in the mouse cumulus-oocyte complex is negatively related with oocyte maturation

  • Kim, Yun-Sun;Kim, Eun-Young;Moon, Ji-Sook;Yoon, Tae-Ki;Lee, Woo-Sik;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • 제38권4호
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    • pp.193-202
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    • 2011
  • Objective: We found previously that $interferon$ $regulatory$ factor ($Irf$)-1 is a germinal vesicle (GV)-selective gene that highly expressed in GV as compared to metaphase II oocytes. To our knowledge, the function of $Irf-1$ in oocytes has yet to be examined. The present study was conducted to determine the relationship between retinoic acid (RA) and RA-mediated expression of $Irf-1$ and the mouse oocyte maturation. Methods: Immature cumulus-oocyte-complexes (COCs) were collected from 17-day-old female mice and cultured $in$ $vitro$ for 16 hours in the presence of varying concentrations of RA (0-10 ${\mu}M$). Rate of oocyte maturation and activation was measured. Gene expression was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and cytokine secretion in the medium was measured by Bio-Plex analysis. Apoptosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: The rates of oocyte maturation to metaphase II and oocyte activation increased significantly with RA treatment (10 nM-1 ${\mu}M$). With 100 nM RA treatment, lowest level of $Irf-1$ mRNA and cumulus cell's apoptosis was found. Among 23 cytokines measured by Bio-Plex system, the substantial changes in secretion of tumor necrosis factor-${\alpha}$, macrophage inflammatory protein-$1{\beta}$, eotaxin and interleukin-12 (p40) from COCs in response to RA were detected. Conclusion: We concluded that the maturation of oocytes and $Irf-1$ expression are negatively correlated, and RA enhances the developmental competence of mouse immature oocytes $in$ $vitro$ by suppressing apoptosis of cumulus cells. Using a mouse model, results of the present study provide insights into improved culture conditions for $in$ $vitro$ oocyte maturation and relevant cytokine production and secretion in assisted reproductive technology.