• 한국식품영양과학회지
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• 제41권9호
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• pp.1226-1234
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• 2012

본 연구에서는 염증반응을 조절하는 다양한 신호전달체계를 중심으로 분자생물학적 방법을 통해 piceatannol의 항염증 기전을 규명하였다. LPS로 염증반응을 유도한 Raw 264.7 대식세포에서 piceatannol은 iNOS의 발현 억제를 통해 NO의 생성을 감소시키고 염증성 사이토카인(TNF-${\alpha}$, IL-6, IL-$1{\beta}$)의 생성을 감소시켰다. 염증반응을 조절하는 신호전달체계 중 piceatannol은 LPS에 의해 유도된 $I{\kappa}B$의 분해와 p65의 핵으로의 이동을 억제하고, LPS에 의해 유도된 SAPK/JNK의 인산화를 억제하였다. 또한 piceatannol은 LPS와 IL-6(LPS에 의해 증가됨)에 의한 STAT3의 활성화를 억제하였다. 뿐만 아니라 piceatannol은 Nrf2의 핵 내 축적을 야기하고 ARE의 transcriptional activity를 증가시켜 HO-1의 발현을 증가시켰다. 본 연구의 결과, piceatannol은 NF-${\kappa}B$와 AP-1, STAT3 신호전달의 억제를 통해, 그리고 HO-1의 발현 증가를 통해 항염증 효과를 나타내었다(Fig. 8).

• 한국식품영양과학회지
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• 제40권7호
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• pp.1053-1062
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• 2011

건강한 삶에 대한 현대인의 관심이 나날이 고조되고 있으며, 이에 따라 노화와 질병의 예방에 효과가 있는 항산화제의 연구가 활발히 이루어지고 있다. 특히 천연물이나 식품을 소재로 한 식이성 항산화제에 대한 연구는 꾸준히 증가하고 있는 추세이며, 천연물의 소재나 연구 분야의 폭이 매우 넓다. 따라서 다양한 식품 소재의 항산화능을 조사할 수 있는 측정법의 중요성이 크게 부각되고 있다. 현재 약용 식물이나 상용 채소, 과일을 시료로 하여 여러 가지 radical이나 target molecule에 대한 항산화능을 측정할 수 있는 다수의 생체외(in vitro) 측정법이 사용되고 있다. 이에 본 총설에서는 널리 사용되고 있는 항산화능 측정법의 특징을 분석하고 적용 사례를 검토하였다. 식품의 구성 성분은 단일물질이 아닌 복합체로 구성되어 있으므로 하나의 측정법으로 항산화능을 평가할 수가 없다. 그러므로 채소와 과일을 포함한 여러가지 식물성 식품 추출물의 항산화 활성을 정확히 측정하기 위해서는 각 실험에 사용되는 radical과 기전(mechanism), 실험 조건(온도, pH, 실험기기, 시료추출방법, 소요 시간, 비용) 등을 고려하여 적절히 수행되어야 할 것이다. 그러나 이들 측정법의 다양성과 사용 단위의 차이로 인해 각 시료의 항산화능을 객관적으로 비교하는데 어려움이 있는 실정이다. 따라서 향후 항산화능 측정법의 표준화와 표현 단위의 통일이 절실히 요구된다. 또한 in vitro에서 항산화능을 나타내는 채소, 과일 등의 생체 내 활성은 다양한 biomechanism과 polymerphism으로 인해 그 효과를 예측하기가 매우 어렵다. 따라서 in vitro에서의 항산화능 screening 결과를 바탕으로 in vivo에서의 그 효과를 검증하는 연구가 부가적으로 시행되어야 할 것이라 사료된다.

• Journal of Periodontal and Implant Science
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• 제25권3호
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• pp.497-517
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• 1995

The cells associated with normal defense mechanism in inflammation release free oxygen radicals, hydroxy radicals, and various protease, all of which can damage the surrounding cells(fibroblasts) and matrix molecules(collagen). The objective of this study was to evaluate the effects of "scavenger" enzyme, superoxide dismutase(SOD). to periodontal ligament (PDL) cells. Human PDL cells were cultured from the teeth extracted for non-periodontal reason. Cultured PDL cells in vitro were treated with SOD and LPS according to dosage and culture times. Cellular activity was exaimed by Microtitration(MTT) assay. The quantitative expression of cellular proliferation by proliferating cell nuclear antigen(PCNA), collagen type I and fibronectin by indirect immunocytochemically stain in PDL cells were done. The results were as follows: 1. As only SOD treated group at 2 and 3 days, PDL cell activity was significantly increased at more than 150U(P<0.05). 2. When LPS(0.5, $5{\mu}g/m{\ell}$) and SOD(more than 150U) were added together, it was significantly increased than LPS only treated and control groups at 2 days(P<0.05). 3. When LPS($5{\mu}g/m{\ell}$) and SOD(150, 300U) were added together, PCNA index was significantly increased than LPS only treated and control groups at 2 and 3 days(P<0.05). 4. When LPS($5{\mu}g/m{\ell}$) and SOD(150U) were added together, collagen type I was significantly increased than LPS only treated and control groups at 3 days(P<0.05). 5.When LPS($5{\mu}g/m{\ell}$) and SOD(300U) were added together, fibronectin was significantly increased than LPS only treated and control groups at 3 days(P<0.05). On the above the results, the SOD in association with collagen type I, fibonectin, and PCNA may afford biological protection to oxy-radicals that were typically liberated during normal inflammatory response. Thus, the exogenous application of SOD may be effective in sthe treatment of the localized breakdown associated with chronic periodontal disease.

• Molecular & Cellular Toxicology
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• 제2권4호
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• pp.236-243
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• 2006

Microarray analysis of gene expression has become a powerful approach for exploring the biological effects of drugs, particularly at the stage of toxicology and safety assessment. Acetaminophen (APAP) has been known to induce necrosis in liver, but the molecular mechanism involved has not been fully understood. In this study, we investigated gene expression changes of APAP using microarray technology. APAP was orally administered with a single dose of 50 mg/kg or 500 mg/kg into ICR mice and the animals were sacrificed at 6, 24 and 72 h of APAP administration. Serum biochemical markers for liver toxicity were measured to estimate the maximal toxic time and hepatic gene expression was assessed using high-density oligonucleotide microarrays capable of determining the expression profile of >30,000 well-substantiated mouse genes. Significant alterations in gene expression were noted in the liver of APAP-administered mice. The most notable changes in APAP-administered mice were the expression of genes involved in apoptosis, cell cycle, and calcium signaling pathway, cystein metabolism, glutatione metabolism, and MAPK pathway. The majority of the genes upregulated included insulin-like growth factor binding protein 1, heme oxygenase 1, metallothionein 1, S100 calcium binding protein, caspase 4, and P21. The upregulation of apoptosis and cell cycle-related genes were paralleled to response to APAP. Most of the affected gene expressions were returned to control levels after 72 hr. In conclusion, we identified potential hepatotoxicity makers, and these expressions profiling lead to a better understanding of the molecular basis of APAP-induced hapatotoxicity.

• Molecules and Cells
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• 제41권11호
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• pp.979-992
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• 2018

Potato (Solanum tuberosum L.) is the third most important food crop, and breeding drought-tolerant varieties is vital research goal. However, detailed molecular mechanisms in response to drought stress in potatoes are not well known. In this study, we developed EMS-mutagenized potatoes that showed significant tolerance to drought stress compared to the wild-type (WT) 'Desiree' cultivar. In addition, changes to transcripts as a result of drought stress in WT and drought-tolerant (DR) plants were investigated by de novo assembly using the Illumina platform. One-week-old WT and DR plants were treated with -1.8 Mpa polyethylene glycol-8000, and total RNA was prepared from plants harvested at 0, 6, 12, 24, and 48 h for subsequent RNA sequencing. In total, 61,100 transcripts and 5,118 differentially expressed genes (DEGs) displaying up- or down-regulation were identified in pairwise comparisons of WT and DR plants following drought conditions. Transcriptome profiling showed the number of DEGs with up-regulation and down-regulation at 909, 977, 1181, 1225 and 826 between WT and DR plants at 0, 6, 12, 24, and 48 h, respectively. Results of KEGG enrichment showed that the drought tolerance mechanism of the DR plant can mainly be explained by two aspects, the 'photosynthetic-antenna protein' and 'protein processing of the endoplasmic reticulum'. We also divided eight expression patterns in four pairwise comparisons of DR plants (DR0 vs DR6, DR12, DR24, DR48) under PEG treatment. Our comprehensive transcriptome data will further enhance our understanding of the mechanisms regulating drought tolerance in tetraploid potato cultivars.

• 한국식품과학회지
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• 제45권2호
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• pp.137-141
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• 2013

플라보노이드는 식물의 주요 2차 대사산물 중 하나로 자외선 차단, 식물의 수분을 위한 곤충 유인 등 외부환경에 적응하는데 이로운 역할을 한다. 특히 플라보노이드는 항산화 효과가 우수한 것으로 알려져 노화방지와 생활습관 질병예방에 유용한 건강기능식품소재로 각광받고 있다. 하지만 플라보노이드의 생체이용률은 매우 낮으며 이러한 플라보노이드 흡수과정에 관한 생물학적기전은 최근에 조금씩 밝혀지기 시작하고 있다. 플라보노이드의 수송기전에는 세포 내에서 일어나는 소포체 매개 수송과 세포막 및 소기관 표면 단백질에 의한 막 수송체 매개 수송이 있다. 소포체 매개 수송의 경우 cellular trafficking에 의한 일련의 소포체 유래 vesicle의 융합 반응을 거쳐 식물 세포의 경우 액포 내에 플라보노이드가 축적되거나 세포 외부로 배출된다. 표면 단백질에 의해 플라보노이드의 세포막 흡수가 일어나게 되는데 ATP를 사용한 능동수송, 막 전위를 이용한 2차 수송에 관여하는 다수의 수송체들이 관여하는 것으로 보인다. 다양한 종류의 플라보노이드가 존재하는 만큼 플라보노이드 수송체도 다양하며 어쩌면 모든 플라보노이드의 특이적 수송체를 규명하는 것은 불가능 할 지도 모른다. 하지만 식품에 다량 존재하는 주요 플라보노이드를 모델 화합물로 이용한 연구를 수행하면 이에 관련된 주요 수송체 단백질과 관련 메커니즘에 대해 깊이 이해할 수 있고 이를 통해 생체 이용률을 향상시키는 방법을 생각해 볼 수 있으며 특히 낮은 혈중 농도 조건에서도 조직 세포 내에 플라보노이드 축적을 통해 건강 기능성을 최적화하는 노력을 기울이는데 적절한 과학적 방법을 제시해 줄 수 있을 것으로 기대한다.

• The Plant Pathology Journal
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• 제24권3호
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• pp.296-304
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• 2008

The transient and rapid expression system of a foreign protein in planta is a very useful technique in biotechnology application. We have investigated optimum condition of Agrobacterium-infiltration technique in which expression level of foreign proteins were maximized without detrimental effects on plants using GFP and Cucumber mosaic virus 2b protein, which is known as an enhancer of gene expression and a suppressor of post-transcriptional gene silencing(PTGS). The optimum expression level of both RNA and protein of GFP with minimum leaf impairment was obtained at $OD_{600}$=0.2 of Agrobactrium inocula. The steady-state levels of GFP RNA and protein generally peaked at 3 and 7 days post-infiltration(dpi), respectively. In the presence of 2b, both the magnitude and duration of GFP expression was highly increased and we could detect GFP level until 17 dpi. On the other hands, the 2b-mediated higher accumulation of foreign proteins resulted in the repression of normal leaf growth, possibly due to the limitation of supply of energy or materials required for growth maintenance. Using this Agrobacterium-infiltration system with 2b and GFP, we tested a hypothesis for the threshold model of PTGS initiation. Four GFP transgenic lines of N. benthamiana, which shows different expression level of GFP were tested to determine the threshold level for PTGS initiation. Agrobacterium-infiltration of GFP into those GFP-transgenic plants resulted in the co-silencing of the transgenic GFP. It was found that very low concentration of Agrobacterium with GFP and GFP+2b($OD_{600}$=0.002-0.02) which could not phenotypically induce an additive GFP expression, was enough to trigger PTGS pathway in all GFP transgenic plants. This strongly indicates that each GFP-transgenic plant should be expressing the transgenic GFP at its own pre-determined level and there was no buffer zone of additive GFP-expression to the threshold. In other words, the PTGS seems to be immediately activated as a self-defensive mechanism if an internal balance of gene expression is broken.

• 대한한방내과학회지
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• 제31권1호
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• pp.128-141
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• 2010

Purpose : The object of this study was to observe the suppressive effects of Yijintang-gamibang (YJGMB), Yijintang with Atractylodis Rhizoma, Massa Medicata Fermentata, Hordei Fructus Germiniatus, and Coptidis Rhizoma. YJGMB has been traditionally used in Korean medicine for treating various digestive diseases. We tested it on the rat reflux esophagitis (RE) induced by pylorus and forestomach ligation in rats as compared with omeprazole, a well-known proton pump inhibitor. Method : Three different dosages of YJGMB 200, 100 and 50mg/kg, were orally pretreated once a day for 28 days before pylorus and forestomach ligation. Seven groups, each of 8 rats per group were used in the study. Six hours after pylorus and forestomach ligation, changes of the stomach and esophagus lesion areas, gastric volumes, acid and pepsin outputs, invasive lesion percentages, fundic mucosa and total thicknesses were measured as histomorphometry. The results were compared with omeprazole, antioxidant and proton pump inhibitor, and 30 and 10mg/kg treated groups in which the effects on RE were already confirmed. Results : As results of pylorus and forestomach ligation, marked increases of esophageal and gastric mucosa lesion areas, gastric volumes, acid outputs, pepsin outputs were observed with histopathological changes of RE, such as hemorrhages, ulcerative lesions and edematous changes on the fundic mucosa. However, these pylorus and forestomach ligation-induced RE were dose-dependently inhibited by treatment of 200, 100 and 50mg/kg of YJGMB. YJGMB 200mg/kg showed similar protective effects as compared with 30mg/kg of omeprazole in the present study, and more favorable effects were observed in 50mg/kg of YJGMB treated rats as compared with omeprazole 10mg/kg in the present study. Conclusion : The results obtained in this study suggest that YJGMB has favorable protective effects on the RE induced by pylorus and forestomach ligation. Therefore, it is expected that YJGMB will also show favorable effects on RE corresponding well to the suggestion of traditional Korean medicine. However, more detailed mechanism studies should be conducted in future with the screening of the biological active chemical compounds in herbs.

• 동의생리병리학회지
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• 제26권5호
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• pp.687-692
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• 2012

Schizandra chinensis extract has been known to possess a variety of efficacy including antitumor. However, it remains unclear how schizandrin, which is a major biological active ingredient of Schizandra chinensis, exerts antitumor effect. This study was designed to investigate the mechanism by which schizandrin inhibits tumor growth and metastasis. In in vivo test using tumor model mice injected with B16BL6 cell line, mice treated with 10 and 100 ${\mu}g/ml$ schizandrin showed a significant inhibition by $73.79{\pm}6.43%$ and $90.46{\pm}1.72%$, respectively, compared with positive tumor controls. Schizandrin did not exert a significant toxicity for the normal cells (HUVECs) and tumor cell lines (A549, B16BL6, Du145, Huh7). Treatment with schizandrin at 10 and 100 ${\mu}g$/head significantly inhibited the tumor-induced angiogenesis by $68.04{\pm}32.21%$ and $103.8{\pm}34.99%$ compared with the positive control group, respectively. Using in vivo lung metastasis model, tumor metastasis assay revealed that 10 and 100 ${\mu}g$/head schizandrin significantly decreased the metastatic lung tumor by $37.51{\pm}8.15%$ and $75.53{\pm}4.38%$ compared with positive controls, respectively. On the other hand, schizandrin did not affect the adherence of B16BL6 cell line to extracellular matrix protein. These results demonstrate that schizandrin exerts inhibitory effect on tumor growth and metastasis by inhibiting angiogenesis. This study thus suggest that schizandrin may be a candidate molecule target for cancer drug development.

• Journal of Periodontal and Implant Science
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• 제26권1호
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• pp.216-229
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• 1996

Interferon(IFN) is a sort of glycoproteins that are produced by activated lymphocyte, monocyte and fibroblast. IFN has anti-viral effects, immuno-defensive mechanism and regulating properties to the several kinds of cells that includes affect on the bone formation and resorption. The effect of IFN on the osteoclast & other tissue cells has been studied in a number of researchers with the limited reports on the osteoblast. The purpose of this study was to evaluate the effects of IFN on the osteoblastic function. The MC3T3/El cell(Mouse osteoblast) was incubated in ${\alpha}-minimum$ essential medium containing 10% FBS. To detect the cytotoxic effect of $IFN-{\gamma}$ on osteoblast, the cells were cultured in 96-well plate to which $IFN-{\gamma}$ of various concentrations were added for 2 days. After staining with trypan blue, total cells and living cells were counted under microscope. To determine the activity of alkaline phosphataset(ALP), various concentrations of $IFN-{\gamma}$ were treated to culture medium, and biochemical assay was performed. $IFN-{\gamma}$ and $IFN-{\gamma}$ plus cycloheximide were added to culture medium separately and then ALP activity were determined. To detect the effect of the $IFN-{\gamma}$ on the bone formation of osteoblast, long-term culture was performed, and calcified nodule formation were observed using von Kossa's staining. After the addition of $IFN-{\gamma}$ with various concentrations to the medium, no cytotoxic effect of $IFN-{\gamma}$ was detected at any concentration. The significant increase in ALP activity of osteoblast were found the concentration of $IFN-{\gamma}$ 500-2500U/ml and the culture time of 24-48 hours respectively. The enhancement of ALP activity by $IFN-{\gamma}$ of osteoblast was decreased significantly by the treatment of cycloheximide. After long-term culture of osteoblast, the nodule formation was found to be increased in number and density by the addition of 500 U/ml $IFN-{\gamma}$. These results suggest that $IFN-{\gamma}$ was affected on the bone formation of osteoblast. Forthemore this kind of study or $IFN-{\gamma}$ to osteoblast will be held continuously.