• Preventive Nutrition and Food Science
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• 제21권3호
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• pp.236-244
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• 2016

In this study, we compared the anti-inflammatory activity of Pinus koraiensis cone bark extracts prepared by conventional extraction and microwave-assisted extraction (MAE). Water extracts and 50% ethanol extracts prepared using MAE were applied to RAW 264.7 cell at 5, 10, 25, and $50{\mu}g/mL$ of concentrations, and tested for cytoxicity. The group treated with $50{\mu}g/mL$ of 50% ethanol extracts showed toxicity. In order to investigate the inhibition of nitric oxide (NO) production in RAW 264.7 cells, extracts of water and ethanol were treated with 5, 10, and $25{\mu}g/mL$ concentrations. The inhibitory activity of water and 50% ethanol extracts groups were determined as 40% and 60% at $25{\mu}g/mL$ concentration, respectively. We found concentration dependent decreases on inducible NO synthase. The inhibitory effect against forming inflammatory cytokines, prostaglandin $E_2$, tumor necrosis factor-${\alpha}$, interleukin (IL)-6, and IL-$1{\beta}$, was also superior in the $25{\mu}g/mL$ treated group than the control group. According to these results, the water extracts and 50% ethanol extracts both inhibited inflammatory mediators by reducing the inflammatory response. Therefore, The MAE extracts of P. koraiensis cone bark can be developed as a functional ingredient with anti-inflammatory activity.

• 한국식품저장유통학회지
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• 제23권1호
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• pp.20-26
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• 2016

본 연구에서 92종의 한약재로부터 물과 ethanol을 용매추출물에 대하여 HAase의 저해에 의한 항염증 효과를 측정한 결과 물 추출물에서는 E. officinalis(86.8%), T. orientalis(80.8%), C. semen(66.5%), M. azedarach(74.7%), S. pubescen (61.3%), S. chinensis(49.15%) 등이 높게 나타났다. Ethanol 추출물에서는 A. altissima와 S. chinensis 추출물이 90% 이상의 높은 항염증 활성을 나타내었다. 이들 중 물과 에탄올 추출물에서 모두 항염증 활성이 높게 측정된 S. chinensis가 선발되었다. 선별된 삼백초의 유효성분을 보다 효율적으로 추출할 방안을 확립하기 위하여 인체에 유해하지 않은 용매로 물과 ethanol을 선택하여 추출 최적조건을 살펴보았다. Phenolic 성분의 추출을 위한 최적조건은 50% ethanol을 사용하여 12시간 추출이 최적이었다. Phenolic 성분의 추출을 위해서는 ethanol이 물보다 더 효율적이었다. 최적조건에서 추출된 삼백초 추출물의 염증억제 활성은 $100{\sim}250{\mu}g/mL$ phenolic 농도로 첨가했을 때 70~80%의 염증 억제 효과를 나타내었으나, $500{\mu}g/mL$ 이상의 농도로 첨가 시 오히려 염증 억제활성이 감소하는 경향을 나타내었다. 삼백초를 초미세 분쇄 하였을 때 추출 수율과 염증억제 활성이 증가하는 것을 확인하였다. 따라서 초미세 분쇄 기술은 한약재로부터 생리활성물질 추출수율의 증가를 목적으로 적용할 수 있는 기술로 개발이 가능할 것으로 판단되었다.

• Journal of Applied Biological Chemistry
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• 제61권2호
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• pp.141-150
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• 2018

본 연구에서는 층꽃나무와 L. plantarum으로 발효한 층꽃나무를 각각 80% ethanol로 추출하여 추출물들이 LPS로 유도된 Raw 264.7 cell의 염증반응에 미치는 영향을 비교 검증하여 항염증 소재 개발 가능성을 검토하였다. HPLC를 이용하여 L. plantarum에 의한 층꽃나무 발효 추출물의 유용성분 변화를 확인한 결과, 발효를 통해 유용성분의 profile 변화가 있는 것을 확인할 수 있었다. Raw 264.7 cell에서 세포 독성을 측정하기 위해 MTT assay를 실시한 결과, 층꽃나무 80% ethanol 추출물은 5, 10, $15{\mu}g/mL$의 농도에서 발효 층꽃나무 80% ethanol 추출물의 경우 10, 20, 30, $40{\mu}g/mL$의 농도에서 90.0% 이상의 세포 생존율을 나타내었다. 항염증 효능을 검정하기 위해 iNOS 단백질 발현량, COX-2 단백질 발현량, NO 생성, $PGE_2$ 생성, pro-inflammatory cytokine 발현량을 측정하였다. NO 생합성 효소인 iNOS 단백질의 발현량을 측정한 결과, 층꽃나무와 발효 층꽃나무 80% ethanol 추출물은 각각 15, $40{\mu}g/mL$의 농도에서 약 50.0% 가까운 발현 억제 효과를 나타내었으며, 농도 의존적으로 감소하는 것을 확인할 수 있었다. $PGE_2$ 생합성 효소인 COX-2 단백질의 발현량을 측정한 결과, 층꽃나무 80% ethanol 추출물은 $15{\mu}g/mL$ 농도에서 50.0%, 발효 층꽃나무 80% ethanol 추출물은 $40{\mu}g/mL$ 농도에서 83.0%의 발현 억제 효과를 보여주었다. NO 생성 억제 효과를 측정한 결과 층꽃나무 80% ethanal 추출물은 $15{\mu}g/mL$ 농도에서 62.0%, 발효 층꽃나무는 $40{\mu}g/mL$ 농도에서 81.0%로 control군과 비교하였을 때 NO 생성이 크게 억제되었다. $PGE_2$ 생성 억제 효과를 측정한 결과, 층꽃나무와 발효 층꽃나무 80% ethanal 추출물은 각각 15, $30{\mu}g/mL$의 농도에서 약 70.0%의 발현 억제 효과를 나타내었다. 또한, pro-inflammatory cytokine의 경우, 층꽃나무 80% ethanol 추출물 $15{\mu}g/mL$ 농도에서 $TNF-{\alpha}$는 43.6%, IL-6는 64.3%, $IL-1{\beta}$는 58.7%의 저해율을 나타냈으며, 발효 층꽃나무 80% ethanol 추출물 $40{\mu}g/mL$ 농도에서 $TNF-{\alpha}$는 75.4%, IL-6는 64.3%, $IL-1{\beta}$는 37.7%의 발현 억제 효과를 나타내었다. 이상의 결과를 통해 층꽃나무는 매우 우수한 항염증 효과를 나타내는 소재임을 확인할 수 있었으며, L. plantarum균을 이용한 발효를 통해 층꽃나무가 가진 세포 독성을 낮추어 안전성하고 우수한 효능을 가진 새로운 항염증제로 개발 가능한 소재로 활용될 수 있을 것으로 기대되었다.

• Journal of Applied Biological Chemistry
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• 제58권2호
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• pp.139-143
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• 2015

본 연구는 다양한 효능을 지닌 초과(Amomum tsao-ko Crevost et Lemaire)의 안전한 이용을 위한 독성평가로 식품의약품안전처 고시 제2014-6호 '의약품등의 독성시험기준'에 맞는 독성시험법에 따라 Balb/c mouse를 이용하여 3주간 반복경구투여를 통해 초과의 안전성을 확인하고자 하였다. 3주간 반복 경구투여 후 체중, 장기중량 측정, 혈액분석 및 혈액생화학 검사를 실시하여 안전성을 확인 한 결과, 초과에 의한 특별한 증상이나 체중, 장기중량의 변화는 관찰되지 않았으며, 복대동맥으로부터 채혈한 혈액을 통한 혈구분석결과에서도 대조군과 초과 추출물 투여군 간의 통계적인 유의성을 관찰 할 수 없었다. 또한 혈청을 이용하여 간기능(GOT, GPT, LDH, ALB, TP-S, T-BIL, D-BIL), 신장기능(BUN, CRE), 지질영양 관련(TG), 전해질 관련(I.P) 지표들의 생화학분석을 수행한 결과, 대조군과 유사하게 모두 정상 범위 내의 결과를 나타내었다. 이러한 결과를 통하여 초과 추출물의 최대무독성용량은 최고 투여량인 2000 mg/kg 이상으로 판단되며, 본 연구결과는 초과의 기능성 식품, 화장품, 의약품 등 다양한 소재로서의 활용에 안전성 관련 기초자료로 이용될 수 있을 것으로 사료된다.

• 생명과학회지
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• 제28권12호
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• pp.1438-1447
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• 2018

본 연구는 추출 용매별 땅두릅 추출물과 Lactobacillus plantarum으로 발효한 후 용매별로 추출한 땅두릅 발효추출물을 이용하여 항염증 효과를 측정한 연구이다. 땅두릅은 중추신경 계통에 대한 흥분작용이 있고 혈압강하 작용이 알려져 있으며 관절염, 감기, 신경통, 류마티스, 피부가려움증 등에 쓰인다. LPS (lipopolysaccharide)로 염증을 유도한 마우스 유래 macrophage에서의 NO 생성 및 염증관련인자의 발현에 미치는 영향을 검토하여 항염증 소재로서의 이용 가능성을 확인하였다. 건조 땅두릅을 water, ethanol, hexane, ethyl acetate, butanol을 이용하여 추출한 추출물들과 L. plantarum으로 발효한 후 추출한 추출물들에서 RAW264.7 대식세포에 대한 독성 여부를 측정하였고, 세포에 대한 독성이 나타나지 않는 농도에서 추출물의 항염증 활성을 확인하였다. 본 실험에 사용된 추출물은 LPS로 유도된 NO 생성을 유의적으로 억제하였으며, 주요 염증 유발인자인 COX-2와 iNOS의 발현 또한 유의적으로 억제하는 효과를 나타냈다. 염증관련 cytokine인 $IL-1{\beta}$, IL-6 및 $TNF-{\alpha}$의 생성량 또한 유의적으로 감소시켰다. 이상의 결과로부터 땅두릅을 L. plantarum으로 발효한 후 water, ethanol, butanol로 추출하였을 때 염증 억제 효과가 있는 기능성 식품 소재로서의 개발 가능성이 있을 것으로 사료된다.

• Nutrition Research and Practice
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• 제8권6호
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• pp.613-617
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• 2014

BACKGROUND/OBJECTIVES: Adipogenesis is part of the cell differentiation process in which undifferentiated fibroblasts (pre-adipocytes) become mature adipocytes with the accumulation of lipid droplets and subsequent cell morphological changes. Several transcription factors and food components have been suggested to be involved in adipogenesis. The aim of this study was to determine whether mulberry leaf ethanol extract (MLEE) affects adipogenesis in 3T3-L1 adipocytes. MATERIALS/METHODS: The 3T3-L1 adipocytes were treated with different doses of MLEE for 8 days starting 2 days post-confluence. Cell viability, fat accumulation, and adipogenesis-related factors including CCAAT-enhancer-binding protein alpha ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), $PPAR{\gamma}$ coactivator 1 alpha (PGC-$1{\alpha}$), fatty acid synthase (FAS), and adiponectin were analyzed. RESULTS: Results showed that MLEE treatments at 10, 25, 50, and $100{\mu}g/ml$ had no effect on cell morphology and viability. Without evident toxicity, all MLEE treated cells had lower fat accumulation compared with control as shown by lower absorbances of Oil Red O stain. MLEE at 50 and $100{\mu}g/ml$ significantly reduced protein levels of $PPAR{\gamma}$, PGC-$1{\alpha}$, FAS, and adiponectin in differentiated adipocytes. Furthermore, protein level of $C/EBP{\alpha}$ was significantly decreased by the treatment of $100{\mu}g/ml$ MLEE. CONCLUSION: These results demonstrate that MLEE treatment has an anti-adipogenic effect in differentiated adipocytes without toxicity, suggesting its potential as an anti-obesity therapeutic.

• 한국식품과학회지
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• 제9권4호
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• pp.277-283
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• 1977

1. 증미(고체) 배양에서는 배양주(Type cultures)보다 분리주가 월등하게 많은 량의 색소를 생성하였다. 2. Lin씨(氏)의 Submerged culture에서는 IAM 8004(Monascus purbigerus)의 경우 carbon source로서 Rice powder 대신에 Wheat bran 1%, Starch 2%와 Corn meal 3%의 것이 색소의 생성량이 많았고, 분리주는 Carbon source를 Rice bran 1%, Starch 2%로 한 것이 가장 좋았다. 3. 사천씨(四川氏) 배지에서는 3%의 탈지대두분을 첨가했을 때 대체적으로 각 균주모두가 더 많은 색소의 생성을 보였다. 4. 추출한 색소액은 pH, 열처리등에 모두 안정하고 각종용매에 대한 추출성도 좋아서 많은 종류의 식품착색에이용될 수 있을 것으로 생각되나 내광성이 약한것이큰결점으로 보인다. 5. 본색소의 안전성은 Mouse의 경우 $LD_{50}$이 체중20g당 0.2359g으로서 안전한 편이며 더구나 시료를 25%의 에틸알코올 추출액으로 투여했기 때문에 에틸알코올의 독성을 감안한다면 색소자체는 극히 안전한 것으로 사려된다. 본연구는 산학협동재단의 학술연구비로서 이루워 진 것이다.

• 대한한방내과학회지
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• 제39권3호
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• pp.313-322
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• 2018

Objectives: Wild ginseng pharmacopuncture is widely used in oriental medicine. However, there is no standard method for efficiently extracting the active ingredient. In this study, in order to determine an efficient extraction method, wild ginseng was extracted by the distillation and 70% ethanol reflux methods, respectively. In comparing each extract, the index compounds were analyzed, and antioxidant activity was measured. Methods: The index compounds, ginsenoside Rg1 and ginsenoside Rb1, were detected using high performance liquid chromatography (HPLC). Antioxidative activities of total phenolic compounds, DPPH (${\alpha}$, ${\alpha}$-diphenyl-${\beta}$-picrylhydrazyl), ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) and FRAP (ferric reducing antioxidant power) were measured to compare their bioactivities. Since saponin is known to be hemolytic, the hemolytic activity of each extract was compared. Results: The index compounds were analyzed. Nothing was detected in the wild ginseng distilled extracts (WGDE). In the wild ginseng 70% ethanol reflux extracts (WGEE), ginsenoside Rg1 was 3.66 mg/g, and ginsenoside Rb1 was 16.70 mg/g. WGEE showed higher levels than WGDE in all antioxidative activities. In the hemolytic test, the extracts showed almost no toxicity, but WGEE showed lower toxicity than WGDE. Conclusions: In this study, it was concluded that WGEE is more advantageous than WGDE in the detection of index compounds and bioactivity. However, additional studies of additional extraction methods and other bioactivity tests are needed.

• Molecular & Cellular Toxicology
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• 제1권2호
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• pp.137-141
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• 2005

The critical role of folate in the remethylation pathway for methionine synthesis from homocysteine has been well documented. Hyperhomocysteinemia resulting from inadequate folate nutrition has been implicated in increased incidence of macrovascular diseases, colorectal cancer, neural tube defects, etc. Chronic exposure to ethanol impairs folate nutrition and one-carbon metabolism in the liver, which often results in fatty liver due to a defective remetylation process. This study was carried out to investigate the chronic effects of moderate levels of alcohol and dietary folate on plasma homocysteine levels, and on histopathology and biochemical functions of the liver. Rats were raised on experimental diets with three levels of folate (0, 2, 8 mg/kg diet), and 50% ethanol (1.8 ml/kg body weight) was administered intragastically by intubation tubes three times a week for 10 weeks. Plasma homocysteine concentrations were found to be significantly influenced by dietary folate intake and alcohol administration. Among all treatment groups, plasma homocysteine levels were the highest in the animals receiving a combined treatment of folate deficient diet and alcohol administration. Plasma homocysteine concentrations were negatively correlated with folate concentration in the plasma (p<0.01) and liver (p<0.05). Among alcohol treated rats, increase in plasma homocysteine values due to macrovascular and microvascular fatty changes and spotted necrosis were observed more frequently in folate-deficient animals diet than those on folate-adequate and folate supplemented diets in alcohol-treated rats. These results indicate that folate supplementation above the recommended level might be beneficial in the prevention of alcohol-related hyperhomocysteinemia and abnormal histologic changes in the liver.

• Toxicological Research
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• 제12권2호
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• pp.203-211
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• 1996

Methanol has been widely used as an industrial solvent and environmental exposure to methanol would be expected to be increasing. In humans, methanol causes metabolic acidosis and damage to ocular system, and can lead to death in severe and untreated case. Clinical symptoms are attributed to accumulation of forrnic acid which is a metabolic product of methanol. In humans and primates, formic acid is accumulated after methanol intake but not in rodents due to the rapid metabolism of methanol. Neverthless, the developmental and reproductive toxicity were reported in rodents. Previous reports showed that perinatal exposure to ethanol produces a variety of damage in human central nervous system by direct neurotoxicity. This suggests that the mechanism of toxic symptoms by methanol in rodents might mimic that of ethanol in human. In the present study I hypothesized that methanol can also induce toxicity in neuronal cells. For the study, primary culture of rat hippocampal neurons and glias were empolyed. Hippocampal cells were prepared from the embryonic day-17 fetuses and maintained up to 7 days. Effect of methanol (10, 100, 500 and 1000 mM) on neurite outgrowth and cell viability was investigated at 0, 18 and 24 hours following methanol treatment. To study the changes in proliferation of glial cells, protein content was measured at 7 days. Neuronal cell viability in culture was not altered during 0-24 hours after methanol treatment. 10 and 100 mM methanol treatment significantly enhanced neurite outgrowth between 18-24 hours. 7-day exposure to 10 or 100 mM methanol significantly increased protein contents but that to 1000 mM methanol decreased in culture. In conclusion, methanol may have a variety of effects on growing and differentiation of neurons and glial cells in hippocampus. Treatment with low concentration of methanol caused that neurite outgrowth was enhanced during 18-24 hours and the numbers of glial cell were increased for 7 days. High concentration of methanol brought about decreased protein contents. At present, the mechanism responsible for the methanol- induced enhancement of neurite outgrowth is not clear. Further studies are required to delineate the mechanism possibly by employing molecular biological techniques.