• 대한한방부인과학회지
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• 제22권1호
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• pp.1-14
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• 2009

Purpose: Oxidative stress was thought to play a critical role in neurodegenerative disease. Many in vivo and in vitro reports explained the possible pathway of human aging. But in therapeutic aspects, there was no clear answers to prevent aging associated with neural diseases. In this study, we investigated the antioxidant and neuroprotective effects of the Insamyangyung-tang (IYT). Methods: To estimate the antioxidant effects, we carried out 1.1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, 2,2'-azinobis-(3- ethylbenzothiazoline-6- sulfonic acid (ABTS) radical cation decolorization assay, and measurement of total polyphenolic content. To evaluate neuroprotective effect of IYT in vitro. We performed thiazolyl blue tetrazolium bromide (MTT) assay, reactive oxygen species (ROS) creation in SH-SY5Y. Tyrosine hydroxylase (TH) immunocytochemistry, nitric oxide (NO) assay, and TNF-${\alpha}$ assay in primary rat mesencephalic dopaminergic neurons. Results: The $IC_{50}$ values were $571.6{\mu}g/m{\ell}$ and $202.3{\mu}g/m{\ell}$ in DPPH and ABTS assay respectively. Total polyphenolic content was 1.05%. In SH-SY5Y culture, IYT significantly increased the decreased cell viability by 6-OHDA at the concentrations of $10{\mu}g/m{\ell}$ in pre-treatment group, $10-100{\mu}g/m{\ell}$ in post-treatment group, and $100{\mu}g/m{\ell}$ in co-treatment group. The production of ROS induced by 6-OHDA was significantly inhibited in IYT treated group. In mesencephalic dopaminergic cell culture, the IYT group reduced the dopaminergic cell loss against 6-OHDA toxicity and the production of No and TNF-${\alpha}$ at the concentration of $0.2{\mu}g/m{\ell}$. Conclusion: These results showed that IYT has antioxidant and neuroprotectctive effects in the dopaminergic cells through decreasing the production of ROS, NO and TNF-${\alpha}$ which can cause many neurodegenerative changes in brain cell.

• Bulletin of the Korean Chemical Society
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• 제34권9호
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• pp.2616-2622
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• 2013

A novel zwitterionic monomer 3-[(2-acrylamido)dimethylammonio]propanesulfonate (DMADAS) was designed and synthesized in this study. Then it was polymerized with acrylamide (AM) by free radical polymerization in 0.5 mol/L NaCl solution with ammonium persulfate ($(NH_4)_2S_2O_8$) and sodium sulfate ($NaHSO_3$) as initiator. The structure and composition of DMADAS and acrylamide-3-[(2-acrylamido)-dimethylammonio]propanesulfonate copolymer (P-AM-DMADAS) were characterized by FT-IR spectroscopy, $^1H$ NMR and elemental analyses. Isoelectric point (IEP) of P-AM-DMADAS was tested by nanoparticle size and potential analyzer. Solution properties of copolymer were studied by reduced viscosity. Antipolyelectrolyte behavior was observed and was found to be enhanced with increasing DMADAS content in copolymer. The results showed that the viscosity of P-AM-DMADAS is 5.472 dl/g in pure water. Electrolyte was added, which weakened the mutual attraction between sulfonic acid group and quaternary ammonium group. The conformation became loose, which led to the increase of reduced viscosity. The ability of monovalent and divalent cation influencing the viscosity of zwitterionic copolymer obeyed the following sequence: $Li^+$ < $Na^+$ < $K^+$, $Mg^{2+}$ < $Ca^{2+}$ < $Ba^{2+}$, and that of anion is in the order: $Cl^-$ < $Br^-$ < $I^-$, $CO{_3}^{2-}$ > $SO{_3}^{2-}{\approx}SO{_4}^{2-}$.

• 동의생리병리학회지
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• 제27권4호
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• pp.391-399
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• 2013

The aim of the present study was to investigate the antioxidant efficacies between extracts of Prunus yedoensis(PY) and Betulae platyphyllae var. japonica(BP). HPLC pattern was different between barks and extract solvents. Content of total phenolic compound was the highest in ethanol extract of BP(382.201 mg/g ext.) and its content was 1.9 times higher than that from the water extract of PY. Total antioxidant efficacy also was the highest in ethanol extract of BP (292 copper reducing equivalents). Nitric oxide scavenging activity was almost 70% in ethanol extract of BP treated 200 ug/ml and it was higher than positive control(ascorbic acid). DPPH radical scavenging ability was up by 80% in all samples. ABTS cation decolorization from each barks was activated over 85% in all samples at 100 ug/ml concentration, especially, the activity was the highest (94.4%) in ethanol extract of BP. Hydrogen peroxide scavenging activities were also highest (45%) in ethanol extract of BP at 200 ug/ml concentration and were as high as positive control. Stimulation of the macrophages RAW 264.7 cells with lipopolysaccharide (LPS) increased intracellular ROS levels and ethanol extract of BP at 200 ug/ml concentration reduced ROS levels up to 41 %. The results indicated that the barks of PY and BP has potent antioxidant activities and ethanol extract of BP of them has the highest antioxidant activities.

• Journal of Applied Biological Chemistry
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• 제60권2호
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• pp.133-136
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• 2017

Fermentation plays a vital role in the nutritional enrichment of food. Korea has a long tradition of adding fermented food to the daily diet and jeotgal is one of the common fermented and salted foods in Korean cuisine. In our study, we added soymilk as an additive to squid jeotgal to improve its functionality. We mixed different concentrations of soymilk (2, 5, and 10 mg/g) with squid jeotgal samples, fermented them for one week, and then tested their antioxidant and anticancer activities to compare with those of squid jeotgal samples without soymilk additive. To investigate the anticancer characteristics, glutathione-S-transferase (GST)-pi enzyme assay was used. To test the antioxidant activities, various assays were performed, including 2,2-diphenyl-1-picryl hydrazyl free radical scavenging activity, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium saltradical cation scavenging assay, and reducing power assay. Samples fermented with a small amount of soymilk showed excellent anticancer activity. The addition of only 2 mg/g of soymilk to squid jeotgal inhibited the activity of GST-pi by almost 50% when compared with the sample with no addition. Moreover, no undesirable bitterness or astringency was noticed. Our results could help to improve the current food status of squid jeotgal and it could be used to reduce the risk of chronic disease along with its basic nutritional function.

• Food Science and Biotechnology
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• 제16권5호
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• pp.822-827
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• 2007

The aroma characteristics and antioxidative activity of volatile compounds in heat-treated garlic (Allium sativum L.) were evaluated. The garlic was heated to various temperatures (100, 110, 120, and $130^{\circ}C$) for different lengths of time (1, 2, and 3 hr). The volatile compounds of heated garlic were extracted by simultaneous steam distillation extraction (SDE). Aroma compound profiles were analyzed by gas chromatography/mass spectrometry (GC/MS) and antioxidative activity was measured by 2,2-diphenyl-2-picrylhydrazyl (DPPH) assay and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) cation decolorization assay. The major aroma compounds were sulfur compounds such as dimethyl disulfide, 2-propen-1-ol, methyl-2-propenyl disulfide, dimethyl trisulfide, diallyl disulfide, methyl-2-propenyl trisulfide, and di-2-propenyl trisulfide. DPPH radical scavenging activity (EDA, %) and the ascorbic acid equivalent antioxidant activity (AEAC) of volatile compounds in heated garlic increased significantly with the increase of temperature and time (p<0.001). The EDA (%) and AEAC of raw garlic were 26.8%/10 mg garlic and 39.05 mg ascorbic acid equivalent per g sample. After heat treatment, the highest values were 40.50%/10 mg garlic for EDA (%) and 46.43 mg ascorbic acid equivalent per g sample for ABTS.

• Preventive Nutrition and Food Science
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• 제20권1호
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• pp.22-28
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• 2015

Phenolic rich ethyl acetate fraction (EAF) from lotus leaves was prepared and its bioactive components, antioxidant and cytoprotective effects were investigated. EAF showed high total phenolic content and flavonoid content and contained rutin ($11,331.3{\pm}4.5mg/100g\;EAF$), catechin ($10,853.8{\pm}5.8mg/100g\;EAF$), sinapic acid ($1,961.3{\pm}5.6mg/100g\;EAF$), chlorogenic acid ($631.9{\pm}2.3mg/100g\;EAF$), syringic acid ($512.3{\pm}2.5mg/100g\;EAF$), and quercetin ($415.0{\pm}2.1mg/100g\;EAF$). EAF exerted the $IC_{50}$ of $4.46{\mu}g/mL$ and $5.35{\mu}g/mL$ toward DPPH and ABTS cation radicals, respectively, and showed strong reducing power, which was better than that of ascorbic acid, a positive control. Additionally, EAF protected hydroxyl radical-induced DNA damage indicated by the conversion of supercoiled pBR322 plasmid DNA to the open circular form and inhibited lipid peroxidation of polyunsaturated fatty acid in a linoleic acid emulsion. In cultured hepatocytes, EAF exerted a cytoprotective effect against oxidative stress by inhibiting intracellular reactive oxygen species formation and membrane lipid peroxidation. In addition, depletion of glutathione under oxidative stress was remarkably restored by treatment with EAF. The results suggest that EAF have great potential to be used against oxidative stress-induced health conditions.

• 대한본초학회지
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• 제23권3호
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• pp.19-25
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• 2008

Objectives : This study was performed to evaluate the neuroprotective effect of water extracts from Thujae Semen(TSW) in PC12 cells. Methods : We performed 2,2-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging assay, 2,2-azinobis- (3-ethyl-benzothiazoline-6-sulfonic acid(ABTS) cation scavenging assay, and determination of total polyphenolic content to examine the antioxidant effects of TSW. We also carried out 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay(MTT), reactve oxygen species(ROS) assay, and nitric oxide(NO) assay to examine neuroprotective effects against 6-hydroxydopamine(6-OHDA) in PC12 cells. Results : TSW showed $IC_{50}$ values of 404.3 and 219.9 ${\mu}g/mL$ in DPPH and in ABTS assays, respectively. TSW showed 9.74 ${\mu}g/mL$ of total polyphenol contents. TSW incresed cell viability in a dose dependent manner and it showed protective effect against 6-OHDA neurotoxicity at the concentration of 25-200 ${\mu}g/mL$. Moreover, it recovered 6-OHDA induced cell death at the same concentrations. The extract showed a dose dependent reduction of ROS and NO generation by 6-OHDA. Conclusions : We concluded that TSW has neuroprotective effect against 6-OHDA-induced toxicity in PC12 cells through ROS and NO inhibition.

• 대한본초학회지
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• 제24권2호
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• pp.87-92
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• 2009

Objectives : This study was performed to evaluate the neuroprotective effect of water extracts from Nelumbinis semen (NSW) in dopaminergic cells. Methods : We performed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azinobis3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) cation scavenging assay, and determination of total polyphenolic content to examine the antioxidant effects of NSW. We also evaluated the neuroprotective effects against 6-hydroxydopamine (6-OHDA)-induced toxicity using 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazolium bromide assay (MIT) assay, trypan blue cytotoxicity assay, and nitric oxide assay in SH-SY5Y cells and tyrosine hydroxylase (TH) immunohistochemistry in primary rat dopaminergic neurons. Results : NSW showed $IC_{50}$ values of 184.80 and 92.90 ${\mu}$g/mL in DPPH and in ABTS assays, respectively. NSW showed 1.05% of total polyphenol contents. NSW showed protective effect against 6-0HDA-induced neurotoxicity whereas no influence on cell viability at the concentration of 1${\sim}$50 ${\mu}$g/mL. NSW reduced NO generation while 6-OHDA produced it. Moreover, it protected rat dopaminergic neurons against 6-0HDA at a dose of 1 ${\mu}$g/mL. Conclusions : These results indicated that NSW has neuroprotective effect against 6-0HDA-induced neurotoxicity through antioxidant activity in dopaminergic cell culture.

• 한국미생물·생명공학회지
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• 제44권4호
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• pp.452-460
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• 2016

함초를 이용한 고부가가치 식의약 소재 개발 연구의 일환으로, 성숙 함초로부터 씨를 회수하고 이의 성분분석 및 유용 생리활성을 평가하였다. 함초 씨의 열수 추출효율은 29.6% 이었으며, 추출물의 total polyphenol (TP) 및 total flavonoid (TF) 함량은 각각 25.7 및 11.5 mg/g을 나타내어 성숙기 함초와 유사한 함량을 보였다. Hexane, ethylacetate (EA), butanol을 이용한 순차적 유기용매 분획물 중 EA 분획이 158.3 및 136.2 mg/g의 가장 높은 TP 및 TF 함량을 보였으며, 특이하게 물 잔류물 및 butanol 분획물보다 높은 총당 함량(228.3 mg/g)을 나타내었다. 함초 씨 시료의 항균 활성 평가 결과, EA 분획에서 그람양성 세균에 대한 광범위한 항세균 활성이 나타났으며, butanol 분획에서도 Staphylococcus epidermidis에 대한 강력한 항균활성을 확인하였다. 한편, 함초 씨 열수 추출물은 우수한 radical 소거능과 환원력을 나타내었으며, 가장 강력한 항산화 활성은 EA 분획에서 나타났다. EA 분획의 DPPH 음이온 소거능, ABTS 양이온 소거능 및 Nitrite 소거능에 대한 $RC_{50}$은 각각 57.0, 29.0 및 $28.9{\mu}g/ml$로 확인되어, vitamin C의 10.7, 4.0 및 $18.0{\mu}g/ml$보다 약하지만 우수한 항산화능을 확인하였다. 항혈전 활성 평가결과, EA 분획은 5 mg/ml 농도에서 thrombin time을 무첨가구에 비해 15배 이상, 7 mg/ml 농도에서 activated partial thromboplastin time을 15배 이상 연장시켜 아스피린에 필적하는 강력한 항응고 활성을 나타내었다. 반면 함초씨 추출물과 분획물(0.25 mg/ml)은 모두 혈소판 응집저해 활성은 나타나지 않았으며, 1 mg/ml 농도까지 인간 적혈구 용혈활성도 나타나지 않았다. 상기의 연구결과는 함초 씨 추출물의 EA 분획의 활성물질을 정제하여 항세균 및 항혈전제 개발이 가능함을 제시하고 있다.

• 생명과학회지
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• 제21권6호
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• pp.858-864
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• 2011

호두과육으로부터 용매별로 추출한 추출액중의 phenolic compound 함량은 열수, 에탄올, 메탄올, 아세톤 추출물 순서로 24.3, 34.4, 32.5, 15.1 mg/g의 함량을 나타내어 에탄올 추출물의 phenolic compound 함량이 가장 높았으며, 인체에 무해한 추출 용매인 열수 및 에탄올 농도별 추출물의 phenolic compound 비교한 결과 60% 에탄올 추출물에서 34.7 mg/g과 열수 추출물에서 24.6 mg/g 함량의 phenolic compound를 나타내었다. 호두추출물의 항산화 활성 중 전자공여능의 경우 열수추출물에서 78.1%, 60% 에탄올추출물에서 80.6%의 활성을 나타내었으며, ABTS radical cation decolorization 측정에서는 열수추출물과 60% 에탄올추출물 각각 98.1%와 98.3%의 높은 항산화 활성을 나타내었다. Antioxidant protection factor (PF)의 경우는 열수추출물과 60% 에탄올추출물에서 각각 $1.1{\pm}0.2$ PF와 $1.1{\pm}0.4$ PF로 높지는 않았으며, TBARs inhibitory activity 측정에서는 열수추출물과 60% 에탄올추출물에서 각각 60%와 75%로 비교적 높은 항산화력을 가졌음이 확인되었다. 호두추출물의 항염증효과를 살펴본 결과 100 ${\mu}g/ml$의 농도에서 50%의 NO발현이 억제되었고, western blot에 의한 iNOS 및 COX-2의 억제효과는 대조군에 비해 100 ${\mu}g/ml$의 농도에서 10% 정도의 낮은 저해 효과를 나타내었다. Tyrosinase의 저해 활성의 경우 60% 에탄올추출물에서 43%의 저해율을 보였으며, 수렴효과의 경우 60% 에탄올추출물에서 55%의 억제효과를 나타내어, 호두추출물은 미백 및 주름개선용 화장품소재로서 개발이 가능하다고 판단되었다. 본 연구는 고령친화산업 기업지원센터의 연구비 지원에 의하여 수행되었으며, 이에 감사드립니다.