• Natural Product Sciences
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• 제19권1호
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• pp.36-48
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• 2013

The aim of the present study was to investigate the effects of several fractions obtained from methylene chloride ($CH_2Cl_2$) extract of self-fermented pine needle (SFPNE) on the acetylcholine (ACh)-evoked CA release from the isolated perfused model of the rat adrenal medulla and to establish the mechanism of the most active fraction (Fr.)-induced inhibitory action on the CA release. We obtained 6 fractions from $CH_2Cl_2$ extract of self-fermented pine needle. For the ACh (5.32 mM)-evoked CA release, the following rank order of inhibitory potency was obtained: Fr.4-5 > Fr.8-11 ${\gg}$ Fr.3 > Fr.6 = Fr.7 > Fr.1-2. Fr. 4 - 5 (60 ${\mu}g/mL$) perfused into an adrenal vein for 90 min produced relatively time-dependent inhibition of the CA secretory responses to ACh (5.32 mM), DMPP (100 ${\mu}M$), McN-A-343 (100 ${\mu}M$) and high $K^+$ (56 mM). Fr. 4 - 5 itself did not affect basal CA secretion. Also, in the presence of Fr. 4 - 5 (60 ${\mu}g/mL$), the CA secretory responses to angiotensin II (AngII, 0.1 ${\mu}M$), veratridine (50 ${\mu}M$), Bay-K-8644 (10 ${\mu}M$), and cyclopiazonic acid (10 ${\mu}M$) were significantly reduced, respectively. In the simultaneous presence of Fr. 4 - 5 (60 ${\mu}g/mL$) and L-NAME (30 ${\mu}M$), the inhibitory responses of Fr. 4 - 5 on the CA secretion evoked by ACh, DMPP, high $K^+$, AngII, Bay-K-8644 and veratridine were considerably recovered to the extent of the corresponding control secretion compared with that of Fr. 4 - 5-treatment alone. The level of NO released from adrenal medulla after the treatment of Fr. 4 - 5 (60 ${\mu}g/mL$) was greatly elevated compared with the basal level. Taken together, these results demonstrate that Fr. 4 - 5 inhibits the CA secretion from the isolated perfused rat adrenal medulla evoked by stimulation of cholinergic receptors as well as by direct membrane-depolarization. It seems that this inhibitory effect of Fr. 4 - 5 is mediated by blocking the influx of $Ca^{2+}$ and $Na^+$ into the adrenomedullary chromaffin cells as well as by inhibition of $Ca^{2+}$ release from the cytoplasmic calcium store, which is evoked at least partly through the increased NO production due to the activation of NO synthase. Based on these results, it is also thought that Fr. 4 - 5 isolated from $CH_2Cl_2$ extract of pine needle may contain beneficial antihypertensive components to prevent or treat hypertension.

• 동의생리병리학회지
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• 제27권5호
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• pp.602-610
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• 2013

This study was aimed to evaluate the cavernosal relaxation effect of Crataegii fructus(CF) in the contracted rabbit penile corpus cavernosum by agonists.In order to study the effect of CF on the vasoconstriction of rabbit penile corpus cavernosum, isolated rabbit penile corpus cavernosum tissues were used for the experiment using organ baths containing Krebs solution.To investigate the cavernosal relaxation of CF, CF extract at $0.01{\sim}3.0mg/m{\ell}$ was added after penile corpus cavernosum were contracted by norepinephrine(NE) $1{\mu}M$. To analyze the mechanism of CF's vasorelaxation, CF extract infused into contracted penile tissues by NE after each treatment of indomethacin(IM), $N{\omega}$-nitro-L-arginine(L-NNA), methylene blue(MB), tetraethylammonium chloride(TEA).To study the effect of CF on influx of extracellular calcium chloride($Ca^{2+}$) in penile tissues, in $Ca^{2+}$-free krebs solution, $Ca^{2+}$ 1 mM infused into contracted penile tissues by NE after pretreatment of CF. Cytotoxic activity of CF on human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, and nitric oxide(NO) prodution was measured by Griess reagent. CF relaxed cavernosal strip with endothelium contracted by NE, but in the strips without endothelium, CF-induced relaxation was significantly inhibited. The pretreatment of L-NNA, MB, TEA decreased significantly on the cavernosal relaxation than not-treatment of them. But the pretreatment of IM had no significant effect on the cavernosal relaxation. In $Ca^{2+}$-free krebs solution, when $Ca^{2+}$ infused into contracted penile tissues by NE, pretreatment of CF inhibit contraction induced by adding $Ca^{2+}$.NO production wasn't increased by treatment of CF on HUVEC. This findings showed that CF is effective for the relaxation of rabbit penile corpus cavernosum, and we suggest that CF relax rabbit corpus cavernosal smooth muscle through multiple action mechanisms that include increasing the release of nitric oxide from corporal sinusoidal endothelium, inhibition of $Ca^{2+}$ mobilization into cytosol from the extracellular fluid, and maybe a hyperpolarizing action.

• 한국환경농학회지
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• 제37권2호
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• pp.117-124
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• 2018

본 연구에서는 HPLC-UVD/MS를 이용하여 농산물 중 oxanthiin 살균제인 oxycarboxin의 잔류 분석법을 확립하였다. 대표 농산물은 현미, 콩, 배추, 고추 및 사과를 선정하였고 acetone을 가하여 추출된 oxycarboxin 성분을 dichloromethane 액-액 분배법과 Florisil 흡착크로마토그래피법으로 정제하여 HPLC-UVD/MS 분석대상 시료로 사용하였다. Oxycarboxin의 정량적 분석을 위한 최적 분석 조건을 확립하였으며, 정량한계(LOQ)는 0.04 mg/kg 이었다. 각 대표 농산물에 대해 정량한계, 정량한계의 10배 및 50배 수준에서 회수율을 검토한 결과 모든 처리농도에서 78.3~96.1% 수준을 나타내었으며, 반복 간 변이계수(CV)는 최대 4.7%를 나타내어 잔류분석 기준인 회수율 70~120% 및 분석오차 10% 이내를 충족시키는 만족한 결과를 도출하였으며, LC/MS SIM을 이용하여 실제 농산물 시료에 적용하여 재확인 하였다. 이상의 결과로 신규 oxycarboxin의 HPLC-UVD/MS 분석법은 검출한계, 회수율 및 분석오차 면에서 국제적 분석기준을 만족하는 신뢰성이 확보된 정량 분석법으로 사용 가능할 것이다.

• 생약학회지
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• 제46권3호
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• pp.234-242
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• 2015

Allium hookeri (Liliaceae), native to India is widely cultivated in Korea lately as a medicinal plant. This study was performed to evaluate the antioxidant and lifespan extending effects of the root of A. hookeri. Methanol extract from the root of A. hookeri was successively partitioned as methylene chloride, ethyl acetate, n-butanol and H₂O soluble fractions. Antioxidant effects of the fractions were measured by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH), and superoxide quenching activities. Lifespan extending effects of the fractions were investigated using Caenorhabditis elegans model system. In addition, stress resistant, superoxide dismutase (SOD) and catalase activities, and intracellular reactive oxygen species (ROS) levels were also studied. Furthermore, we had investigated aging-related factors such as reproduction, food intake, growth and movement of C. elegans to find any sample-induced significant change. Our results revealed that ethyl acetate soluble fraction from the root of A. hookeri showed the most potent antioxidant and lifespan extending effects, and this fraction elevated the most potent SOD and catalase activities of worms, and reduce intracellular ROS accumulation.

• 생약학회지
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• 제46권3호
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• pp.214-222
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• 2015

Recently, many studies have focused on the aging and oxidative stress. Several papers reported that Vigna angularis has various biological properties including antiaging, antioxidative and anti-inflammatory activities. Methanol extract from the red bean sprouts was successively partitioned as n-hexane, methylene chloride, ethyl acetate, n-butanol and H₂O soluble fractions. We had studied lifespan extending and stress resistant effects of the fractions using Caenorhabditis elegans. Superoxide dismutase (SOD), catalase activities, and intracellular reactive oxygen species (ROS) levels were also investigated. Moreover, we had studied to find any significant change in aging-related factors such as reproduction, food intake, growth and movement of C. elegans. Our results represent that ethyl acetate fraction showed the most potent lifespan extending and stress resistant effects, and this fraction was able to elevate SOD and catalase activities of worms, and reduce intracellular ROS accumulation.

• 생약학회지
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• 제48권1호
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• pp.25-30
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• 2017

This study was performed to evaluate the lifespan extending effects of the dried ginger (Zingiber officnale Roscoe, Zingiberaceae). Ethanol extract of the dried ginger was successively partitioned as methylene chloride, ethyl acetate, n-butanol and $H_2O$ soluble fractions, and lifespan-extending effects of the fractions were investigated using Caenorhabditis elegans model system. Ethyl acetate-soluble fraction showed the most potent longevity property. Furthermore, ethyl acetate-soluble fraction elevated thermal stress tolerance, and increased expression of stress resistance protein. Further studies were performed to investigate several aging-related factors such as reproduction, food intake, growth and movement of C. elegans. The results revealed that there were no significant changes in aging-related factors including reproduction, food intake and growth, however, ethyl acetate-soluble fraction treatment led to an up-regulation of locomotory ability of aged worms, suggesting ethyl acetate-soluble fraction affected healthspan as well as lifespan of nematode.

• Korean Chemical Engineering Research
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• 제52권4호
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• pp.497-502
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• 2014

바이오매스 유래 타르 및 왁스 성분은 파클리탁셀의 분리정제에 많은 악영향을 미치기 때문에 반드시 제거되어야 한다. 흡착제 처리는 식물세포배양으로부터 유래된 타르 및 왁스 성분을 매우 간단하고 효과적으로 제거할 수 있는 방법이다. 본 연구에서는 전처리 단계에서 상용흡착제 실로퓨트를 이용한 흡착제 처리 공정에서 주요 공정 변수들(온도, 시간, 용매, 건조시료/실로퓨트 비율)을 최적화하였다. 최적의 흡착제 처리 온도, 시간, 용매, 건조시료/실로퓨트 비율은 각각 $30^{\circ}C$, 15분, 메틸렌 클로라이드, 1:1(w/w)이었다. 이러한 결과는 TGA와 HPLC 분석을 통한 유기물의 흡착 양상으로도 확인하였다. 흡착제 처리 단계에서 순도 증가는 미미하였으나 타르 및 왁스 성분의 제거로 인한 후속공정의 공정 가능성과 편리성에 상당히 영향을 미쳤다.

• Preventive Nutrition and Food Science
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• 제16권1호
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• pp.18-23
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• 2011

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by neuronal loss and extracellular senile plaques containing $\beta$-amyloid peptide (A$\beta$). The deposition of the A$\beta$ peptide following proteolytic processing of amyloid precursor protein (APP) by $\beta$-secretase (BACE1) and $\gamma$-secretase is a critical feature in the progression of AD. Among the plant extracts tested, the ethanol extract of Petasites japonicus leaves showed novel protective effect on B103 neuroblastoma cells against neurotoxicity induced by A$\beta$, as well as a strong suppressive effect on BACE1 activity. Ethanol extracts of P. japonicus leaves were sequentially extracted with methylene chloride, ethyl acetate and butanol and evaluated for potential to inhibit BACE1, as well as to suppress A$\beta$-induced neurotoxicity. Exposure to A$\beta$ significantly reduced cell viability and increased apoptotic cell death. However, pretreatment with ethyl acetate fraction of P. japonicus leaves prior to A$\beta$ (50 ${\mu}M$) significantly increased cell viability (p<0.01). In parallel, cell apoptosis triggered by A$\beta$ was also dramatically inhibited by ethyl acetate fraction of P. japonicus leaves. Moreover, the ethyl acetate fraction suppressed caspase-3 activity to the basal level at 30 ppm. Taken together, these results demonstrated that P. japonicus leaves appear to be a useful source for the inhibition and/or prevention of AD by suppression of BACE1 activity and attenuation of A$\beta$ induced neurocytotoxicity.

• 생약학회지
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• 제47권2호
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• pp.179-185
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• 2016

Ginger (Zingiber officnale Roscoe, Zingiberaceae), which is a well-known food seasoning, has been used as a traditional medicine for the treatment of gastrointestinal disorder, vomiting and cough in Korea, China and Japan. Ethanol extract from the dried ginger (DG) was successively partitioned as methylene chloride, ethyl acetate, n-butanol and $H_2O$ soluble fractions. Among those fractions the ethyl acetate soluble fraction (EDG) showed the most potent DPPH radical scavenging and superoxide quenching activities. To know the effect of antioxidant activities of EDG, we tested the activities of superoxide dismutase (SOD) and catalase together with oxidative stress tolerance and intracellular ROS level in Caenorhabditis elegans. To investigate whether EDG-mediated increased stress tolerance was due to regulation of stress-response gene, we quantified SOD-3 expression using transgenic strain including CF1553. Consequently, EDG elevated SOD and catalase activities of C. elegans, reduced intracellular ROS accumulation in a dose-dependent manner. Moreover, EDG-treated CF1553 worms exhibited significantly higher SOD-3::GFP intensity.

• Nutritional Sciences
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• 제9권3호
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• pp.159-166
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• 2006

This study was performed to investigate antioxidative and anti-aging action of extracts from Sancho (Zanthoxylum schinifolium) leaves. Two extracts were obtained by 80% methanol extraction followed by subsequent fractionation with methylene chloride (MC) and n-butanol (B) and fed at one or three levels to rats on normal level (5%) of fat (control) and high fat(20%) in diets. Male Sprague-Dawley rats weighing about 100 g were divided into ten groups such as control diet group(C), control diet+0.50%B group (CB), control diet+0.50%MC group (CMC), high-fat diet group (HF), high-fat diet+0.25%B group (HBL), high-fat diet+0.50%B group (HBM), high-fat diet+0.75%B group (HBH), high-fat diet+0.25%MC group (HMCL), high-fat diet+0.50%MC group (HMCM) and high-fat diet+ 0.75%MC group (HMCH) and fed each diet for four weeks. The effects of the extracts on antioxidant enzyme activities and indices of lipid peroxidation and aging were seen only in high fat diet groups. Hepatic superoxide dismutase and aryleaterase activities were not changed by Sancho extracts. But glutathione peroxidase, catalase and paraxonase activities were significantly restored by both MC and B at the level of 0.75% lipid peroxide which was increased by high fat diet was significantly reduced by B and MC at the level of 0.25% and over. Lipofuscin fluorescence and cabonyl value were increased by high fat diet were reduced by B and MC at the level of 0.5% and over. It is concluded that the Sancho extracts can be utilized as functional ingredients of health foods for reducing oxidative stress.