• 대한한의학회지
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• 제21권3호
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• pp.174-185
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• 2000

Objectives : This study was carried out to evaluate the effects of five fractions on cell viability, cell cycle progression and apoptosis. Methods : This study employed MTT assay, Cell cycle analysis, Cpp32 protease assay, DNA fragmentation assay and Quantitative RT-PCR analysis. Results : In MTT assay, the butanol fraction of Injinsaryung-San has showed magnificent viability, while the $H_2O$ fraction and ethylacetate fraction also showed higher viability than the control group. The $H_2O$ fraction of Injinsaryung-San has showed magnificent viability, and butanol fraction and ethylacetate fraction of Injinsaryung-San with etoposide have also showed higher viability than the only etoposide group. Cell cycle analysis showed that each fraction of Injinsaryung-San had no significant effect on the cell cycle. DNA fragmentation assay showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction carried inhibitory effects on apoptosis induction. Cpp32 protease activity assay showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction decreased Cpp32 protease activity, with the butanol fraction displaying greater effects. Quantitative RT-PCR showed that the butanol fraction, $H_2O$ fraction and ethylacetate fraction suppressed Fas and Bax genes, the butanol fraction increased BcI-2 gene, however no effect on Cpp32. Conclusions : The data shows that the butanol fraction of Injinsaryung-San increases the hepatocyte viability and has the heptocelluar protective effect by the suppression of apoptosis through gene regulation.

• Biomolecules & Therapeutics
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• 제5권4호
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• pp.331-335
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• 1997

In a preliminary screening of plant extracts for the antigastritic and antiulcer actions in rats, the methanol extracts of Taheebo showed positive activity in HCI . ethanol-induced gastric lesion. Among the systematic fractions of hexane, chloroform, butanol and water, the most potent $H_2O$ fraction reduced significantly HCI . ethanol-induced gastric lesion at the oral dose of 300 mg/kg. In pylorus ligated rats chloroform and butanol fraction showed decreases in the volume of gastric secretion and acid output of which effects were stronger in chloroform fraction. Further assays with hexane butanol and $H_2O$ fraction disclosed that it significantly suppressed the aspirin-induced ulcer. The butanol fraction reduced significantly acetic acid induced ulcer at the dose of 400 mg/kg. The butanol and $H_2O$ fraction reduced the malondialdehyde level in HCI . ethanol-induced gastric lesion. In pylorus ligated rats, chloroform and butanol fraction reduced the malondialdehyde level and in aspirin-induced ulcer, chloroform fraction reduced that levle. These results might suggest that the butanol and $H_2O$ fraction of Taheebo had inhibitory action in gastric lesion and ulceration through inhibition of gastric acid secretion and the decrease malondialdehyde level.

• Journal of the Korean Wood Science and Technology
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• 제43권5호
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• pp.591-599
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• 2015

Cercidiphyllum japonicum leaves were collected, air-dried and extracted with 70% aqueous acetone, then concentrated and sequentially fractionated using n-hexane, methylene chloride ($CH_2Cl_2$), ethylacetate (EtOAc), and $H_2O$. A portion of EtOAc fraction (10 g) was chromatographed on a Sephadex LH-20 column, by the successively elution with various aqueous $MeOH-H_2O$ (1:9, fraction $1-2{\rightarrow}3:7$, fraction $3-5{\rightarrow}1:1$, fraction $6-9{\rightarrow}7:3$, fraction $10-13{\rightarrow}9:1$, fraction 14-16). Compound 2 was isolated from fraction 6 and compound 1 was separated from fraction 11 and 12. Compound 3 and 4 were purified from fraction 13. The isolated compounds were elucidated as quercetin-3-O-${\alpha}$-L-rhamnopyranoside (1), chlorogenic acid (2), quercetin-3-O-${\alpha}$-L-arabinofuranoside (3) and quercetin-3-O-${\beta}$-D-xylopyranoside (4) by the spectral and literature data, and by comparison with the authentic samples. These compounds were reported, for the first time, from the extracts of C. japonicum leaves. Also chlorogenic acid (2) has never been reported before in domestic tree species and can be used as an index compound for C. japonicum.

• 한국식품위생안전성학회지
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• 제11권4호
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• pp.235-241
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• 1996

The rhamnan sulfate extracted from green algae seaweed, Monostroma nitidum was characterized as activity in vitro culture assay with macrophages from mice. Rhamnan sulfate indicated that F-4-3 fraction enhanced glucose consumption, as well as the production of nitrogen dioxide and tumor necrosis factor(TNF). F-4-3 fraction was also augmented IL-1 secretion from those macrophages. Effects of the pretreatment of peritoneal macrophages with rhamnan sulfate F-4-3 fraction and several polysaccharides as relative standard on the production of H2O2 induced with unopsonized zymosan A were examined. Pretreatment with polysaccharides inhibited the zymosan A mediated H2O2 production by macrophages. The phorbol myristate acetate (PMA) mediated H2O2 production was not affected by the pretreament. These result suggested that pretreatment of rhamnan sulfate interfered with the interaction of macrophages zymosan A. Rhamnan sulfate inhibited zymosan A mediated production of H**O** by macrophages and F-4-3 Fraction was also activator of macrophages.

• 한국식품영양과학회지
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• 제30권1호
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• pp.97-101
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• 2001

This study was done to investigate the protective effects of Diospyrus kaki Folium on carbon tetrachloride intoxicated rats. Male Sprague-Dawley rats (200~230g) was used. Experimental groups were divided into normal group and $CCl_{4}$-treated group and then $CCl_{4}$-treated groups were divided into 7 groups: only $CCl_{4}$-treated group, methanol extract pretreated group, butanol fraction pretreated group and $H_{2}O$ fraction pretreated group, $CCl_{4}$ treated groups were injected with $CCl_{4}$ 0.6 mg/kg (ip). The activities of ananine aminotransferasein hexane, chloroform and $H_{2}O$ fraction pretreated groups were significantly decreased (p<0.05) compared to the only $CCl_{4}$-treated group and also activity of aspartate aminotransferase and hepatic malondialdehyde content in $H_{2}O$ fraction pretreated group were significantly decreased (p<0.05) compared to the only $CCl_{4}$-treated group. The activity of alkaline phosphatase in chloroform and $H_{2}O$ fraction pretreated groups were decreased compared to the only $CCl_{4}$-treated group but the difference was not statistically significant. The activity of ${\gamma}$-glutamyltranspeptidase in hexane, chloroform and ethylacetate fraction pretreated groups were significantly decreased (p<0.05) compared to the only $CCl_{4}$-treated group. The contents of cholesterol and triglyceride were not influenced in any fraction pretreated groups, but HDL-cholesterol content in hexane and chloroform fraction pretreated groups were significantly increased (p<0.05) compared to the only $CCl_{4}$-treated group. Therefore, these results demonstrate a possible hepatoprotective role of $H_{2}O$ fraction of Diospyrus kaki Folium methanol extract in carbon tetrachloride intoxicated rats.

• 한국작물학회지
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• 제59권4호
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• pp.435-444
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• 2014

본 연구는 옥수수수염에 함유된 flavonoid 계열의 물질인 maysin, luteolin 7-O-neohesperidoside, luteolin 3'-methyl ether 7-glucuronosyl-($1{\rightarrow}2$)-glucuronide 및 polyphenol성 물질인 chlorogenic acid을 추출 및 분리하는 방법에 관한 것으로, 얻어진 결과를 요약하면 다음과 같다. 1. 수정되지 않은 옥수수수염의 메이신 함량은 출사 후 3일에 함량이 최대치에 도달하고, 그 후 감소되는 것으로 나타났으며, 방임수분된 옥수수수염의 메이신 함량은 출사후 지속적으로 감소되는 것으로 나타나 화분의 수분 여부에 따라 메이신 함량에 많은 차이가 있음을 알 수 있었다. 2. 채취한 옥수수수염에 에탄올을 가하여 9일간 상온에서 추출 후 엽록소, 지질 및 당질 등을 제거시키고, $C_{18}$ column chromatography를 수행하여 분취물 I, II, III, IV를 얻었다. 3. 분취물의 흡광도를 분석한 결과 분취물 I은 327 nm 및 239 nm에서 최대 흡수 파장(${\lambda}_{max}$)을 나타내었고, 분취물 II는 339 nm 및 274 nm, 분취물 III는 345 nm 및 277 nm, 분취물 IV는 352 nm, 270 nm, 및 257 nm 에서 최대 흡수파장을 나타내었다. 4. 분취물 I은 m/z $355[M+H]^+$인 chlorogenic acid(3-(3,4-dihydroxycinnamoyl)quinic acid, $C_{16}H_{18}O_9$)이었으며, 분취물 II는 chlorogenic acid와 m/z $653[M+H]^+$인 luteolin 3'-methyl ether 7-glucuronosyl-($1{\rightarrow}2$)-glucuronide ($C_{28}H_{28}O_{18}$)를 함유한 혼합물, 분취물 III는 chlorogenic acid와 m/z $595[M+H]^+$인 luteolin 7-O-neohesperidoside ($C_{27}H_{30}O_{15}$) 및 luteolin 3'-methyl ether 7-glucuronosyl-($1{\rightarrow}2$)-glucuronide를 함유한 혼합물이었고, 분취물 IV는 m/z $577[M+H]^+$인 maysin ($C_{27}H_{28}O_{14}$, 2"-O-${\alpha}$-L-rhamnosyl-6-C-(6-deoxy-xylo-hexose-4-ulosyl)luteolin 임을 각각 확인 하였다. 5. 옥수수수염의 알코올 추출물로부터 분리된 분취물 I은 옥수수수염 생체 100 g 당 (수분함량 약 92%) 35 mg/100 g, 분취물 II는 48 mg/100 g, 분취물 III는 46 mg/100 g, 분취물 IV는 138 mg/100 g을 얻을 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1090-1097
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• 2017

Rhus verniciflua Stokes (RVS), an herbal medicine found in East Asia, was extracted and further fractionated to investigate its antioxidant capacity and neuroprotective effects. The RVS ethyl acetate (EtOAc) fraction had the highest level of total phenolics and antioxidant capacity among all solvent fractions tested. Pretreatment of PC-12 cells with the EtOAc fraction effectively attenuated $H_2O_2$-induced oxidative damage. Furthermore, the EtOAc fraction significantly attenuated caspase-3 activity, resulting in inhibition of $H_2O_2$-induced apoptosis. We identified and quantified fustin, sulfuretin, and butein in the EtOAc fraction using accurate mass quadrupole time-of-flight mass spectrometry and reversed-phase high-performance liquid chromatography. The intracellular antioxidant capacity and superoxide dismutase (SOD) activity were significantly increased in PC-12 cells treated with the EtOAc fraction and with individual flavonoids. When cells were pretreated with the EtOAc fraction or individual flavonoids and then co-incubated with diethyldithiocarbamic acid (an inhibitor of SOD activity), cell viability against $H_2O_2$-induced oxidative stress was attenuated. These results suggest that the RVS EtOAc fraction and its flavonoid constituents protect PC-12 cells against $H_2O_2$-induced neurotoxicity through their antioxidant properties.

• 대한한의학회지
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• 제21권1호
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• pp.53-61
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• 2000

Objectives: This study was camed out to examine the effect of five fractions of aqueous extract from Artemisia capillaris THUNB(ACT), on TGF, 1-induced apoptosis, cell viability, cell cycle progression and mRNA expression of apoptosis-related genes in human hepatocyte cell line HepG2. Methods: This study employed Tryphan blue exclusion assay, DNA fragmentation assay, Cpp32 protease activity assay and Quantitative RT-PCR analysis. Results: In the Tryphan blue exclusion assay, the butanol fraction of ACT with $TGF{\beta}$, l showed magnificent (Nice word, ut is it appropriate in a medical abstract\ulcorner) viability and the H2O fraction of ACT with $TGF{\beta}$, l also showed higher viability than only $TGF{\beta}$, l-treated group. DNA fragmentation assay showed that the butanol fraction and the H2O fraction carried inhibitory effects on apoptosis induction, with the butanol fraction displaying greater effects. The Cpp32 protease activity assay showed that the butanol fraction decreased Cpp32 protease activity. The H2O fraction of ACT had no significant effect on the Cpp32 protease activity. Quantitative RT-PCR showed that the butanol fraction suppressed Bax, p 15/INK4B, p21/Waf1, PAI-1 and increased Bcl-2 gene. Conclusions: The data shows that butanol fraction of ACT increases the hepatocyte viability and has the hepatocellular protective effect by the suppression of $TGF{\beta}$, l induced-apoptosis through gene regulation.

• Fisheries and Aquatic Sciences
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• 제13권1호
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• pp.26-35
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• 2010

In our previous study, we preliminarily demonstrated that Celluclast and Neutrase extracts exhibited the strongest $H_2O_2$-scavenging activities among five carbohydrases (Viscozyme, Celluclast, Termamyl, Ultraflo and AMG) and five proteases (Kojizyme, Alcalse, Flavourzyme Protamex and Neutrase) extracts. Thus, Celluclast and Neutrase extracts were selected for use in further experiments and were separated into four different molecular weight fractions (<5, 5-10, 10-30 and >30 kDa). Among them, the 5-10 kDa fraction showed the highest $H_2O_2$-scavenging activity. The 5-10 kDa fraction also strongly enhanced cell viability against $H_2O_2$-induced oxidative damage. Furthermore, the fraction reduced the proportion of apoptotic cells induced by $H_2O_2$, as demonstrated by decreased sub-G1 hypodiploid cells and decreased apoptotic body formation by flow cytometry. These results indicated that the 5-10 kDa fraction of the Celluclast and Neutrase extracts from S. coreanum exhibited strong antioxidant activity over $H_2O_2$-mediated cell damage in vitro.

• Journal of the Korean Wood Science and Technology
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• 제37권5호
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• pp.480-485
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• 2009

Juglans mandshurica bark was collected and extracted with 70% aqueous acetone. The extracts were concentrated and then sequentially fractionated using n-hexane, $CH_2Cl_2$, EtOAc, and $H_2O$ to be freeze dried. A portion of $H_2O$ fraction was chromatographed on a Sephadex LH-20 column using aqueous methanol to isolate (S)-2,3-HHDP-D-glucopyranoside (1). Some of EtOAc fraction was also treated on a Sephadex LH-20 column using aqueous methanol and EtOH-hexane mixture to purify pedunculagin (2). The structures of these two epimeric ellagitannins were elucidated by spectroscopic tools, NMR and MS.