• 원예과학기술지
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• 제35권2호
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• pp.252-264
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• 2017

Tea is one of the most consumed beverages worldwide and the relatively high levels polyphenols is benefit for health. In this study, we developed an efficient system for proliferation of callus from 'Anji Baicha', a cultivar of tea (Camellia sinensis). Callus tissue was initially induced by culturing leaf explants on medium containing different plant growth regulators. For callus induction, thidiazuron (TDZ) was more effective than 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}-naphthalene$ acetic acid (NAA), and $N^6-benzyladenine$ (BA). The frequency of callus induction from leaf explants reached 90.21% on $1.0mg{\cdot}L^{-1}$ TDZ and the developed callus was reddish and friable. We also tested the effect of different concentrations of NAA, 2,4-D, indole 3-acetic acid (IAA), BA, and TDZ, alone and in combinations, on callus proliferation. Medium supplemented with TDZ in combination with IAA was suitable for callus proliferation and accumulation of tea polyphenols. The growth index value and tea polyphenol content of callus cultured on MS medium containing $0.5mg{\cdot}L^{-1}$ TDZ and $1.0mg{\cdot}L^{-1}$ IAA was maximally 1,351% and 23.24%, respectively, and the relative abundance of epicatechin was as high as 17.449%. We also measured the antioxidant activity of all samples and the callus with the highest tea polyphenol content also exhibited high potential radical scavenging activity.

• 동의생리병리학회지
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• 제31권3호
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• pp.159-166
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• 2017

Through this study, the authors investigated the anti-steatosis effects of the Amomum cardamomum ethyl acetate fraction in free fatty acids (FFAs)-induced human hepatocellular carcinoma HepG2 cells. The ethyl acetate fraction of Amomum cardamomum (ACEA) was extracted with 70% ethanol and then the extract was evaporated using a rotary evaporator prior to sequential fractionation. Human hepatocellular carcinoma were treated with different concentrations of ACEA in the presence and absence of FFAs. To demonstrate the reactive oxygen species (ROS) scavenging activity, DCFDA level was analyzed by using in vitro assay system. Cell viability, lipid accumulation, intracellular triglycerides, malondialdehyde (MDA), liver steatosis related signaling molecules and inflammatory cytokines such as interleukin (IL)-6, 8, tumor necrosis factor-alpha ($TNF-{\alpha}$) were also investigated. As results, ACEA inhibited the FFAs-induced ROS, lipid accumulation, intracellular triglycerides, and MDA in a dose dependent manner. Treatment of human hepatocellular cells with ACEA induced the phosphorylation of 5' adenosine monophosphate-activated protein kinase (AMPK) and carnitine palmitoyltransferase I (CPT1) expression using western blot analysis. ACEA also potently suppressed the FFAs-induced inflammatory cytokines including IL-6, IL-8 and $TNF-{\alpha}$. These results suggest that the ethyl acetate fraction of Amomum cardamoum extract own inhibitory effects of liver steatosis by inhibiting ROS, lipid accumulation, intracellular triglycerides, MDA through AMPK signaling and anti-inflammatory actions.

• 한국환경보건학회지
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• 제28권2호
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• pp.81-88
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• 2002

Effect of cyclohexanone treatment on the activities oxygen free radical and cyclohexanone metabolizing enzyme in acute liver damaged rats, was investigated. Acute liver damage was induced in rats with pretreatment of 50% $CCl_4$ in olive oil(0.1ml/100g body wt) intraperitoneally 3 times every other day. Cyclohexanone(1.56g/kg body wt, i.p.) was administered to the animals 24 hours after the last Pretreatment of CC1$_4$. Rats were sacrificed at 4 hours after injection of cyclohexanone. On the basis of liver weight/body weight(％), serum levels alanine aminotransferase activity and hepatic protein content, cyclohexanone treatment to acute liver damaged animals led to the more enhanced liver damage. On the other hand, injection of cyclohexanone to the rats led to the increased activities of hepatic cytochrome P-450 dependent aniline hydroxylase and xanthine oxidase. Furthermore, by treatment of cyclohexanone to the acute liver damaged rats hepatic xanthine oxidase activity was more increased than the $CCl_4$ treated rats. In case of oxygen free radical scavenging system, the hepatic glutathione content and the activities of hepatic glutathione S-transferase, catalase, superoxide dismutase were generally increased by injection of cyclohexanone to rats, and the hepatic glutathione content, catalase and alcohol dehydrogenase activities were more decreased in liver damaged rats by the treatment of cyclohexanone. In conclusion, the cyclohexanone treatment to acute liver damaged rats led to enhancement of liver damage that may be due to oxygen free radical together with cyclohexanone.

• Asian-Australasian Journal of Animal Sciences
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• 제25권6호
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• pp.818-823
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• 2012

Tall fescue (Festuca arundinacea Schreb.) is an important cool season forage plant that is not well suited to extreme heat, salts, or heavy metals. To develop transgenic tall fescue plants with enhanced tolerance to abiotic stress, we introduced an alfalfa Hsp23 gene expression vector construct through Agrobacterium-mediated transformation. Integration and expression of the transgene were confirmed by polymerase chain reaction, northern blot, and western blot analyses. Under normal growth conditions, there was no significant difference in the growth of the transgenic plants and the non-transgenic controls. However, when exposed to various stresses such as salt or arsenic, transgenic plants showed a significantly lower accumulation of hydrogen peroxide and thiobarbituric acid reactive substances than control plants. The reduced accumulation of thiobarbituric acid reactive substances indicates that the transgenic plants possessed a more efficient reactive oxygen species-scavenging system. We speculate that the high levels of MsHsp23 proteins in the transgenic plants protect leaves from oxidative damage through chaperon and antioxidant activities. These results suggest that MsHsp23 confers abiotic stress tolerance in transgenic tall fescue and may be useful in developing stress tolerance in other crops.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.2015-2020
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• 2012

Oleanolic acid (OA) is a naturally occurring triterpenoid in food materials and is a component of the leaves and roots of Olea europaea, Viscum album L., Aralia chinensis L. and more than 120 other plant species. There are several reports validating its antitumor activity against different cancer cells apart from its hepatoprotective activity. However, antitumor activity against skin cancer has not beed studied well thus far. Hence the present study of effects of OA against HaCaT (immortalized keratinocyte) cells - a cell-based epithelial model system for toxicity/ethnopharmacology-based studies - was conducted. Radical scavenging activity ($DPPH{\cdot}$) and FRAP were determined spectrophotometrically. Proliferation was assessed by XTT assay at 24, 48 and 72 hrs with exposure to various concentrations (12.5-200 ${\mu}M$) of OA. Apoptotic induction potential of OA was demonstrated using a cellular DNA fragmentation ELISA method. Morphological studies were also carried out to elucidate its antitumor potential. The results revealed that OA induces apoptosis by altering cellular morphology as well as DNA integrity in HaCaT cells in a dose-dependent manner, with comparatively low cytotoxicity. The moderate toxicity observed in HaCaT cells, with induction of apoptosis, possibly suggests greater involvement of programmed-cell death-mediated mechanisms. We conclude that OA has relatively low toxicity and has the potential to induce apoptosis in HaCaT cells and hence provides a substantial and sound scientific basis for further validation studies.

• 대한본초학회지
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• 제23권4호
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• pp.121-134
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• 2008

Objectives: Melia toosendan(MT) has been used as a traditional Korean herbal medicine, and today it is used as a medication for colic, side aches, heartache and other disorders of liver. The aim of this study was to determine whether fractionated extracts of MT inhibit free radical generation such as DPPH radical, superoxide radical and nitric oxide, production of nitrite, an index of NO, PGE2, iNOS, COX-2 and proinflammatory cytokines in lipopolysaccharide(LPS)-treated RAW 264.7 macrophages. Methods: MT extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of MT onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt(MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide(NO) production was measured by Griess reagent system. The levels of iNOS and COX-2 expression were confirmed by western blot. And pro inflammatory cytokines were measured by ELISA kit. Results: Our results indicated that fractionated extracts, especially dichloromethane and ethyl acetate extracts, significantly inhibited free radical generation, the LPS-induced $H_2O_2$, NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-1${\beta}$ and IL-6 formation in macrophages. Conclusions: These results indicate that dichloromethane and ethyl acetate extracts of MT have potential as an agent of chronic inflammatory diseases.

• 동의생리병리학회지
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• 제22권5호
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• pp.1240-1249
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• 2008

Kamigingansikpung-tang(KGST) has been used for many years as a therapeutic agent for acute stage of cerebrovascular disease and hypertension in oriental medicine. But the effect of KGST on hypertension and vascular system is not well-known. This study was done to investigate the effects of KGST on hypertension. The results were obtained as follow: KGST showed scavenging activity on DPPH free radical. KGST showed the inhibitory effect on ROS and ACE, and the accelerated SOD activity. KGST significantly decreased the blood pressure and pulse in DOCA-salt induced hypertensive rat. KGST significantly decreased the levels of aldosterone in DOCA-salt induced hypertensive rat. KGST significantly decreased the levels of dopamine, epinephrine in DOCA-salt induced hypertensive rat. KGST significantly decreased the levels of potassium(K+) and chloride(Cl-) in DOCA-salt induced hypertensive rat. KGST significantly decreased the levels of uric acid and creatine in DOCA-salt induced hypertensive rat. KGST has an effect on inhibiting cell damage of the heart, liver, kidney, and adrenal gland. results suggest that KGST might be effective in treatment and prevention of hypertension.

• Biomolecules & Therapeutics
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• 제21권4호
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• pp.270-276
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• 2013

Fucoxanthin is an important carotenoid derived from edible brown seaweeds and is used in indigenous herbal medicines. The aim of the present study was to examine the cytoprotective effects of fucoxanthin against hydrogen peroxide-induced cell damage. Fucoxanthin decreased the level of intracellular reactive oxygen species, as assessed by fluorescence spectrometry performed after staining cultured human HaCaT keratinocytes with 2',7'-dichlorodihydrofluorescein diacetate. In addition, electron spin resonance spectrometry showed that fucoxanthin scavenged hydroxyl radical generated by the Fenton reaction in a cell-free system. Fucoxanthin also inhibited comet tail formation and phospho-histone H2A.X expression, suggesting that it prevents hydrogen peroxide-induced cellular DNA damage. Furthermore, the compound reduced the number of apoptotic bodies stained with Hoechst 33342, indicating that it protected keratinocytes against hydrogen peroxide-induced apoptotic cell death. Finally, fucoxanthin prevented the loss of mitochondrial membrane potential. These protective actions were accompanied by the down-regulation of apoptosis-promoting mediators (i.e., B-cell lymphoma-2-associated ${\times}$ protein, caspase-9, and caspase-3) and the up-regulation of an apoptosis inhibitor (B-cell lymphoma-2). Taken together, the results of this study suggest that fucoxanthin defends keratinocytes against oxidative damage by scavenging ROS and inhibiting apoptosis.

• Natural Product Sciences
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• 제15권1호
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• pp.37-43
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• 2009

Oxidative stress is caused by an imbalance between the production of reactive oxygen and an ability of a biological system, to readily detoxify the reactive intermediates or easily repair the resulting damage. It has been suggested that developmental alloxan-induced liver damage is mediated through increases in oxidative stress. The anti-diabetic effect and antioxidant activity of Phaleria macrocarpa (PM) fractions were investigated in alloxan-induced diabetic rats. After two weeks administration of PM, the liver antioxidant enzyme and hyperglycemic state were evaluated. The results showed that oral administration of PM treatments reduced blood glucose levels in diabetic rats by oral administration (P < 0.05). Serum glutamic-oxaloacetic transaminase (sGOT) and serum glutamic-pyruvate-transaminase (sGPT) were also diminished by PM supplementation. The superoxide dismutase (SOD), catalase (CAT) and glutathione-peroxidase (GPx) activities, and glutathione (GSH) level in the alloxan-induced diabetic rats were significantly decreased (P < 0.05) compared to those in the normal rats but were restored by PM treatments. PM fractions also repressed the level of malondialdehyde (MDA) in the liver. Glutathione reductase (GR), glutathione-S-transferase (GST) and $\gamma$-glutamylcysteine synthase (GCS) were also reduced in alloxan-induced diabetic rats. PM fractions could restore the GR and GST activities, but the GCS activity was not affected in rat livers. From the results of the present study, the diabetic effect of the butanol fraction of PM against alloxan-induced diabetic rats was concluded to be mediated either by preventing the decline of hepatic antioxidant status or due to its indirect radical scavenging capacity.

• 생약학회지
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• 제47권1호
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• pp.55-61
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• 2016

Antioxidant effects and nuclear factor E2-related factor-2 (Nrf-2) signal pathway of methanol extract and 4 fractions [n-hexane, methylene chloride, ethyl acetate (EtOAc), and n-butanol fractions] from Petasites japonicus were investigated. The EtOAc fraction showed highest polyphenol and flavonoid contents among other fractions. In addition, EtOAc fraction showed stronger scavenging activity against superoxide anion radical than other fractions. Furthermore, we investigated antioxidants effects of the EtOAc fraction under cellular system using $LLC-PK_1$ cells. The EtOAc fraction dose-dependently increased the antioxidant protein expressions of heme oxygenase 1 (HO-1) and thioredoxin reductase 1 (TrxR1) known to be involved in oxidative stress, through activation of Nrf-2. The treatment of EtOAc fraction ($100{\mu}g/mL$) led to the elevation of the high expression of Nrf-2-dependent factor such as HO-1 and TrxR1. These results indicated that the EtOAc fraction of P. japonicus showed high antioxidant activity by regulation of Nrf-2 signaling pathway.