• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.9
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• pp.1257-1264
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• 2016

Extracts from Artemisia annua $Linn\acute{e}$ (AAE) have various functions (anti-malaria, anti-virus, and anti-oxidant). However, the mechanism of the effects of AAE is not well known. Thus, we determined the apoptotic effects of AAE in AGS human gastric carcinoma cells. In this study, we suggested that AAE may exert cancer cell apoptosis through the Akt/mammalian target of rapamycin (mTOR)/glycogen synthase kinase (GSK)-$3{\beta}$ signal pathway and mitochondria-mediated apoptotic proteins. Activation by Akt phosphorylation resulted in cell proliferation through phosphorylation of tuberous sclerosis complex 2 (TSC2), mTOR, and GSK-$3{\beta}$. Thus, de-phosphorylation of Akt inhibited cell proliferation and induced apoptosis through inhibition of Akt, mTOR, phosphorylation of GSK-$3{\beta}$ at serine9, and control of Bcl-2 family members. Inhibition of GSK-$3{\beta}$ attenuated loss of mitochondrial membrane potential and release of cytochrome C. Bax and pro-apoptotic proteins were activated by their translocation into mitochondria from the cytosol. Translocation of Bax induced outer membrane transmission and generated apoptosis through cytochrome C release and caspase activity. We also measured 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase assay, Hoechst 33342 staining, Annexin V-PI staining, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide staining, and Western blotting. Accordingly, our study showed that AAE treatment to AGS cells resulted in inhibition of Akt, TSC2, GSK-$3{\beta}$-phosphorylated, Bim, Bcl-2, and pro-caspase 3 as well as activation of Bax and Bak expression. These results indicate that AAE induced apoptosis via a mitochondrial event through regulation of the Akt/mTOR/GSK-$3{\beta}$ signaling pathways.

• Food Science and Preservation
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• v.24 no.2
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• pp.254-265
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• 2017

For the production of ${\gamma}-aminobutyric$ acid (GABA) contents increased Seomaeyaksuk fermentant, 0, 0.25, 0.5 and 1.0% of monosodium galutamate (MSG) was added in Seomaeyaksuk water extract 15% contained Sikhye and inoculated with Lactobacillus brevis (KI271266). Physicochemical properties were sampling and analyzed at each 1 day during 3 days fermentation. Goes on fermentation periods, the turbidity was gradually reduced, but lightness and the yellowness was increased, redness was decreased. Soluble solid was also decreased. The reducing sugars content were also decreased during fermentation. pH was 4.65-4.83 before fermentation but it was lower 3.15-3.68 after three days fermentation. The GABA contents increased by fermentation periods and it was the highest in MSG 1.0% added sample (354.38 mg/L). Fructose, glucose and sucrose contents were 50-67% decreased at three days fermentation than before fermentation. Among the organic acids, propionic acid, oxalic acid, citric acid and fumaric acid contents were decreased and lactic acid, acetic acid and succinic acid were increased during fermentation periods. Contents of total polyphenol and DPPH radical scavenging activity were the highest in MSG 0.5% added sample. From these results, we confirmed that increasing of GABA content when the manufacturing of Seomaeyaksuk lactic acid fermentation product, is possibile by addition of MSG without affecting physicochemical characteristics.

• Journal of Life Science
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• v.26 no.7
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• pp.764-771
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• 2016

Extracts from Artemisia annua Linné (AAE) have been known to possess various functions, including anti-bacterial, anti-virus, and anti-oxidant effects. However, the mechanism of those effects of AAE is not well-known. The aim of this study was to analyze the inhibitory effects of AAE on cell proliferation of the human hepatoma cell line (Hep3B) and to examine its effects on apoptosis. Activation by phosphorylation of Akt is cell proliferation through the phosphorylation of TSC2, mTOR, and GSK-3β. We suggested that AAE may exert cancer cell apoptosis through Akt/mTOR/GSK-3β signal pathways and mitochondria-mediated apoptotic proteins. For this, we examined the effects of extracts of AAE on cell proliferation according to treatment concentration. Treatment with AAE not only reduced cell viability, but also resulted in the induced release of lactate dehydrogenase (LDH). These results were determined with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a lactate dehydrogenase (LDH) assay. Furthermore, we determined the effects of apoptosis through Hoechst 33342 staining, annexinⅤ-propidium iodide (PI) staining, 5,5′, 6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide (JC-1) staining, and Western blotting. Our study showed that the treatment of liver cancer cells with AAE resulted in the inhibition of Akt, TSC2, GSK-3β-phosphorylated, Bcl-2, and pro-caspase 3 and the activation of Bim, Bax, Bak, and cleaved PARP expressions. These results indicate that AAE induced apoptosis by means of a mitochondrial event through the regulate of Akt/mTOR/GSK-3β signaling pathways.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.6
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• pp.869-876
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• 2013

To develop functional products based on black garlic, a black garlic extract (BG) of 7 brix, a gaeddongssuk extract (GS) of 0.7 brix and two types of mixtures (MBS-I, black garlic 7 brix : gaeddongssuk 0.7 brix; MBS-II, black garlic 14 brix : gaeddongssuk 1.4 brix, 93:7, v/v) were supplemented to rats training on a treadmill for 4 weeks. Body weight from the training did not decrease during the experimental period. Serum albumin content significantly increased in the groups fed an experimental diet compared to the control. The BUN content significantly decreased in BG and MBS-II groups compared to the control. AST and ALP activities significantly decreased in the groups fed an experimental diet compared to the control. Serum triglyceride and total cholesterol levels in MBS-I and MBS-II groups significantly decreased compared to the control. Lipid levels in the serum and liver tissue were not significantly different between the MBS-I and MBS-II groups. The lipid peroxide content in the serum and liver tissue was significantly reduced in the groups fed all extracts compared to the control; the serum and liver lipid contents was lowest in the MBS-I and MBS-II groups, respectively. Hepatic catalase activity in the GS and MBS groups increased by 1.8~2.3 times compared to the control. SOD and GSH-px activities significantly increased from treatment with the extracts by 1.3~1.5 times and 1.2~1.7 times, respectively. These results indicate that a mixture of BG and GS extracts has higher biological activity than a single supplementation of BG or GS extract. Therefore, the addition of gaeddongssuk to black garlic (MBS-I and MBS-II) is effective as a defense material against oxidative stress. MBS-I may be especially effective for its biological activities.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.195-199
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• 2003

The effect of ultrasonification on immune-stimulatory activities of extracts Acanthopanax senticosus Harms, Ephedra sinica Stapf, Rubus coreanus Miq and Artemisia capillaris Thunb was observed by the growth of human immune cells. The extracts yield make a comparative study of water extracts and water extract with ultrasonification at $40^{\circ}C,$ $60^{\circ}C$ and $100^{\circ}C$. The yield is highest in the water extracts with ultrasonification at $60^{\circ}C$ The growth of human T-cell was increased up to $13.8{\times}10^{-4}$ viable cells/ml in adding ultrasonification extracts. The treated extracts increased up to $10.5{\times}10^{-4}$ pg/cells that TNF-${\alpha}$ specific secretion of human T-cell. These results suggested that biological activities of water extracts with ultrasonification have more strong than water extracts in human immune system.

• Korean Journal of Organic Agriculture
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• v.23 no.2
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• pp.373-390
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• 2015

This study investigated the effect of coated feed using mixed oil on rumen fermentation characteristics. Two experiments were conducted based on materials that were mixed. First, cashew nut and soybean oils were mixed with white mineral oil. And second, different plant extracts were mixed with white mineral oil. At first experiment, inclusion levels of mixed oil on diet (0.03%, 0.1%, 0.5%) were applied as variables. A coated diet was fermented with rumen inoculum according to in vitro rumen fermentation and its parameters were investigated. In the result of first experiment, no negative effects on rumen pH were found. Significantly decreased dry matter digestibility was detected at 0.5% treatment (P<0.05). Total gas productions in control and 0.03% were significantly greater than those of others (P<0.05). Significantly reduced methane productions were found in all treatments compared to the control (P<0.05). Inclusion of mixed oil did not affect on ammonia production. Total volatile fatty acid production was also not influenced by coating with mixed oils. Rumen fermentation parameters were greatly changed according to introduced plant extracts at second experiment. The significantly lowest and greatest ammonia productions were found at treatments with Ixeris dentata and Plantago asiatica, respectively (P<0.05). The significantly greatest acetate and propionate productions were detected at treatments with Crucuma longa and Zizyphus vulgaris 1, respectively (P<0.05). All treatments, except Chrysanthemum idicum, Euyale ferox seed, Moringa leaf and fruit and Zizyphus vulgaris 1, showed significantly increased total volatile fatty acid production compared to the control (P<0.05). Only Paeonia lactiflora showed significantly lesser gas production than the control (P<0.05). In methane production, Ceramium, Zizyphus, Paeonia, Agrimonia, Torilis, Mugwort, Foeniculum, Euphorbia, Taraxacum, Artemisia, Momordica, Curcuma and Moringa reduced methane significantly compared to the control (P<0.05).

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.5
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• pp.1081-1086
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• 2007

This study was performed to investigate the medicinal effects of the herbal combination extracts-2 (HCE-2), consisting of Artemisia capillaris Thunb., Lonicera japonica Thunb., Prunella vulgaris var. lilacina, and Hovenia dulcis Thunb. on the alcohol-induced liver injury in rats. The rats were randomly divided into four groups: normal group (n =6), non-treated control group (n =6), saline-treated group (n =6) and the herbal combination extract (HCE-2)-treated group (n =6). The rats in the alcohol-loaded groups were orally administered with ethanol at a daily dose of 4 g/kg-body weight for 5 weeks. Thirty minutes before the ethanol injection, saline or herbal combination extracts was administered by using a gastrogavage. Blood and liver tissue samples were taken out from the hearts and livers of the rats, respectively, on 15th and 38th days. The activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured using an enzyme-linked immunosorbent assay (ELISA). We also investigated the protective effect of the herbal combination extracts by Hematoxylin-Eosin staining on histological sections of rat liver. In this study, the oral administration of the herbal combination extracts significantly reduced the serum levels of AST and ALT, which had been raised by alcohol-induced liver injury. Histological analysis and apparent observation of liver also showed the preventive effect of the herbal combination extracts in a chronic alcohol-induced rat model. Theses results revealed that the herbal combination extracts effectively prevented hepatic damage consequent to the chronic exposure to repetitive administration of ethanol and could be used as a primary resource of a health beverage or herbal medicine, alleviating the alcohol-induced hepatic injury and hangover symptoms.

• Proceedings of the Korean Society of Toxicology Conference
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• 2002.05b
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• pp.88.2-98
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• 2002

A wide arry of naturally occurring substances particularly those present in dietary and medicinal plants, have been reported to possess substantial cancer chemopreventive properties. Certain phytochemicals retain strong antioxidative and anti-inflammatory properties which appear to contribute to their chemopreventive or chemoprotective activities. Inducible cyclooxygenase(COX-2) and nitric oxide synthase (iNOS) are important enzymes that mediate inflammatory processes. There is some evidence that expression of both COX-2 and iNOS is co-regulated by the eukaryotic transcription factor NF-$textsc{k}$B. Increased expression of COX-2 and/or iNOS has been associated with pathophysiology of certain types of human cancers as well as inflammatory diseases. Since inflammation is closely linked to tumor promotion, substances with potent anti-inflammatory activies are anticipated to exert chemopreventive effects on carcinogenesis, particularly in the promotion stage. An example is curcumin, a yellow pigment of turmeric (Curcuma longa L., Zingiberaceae), that strongly occurring diaryl heptanoids structurally related to curcumin have substantial anti-tumor promotional activities in two-stage mouse skin carcinogenesis. Thus, yakuchinone A [1-(4'-hydroxy-3'-methoxyphenyl)-7-phenyl-3heptanone] and yakuchinone B [1-(4'-hydroxy-3'methoxyphenyl)-7-phenylhept-1-en-3-one] present in Alpinia oxyphylla Miquel (Zingiberacease) attenuate phorbol ester-induced inflammation and papilloma formation in female ICR mice. These diarylheptanoids also suppressed phorbol ester-induced activation of epdermal ornithine decarboxylase and its mRNA expression when applied onto shaven backs of mice. Yakuchinone A and B as well as curcumin inhibited phorbol ester-induced expression of COX-2 and iNOS and their mRNA in mouse skin via inactivation of NF-$textsc{k}$B. Capsaicin, a major pungent ingredient of red pepper also attenuated phorbol ester-induced NF-$textsc{k}$B activation. Similar suppression of COX-2 and iNOS and down-regulation of NF-$textsc{k}$B activation for its DNA binding were observed with the ginsenosied Rg3 and the ethanol extract of Artemisia asiatica. We have also found that certain anti-inflammatory phytochemicals exert inhibitory effects on phorbol ester-induced COX-2 expression and NF-$textsc{k}$B activation in immortalized human breast epithelial (MCF-10A) cells in culture. One of the plausible mechanisms undelying inhibition by aforementioned phytochemicals of phorbol ester-induced NF-$textsc{k}$B activation involves interference with degragation of the inhibitory unit, I$textsc{k}$Ba, which blocks subsequent nuclear translocation of the functionally active p65 subunit of NF-$textsc{k}$B. the activation of epidermal NF-$textsc{k}$B by phorbol ester and subsequent induction of COX-2 hence appear to play an important role in intracellular signaling pathwasy leading to tumor promotion and targeted inhibition of NF-$textsc{k}$B may provide a new promising cancer chemopreventive strategy.

• Applied Biological Chemistry
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• v.38 no.4
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• pp.364-369
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• 1995

Anticarcinogenic activity of Moutan radix for mouse ascites cancer induced by mouse Sarcoma 180 (S-180) cells was investigated. Methanol extract of Moutan radix including other folk medicinal plants (Taxus cuspidata, Curcuma longa, Artemisia capillaris, Ligrstri fructus, and Liriope platyphylla) used to remedy or cure many chronic human diseases like cancer was fractionated into hexane, chloroform ($CHCl_3$), ethylacetate (EtOAc), and butanol (BuOH) fractions. Anticarcinogenic activity of the fractions, exhibited a strong cytotoxicity for L1210 and S-180 cells, was examined for mouse ascites cancer induced by S-180 cells. Male ICR mice (7 mice/treatment, $5{\sim}6$ weeks of age, $23{\pm}1\;g$ were injected i.p. with S-180 cells ($1{\times}10^{7}\;cell/1\;ml$ PBS). One day later, each mouse was given 0.1 ml of 10% DMSO containing sample ($30\;{\mu}g/g$ body weight) every day for 10 consecutive days. Control mice were only given 0.1ml S-180 cells and 0.1 ml 10% DMSO. Mice treated with EtOAc fraction of Moutan radix showed 28.7 days of life, which is 167% of control mice's life. Based on the dose-dependant experiment mice treated with $30\;{\mu}g$ showed longer life relative to mice treated with ootherr doses (5, 15, $60\;{\mu}g$), and mice treated with $60\;{\mu}g$ exhibited toxic symptoms. Body weight of mice treated with Moutan radix was significantly reduced relative to that of control mice (p<0.05). GC-MS analysis in conjunction with silica-gel column chromatography revealed that the EtOAc fraction contained 2-methoxylphenol, benzoic acid, 1-(4-hydroxy-3-methoxyphenyl)ethanone, 8-methyl-2,4(1H,3H)pteridinedione and 2,5-furan-dicarboxylic dimethyl ester as regards to the anticarcinogenic property of the EtOAc fraction. These results suggest that Moutan radix might be included as an anticarcinogenic medicinal plant for treatment of ascites cancer.

• Food Science and Preservation
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• v.20 no.4
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• pp.546-553
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• 2013

In this study, we examined the antioxidant activity of the Sumaeyaksuk (Artemisia argyi) tea extracts from different pre-treatment and extraction methods. Sumaeyaksuk was sun-dried for 3.5 days (control, RC) and aged at a temperature of $60^{\circ}C$ for 3.5 days (HA), 7 days (HB), and 14 days (HC), respectively. Each sample was extracted in $60^{\circ}C$ and $95^{\circ}C$ hot water for 2 minutes. The soluble solids content of HA from the $60^{\circ}C$and $95^{\circ}C$ hot water extraction were $0.52{\pm}0.18%$ and $0.92{\pm}0.18%$, respectively. The soluble solids content was increased by the higher extraction temperature. The reducing sugar content of RC was $9.55{\pm}0.18mg/g$ in the $95^{\circ}C$ extraction, which was significantly higher than in the $60^{\circ}C$ extracted sample. However, the reducing sugar content did not show a remarkable difference based on aging periods. The total phenolic compound content of the $95^{\circ}C$ extracted samples was $3.36{\pm}0.13{\sim}9.88{\pm}0.23mg/g$, which was significantly higher than that of the $60^{\circ}C$ extracted sample. The ABTS radical scavenging activity of the $60^{\circ}C$ extracted RA and HA samples were 35.63% and 95.10%, respectively. Moreover, the radical scavenging activity increased to 63.35% and 96.78%, respectively, in the $95^{\circ}C$ extracted samples. As a result of the high temperature, the extracted sample showed an increase in the FRAP. In the RC sample, the FRAP was two times higher in the $95^{\circ}C$ extracted sample ($181.28{\pm}2.90{\mu}M$) than in the $60^{\circ}C$ extracted sample ($83.88{\pm}0.43{\mu}M$).