• Korean Journal of Medicinal Crop Science
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• v.19 no.2
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• pp.103-110
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• 2011

This study was performed to enhance anticancer activities of Lithospermum erythrorhizon by eluting high amount of shikonin through ultra high pressure process. Extraction yield was increased up to 5~10% by ultra high pressure process, compare to the normal extraction processes such as water solvent extraction, 70% ethyl alcohol solvent extraction. The cytotoxicity of the extracts ($1.0{\mu}g/m{\ell}$) from ultra high pressure process was showed the lowest cytotoxicity 13.4% for human lung cell (HEL299). The anticancer activities showed 80~85% by adding $1.0{\mu}g/m{\ell}$ of the extracts from ultra high pressure process in several cancer cell lines such as AGS, Hep3B, MCF-7 and HeLa cells. Among them, MCF-7 cell of the endocrine system was highest inhibited than other cells. The anticancer activities of the extracts from ultra high pressure extraction process showed 10~15%, which was higher than the extracts from normal extraction processes. From HPLC analysis of the extracts, the contents of shikonin in the extracts from ultra high pressure process was 11.42% (w/w), which was 20% higher than others. This results indicate that ultra high pressure process could increase the extraction yield of shikonin and other contents, which resulted in higher anticancer activities.

• Korean Journal of Medicinal Crop Science
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• v.12 no.4
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• pp.309-314
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• 2004

The cytotoxic effects of water and ethanol extracts of Echinacea purpurea (L.) (EP) and chloroform, ethyl acetate, butanol and aqueous fractions from each extract of EP were examined. Every extract and fraction of EP inhibited the growth of human hepatocarcinoma, human gastric cancer cell, human breast cancer cells and human lung carcunoma in concentration-dependent manners over a concentration range of $0.05{\sim}1.0\;mg/ml$. Most extracts and fractions with the concentraction of 1 mg/ml showed strong inhibition of more than 70% for every cancer cell. Only aqueous fractions of each extract showed very weak inhibitons of 12 to 25% on the growth of human normal lung cell with the concentration of 1 mg/ml. Overall selectivity of the extrats and fractions on the four human cancer cell lines was over 2.5. These results indicate that EP has a very potent selective toxicity for cancer cells.

• Food Science and Biotechnology
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• v.18 no.1
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• pp.172-178
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• 2009

Fermented ginseng (FG) is an ethanol extract of ginseng radix processed with $\beta$-galactosidase. It was hypothesized that FG may exert anti-hyperlipidemic and anti-diabetic activities through modulating AMP-activated protein kinase (AMPK) in HepG2 human hepatoma cells. In this study, we showed that AMPK phosphorylation was stimulated by FG. These effects were abolished by pretreatment with an AMPK inhibitor, compound C. In addition, FG regulated the expression of genes associated with lipogenesis and lipolysis, thus causing suppression of hepatic triglyceride accumulation. In vivo study using db/db mice, FG reduced fasting plasma glucose, HbAlc, and insulin resistance index, when compared to diabetic control. FG also increased the phospho-AMPK and glucose transporter 4 (GLUT4) expressions in liver and skeletal muscle, respectively. In liver, expressions of lipogenic gene were decreased whereas expressions of lipolytic genes were induced, when compared to diabetic control. Taken together, we may suggest that FG ameliorates hyperglycemia and hyperlipidemia through activation of AMPK and could be developed as a health functional food or therapeutic agent for type 2 diabetic patients.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.42 no.3
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• pp.269-278
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• 2016

In searching for novel agents for skin anti-aging from natural resources, we found that the extract of the fruiting bodies of Ramaria formosa (R. formosa) had significant antioxidant and human neutrophil elastase (HNE) inhibitory activities. R. formosa extract exhibited a considerable DPPH radical scavenging activity with an antioxidant content of 117.0mg/mL (ascorbic acid equivalents) at the concentration of $500{\mu}g/mL$. The capacity of R. formosa extract to scavenge peroxy radicals measured by ORAC assay also showed dose-dependent antioxidant effect with $ORAC_{Roo}$ (trolox equivalents, $1{\mu}M$) values of 0.8, 5.2, and 7.8 at the concentrations of 1, 10, and $20{\mu}g/mL$. The cellular antioxidant capacity of R. formosa extract was investigated by assaying the cellular fluorescence intensity using dichlorodihydrofluorescein (DCF). The cellular oxidative stress induced by AAPH, $Cu^{2+}$ or $H_2O_2$ in HepG2 cells was significantly attenuated by more than 30% at $20{\mu}g/mL$ of R. formosa extract. HNE activity was reduced by treatment with R. formosa extract in a dose-dependent manner, and the $ED_{50}$ value for the ethanol extract of R. formosa was $42.9{\mu}g/mL$. R. formosa extract did not exhibited antimicrobial activity against four microorganisms including Bacillus subtilis (B. subtilis), Escherichia coli (E. coli), Candida albicans (C. albicans), Aspergillus oryzae (A. oryzae). Furthermore, the extract did not affect the inflammatory cytokine production of interleukin-10 and interferon-${\gamma}$ in NK92 cells. From the above results, we found that R. formosa extract has considerable antioxidant and elastase inhibitory effects, and does not stimulate immune cells. These findings suggest that R. formosa extract may be used as a bioactive component in cosmetic composition.

• Biomolecules & Therapeutics
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• v.27 no.2
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• pp.193-200
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• 2019

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.

• Journal of Ginseng Research
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• v.41 no.2
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• pp.169-179
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• 2017

Background: Ginsenosides are the main pharmacological components of Panax ginseng root, which are thought to be primarily responsible for the suppressing effect on oxidative stress. Methods: 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and oxygen radical absorption capacity were applied to evaluate the antioxidant activities of the ginsenosides. Human embryonic kidney 293 (HEK-293) cells were incubated with ginsenosides extracted by pulsed electric field (PEF) and solvent cold soak extraction (SCSE) for 24 h and then the injury was induced by $40{\mu}M$ $H_2O_2$. The cell viability and surface morphology of HEK-293 cells were studied using MTS assay and scanning electron microscopy, respectively. Dichloro-dihydro-fluorescein diacetate fluorescent probe assay was used to measure the level of intracellular reactive oxygen species. The intracellular antioxidant activities of ginsenosides were evaluated by cellular antioxidant activity assay in HepG2 cells. Results: The PEF extracts displayed the higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and stronger oxygen radical absorption capacity (with an oxygen radical absorption capacity value of $14.48{\pm}4.04{\mu}M\;TE\;per\;{\mu}g/mL$). The HEK-293 cell model also suggested that the protective effect of PEF extracts was dose-dependently greater than SCSE extracts. Dichloro-dihydro-fluorescein diacetate assay further proved that PEF extracts are more active (8% higher than SCSE extracts) in reducing intracellular reactive oxygen species accumulation. In addition, scanning electron microscopy images showed that the HEK-293 cells, which were treated with PEF extracts, maintained more intact surface morphology. Cellular antioxidant activity values indicated that ginsenosides extracted by PEF had stronger cellular antioxidant activity than SCSE ginsenosides extracts. Conclusion: The present study demonstrated the antioxidative effect of ginsenosides extracted by PEF in vitro. Furthermore, rather than SCSE, PEF may be more useful as an alternative extraction technique for the extraction of ginsenosides with enhanced antioxidant activity.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.149-154
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• 2011

Objectives : Myocardial infarction is caused by heart cell death in a region where coronary arteries supplying blood to the region are occluded. In the present study, we determined whether ethanol extract of Cortex fraxini (HY5053) could attenuate heart injury by inhibiting apoptosis. Methods : Improvement of survival of HepG2 cells, a human hepatocellular carcinoma cell line, and reduction of apoptosis under hypoxic conditions (3% $O_2$) were assessed by trypan blue staining and DNA fragmentation assay, respectively. To assess the impact of HY5053 on the heart injury, Sprague-Dawley rats underwent 1 day of the left anterior descending coronary artery occlusion. HY5053 was given by intraperitoneal injection (200 mg/kg) 1 hr prior to the occlusion. Subsequently, the heart were harvested, excised into 4 slices, and the slices were stained with 2,3,5-triphenyl tetrazolium chloride. Finally, the extent of heart injury represented as ischemic index (%) was assessed. Results : Addition of HY5053 (400 ${\mu}g$/mL) into the culture medium for 1 day under ischemic conditions improved the cell survival by 50%, compared with control (0 ${\mu}g$/mL), consequently delayed apoptosis in 6 hr difference. Also, HY5053 (200 mg/kg) reduced the ischemic index by 44%, compared with control (0 mg/kg). Conclusions : These findings suggested that HY5053 attenuated myocardial infarction by inhibiting apoptosis. Thus, Cortex fraxini could be developed as a novel cardioprotectant to complement a currently available treatment, coronary angioplasty.

• Preventive Nutrition and Food Science
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• v.16 no.3
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• pp.217-223
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• 2011

Ischemic stroke constitutes about 80% of all stroke incidences. It is characterized by brain cell death in a region where cerebral arteries supplying blood are occluded. Under these ischemic conditions, apoptosis is responsible for the cell death, at least in part. Goat's-beard (Aruncus dioicus var. kamtschaticus) is a perennial plant that grows naturally in the alpine regions of Korea. In the present study, we first determined whether water extract of goat's-beard (HY1646) and some of its fractions prepared by partitioning with organic solvents could improve the viability of human hepatocellular carcinoma cells (HepG2) cultured under hypoxic condition by blocking apoptotic pathways. Based on the in vitro findings, we subsequently investigated whether HY1646 and the ethyl acetate fraction (EA) selected from cell culture-based screening could attenuate brain injury in a rat middle cerebral artery occlusion (MCAO) model of ischemia (2 hr), followed by 22 hours of reperfusion. The cell number was sustained close to that initially plated in the presence of HY1646 even after 24 hr of cell culture under hypoxic condition (3% $O_2$), at which time the cell number reached almost zero in the absence of HY1646. This improvement in cell viability was attributed to the delay in apoptosis, identified by the formation of DNA ladder in gel electrophoresis. Of fractions soluble in hexane, ethyl acetate (EA) and butanol, EA was chosen for the animal experiments because EA demonstrated the best cell viability at the lowest concentration (10 ${\mu}g$/mL). HY1646 (200 mg/kg) and EA (10 and 20 mg/kg) significantly reduced infarct size, an index of brain injury, by 16.6, 40.0 and 61.0%, respectively, as assessed by 2,3,5-triphenyl tetrazolium chloride staining. The findings suggest that prophylactic intake of goat's beard might be beneficial for preventing ischemic stroke.

• Microbiology and Biotechnology Letters
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• v.42 no.1
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• pp.73-81
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• 2014

Salsola collina, also known as Russian thistle, is widely distributed in and around waste facilities, roadsides, and drought and semi-drought areas, and is used as a traditional folk remedy in Chinese medicine for the treatment of hypertension. In this study, we have evaluated the anti-oxidative and anti-cancer activities of the ethanol extract of S. collina Pall. (EESC), and the molecular mechanisms of its anti-cancer effects on human colon carcinoma HT29 cells. EESC exhibited anti-oxidative activity through DPPH radical scavenging capacity and showed cytotoxic activity in a dose-dependent manner in HT29 cells. After EESC treatment, HT29 cells altered their morphology, becoming smaller and irregular in shape. EESC also induced cell accumulation in the G2/M phase in a dose-dependent manner, accompanied by a decrease of cell population in the G1 phase. The G2/M arrest by EESC was associated with the increased expression of cyclin-dependent kinase (CDK) inhibitor p21 and Wee1 kinase, which phosphorylates, or inactivates, Cdc2. EESC treatment induced the phosphorylation of Cdc2 and Cdc25C, and inhibited cyclin A and Cdc25C protein expression. In addition, S arrest was induced by the highest concentration of EESC treatment, associated with a decrease of cyclin A and Cdk2 expression. These findings suggest that EESC may possess remarkable anti-oxidative activity and exert an anti-cancer effect in HT29 cells by cell cycle regulation.

• Journal of Microbiology
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• v.34 no.1
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• pp.25-29
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• 1996

Since sequencing of randomly selected cDNA clones has been known to be a powerful approach to obtain information on gene expression pattern in specific cells or tissues, we have analyzed a 3'-directed cDNA library of vegetative mycelia of A. nidulans by single-pass sequencing of hundreds of randomly selected clones. Sequencing of 292 cDNA clones yielded 209 gene signatures (GSs) probably representing highly or lesser expressed genes in the vegetative mycelia. Among the 209 GSs, 25 (79 cDNA clones) appeared more than once and 184 only once. One GS appeared at a highest frequency of 6 times, 2 GSs5 times, 4 GSs 4 times, a GSs 3 times and 16 GSs twice. About 6.6% GSs comprizing of 13 GSs showed alternative polyadenylation. Among 23 redundant GSs, three were common in both mycelia and sexual organs, and 22 were probably mycelia-specific. Out of 209 GSs, 36 were identified in GenBank showing of 70% or greater similaritis. Only six GSs were for A. nidulans genes, and 13 GSs were of DNA or genes encoding cytoplasmic or organellar proteins. This pattern is similar to those in the human HepG2 cell line and in human colonic mucosa, although very few genes for nuclear proteins and for protein synthesis were in A. nidulans.