• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.185-193
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• 2008

We evaluated the response to salt stress of two different ginseng lines, STG3134 and STG3159, which are sensitive and tolerant, respectively, to salt treatment. Plants were exposed to a 5 dS/m salt solution, and chlorophyll fluorescence was measured. STG3134 ginseng was more sensitive than STG3159 to salt stress. To characterize the cellular response to salt stress in the two different lines, changes in protein expression were investigated using a proteomic approach. Total protein was extracted from detached salt-treated leaves of STG3134 and STG3159 ginseng, and then separated by two-dimensional polyacrylamide gel electrophoresis(2-DE). Approximately 468 protein spots were detected by 2-DE and Coommassie brilliant blue staining. Twenty-two proteins were found to be reproducibly up- or down-regulated in response to salt stress. Among these proteins, twelve were identified using MALDI-TOF MS and ESI-Q-TOF and classified into several functional groups: photosynthesis-related proteins(oxygen-evolving enhancer proteins 1 and 2, rubisco and rubisco activase), detoxification proteins(polyphenol oxidase) and defense proteins($\beta$-1,3-glucanase, ribonuclease-like storage protein, and isoflavone reductase-like protein). The protein levels of ribonuclease-like storage protein, which was highly induced in STG3159 ginseng as compared to STG3134, correlated tightly with mRNA transcript levels, as assessed by reverse-transcription(RT)-PCR. Our results indicate that salinity induces changes in the expression levels of specific proteins in the leaves of ginseng plants. These changes may, in turn, playa role in plant adaptation to saline conditions.

• Nutrition Research and Practice
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• v.9 no.1
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• pp.22-29
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• 2015

BACKGROUND/OBJECTIVES: Recently, anthocyanins have been reported to have various biological activities. Furthermore, anthocyanin-rich purple corn extract (PCE) ameliorated insulin resistance and reduced diabetes-associated mesanginal fibrosis and inflammation, suggesting that it may have benefits for the prevention of diabetes and diabetes complications. In this study, we determined the anthocyanins and non-anthocyanin component of PCE by HPLC-ESI-MS and investigated its anti-diabetic activity and mechanisms using C57BL/KsJ db/db mice. MATERIALS/METHODS: The db/db mice were divided into four groups: diabetic control group (DC), 10 or 50 mg/kg PCE (PCE 10 or PCE 50), or 10 mg/kg pinitol (pinitol 10) and treated with drugs once per day for 8 weeks. During the experiment, body weight and blood glucose levels were measured every week. At the end of treatment, we measured several diabetic parameters. RESULTS: Compared to the DC group, Fasting blood glucose levels were 68% lower in PCE 50 group and 51% lower in the pinitol 10 group. Furthermore, the PCE 50 group showed 2-fold increased C-peptide and adiponectin levels and 20% decreased HbA1c levels, than in the DC group. In pancreatic islets morphology, the PCE- or pinitol-treated mice showed significant prevention of pancreatic ${\beta}$-cell damage and higher insulin content. Microarray analyses results indicating that gene and protein expressions associated with glycolysis and fatty acid metabolism in liver and fat tissues. In addition, purple corn extract increased the phosphorylation of AMP-activated protein kinase (AMPK) and decreased phosphoenolpyruvate carboxykinase (PEPCK), glucose 6-phosphatase (G6pase) genes in liver, and also increased glucose transporter 4 (GLUT4) expressions in skeletal muscle. CONCLUSIONS: Our results suggested that PCE exerted anti-diabetic effects through protection of pancreatic ${\beta}$-cells, increase of insulin secretion and AMPK activation in the liver of C57BL/KsJ db/db mice.

• The Korean Journal of Food And Nutrition
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• v.24 no.3
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• pp.362-366
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• 2011

The objective of this study was to compare the anticancer activity of glucosinolates against four different cancer cells; SNU-251, SNU-354, SNU-C4, MCF-7 and to determine the amounts of glucosinolates in mustard leaves. Green and red mustard leaves were cultivated on the field of Rural Development Administration from May to July, 2010. After the leaves were harvest and lypophilized, the fine powder was extracted with boiling 70% methanol(v/v) for the measurement of anticancer activity and then applied to the cancer cells obtained from Korean Cell Line Bank(KCLB). The anticancer activity of red leaf mustard was higher than that of green mustard leaf, particularly in SNU-251 for 24 hr and in SNU-C4 for 48 hr. In both green and red mustard leaf, sinigrin was the most abundant glucosinolte and was determined as 4.71 and 3.06 mg/g dry wt., respectively, but glucoiberin only presented 0.14 mg/g dry wt. with minor amounts.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.05a
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• pp.14-14
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• 2010

Plastid proteomics are essential organelles present in virtually all cells in plants and green algae. Plastids are responsible for the synthesis and storage of key molecules required for the basic architecture and functions of plant cells. The proteome of plastid, and in particular of chloroplast, have received significant amounts of attention in recent years. Various fractionation and mass spectrometry (MS) techniques have been applied to catalogue the chloroplast proteome and its sub-organelles compartments. To better understanding the function of the lumenal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine-SDS-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, TMHMM, and TOPPRED) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

• The Korean Journal of Pesticide Science
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• v.16 no.4
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• pp.267-272
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• 2012

Vitex trifolia L. is a full grown fruit of Vitex rotundifolia L. (Verbenaceae). It has been used for treating headache, dizziness, toothache and removal of fever as a traditional medicine in Korea. V. trifolia (500 g) were extracted three times with 80% aqueous MeOH at room temperature. The MeOH extract (38 g) was successively partitioned with n-hexane, EtOAc, n-BuOH and $H_2O$. Both n-hexane and EtOAc fractions showed more than 80% antifungal activity in vivo against several plant pathogens at 2000 ppm. Successive repeated silica gel, Sephadex LH-20, preparative TLC, and preparative HPLC of these fractions led to isolation of three compounds. Using mainly mass spectroscopy and nuclear magnatic resonance (NMR) spectroscopy, their chemical structures were determined as agnuside(1), chrysosplenol B(2), and artemetin(3). Compound 2 was isolated for the first time from V. trifolia. Study on in vitro and in vivo antifungal activities of the isolated compounds is in progress.

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1039-1044
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• 2003

We investigated the changes in saponins during the cultivation of soybean sprout. Crude saponin content was 4.15mg/g in germinated soybean and reached its peark (5.33mg/g) in soybean sprout cultivated for six days. Saponin content in the cotyledon, stem, and root of the soybean sprout cultivated for six days were 4.17, 7.46, and 7.45mg/g, respectively. Soyasaponins extracted from the soybean sprout were analyzed with LC-electrospray ionization (ESI)-mass spectrometry, in which a reverse phase $C_18$ column was used for separation of saponins. In the soybeen sprout, group B saponin, I, II, III, IV, and V increased 7, 2, 1.4, 8.7, and 3.3 fold, respectively, compared to those in the soybean seed. Group B saponin I, II, III, IV, and V in the stem of the soybean sprout were 10.53, 1.45, 10.49, 5.72 and 8.14 fold the level of those in the cotyldon, respectively. In the root, the contents of group B saponin I, III, IV, and V were 5.54, 2.77, 4.86 and 9.73 fold, respectively, higher than those in cotyledon, but the content of group B saponin 2 was 2.96 fold less than that in cotyledon. These results indicate that the biosyntheses of group B saponins are differentially regulated in growing soybean sprout.

• The Korean Journal of Food And Nutrition
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• v.27 no.3
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• pp.532-537
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• 2014

Cortex Phellodendri (CP) is derived from the dried bark of Phellodendron amurense. It has been widely used as a drug in traditional Korea medicine for treating diarrhea, jaundice, swelling pains in the knees and feet, urinary tract infections, and infections of the body surface. Many analytical methods have been used to study oriental herbal medicines, such as thin-layer chromatography, column liquid chromatography, and high performance liquid chromatography (HPLC). In this study, preparative centrifugal partition chromatography (CPC) was successfully carried out in order to separate pure compounds from a CP methanol extract. The optimum two-phase CPC solvent system was composed of n-butanol: acetic acid: water (4:1:5 v/v/v). The flow rate of the mobile phase was 3 mL/min in ascending mode with rotation at 1,000 rpm. The CPC-separated fraction and purification procedures were carried out by preparatory HPLC. The $^1H$ NMR spectrum revealed that the resonances at ${\delta}$ 4.10 and 4.20 ppm corresponded to three protons ($-OCH_3$), whereas those at ${\delta}$ 6.10 ppm corresponded to two protons ($-OCH_2O-$). Further, two aromatic protons (H-11 and H-12) conveys a doublet-doublet pattern. The H-11 doublet and H-12 doublet appear at ${\delta}$ 7.98 and 8.11, respectively. The $^{13}C$ NMR. spectrum showed a tetrasubstituted with a methylenedioxy group at C2 and C3, and two methoxy groups at C9 and C10. The chemical structure of the berberine was identified by $^1H$, $^{13}C$-nuclear magnetic resonance and electrospray ionization-mass spectroscopy spectral data analysis.

• Herbal Formula Science
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• v.27 no.4
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• pp.269-284
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• 2019

This study was to investigate whether cultivated ginseng (CG), cultivated wild simulated ginseng (CWG) and cultivated red ginseng (CRG) extracts influences on the obesity. The saponin contents of 3 kinds of ginsengs were analysed by HPLC-ESI-TOF-MS. Total saponin contents were determined in CG on the most contents and since red ginseng has the highest PD (protopanaxadiol type) / PT (protopanaxatriol type) ratio, there may be differences between ginseng, wild ginseng, and red ginseng with respect to their pharmacological effects. Male C57BL/6J mice were fed a normal diet(N), HFD (60% Kcal fat, C), HFD with CG, CWG and CRG extracts (800 mg/kg) for 5 weeks. We observed change of total body weight, degree of hepatic lipid accumulation and immunohistochemical change of GLP-1 and insulin-secreting cells. Also this study attempts to use the physiological analysis method to analyze the changes of blood lipids, insulin and leptin concentration. The change of body weight and size of accumulated lipid droplets in liver lobules decreased in all of the experimental groups than the control(C) group. In the pancreas, the immunohistochemical density of insulin-secreting cells were significantly stronger in the CWG and CRG than C group. The levels of serum insulin and leptin significantly decreased 55.6%, 54.3% respectively in CWG and CRG. The changes of triglyceride, total cholesterol in serum decreased in CRG than the C group. Obesity related CG, CWG and CRG extracts might have contribute to improvement of obesity by regulating the levels of blood lipids and biochemical indicator of fat accumulation.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1665-1672
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• 2012

An 80% ethanol extract of Hizikia fusiforme was obtained and followed by successive fractionation using the organic solvents n-hexane, ethyl acetate, and n-butanol to identify the antioxidative substance. The aqueous part of the nbutanol fractionation step, showing high antioxidative activity, was subjected to reverse-phase liquid chromatography. As a result, a substance purified from a BB-2 fraction showed high antioxidative activity. The m/z 419 [M+H] molecular ion peak in the fraction was observed by the analysis of the ESI-LC/MS spectrum. By the analysis of 1H NMR (500 MHz, DMSO-$d_6$) and $^{13}C$ NMR (125 MHz, DMSO-$d_6$) spectra, a unique compound of the fraction was biochemically identified as a 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5HHMF). We also investigated the effect of 5HHMF on human gastric AGS carcinoma cells. Western blot analysis suggested that the flavone substantially increased the levels of the death receptor-associated apoptosis mediators Fas, Fas L, FADD, TRADD, and DR4 in a concentration-dependent manner. The levels of Fas, Fas L, TRADD, and DR4 in the cells treated with 5HHMF ($5{\mu}g/ml$) were approximately 26.4-, 12.8-, 6.7-, and 9.8-times higher than those of non-treated cells, respectively. Of note, the level of FADD protein in the cells exposed to 5HHMF ($1{\mu}g/ml$) increased approximately 9.6-times. In addition, the cleavage of caspase-3, -8, and -9 in cultured AGS cells treated with 5HHMF was significantly confirmed. Therefore, our results suggest that 5HHMF from H. fusiforme is involved in the induction of death receptor-associated apoptosis mediators in human gastric AGS carcinoma cells.

• Korean Journal of Environmental Agriculture
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• v.34 no.1
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• pp.57-63
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• 2015

BACKGROUND: Momordica charantia (MC) have been used in various parts of the world to treat diabetes. Despite the highest effects of phytochemicals than any other part of the plant, the main components of seed of MC grown in Korea and their properties have not been studied extensively. METHODS AND RESULTS: The seeds of MC were extracted into five different polarity solvents: 30%, 50%, 70%, 95% ethanol and distilled water. The 95% ethanol extract showed the most potent inhibition ($IC_{50}=88.7{\mu}g/mL$) against ${\alpha}$-glucosidase. To investigate the compounds responsible for this effect, activity guided fractionation of MC seeds by chromatography yielded two phytochemicals which were confirmed as vicine (1) and ${\alpha}$-eleostearic acid (2) based on their NMR and ESI-MS spectroscopic data. Among them, ${\alpha}$-eleostearic acid (2) possessed potent ${\alpha}$-glucosidase inhibitory activities with $IC_{50}$ values from $32.4{\mu}g/mL$. CONCLUSION: Collectively, the results from this present study strongly suggest that both extract and ${\alpha}$-eleostearic acid (2) have potent ${\alpha}$-glucosidase inhibitory activity. Furthermore, ${\alpha}$-eleostearic acid (2) as the most active ${\alpha}$-glucosidase inhibitor was proven to be present in high quantities in the MC seeds by a HPLC chromatogram.