• BMB Reports
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• v.39 no.4
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• pp.432-440
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• 2006

A tuber lectin from Arisaema jacquemontii Blume belonging to family Araceae was purified by employing a single step affinity chromatography using column of asialofetuin-linked amino activated silica beads and the bound lectin was eluted with 100 mM glycine-HCl buffer pH 2.5. The purified A. jacquemontii lectin (AJL) showed a single protein band with an apparent molecular mass of 13.4 kDa when submitted to SDS-polyacrylamide gel electrophoresis under reducing as well as non-reducing conditions. The native molecular mass of AJL determined by gel filtration on a Biogel P-200 column was 52 kDa and its carbohydrate content was estimated to be 3.40%. Thus AJL is a tetrameric glycoprotein. The purified lectin agglutinated erythrocytes from rabbit but not from human. Its activity was not inhibited by any of the mono- and disaccharides tested except N-acetyl-D-lactosamine having minimal inhibitory sugar concentration (MIC) 25 mM. Among the glycoproteins tested only asialofetuin was found to be inhibitory (MIC $125\;{\mu}g/mL$). A single band was obtained in native PAGE at pH 4.5 while PAGE at pH 8.3 showed two bands. Isoelectric focusing of AJL gave multiple bands in the pI range of 4.6-5.5. When incorporated in artificial diet AJL significantly affected the development of Bactrocera cucurbitae (Coquillett) larvae indicating the possibility of using this lectin in a biotechnological strategy for insect management of cucurbits. Larvae fed on artificial diet containing sub-lethal dose of AJL showed a significant decrease in acid phosphatase and alkaline phosphatase activity while esterase activity markedly increased as compared to larvae fed on diet without lectin. Out of various human cancer cell lines employed in sulphorhodamine B (SRB) assay, this lectin was found to have appreciable inhibitory effect on the in vitro proliferation of HCT-15, HOP-62, SW-620, HT-29, IMR-32, SKOV-3, Colo-205, PC-3, HEP-2 and A-549 cancer cell lines by 82, 77, 73, 70, 41, 41, 37, 29, 21 and 21% respectively.

• The Journal of Internal Korean Medicine
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• v.25 no.2
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• pp.202-213
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• 2004

Object : Objective: This study was conducted to investigate the anti-cancer effects of Mylabris phalerata (반모) in some kinds of cancer cells. Materials and Methods: Some kinds of cancer cells lines were treated. We used nine kinds of cancer cell lines, such as stomach cancer cells (Kato), lung cancer cells (Calu-1, NCI-H 1395), urinary bladder cancer cells (HS789T), bone cancer cells (Saos-2), brain cancer cells (SK-N-MC), liver cancer cells (Hep-G2), skin cancer cells (Mo-1) and prostate cancer cells (PC-3) with the water decoction of Mylabris phalerata. The histological changes of all cell lines in the media (RPMI-1640) containing the decoction of Mylabris phalerata were observed and we examined cell death assay by trypan blue exclusion testing was examined. Finally, the change of mitochondrial membrane potential was measurd and the inhibitory effect of Mylabris phalerata on cell increase was examined by analyzing the cell cycle. Results: In histologic change all cancer cell lines showed withdrawn and floating appearance that is typical in cellular impairment. Most of the cell lines showed over 50% death rate after 24 hours in trypan blue exclusion tests. Especially the stomach, urinary bladder. brain and liver cell lines showed over 30% death rate after 12 hours. All cell lines treated with Mylabris phalerata were less stained than the control group and the mitochondrial membrane potential in the Mylabris phalerata treated cell lines was markedly lower than that in the control group. The measurement of DNA quantity in all cell lines showed the disappearance of the peak and the thickened left axis, which suggests that all cellular DNA degraded. Conclusion: Mylabris phalerata had cytotoxicity on various kinds of cancer cell lines and the mechanism of that was the impairment of mitochondria by the breakdown of the mitochondrial cell membrane. We propose that this is in part attributable to the destruction of DNA in cancer cells.

• Korean Journal of Food Science and Technology
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• v.49 no.6
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• pp.617-624
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• 2017

Chocolate, one of the most popular confectioneries in the world, is known for its aromatic flavor and high antioxidant activities. In this study, we investigated the effects of fermentation with commercially available lactic acid bacteria, ABT-5, on the flavor and antioxidant activities of dark chocolate. During 24 h fermentation, pH decreased from 5.52 to 3.97 and total acidity increased from 0.51 to 1.85%, whereas total polyphenol and flavonoid contents as well as DPPH and ABTS radical scavenging activities remained unchanged. Furthermore, compared with control HepG2 cells treated with unfermented dark chocolate, those treated with the fermented dark chocolate showed significantly lower levels of reactive oxygen species and higher viability under $H_2O_2-induced$ oxidative stress. Finally, GC-MS and headspace GC-MS analysis detected 4-hydroxy-2,5-dimethyl-3(2H)-furanone and 2-furanmethanol, known to enhance flavor, in the fermented dark chocolate. Collectively, these results suggest that ABT-5-fermented dark chocolate could be utilized for developing value-added dark chocolate products.

• Journal of the Korean Applied Science and Technology
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• v.35 no.1
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• pp.12-19
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• 2018

Perilla frutescens (P. frutescens) is one of evergreen shrubs belonging to the Labiatae and is grown wildly in Korea. This study was carried out to evaluate the anti-oxidative effects of Perilla frutescens Extracts by Extraction Methods (water, heating and sonication). Anti-oxidative effects were measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and total phenol content. Cell viability and hepatoprotective effects were identified by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay. Among various extracts, P. rutescens extracts by sonication showed the highest DPPH radical scavenging activity at $5000{\mu}g/mL$. Total phenolic content in P. frutescens extracts by sonication was $51.60{\pm}1.06mg\;GAE/g$ extract. However, P. frutescens extracts did not show hepatoprotective effects. This study identified anti-oxidative effects of P. frutescens extracts by sonication, and it would be necessary to perform further studies of P. frutescens extracts by sonication.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.5
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• pp.434-441
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• 2009

This study was conducted to compare the biological activities of 7 melania snails from the family Pleuroceridae (Semisulcospira coreana, Koreanomelania nodifila, Semisulcospira forticosta, Koreoleptoxis globus ovalis, Semisulcospira libertina, Semisulcospira tegulata and Semisulcospira gottschei) in Korea. Among the 7 species, S. coreana, Korean. nodifila, S. forticosta and S. gottschei showed over 80% cytotoxicities on three cancer cell lines (SNU-1, A549 and Hep 3B) compared to the non-treatment, whereas S. libertina and S. tegulata showed almost no growth inhibition activities on the same cancer cell lines. In relation to ACE inhibition activity, only S. coreana, Korean. nodifila, and S. forticosta showed over 60% ACE inhibition activities, whereas other melania snails exhibited inhibition activities of lower than 25%. DPPH radical scavenging activities were also determined, and used to categories melania snails into three groups based on Duncan's multiple range test at P<0.05. The amount of TNF-${\alpha}$ produced by in vitro mouse peritoneal macrophage was determined according to bioactivity on L-929 cells. Three melania snails, S. coreana, Korean. nodifila and S. gottschei, exhibited 95.2%, 89.7% and 93.7% cell death(%) on L-929 cells, respectively. Glucose-6-phosphate dehydrogenase inhibitory activity was also obtained in the extract of S. coreana (31.9%) and Korean. nodifila (28.1%), showing that these extracts can be used as supplemental dietary health foods. In conclusion, we believe that the extracts of melania snails should be given due consideration in functional health food development.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2003.04a
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• pp.22-39
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• 2003

Biological activity of phenolic compounds in seeds and leaves of safflower (Carthamu tinctorius L.) were evaluated using several in vitro and in vivo assays. Six phenolic constituents were isolated from the seeds and identified as N-feruloylserotonia, N- (p-coumaroyl)serotonin, matairesinol, 8′-hydroxyarctigenin, acacetin 7-O-$\beta$-D-glucoside (tilianine) and acacetin. Six phenolic compounds exhibited considerable antioxidative activity, and especially two serotonins showed potent DPPH radical scavenging activity and antiperoxidative activity against rat liver microsomal lipid peroxidation induced by the hydroxyl radical generated via a Fenton-type reaction. Additionally, six phenolic compounds possessed comparable cytotoxicity against three cancer cells, Hela cell, MCF-7 and HepG2 cell, and particularly acacetin and its glycosides had the most potent cytotoxicity. Moreover, we found that feeding safflower seeds attenuated bone loss, and lowered levels of plasma and liver lipids in ovariectomized rats. Serotonins, lignans and flavones stimulated proliferation of the osteoblast-like cells in a dose-dependent manner (10$^{-15}$ ~10$^{-6}$ M), as potently as E$_2$ (17$\beta$-estradiol). Particularly, serotonins were mainly responsible for bone-protecting and lipid lowering effects in ovariectomized rats. Meanwhile, eight flavonoids, including a novel quercetin-7-O-(6"-O-acetyl)-$\beta$-D-glucopyranoside and seven kown flavonoids, luteolin quercetin, luteolin 7-O-$\beta$-D-glucopyranoside, luteolin-7-O-(6"-O-acetyl)-$\beta$-D-gluco-pyranoside, quercetin 7-O- -glucopyranoside, acacetin 7-O-$\beta$-D-glucuronide and apigenin-6-C-$\beta$-D-glucopyranosyl-8-C-$\beta$-D-glucopyranoside were first isolated and identified from safflower leaf. Among these flavonoids, luteolin-acetyl-glucoside and $\beta$quercetin- acetyl-glucoside showed potent antioxidative activities against 2-deoxyribose degradation and lipid peroxidation in rat liver microsomes. Luteolin, quercetin and their corresponding glycosides also exhibited strong antioxidative activity, while acacetin glucuronide and apigenin-6, 8-di-C-glucoside were relatively less active. Finally, changes in phenolic compositions were also determined by HPLC in the safflower seed and leaf during growth stages and roasting process to produce standardized supplement powerds. These results suggest that phenolic compounds in the roasted safflower seed and leaf may be useful as potential sources of therapeutic agents against several pathological disorders such as carcinogenesis, atherosclerosis and osteoporosis.

• The Korea Journal of Herbology
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• v.30 no.4
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• pp.57-64
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• 2015

Objectives : Oxidative stress is one of the most causes of hepatocyte injury. Gleditsia spina, the thorns ofGleditsia sinensisLam., has been known for its anti-cancer and anti-inflammatory effects in Korean medicine. The present study investigated hepatoprotective effect of Gleditsia spina water extract (GSE) against oxidative stress induced by arachidonic acid (AA) + iron in HepG2 cells.Methods : To investigate cytoprotective effect of GSE, cells were pretreated with GSE and then subsequently exposed to 10 μM AA for 12 h, followed by 5 μM iron. Cell viability was monitored by MTT assay, and expression of apoptosis-related proteins was examined by immunoblot analysis. To identify responsible molecular mechanisms, reactive oxygen species (ROS) production, GSH contents, and mitochondrial membrane potential were measured. In addition, effect of GSE on nuclear factor erythroid 2-related factor 2 (Nrf2) activation was determined by immunoblot and antioxidant response element (ARE)-driven reporter gene assays.Results : GSE pretreatment prevented AA + iron-mediated cytotoxicity in concentration dependent manner. In addition, ROS production, glutathione depletion, and mitochondrial impairment by AA + iron were significantly inhibited by GSE. Furthermore, GSE promoted translocation of Nrf2 to nucleus, which acts as essential transcription factor for induction of antioxidant genes. Increased nuclear Nrf2 that caused by GSE treatment promoted transcriptional activity of ARE. Finally, GSE up-regulated sestrin-2 which was widely recognized as target gene of Nrf2.Conclusions : This study demonstrates that GSE protects hepatocytes from oxidative stress via activation of Nrf2 signaling pathway.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.1
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• pp.10-17
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• 2017

The natural preservative "complex Scutellaria baicalensis extract (BHC)" contains Scutellaria baicalensis, Glycyrrhiza uralensis (liquorice), Zizyphus jujube (jujube), and Astragalus propinquus (milk vetch root). BHC has been used as a natural preservative for more than 10 years to increase storage duration and quality of food with strong antibacterial activity. BHC has been added into functional foods as a subsidiary ingredient. However, no studies have been performed to test whether or not BHC affects the activity of main functional ingredients. In this study, we tested whether or not BHC has any effect on the hepatoprotective activity of lactic acid-fermented garlic extract (LAFGE) when formulated in a clinical test supplement. $H_2O_2-induced$ oxidative damage in HepG2 cells was not attenuated by BHC, indicating that BHC had no influence on the protective effect of LAFGE against oxidative damage. Furthermore, BHC had no effect on the hepatoprotective effect of LAFGE against acetaminophen-induced acute liver injury in rats, as indicated by no changes in alanine transaminase and aspartate transaminase levels. In conclusion, BHC, formulated in the clinical test supplement with LAFGE, had no effect on hepatoprotective activity, indicating BHC could be considered as a suitable natural preservative for liquefied functional food materials.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.12
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• pp.1672-1678
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• 2009

This study was carried out to investigate the total polyphenol, total flavonoid content, antioxidant activity, angiotensin I-converting enzyme (ACE) inhibitory activity, $\alpha$-glucosidase inhibitory activity, and antiproliferation activity of the citron seed. The citron seed were separated to hull and embryo, and extracted with n-hexane and 70% ethanol. Antioxidant activity of ethanol extract was higher than that of n-hexane extract. IC50 value for DPPH radical scavenging activity of ethanol extract of hull (CSE1) and embryo (CSE2) were 3.18 and 8.43 mg/mL, and those of total antioxidant activity were 19.96 and 11.28 mg AA eq/g, respectively. ACE inhibitory activity and $\alpha$-glucosidase inhibitory activity on CSE1 showed the highest values of 31.61 and 45.17%, respectively. Antiproliferation effects on the MCF7, HepG2, H460, HCT-116, and PC3 cell line showed the highest values of 14.09, 19.12, 12.29, 9.78, and 9.12% in extract concentration of 5 mg/mL, respectively. These results suggested that citron seed can be used for development of functional food material which have biological activities.

• Molecular & Cellular Toxicology
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• v.5 no.4
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• pp.298-303
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• 2009

(-)-Epigallocatechin-3-gallate (EGCG), a major flavonoid in green tea has multiple health benefits including chemoprevention, anti-inflammatory, anti-diabetic, and anti-obesity effects. In connection with these effects, EGCG can be a candidate to help the treatment of metabolic diseases. Metformin is a widely used anti-diabetic drug regulating cellular energy homeostasis via AMP-activated protein kinase (AMPK) activation. Therefore, the combination of metformin with EGCG may have additive or synergistic effects on treatment of type 2 diabetes. Nevertheless, there is no report for the pharmacokinetic and/or pharmacodynamic interaction of EGCG with metformin. Here, we evaluated the pharmacokinetic and pharmacodynamic interaction between metformin and EGCG in rats. Pharmacokinetics parameters of metformin were measured after oral administration of metformin in rats pre-treated with EGCG (10 mg/kg) or saline for 7 days. The results showed that there is no significant difference in pharmacokinetic parameters between saline control and EGCG-treated group. In addition, the hepatic AMPK activation by metformin in EGCG-treated rats was also similar to the control. The lack of additive effects of EGCG on AMPK activation or intracellular uptake of metformin was also evaluated in cells in the presence or absence of EGCG. Treatment of HepG2 cells with EGCG inhibited the metformin-induced AMPK activation. Combined results suggested that EGCG has no effect on the pharmacokinetics of metformin but may contribute to metformin action.