• Journal of Nutrition and Health
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• v.47 no.1
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• pp.1-11
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• 2014

Purpose: Several studies have proven that EGCG, the primary green tea catechin, and glucosamine-6-phosphate (PGlc) reduce triglyceride contents in 3T3-L1 adipocytes. The objective of this study is to evaluate the combination effect of EGCG and PGlc on decline of accumulated fat in differentiated 3T3-L1 adipocytes. Methods: EGCG and PGlc were administered for 6 day for differentiation of 3T3-L1 adipocytes. Cell viability was measured using the CCK assay kit. In addition, TG accumulation in culture 3T3-L1 adipocytes was investigated by Oil Red O staining. We examined the expres-sion level of several genes and proteins associated with adipogenesis and lipolysis using real-time RT-PCR and Western blot analysis. A flow cytometer Calibar was used to assess the effect of EGCG and PGluco on cell-cycle progression of differentiating 3T3-L1 cells. Results: Intracelluar lipid accumulation was significantly decreased by combination treatment with EGCG $60{\mu}M$ and PGlc $200{\mu}g/m$ compared with control and EGCG treatment alone. In addition, use of combination treatment resulted in directly decreased expression of $PPAR{\gamma}$, $C/EBP{\alpha}$, and SREBP1. In addition, it inhibited adipocyte differentiation and adipogenesis through downstream regulation of adipogenic target genes such as FAS, ACSL1, and LPL, and the inhibitory action of EGCG and PGlc was found to inhibit the mitotic clonal expansion (MCE) process as evidenced by impaired cell cycle entry into S phase and the S to G2/M phase transition of confluent cells and levels of cell cycle regulating proteins such as cyclin A and CDK2. Conclusion: Combination treatment of EGCG and PGlc inhibited adipocyte differentiation through decreased expression of genes related to adipogenesis and adipogenic and cell cycle arrest in early stage of adipocyte differentiation.

• The Korean Journal of Physiology
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• v.18 no.2
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• pp.163-169
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• 1984

The effect of vanadate on the optimum pH of Na-K-ATPase was investigated. The results were as follows: 1) The optimum PH of Na-K-ATPase was shifted from PH 7.4 to 6.8 at 10 mM K by $5{\times}10^{-6}M$ vanadate. 2) The ratio of Na-K-ATPase activity at pH 6.8 and 7.4 increased with increasing vanadate concentration. 3) Inspite of the presence of $5{\times}10^{-6}M$ vanadate Na-K-ATPase activity at pH 7.4 was higher than that at pH 6.8 below 50 mM $Na^+$, and the ratio of Na-K-ATPase activity at pH 7,4 and 6.8 was higher than that of the control. 4) Na-K-ATPase activity at pH 7.4 was higher than that at pH 6.8 below 7mM $K^+$. 5) Optimum pH of Na-K-ATPase activity was shifted from pH 7.4 to 6.8 by $10^{-5}M$ vanadate at 5 mM $K^+$. 6) $K^+$-pNPPase activity increased with lowering of pH, and the degree of inhibition of $K^+$-pNPPase activity by $10^{-7}$M vanadate was decreased with lowering of pH. These results suggest that vanadate shifts the optimum pH of Na-K-ATPase activity to more acidic PH than PH 7.4. This effect may not be caused by the decrease in the inhibitory potency of vanadate itself to Na-K-ATPase by the change of medium pH, but mainly by the alteration of Na-and K-binding site, which appears in the presence of vanadate only.

• Journal of the Korean Applied Science and Technology
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• v.37 no.1
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• pp.124-132
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• 2020

The purpose of this study was to investigate the biological activities such as cytotoxicity and anti-inflammation using Manjakani (Quercus infectoria Olivier) extract. Manjakani was extracted from hot DW and 80% ethanol. Cell viability was assessed using MTT assay on RAW 264.7 cells. Also, anti-inflammatory activities were measured through changes in the levels of nitric oxide (NO), prostaglandin E₂ (PGE₂), leukotrien B4 (LTB₄), pro-inflammatory cytokines (IL-1β, IL-6 and tumor necrosis factor (TNF)-α) and transcription factor on LPS-induced RAW264.7 cells. The results confirmed that significant cytotoxicity does not appear in the concentration range of 1, 5, and 10 ㎍/㎖ of both extracts of this study. The production of NO was slowed by approximately MDE 37.2% and MEE 43.7% at 10 ㎍/㎖ concentration. Also, level of PGE₂ and LTB₄ was decreased MDE 30.9%/MEE 43.7% and MDE 37.1%/MEE 43.7%. In the case of inflammatory cytokine was reduced to MDE 38.8%/MEE 50.8% for IL-1β, MDE 35.0%/MEE 44.2% for IL-6 and MDE 31.9%/MEE 36.6% for TNF-α at 10 ㎍/㎖ concentration. The mRNA expression of NF-κB, iNOS and COX-2 significantly decreased by MDE 44.0%/MEE 16.0%, MDE 44.0%/MEE 55.0% and MDE 45.0%/MEE 40.0%, respectively, following the 10 ㎍/mL sample treatment when compared to the control. Both extracts were effective in anti-inflammatory activity. In addition, both extracts showed efficient changes of production of NO, PGE₂, LTB₄, pro-inflammatory cytokines and transcription factor. But MEE was found to have a higher inhibitory effect than MDE. In other words, Manjakani was showed significant biological activities showing anti-inflammation without cytotoxicity. These results will be provided as fundamental data for further development of the new health food and therapeutics related to the results above.

• Journal of Life Science
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• v.25 no.1
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• pp.68-74
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• 2015

Recently developed black waxy rice with a giant embryo ('Nunkeunheukchal', BGE) was selected and processed to produce high quality nutritional food. BGE contains high levels of several phytochemicals with antioxidant activities, as well as other reported health beneficial properties. In addition, the giant embryo has high protein, lipid, and amino acids contents. Within the free amino acids, ${\gamma}$-aminobutyric acid (GABA), a major inhibitory neurotransmitter, has long been used for treating the aftereffects of brain injuries and stroke. A method for manufacturing pop-rice and black rice tea by popping process in BGE is provided to increase a taste, nutrition and functionality. The produced 'pop-rice' showed increased protein (11.3%) and lipid (3.7%) contents compared with control variety, IB ('Ilmibyeo'). In addition, melanoidin related products, polyphenol and functional amino acid contents were increased by the popping process. Pop-rice tea made of BGE showed the highest extraction of total sugar, glucose, raffinose and sucrose (4 times higher than brown rice) by hot water. Scavenging activity ($SC_{50}$) of processed BGE rice powder showed strong antioxidative activity of 0.24 mg/ml using DPPH and 1.82 mg/ml using ABTs method. Thereafter, these results suggested that the popping processed rice of BGE could be one of the promising materials for healthy food development.

• Toxicological Research
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• v.33 no.2
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• pp.107-118
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• 2017

Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration ($IC_{50}$) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.

• Korean Journal of Food Science and Technology
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• v.49 no.5
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• pp.550-558
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• 2017

The effect of Artemisia argyi H. under liquid-state fermentation by Monascus purpureus (AAFM) on cognitive impairments has been studied in a mice model of diabetes-associated cognitive decline induced by streptozotocin (STZ). C57BL/6 mice (9 weeks of age, male) were separated into four groups: a normal control, STZ-induced diabetic mouse group (STZ group), Artemisia argyi H. (AA) 10 group (diabetic mouse+AA 10 mg/kg/day), AAFM 10 group (diabetic mouse+AAFM 10 mg/kg/day). Administration of AA and AAFM significantly improved glucose tolerance, as shown by the intraperitoneal glucose tolerance test (IPGTT), and ameliorated cognitive deficit, as shown by the behavioral tests including passive avoidance, Morris water maze, and Y-maze tests. After behavioral tests, the cholinergic system was examined by assessment of the acetylcholine (ACh) level and acetylcholinesterase (AChE) inhibitory activity, and the antioxidant system was also assessed by measuring malondialdehyde (MDA) and superoxide dismutase (SOD) levels in the brain and liver.

• Journal of Life Science
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• v.21 no.11
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• pp.1586-1591
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• 2011

This study was carried out to evaluate the antiobesity effects of Crataegus fructus in 3T3-L1 adipocytes and mice fed a high fat diet (high fat 45% cal). The inhibitory effect of methanol extract from Crataegus fructus on lipid accumulation in 3T3-L1 adipocytes was quantified using Oil red O staining. Compared with the control, lipid accumulation was significantly decreased by 10-25% with treatment with Crataegus fructus extract at a concentration of 600-2,000 ug/ml. Three-week old ICR mice (n=24) were randomly divided into four groups (T0: normal diet, T1: high fat diet, T2: high fat diet and 50 ug of Crataegus fructus extract, T3: high fat diet and 100 ug of Crataegus fructus extract) and were fed an experimental diet for 5 weeks. At the end of the experiment, body weight gain in the T1 group (3.9${\pm}$0.24 g) was higher than that in the T0 group (2.56${\pm}$0.14 g), while body weight gain in the T2 (3.02${\pm}$0.25 g) and T3 (2.58${\pm}$0.16 g) groups was significantly reduced as compared with that of the T1 group. Moreover, liver weight in the T1 (4.8${\pm}$0.17 g) and T2 (4.8${\pm}$0.16 g) groups was significantly higher than that of the T0 (4.05${\pm}$0.16 g) and T3 (4.57${\pm}$0.10 g) groups, while kidney weight was significantly lower than that of the T0 and T3 groups (p<0.05). The levels of total cholesterol and triglyceride in serum in the T2 and T3 groups were significantly decreased compared to the T1 group. These results suggest that Crataegus fructus can be used as functional materials in food and medicine.

• Food Science and Preservation
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• v.24 no.7
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• pp.992-999
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• 2017

The purpose of this study was to investigate the effects of the Bambusae Caulis in liquamen (BCL) on blood circulation in animal models. Previous studies on BCL have shown effects on thrombolytic activity and angiotensin-converting enzyme (ACE) inhibitory activity. In the mouse model, the triglyceride content were 301.5 mg/dL in the high fat diet+BCL II 0.01% group, 289.2 mg/dL in the high fat diet+BCL II 0.05% group, which was significantly lower than the high fat diet group. The total cholesterol content was 311.9 mg/dL in high fat diet+BCL II 0.01% and 293.7 mg/dL in high fat diet+BCL II 0.01% 0.05%, respectively, which was significantly lower than the high fat diet group. The HDL-cholesterol level was 206.0 mg/dL for the high fat diet, 196.6 mg/dL for the high fat diet+BCL II, and 189.2 mg/dL for the high fat diet+BCL II. There was no significant difference between the 0.01% and 0.05% groups. The high-fat diet+0.05% group was significantly improved in the blood flow compare to the high fat diet and the high fat diet+0.01% group. Platelet aggregation inhibition ability was inhibited in the high fat diet+0.01% and 0.05% groups compared to the high fat diet group.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.13-13
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• 2018

The anti-inflammatory (INF) compounds (1-15) were isolated from Agrimonia pilosa Ledeb. (APL) by activity-guided isolation technique. The isolated compounds (1-15) were identified as quercetin-7-O-rhanmoside (1), apigenin-7-O-glycoside (2), kaempferol-7-O-glycoside (3), apigenin-7-O-[6"-(butyl)-glycoside] (4), querceitn (5), kaempferol (6), apigenin (7), apigenin-7-O-[6"-(pentyl)-glycoside] (8), agrimonolide (9), agrimonolide-6-O-glucoside (10), desmethylagrimonolide (11), desmethylagrimonolide-6-O-glucoside (12), luteolin (13), vitexin (14) and isovitexin (15). Flavonoids, compound 2, 3, 11, and 14-15 have been found in APL for the first time. Furthermore, two novel flavone derivatives, compound 4 and 8, have been isolated inceptively in plant. In the no cytotoxicity concentration ranges of $0-20{\mu}M$, nitric oxide (NO) production level of 1-15 was estimated in LPS-treated Raw 264.7 macrophage cells. The flavone aglycones, 7 (apigenin, $IC_{50}=3.69{\pm}0.34{\mu}M$), 13 (luteolin, $IC_{50}=4.62{\pm}0.43{\mu}M$), 6 (kaempferol, $IC_{50}=14.43{\pm}0.23{\mu}M$) and 5 (quercetin, $IC_{50}=19.50{\pm}1.71{\mu}M$), exhibited excellent NO inhibitory (NOI) activity in dose-dependent manner. In the structure activity relationship (SAR) study of apigenin-derivatives (APD), apigenin; Api, apigenin-7-O-glucoside; Api-G, apignenin-7-O-[6"-(butyl)-glycoside]; Api-BG and apignenin-7-O-[6"-(pentyl)-glycoside]; Api-P, from APL on INF activity was investigated. The INF mediators level such as NO, INF-cytokines, NF-KB proteins, iNOS and COX-2 were sharply increased in Raw 264.7 cells by LPS. When pretreatment with APD in INF induced macrophages, NOI activity of Api was most effective than other APD with $IC_{50}$ values of $3.69{\pm}0.77{\mu}M$. And the NOI activity was declined in the following order: Api-BG ($IC_{50}=8.91{\pm}1.18{\mu}M$), Api-PG ($IC_{50}=13.52{\pm}0.85{\mu}M$) and API-G ($IC_{50}=17.30{\pm}0.66{\mu}M$). The NOI activity of two novel compounds, Api-PG and Api-BG were lower than their aglycone; Api, but more effective than Api-G (NOI: Api-PG and Api-BG). And their suppression ability on INF cytokines such as $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 mRNA showed the similar tendency. Therefore, the anti-INF mechanism study of Api-PG and Api-BG on nuclear factor-kappa B ($NF-{\kappa}B$) pathway, representative INF mechanism, was investigated and Api was used as positive control. Api-BF was more effectively prevent the than phosphorylation of $pI{\kappa}B$ kinase (p-IKK) and p65 than Api-PG in Raw 264.7 cells. In contrast, Api-PG and Api-BG were not reduced the phosphorylation of inhibitor of kappa B alpha ($I{\kappa}B{\alpha}$). Moreover, pretreatment with Api-PG and Api-BG, dose-dependently inhibited LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNAs and proteins in macrophage cells, and their expression were correlated with their NOI activity. Therefore, APL can be utilized to health promote agent associated with their AIN metabolites.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.11
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• pp.1528-1534
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• 2009

This study was designed to evaluate the anti-obesity and hypoglycemic effects of Gugija (Lycium chinense Mill) extracts in 3T3-L1 adipocytes. We investigated the $\alpha$-amylase and $\alpha$-glucosidase inhibitory activities of extracts from Gugija. Gugija was extracted by 70% EtOH and 80% MeOH and aqueous, respectively. A single oral dose of Gugija extract inhibited the increase of blood glucose levels significantly at 0, 30, 60, 90 and 120 min and decreased incremental response areas under the glycemic response curve. These results suggest that Gugija 70% EtOH extracts may delay carbohydrate digestion and reduce postprandial hyperglycemia. In addition, triglyceride content in 3T3-L1 adipocytes decreased at higher concentrations of Gugija 70% EtOH extract. Free fatty acid content in 3T3-L1 adipocytes was increased at higher concentrations of Gugija 70% EtOH extract. Also, glucose transporter 4 (GLUT4), the key insulin signaling pathway transcription factor, was remarkably increased by the Gugija 70% EtOH extract when compared to those of control cells in protein expression levels. Therefore, Gugija can be developed as an effective anti-obesity and hypoglycemic agent.