• The Korean Journal of Food And Nutrition
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• v.25 no.1
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• pp.123-131
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• 2012

The objective of the present study was to evaluate the potential antidiabetic and antioxidant effect of the ethanol extract from Chrysanthemum cornarium L. var. spatiosum(CSE) against alloxan-induced oxidative stress in pancreatic ${\beta}$-cells, HIT-T15. In this study, the antidiabetic effect of CSE was examined using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliu bromide(MTT) cell proliferation assay, lactate dehydrogenase(LDH) release assay, $NAD^+$/NADH ratio and insulin secretion. To further investigate whether CSE is involved in the antioxidant activity of alloxan-damaged HIT-T15 cells, its antioxidant effect against alloxan-induced oxidative stress was measured in HIT-T15 cells by determining the levels of antioxidant enzymes including superoxide dismutase(SOD), glutathione S-transferase(GST), glutathione reductase(GR) and glutathione peroxidase(GPx). The results of this analysis showed that alloxan significantly decreased cell viability, increased LDH leakage, and lowered $NAD^+$/NADH ratio and insulin secretion in HIT-T15 cells. However, CSE significantly increased the viability of alloxan-treated cells and lowered LDH leakage. The intracellular NAD+/NADH ratio and insulin secretion were also significantly increased by 1.7-fold and 1.3-fold, respectively, after treatment with 100 ${\mu}g/m{\ell}$ CSE. The HIT-T15 cells treated with alloxan showed significant decreases in the activities of antioxidant enzymes, while CSE significantly elevated the levels of antioxidant enzymes. These findings suggest that CSE could have a protective effect against cytotoxicity and dysfunction of pancreatic cells in the presence of alloxan-induced oxidative stress.

• Korean Journal of Plant Resources
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• v.25 no.2
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• pp.184-192
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• 2012

The present study was designed to examine the potential antidiabetic and antioxidant effect of ethanol extract from $Prunus$ $mume$ fruit (PME) against alloxan-induced oxidative stress in pancreatic ${\beta}$-cells, HIT-T15. To evaluate the antidiabetic effect of PME, 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliu bromide (MTT) cell proliferation assay, lactate dehydrogenase (LDH) release assay, $NAD^+$/NADH ratio and insulin secretion were assessed. We also measured its antioxidant effect against alloxan-induced oxidative stress in the cells by assessing the levels of the antioxidant enzymes including superoxide dismutase (SOD), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx). The results of this analysis showed that alloxan significantly decreased cell viability, increased LDH leakage, and lowered $NAD^+$ /NADH ratio and insulin secretion in HIT-T15 cells. However, PME significantly increased the viability of alloxan-treated cells and lowered LDH leakage. The intracellular $NAD^+$ /NADH ratio and insulin secretion were also increased by 1.5~1.9-fold and 1.4-fold, respectively, after treatment with the PME. The HIT-T15 cells treated with alloxan showed significant decreases in the activities of antioxidant enzymes, while PME significantly elevated the levels of antioxidant enzymes. Based on these results, we suggest that PME could have a protective effect against the cytotoxicity and dysfunction of pancreatic ${\beta}$-cells in the presence of alloxan-induced oxidative stress.

• Biomolecules & Therapeutics
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• v.16 no.2
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• pp.100-105
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• 2008

The sphingolipid metabolites act as lipid mediator for cell proliferation and apoptosis in mammalian cells. In bacteria, sphingolipid metabolism remains unknown. The purpose of this study was to investigate whether sphingolipid metabolism is potential target for fumonisin $B_1$($FB_1$) and desipramine in Sphingomonas chungbukensis, Gram-negative bacteria, by comparing the intracellular contents of bacterial sphingolipids with ones of HIT-T15 ${\beta}$-cells, hamster pancreatic cells. The concentrations of ceramide and dihydroceramide were 18.0 ${\pm}$ 12.0 and 0.025 ${\pm}$ 0.018 nmol/mg protein, respectively, in HIT-T15 cells. However, the concentrations of ceramide and dihydroceramide in the bacterial culture were 2.0 ${\pm}$ 1.2 and 10.6 ${\pm}$ 5.5 nmol/mg protein, respectively. $FB_1$ decreased the level of ceramide from 18.0 to 3.8 nmol/mg protein in HIT-T15 ${\beta}$-cells. However, dihydroceramide content in $FB_1$-treated HIT-T15 cells was slightly decreased compared with the control culture. When S. chungbukensis was treated with either $FB_1$ or desipramine, dihydroceramide level was increased by 5- and 4-fold, respectively, compared with the control bacteria. These results indicate that $FB_1$ and desipramine may act as an activator in bacterial sphingolipid biosynthetic pathway, and bacterial sphingolipid metabolism pathway appears to be different from the pathway of mammalian cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.8
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• pp.1102-1106
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• 2010

This study was carried out to investigate the effect of ethanol extract from citrus peels (CP-Et) against the alloxan-induced oxidative damage on HIT-T15, Hamster pancreatic $\beta$-cell. Total polyphenol and flavonoid contents in CP-Et were $57.00{\pm}2.91\;mg/g$ and $8.11{\pm}2.83\;mg/g$, respectively. Cell toxicity on HIT-T15 by CP-Et (0.125~0.75 mg/mL) was not observed. CP-Et (0.125 mg/mL) increased cell proliferation rate of HIT-T15, which was treated alloxan ($IC_{50}=11.58\;mM$) (cell viability=$80.52{\pm}3.29%$ of normal cell, p<0.05). In comparison with insulin secretion of oxidative damaged HIT-T15, 1.5 fold ($116.93{\pm}2.11\;{\mu}g/mg$ protein) was increased by treatment CP-Et treatment (0.125 mg/mL) in HIT-T15 (p<0.05). These results showed that CP-Et contribute to repairing cells and improvement of insulin expression on oxidative stress pancreatic $\beta$-cell, and also suggested application of CP-Et as a functional food material for type 2 diabetes.

• YAKHAK HOEJI
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• v.41 no.2
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• pp.260-267
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• 1997

To establish prediabetes in vitro model concerning the etiology of IDDM(Insulin Dependent Diabetes Mellitus) in cellular level we have designed prediabetes in vitro models in pa ncreatic beta cells. HIT-T15, RINm5F and isolated rat islets were chosen as pancreatic beta cells, and streptozotocin (STZ) used as diabetogenic agent. Degree of beta cell destruction to establish prediabetic in vitro model was determined by cell proliferation and insulin release using thymidine uptake and radio immuno assay. When HIT-T15 and RINm5F cells were treated with STZ, the degree of cell deterioration was dependent upon the origin and passage number of beta cells, and in the case of isolated islets STZ showed the more sensitivity than above two beta cell lines. The concentration and exposure time of STZ treatment to establish prediabetes in vitro model in beta cell lines and isolated rat islets were 2 ~ 10mM, 30 min. and 1 ~ 5mM, 30 min., respectively.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.1
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• pp.51-55
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• 2004

Sopungsungi-won has been used as a traditional medicine for diabetes and it has been proved to be a potential remedy for type 2 diabetes mellitus. We previously reported that water extract of Sopungsungi-won exhibits anti-diabetic effects both in vivo and in vitro experiments. In the present study, we have chosen to examined anti-apoptotic effect of Rheum undulatum, which is the main component of Sopungsungi-won, on pancreatic ${\beta}-cells$, HIT-T15, against hydrogen peroxide $(H_2O_2)$. oxidative stress. To investigate the anti-apoptotic effect of Rheum undulatum water extract (RUWE) against $H_2O_2-induced$ apoptosis in pancreatic ${\beta}-cell$ line of hamster, HIT-T15, MTT assay, DAPI staining, TUNEL assay, RT-PCR and caspase-3 enzyme assay were performed. The morphological analysis demonstrated that cells treated with $H_2O_2$ exhibited classical apoptotic features, while such changes was reduced in cells pre-treated with RUWE. In addition, RUWE pre-treated cells prior to $H_2O_2$ treatment induced increase of levels of bcl-2 expression and decrease of caspase-3 enzyme activity compared to cells treated with $H_2O_2$ only. These results provide the possibility of usage of RU in patients with progressively deteriorated diabetes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.1
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• pp.31-35
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• 2010

This study was designed to evaluate anti-diabetes effect of Helianthus tuberosus extract (HT) in HIT-T15 cells. There were 5 experimental groups according to treatment NC (0 ${\muL/mL$), HT2 (1.1 ${\muL/mL$), HT3 (1.5 ${\muL/mL$), IN2 (1.8 ${\muL/mL$), IN3 (2.5 ${\muL/mL$). Inulin (IN) was used as a positive control for the Helianthus tuberosus extract groups. Cell viability was significantly increased in the HT3 (1.5 ${\muL/mL$), IN2 (1.8 ${\muL/mL$), IN3 (2.5 ${\muL/mL$) groups, compared with the NC group. There was no significant difference in cytotoxicity among all groups. Cell survival by MTT assay with alloxan was significantly increased in the HT2 (1.1 ${\muL/mL$), HT3 (1.5 ${\muL/mL$) groups, compared with the NC group. Insulin secretion and NAD+/NADH ratio were significantly increased in the HT3 group, compared with the NC group. We found that Helianthus tuberosus extract increased cell viability, had a protective effect on $\beta$-cells, and increased insulin secretion level and $NAD^+$/NADH ratio in HIT-T15 cells. These results suggest that Helianthus tuberosus extract improves the diabetes-related factors.

• Biomolecules & Therapeutics
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• v.6 no.4
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• pp.371-377
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• 1998

The aim of the present study was to investigate the effect of Palmiwon on the type 1-prediabetic models induced by streptozotocin (STZ) in RINm5F cells and HIT-T15 cells. Palmiwon increased the cell proliferation and insulin release when pre- and post-treated for the STZ-exposed pancreatic beta cells. The cell proliferation and insulin release of these beta cells were measured by $^3$H-thymidine uptake and RIA, respectively. We also analyzed nutrients such as sugars, fatty acid and amino acids and minerals containing in Palmiwon using by gas chromatography, amino acid analyzer and AA spectrometer, respectively. Palmiwon seems to have protective and recovery properties on the prediabetic model in cellular level, which were ascribe to various nutrients and minerals containing in Palmiwon. From these results, it could be suggested that Palmiwon may be available as preventive and therapeutic prescription of type 1 diabetes mellitus.

• The Korean Journal of Food And Nutrition
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• v.23 no.2
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• pp.233-239
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• 2010

Phenolic acid concentrates of rice bran(RB-ex) and hydroxycinnamic acids were investigated for their anti-hyperglycemic activities through glucose uptake and glucokinase activity using HepG2 cells and stimulatory effects on insulin secretion using HIT-T15 cells. RB-ex was prepared as an ethylacetate extract after alkaline hydrolysis and hydroxycinnamic acids, found as major compositions of RB-ex, such as ferulic acid(FA), sinapic acid(SA) and p-coumaric acid(p-CA) were investigated to compare with the properties of RB-ex. The properties of glucose uptake in HepG2 cells were examined in the absence of insulin and two different glucose concentrations(5.5 mM and 25 mM). RB-ex and FA showed anti-hyperglycemic activities through the increase of glucose uptake and the stimulation of glucokinase activity in HepG2 cells. RB-ex exhibited higher glucose uptakes with higher glucose concentrations, whereas FA exhibited the same increasing effects on both concentrations of glucose. RB-ex and FA exhibited doubled glucokinase activities relative to control. In the presence of insulin in the 25 mM glucose-containing medium, the levels of glucose uptake were increased in all treatments compared with control. As stimulatory effects of samples on insulin secretion were estimated, RB-ex and FA stimulated insulin secretion at a concentration of 25 ${\mu}g/m{\ell}$ and in particular, FA showed the highest amount of insulin-release in HIT-T15 cells. Antioxidative effects on HIT-T15 cells, RB-ex and hydroxycinnamic acids, excluding p-CA, showed inhibitory activities of 78% to 80% at a concentration of 100 ${\mu}g/m{\ell}$. On the basis of these results, we conclude that RB-ex and FA could help decrease blood glucose levels and prevent the cell damages via antioxidant activity.

• Food Science and Preservation
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• v.18 no.6
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• pp.1002-1008
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• 2011

This study aimed to examine the effect of the Schizandra chinensis fruit and Morus alba leaf on insulin expression in HIT-T15 cells, which is exposed by glucose. The total polyphenol contents of the S. chinensis fruit ethanol extract and the M. alba leaf hot-water extract were $20.11{\pm}0.35$ mg/g and $50.02{\pm}0.62$ mg/mL, respectively. The S. chinensis fruit ethanol extract and the M. alba leaf hot-water extract contained $2.85{\pm}0.15$ and $8.76{\pm}0.43$ mg/g flavonoids, respectively. The antioxidant ability of the M. alba leaf hot-water extract was found to be superior to that of the S. chinensis fruit ethanol extract. Compared to the HIT-T15-treated 10 mM 2-deoxy-D-glucose, the $100{\mu}g/mL$ S. chinensis ethanol extract was found to have a two fold increase in insulin productivity. Moreover, the $100{\mu}g/mL$ M. alba leaf hot-water extract promoted the insulin secretion of high-glucose-damaged HIT-T15 almost ten fold. The above results showed that the S. chinensis fruit ethanol extract and M. alba leaf hot-water extract may improve the insulin productivity of the beta cell with glucose-induced oxidative damage. These data suggest that the S. chinensis fruit ethanol extract and the M. alba leaf hot-water extract can be used as food materials for the regulation of insulin secretion.