• 한국식품영양과학회지
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• 제39권8호
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• pp.1102-1106
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• 2010

본 연구는 진피 에탄올 추출물의 alloxan에 의해 유도된 HIT-T15 세포의 산화적 손상으로부터 세포 생존율 및 인슐린분비 조절능에 대해 조사하였다. 진피 에탄올 추출물의 총 폴리페놀 함량과 총 플라보노이드 함량을 측정한 결과, 각각 $57.00{\pm}2.91\;mg/g$, $8.11{\pm}2.83\;mg/g$으로 나타났으며 alloxan 처리 농도가 증가됨에 따라 HIT-T15 세포의 생존율은 감소하였으며, alloxan 11.58 mM에서 약 50%의 세포 독성이 일어남을 확인하였다. 동결건조 된 진피 에탄올 추출물(CP-Et) 0.125~0.75 mg/mL 농도에서 무처리구의 세포 생존율은 $100.90{\pm}1.51{\sim}97.56{\pm}0.73%$로 나타나 HIT-T15 세포에 대한 CP-Et의 처리 농도는 0.125~0.75 mg/mL로 결정하였다. 0.125, 0.25, 0.5 mg/mL 농도의 CP-Et를 처리한 결과, alloxan에 의해 유도된 산화적 세포손상으로부터 세포 생존율이 각각 $80.52{\pm}3.29$, $74.17{\pm}6.75$, $67.53{\pm}5.8%$로 나타나 유의성 있게 보호되어짐을 확인하였다. CP-Et를 처리한 HIT-T15 세포에서는 산화적 손상에 대한 세포 생존율과는 다르게 0.125 mg/mL 농도에서 $116.93{\pm}2.11{\mu}g/mg$로 인슐린 분비가 대조구와 비교 시 유의적으로 증가되었음을 확인 할 수 있었다. 따라서 진피 에탄올 추출물에 의한 항산화적 방어로 2형 당뇨의 $\beta$-cell 양의 감소와 인슐린 저항성에 대한 개선이 가능함을 제시하며, 그 작용 기작에 대해서는 차후 더 많은 연구가 필요하다 하겠다.

• 한국식품영양학회지
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• 제25권1호
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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.

• 한국자원식물학회지
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• 제25권2호
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• pp.184-192
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• 2012

본 연구는 췌장베타세포인 HIT-T15 세포를 이용하여 매실주정추출물(PME)의 alloxan에 의한 산화스트레스로부터의 세포보호, 인슐린 분비능 및 항산화 효소 활성을 평가하였다. PME는 alloxan에 의해 유발된 산화스트레스로부터 세포를 보호하여 세포생존율을 증가시켰다. PME는 세포막 손상지표인 LDH 방출을 억제하였고 $NAD^+$/NADH ratio를 유의적으로 증가시켜 세포사멸이 억제되어짐을 확인하였다. 또한 alloxan 단독처리군에 비해 250 ${\mu}g$/ml PME 처리군에서 인슐린 분비량이 유의성 있게 증가하였다. Alloxan과 PME를 동시에 처리하여 HIT-T15세포의 항산화효소 활성을 측정했을 때 산화스트레스에 의해 감소되었던 항산화효소 활성이 PME에 의해 보호되는 효과를 확인하였다. 이상의 연구결과로부터 PME는 세포괴사 및 DNA fragmentation을 억제하고 세포내 항산화효소 활성을 증가시켜 alloxan에 의해 유발된 산화스트레스로부터 췌장베타세포를 보호하고 이에 따른 인슐린 분비능 조절 효과가 있는 것으로 생각된다.

• 한국식품영양과학회지
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• 제39권1호
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• pp.31-35
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• 2010

본 실험에서는 hamster $\beta$-cell인 HIT-T15 cell을 이용하여 돼지감자추출물의 생리활성 및 기능을 검증하고자 하였다. 돼지감자추출물을 첨가한 NC(0 ${\muL/mL$), HT2(1.1 ${\muL/mL$), HT3(1.5 ${\muL/mL$)군과 inulin을 첨가한 NC(0 ${\muL/mL$), IN2(1.8 ${\muL/mL$), IN3(2.5 ${\muL/mL$)군으로 나누어 실험하였다. 세포 viability 측정한 결과 시료를 첨가하지 않은 군을 100%로 보았을 때 돼지감자추출물을 첨가한 HT3(1.5 ${\muL/mL$)군과 inulin을 첨가한 IN2(1.8 ${\muL/mL$), IN3(2.5 ${\muL/mL$) 군에서 세포생존율이 유의적으로 증가하였다(p<0.05). 시료처리 후 췌장 $\beta$-세포 파괴를 유도하지 않고 HIT-T15 cell의 cell culture supernatant를 이용하여 cytotoxicity를 측정한 결과 시료를 첨가하지 않은 NC(0${\muL/mL$)군에 비해 모든 군에서 cytotoxicity가 낮게 나타났다. Alloxan(4 mM)으로 $\beta$-세포 파괴를 유도하여 HIT-T15 cell에서 세포보호 효과를 측정한 결과 시료를 첨가하지 않은 NC(0 ${\muL/mL$)군에 비해 돼지감자추출물을 첨가한 HT2(1.1 ${\muL/mL$), HT3(1.5 ${\muL/mL$)군에서 세포생존율이 유의적으로 증가하였다(p<0.05). 또한 췌장 $\beta$-세포 파괴를 유도하여 HIT-T15 cell이 분비한 인슐린 분비능 및 세포 내 $NAD^+$/NADH 함량을 측정한 결과 시료를 첨가하지 않은 NC(0 ${\muL/mL$)군에 비해 돼지감자추출물을 첨가한 HT3(1.5 ${\muL/mL$)군에서 인슐린 분비량과 $NAD^+$/NADH 함량이 유의적으로 증가하였다(p<0.05). 이상의 연구 결과 돼지감자추출물은 HIT-T15 cell의 생존율을 높이고, 세포보호 효과를 가짐으로써 인슐린 분비능 정상화 및 $NAD^+$ 함량을 증가시켜 혈당 조절 및 당뇨에 긍정적 효과가 있을 것으로 사료된다.

• 한국식품저장유통학회지
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• 제18권6호
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• pp.1002-1008
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• 2011

본 연구는 당에 의해 유도된 HIT-T15를 이용하여 항당뇨 소재로 잘 알려져 있는 오미자 및 뽕잎 추출물의 인슐린 분비능에 미치는 영향에 대해 조사하였다. 오미자 발효주정 추출물과 뽕잎 열수 추출물의 총 폴리페놀함량은 각각 $20.11{\pm}0.35$ mg/g 및 $50.02{\pm}0.62$ mg/g이었으며, 총 플라보노이드함량은 $2.85{\pm}0.15$ mg/g 및 $8.76{\pm}0.43$ mg/g 이었다. 전자공여능은 뽕잎 열수 추출물이 $83.21{\pm}3.96%$로 오미자 발효 주정 추출물의 $60.84{\pm}1.86%$보다 높았으며 오미자 및 뽕잎 추출물의 ORAC은 각각 $32.22{\pm}1.46$, $63.54{\pm}2.52\;{\mu}moles$ TE/g였는데, 뽕잎 열수 추출물이 오미자 발효주정 추출물보다 높은 항산화능을 보여주었다. Glucose 10 mM 농도를 처리한 HIT-T15에 오미자 발효주정 추출물 $100{\mu}g/mL$ 첨가시 인슐린 생산량은 2배 정도 증가되었다. 고농도인 16.7 mM의 glucose가 처리된 HIT-T15의 인슐린 생산량($1.54{\pm}0.09{\mu}g/g$ protein)은 glucose 무첨가에서 생산되는 인슐린량($1.06{\pm}0.04{\mu}g/g$ protein)과 유사하였는데 뽕잎 열수추출물 $100{\mu}g/mL$ 첨가에 의해 인슐린 생산성은 대략 6.7배 정도 향상되었다. 따라서 오미자 발효주정 추출물과 뽕잎의 열수 추출물이 고농도의 당으로부터 유도되는 췌장세포의 인슐린 분비를 개선시킬 수 있음을 확인하였다.

• Biomolecules & Therapeutics
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• 제16권2호
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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.

• The Korean Journal of Physiology and Pharmacology
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• 제8권1호
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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.

• 약학회지
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• 제41권2호
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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.

• Biomolecules & Therapeutics
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• 제6권4호
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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.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.95.1-95.1
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• 2003

Sopungsungi-won has been used as a traditional medicine for diabetes and it has been proved evidently as a potential remedy for type 2 diabetes mellitus. Both in vivo and in vitro experiments with water extract of Sopungsungi-won have been reported to exhibit anti-diabetic effects in our previous studies. In the present study, we have chosen Rheum undulatum (RU), which is the main component of Sopungsungi-won, to examine its anti-apoptotic effect on pancreatic b-cells, HIT-T15, against oxidative stress induced by hydrogen peroxide (H$_2$O$_2$). (omitted)