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Kwon, Jung, Kim, Kang, Seo, Song, Yoon, Jeon, Ha, Yoon, Kim, Lee, Noh, Kwon, Kim, Koh, Rhee, Lim, Lee, and Park: Cytoprotective Effect by Antioxidant Activity of Quercetin in INS-1 Cell Line
Original Articles
The Journal of Korean Diabetes Association

The Journal of Korean Diabetes Association 2007; 31(5): 383-390.

Published online: 30 September 2007

DOI: https://doi.org/10.4093/jkda.2007.31.5.383

Cytoprotective Effect by Antioxidant Activity of Quercetin in INS-1 Cell Line

Min Jeong Kwon1, 2, Hye Sook Jung2, Mi Kyung Kim1, Seong Hoon Kang1, Gwang Wook Seo1, Jae Kwang Song1, Tae Yeon Yoon1, Min Kyeong Jeon1, Tae Hwan Ha1, Chang Shin Yoon2, Mi Kyung Kim2, 3, Woo Je Lee1, Jeong Hyun Noh1, Soo Kyung Kwon1, Dong Joon Kim1, Kyung Soo Koh1, Byung Doo Rhee1, Kyung Ho Lim1, Soon Hee Lee1, Jeong Hyun Park1

1Department of Internal Medicine, College of Medicine, Inje University, Korea.

2Molecular Therapy Lab, Paik Memorial Institute for Clinical Research, Inje University, Korea.

3Department of Internal Medicine, Maryknoll General Hospital, Korea.

Received 27 June 2007       Accepted 11 September 2007

Copyright © 2007 Korean Diabetes Association

Abstract

Background

Oxidative stress is induced under diabetic conditions and causes various forms of tissue damages in the patients with diabetes. Recently, pancreatic beta cells are regarded as a putative target of oxidative stress-induced tissue damage, and this seems to explain in part the progressive deterioration of beta cell function in type 2 diabetes. The aim of this study was to examine the potential of Quercetin (QE) to protect INS-1 cells from the H2O2-induced oxidative stress and the effects of QE on the glucose-stimulated insulin secretion in INS-1 cells.

Methods

To study the cell viability, cells were incubated with H2O2 and/or QE at the various concentrations. To confirm the protective effect by QE in response to H2O2, the levels of antioxidant enzymes were assessed by RT-PCR and Western blot, and glutathione peroxidase activities were quantified by spectrophotometrical method. Glucose-stimulated insulin secretion (GSIS) was measured by ELISA.

Results

Cell incubations were performed with 80 µM of H2O2 for 5 hours to induce 40 - 50% of cell death. QE gradually showed protective effect (IC50 = 50 µM) in dose-dependent manner. Superoxide dismutase (SOD) mRNA level in H2O2 + QE group was increased as compared to H2O2 group, but catalase did not changed. And the QE recruited glutathione peroxidase activity against H2O2-induced oxidative injuries in INS-1 cells.

Conclusion

In conclusion, these findings suggest that QE might have protective effect on beta cells by ameliorating oxidative stress and preserving insulin secretory function.

Keywords: Antioxidant effect, Diabetes mellitus, INS-1 cell, Oxidative stress, Quercetin

Figures and Tables

Fig. 1
H2O2-induced cytotoxicity in INS-1 cells. INS-1 cells exposed to various concentrations of H2O2(0, 10, 20, 50, 80 and 100 µM) for 5 hours, and then, cell viability was measured by MTT method.
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Fig. 2
Effect of quercetin on INS-1 cells. INS-1 cells were treated with 80 µM of H2O2(IC50) and various concentration of quercetin (0, 10, 25, 50 and 100 µM) for 5 hours, and then, cell viability was measured with MTT method.
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Fig. 3
The effects of quercetin (QE) on H2O2-induced cell damage in INS-1 cells. Morphology of INS-1 cells of control (A) and treated with 80 µM H2O2 (B) or 80 µM H2O2 + 50 µM QE (C) for 5 hours was observed by optical microscope (×200).
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Fig. 4
The effect of quercetin on SOD and catalase expression in INS-1 cells. A, SOD and catalase mRNA expression in INS-1 cells of control and treated with H2O2 and/or Quercetin for 5 hours was evaluated by RT-PCR; B, Expression of SOD and catalase in INS-1 cells of all groups was detected by western blotting. C, control; QE, quercetin; WB, western blotting. *P < 0.05.
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Fig. 5
The effect of quercetin (QE) on GPx activity. The GPx activity in contol, H2O2 and H2O2 + QE group was measured by Total Glutathione Quantification Kit and determined the change in absorbance per minute. C, control; GPx, Glutathione peroxidase; QE, quercetin.
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Fig. 6
GSIS in Contol, H2O2 and H2O2 + QE Group. Insulin secretion depended on glucose concentration was detected by ELISA in INS-1 cells of control and treated with H2O2 and/or Quercetin for 5 hours. C, control; GSIS, glucose-stimulated insulin secretion; QE, quercetin.
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Table 1
Primers used for RT-PCR
jkda-31-383-i001

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