J Korean Diabetes Assoc.  2007 Sep;31(5):383-390.

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

• Kwon MJ
• Jung HS
• Kim MK
• Kang SH
• Seo GW
• Song JK
• Yoon TY
• Jeon MK
• Ha TH
• Yoon CS
• Kim MK
• Lee WJ
• Noh JH
• Kwon SK
• Kim DJ
• Koh KS
• Rhee BD
• Lim KH
• Lee SH
• Park JH

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 microM of H2O2 for 5 hours to induce 40 - 50% of cell death. QE gradually showed protective effect (IC50 = 50 microM) 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.

Keyword

Antioxidant effect; Diabetes mellitus; INS-1 cell; Oxidative stress; Quercetin

MeSH Terms

Antioxidants
Blotting, Western
Catalase
Cell Death
Cell Line*
Cell Survival
Diabetes Mellitus
Enzyme-Linked Immunosorbent Assay
Glutathione Peroxidase
Humans
Insulin
Insulin-Secreting Cells
Oxidative Stress
Quercetin*
RNA, Messenger
Superoxide Dismutase
Antioxidants
Catalase
Glutathione Peroxidase
Insulin
Quercetin
RNA, Messenger
Superoxide Dismutase

Figure

• 

  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.

• 

  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.

• 

  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).

• 

  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.

• 

  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.

• 

  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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