Diabetes and Metabolism Journal

Korean Diabetes Association (대한당뇨병학회)
권호 표지

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

INS-1 세포에서 항산화 효과를 통한 Quercetin의 세포 보호 효과

Kwon, Min-Jeong;Jung, Hye-Sook;Kim, Mi-Kyung;Kang, Seong-Hoon;Seo, Gwnag-Wook;Song, Jae-Kwang;Yoon, Tae-Yeon;Jeon, Min-Kyeong;Ha, Tae-Hwan;Yoon, Chang-Shin;Kim, Mi-Kyung;Lee, Woo-Je;Noh, Jeong-Hyun;Kwon, Soo-Kyung;Kim, Dong-Joon;Koh, Kyung-Soo;Rhee, Byung-Doo;Lim, Kyung-Ho;Lee, Soon-Hee;Park, Jeong-Hyun
권민정;정혜숙;김미경;강성훈;서광욱;송재광;윤태연;전민경;하태환;윤창신;김미경;이우제;노정현;권수경;김동준;고경수;이병두;임경호;이순희;박정현

  • Published : 20070900
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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. (J Kor Diabetes Assoc 31:383~390, 2007)

배경: 고혈당에 의한 산화 스트레스는 당뇨병환자에서 만성합병증을 보이는 여러 조직 손상의 원인 중의 하나로 알려져 있다. 제1형 당뇨병뿐만 아니라 인슐린저항성을 중심으로 한 제2형 당뇨병 역시 결국 베타세포 기능의 장애로 인해 발병한다. 최근 췌장베타 세포 자체에 대한 산화 스트레스가 당뇨병 발병의 원인일 것이라는 연구가 보고되고 있으며 항산화제의 당뇨병 예방에 대한 역할에 관심이 기울여지고 있다. 우리는 양파, 포도 등에 있는 강력한 항산화제의 일종인 Quercetin (QE)을 사용하여 인슐린을 분비하는 베타세포 모델(INS-1 cell)에서 보호효과를 알아보고자 하였다.방법: H2O2로 산화스트레스를 주고 QE 첨가 정도에 따른 세포 생존능을 MTT를 이용하여 측정하였다. 세포 보호에 관한 작용이 항산화 효과에 의한 것임을 확인하기 위하여 SOD와 catalase를 Western blot으로 확인하고 RT-PCR로 mRNA를 측정하였으며 Total Glutathione Quantification Kit로 glutathione peroxidase 활성을 측정하였다. 기능 보존을 알아보기 위하여 고혈당 자극에 의한 인슐린분비능을 ELISA로 측정하여 확인하였다.결과: 세포 생존능에 있어 QE은 농도에 비례하여 산화스트레스에 대한 보호 효과를 보였다. SOD는 QE 첨가군에서 증가하였지만 catalase는 변화가 없었고 산화스트레스만 받은 INS-1 cell에 비해 QE 첨가군에서 유의하게 glutathione peroxidase 활성의 개선을 보였다. 고혈당 자극에 의한 인슐린분비능 또한 QE 첨가군에서 높게 나타났다.

Keywords

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