Padina arborescens extract protects high glucose-induced apoptosis in pancreatic β cells by reducing oxidative stress
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
BACKGROUND/OBJECTIVES: This study investigated whether Padina arborescens extract (PAE) protects INS-1 pancreatic β cellsagainst glucotoxicity-induced apoptosis. MATERIALS/METHODS: Assays, including cell viability, lipid peroxidation, generation of intracellular ROS, NO production, antioxidantenzyme activity and insulin secretion, were conducted. The expressions of Bax, Bcl-2, and caspase-3 proteins in INS-1 cellswere evaluated by western blot analysis, and apoptosis/necrosis induced by high glucose was determined by analysis of FITC-AnnexinV/PI staining. RESULTS: Treatment with high concentrations of glucose induced INS-1 cell death, but PAE at concentrations of 25, 50 or100 μg/ml significantly increased cell viability. The treatment with PAE dose dependently reduced the lipid peroxidation andincreased the activities of antioxidant enzymes reduced by 30 mM glucose, while intracellular ROS levels increased under conditionsof 30 mM glucose. PAE treatment improved the secretory responsiveness following stimulation with glucose. The results alsodemonstrated that glucotoxicity-induced apoptosis is associated with modulation of the Bax/Bcl-2 ratio. When INS-1 cells werestained with Annexin V/PI, we found that PAE reduced apoptosis by glucotoxicity. CONCLUSIONS: In conclusion, the present study indicates that PAE protects against high glucose-induced apoptosis in pancreaticβ cells by reducing oxidative stress.
Abstract
BACKGROUND/OBJECTIVES: This study investigated whether Padina arborescens extract (PAE) protects INS-1 pancreatic β cellsagainst glucotoxicity-induced apoptosis. MATERIALS/METHODS: Assays, including cell viability, lipid peroxidation, generation of intracellular ROS, NO production, antioxidantenzyme activity and insulin secretion, were conducted. The expressions of Bax, Bcl-2, and caspase-3 proteins in INS-1 cellswere evaluated by western blot analysis, and apoptosis/necrosis induced by high glucose was determined by analysis of FITC-AnnexinV/PI staining. RESULTS: Treatment with high concentrations of glucose induced INS-1 cell death, but PAE at concentrations of 25, 50 or100 μg/ml significantly increased cell viability. The treatment with PAE dose dependently reduced the lipid peroxidation andincreased the activities of antioxidant enzymes reduced by 30 mM glucose, while intracellular ROS levels increased under conditionsof 30 mM glucose. PAE treatment improved the secretory responsiveness following stimulation with glucose. The results alsodemonstrated that glucotoxicity-induced apoptosis is associated with modulation of the Bax/Bcl-2 ratio. When INS-1 cells werestained with Annexin V/PI, we found that PAE reduced apoptosis by glucotoxicity. CONCLUSIONS: In conclusion, the present study indicates that PAE protects against high glucose-induced apoptosis in pancreaticβ cells by reducing oxidative stress.
참고문헌 (38)
- Robertson RP, "Chronic oxidative stress as a central mechanism for glucose toxicity in pancreatic islet beta cells in diabetes", J Biol Chem, 279, pp.42351~42354 (2004)
- Zúrová-Nedelcevová J, "Participation of reactive oxygen species in diabetes-induced endothelial dysfunction", Neuro Endocrinol Lett, 27(Suppl 2), pp.168~171 (2006)
- Mandrup-Poulsen T, "Cytokine and free radicals as effector molecules in the destruction of pancreatic beta cells", Curr Top Microbiol Immunol, 164, pp.169~193 (1990)
- Butler AE, "β-cell deficit and increased β-cell apoptosis in humans with type 2 diabetes", Diabetes, 52, pp.102~110 (2003)
- Brownlee M, "The pathobiology of diabetic complications: a unifying mechanism", Diabetes, 54, pp.1615~1625 (2005)
- Heo SJ, "Antioxidant activities of enzymatic extracts from brown seaweeds", Bioresour Technol, 96, pp.1613~1623 (2005)
- Pal Singh I, "Phloroglucinol compounds of natural origin", Nat Prod Rep, 23, pp.558~591 (2006)
- 박미화, "Protective Effect of Padina arborescens Extract against High Glucose-induced Oxidative Damage in Human Umbilical Vein Endothelial Cells", Preventive Nutrition and Food Science, 18(1), pp.11~17 (2013), 한국식품영양과학회[DOI]
- 박미화, "Hypoglycemic Effect of Padina arborescens Extract in Streptozotocin-induced Diabetic Mice", Preventive Nutrition and Food Science, 17(4), pp.239~244 (2012), 한국식품영양과학회[DOI]
- Fautz R, "Application of the neutral red assay (NR assay) to monolayer cultures of primary hepatocytes: rapid colorimetric viability determination for the unscheduled DNA synthesis test (UDS)", Mutat Res, 253, pp.173~179 (1991)
- Wang H, "Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader", Free Radic Biol Med, 27, pp.612~616 (1999)
- Fraga CG, "Lipid peroxidation measured as thiobarbituric acid-reactive substances in tissue slices: characterization and comparison with homogenates and microsomes", Free Radic Biol Med, 4, pp.155~161 (1988)
- Nath J, "Modulation of human neutrophil inflammatory responses by nitric oxide: studies in unprimed and LPS-primed cells", J Leukoc Biol, 62, pp.805~816 (1997)
- Marklund S, "Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase", Eur J Biochem, 47, pp.469~474 (1974)
- Aebi H, "Catalase in vitro", Methods Enzymol, 105, pp.121~126 (1984)
- Lawrence RA, "Glutathione peroxidase activity in seleniumdeficient rat liver", Biochem Biophys Res Commun, 71, pp.952~958 (1976)
- Green CD, "Elevated insulin secretion from liver X receptor-activated pancreatic β-cells involves increased de novo lipid synthesis and triacylglyceride turnover", Endocrinology, 150, pp.2637~2645 (2009)
- Yamabe N, "Beneficial effect of Corni Fructus, a constituent of Hachimi-jio-gan, on advanced glycation end-product-mediated renal injury in streptozotocintreated diabetic rats", Biol Pharm Bull, 30, pp.520~526 (2007)
- Lenzen S, "Oxidative stress: the vulnerable beta-cell", Biochem Soc Trans, 36, pp.343~347 (2008)
- Robertson RP, "Beta-cell glucose toxicity, lipotoxicity, and chronic oxidative stress in type 2 diabetes", Diabetes, 53(Suppl 1), pp.S119~S124 (2004)
- Tiedge M, "Complementary action of antioxidant enzymes in the protection of bioengineered insulinproducing RINm5F cells against the toxicity of reactive oxygen species", Diabetes, 47, pp.1578~1585 (1998)
- Evans JL, "Oxidative stress and stress-activated signaling pathways: a unifying hypothesis of type 2 diabetes", Endocr Rev, 23, pp.599~622 (2002)
- Del Guerra S, "Gliclazide protects human islet beta-cells from apoptosis induced by intermittent high glucose", Diabetes Metab Res Rev, 23, pp.234~238 (2007)
- Robertson R, "Chronic oxidative stress as a mechanism for glucose toxicity of the beta cell in type 2 diabetes", Cell Biochem Biophys, 48, pp.139~146 (2007)
- Kong Q, "Oxidative damage to RNA: mechanisms, consequences, and diseases", Cell Mol Life Sci, 67, pp.1817~1829 (2010)
- Kappus H, "Oxidative stress in chemical toxicity", Arch Toxicol, 60, pp.144~149 (1987)
- Du X, "Generation of reactive oxygen intermediates, activation of NF-kappaB, and induction of apoptosis in human endothelial cells by glucose: role of nitric oxide synthase?", Free Radic Biol Med, 27, pp.752~763 (1999)
- Messmer UK, "Bcl-2 protects macrophages from nitric oxide-induced apoptosis", J Biol Chem, 271, pp.20192~20197 (1996)
- McDaniel ML, "Cytokines and nitric oxide in islet inflammation and diabetes", Proc Soc Exp Biol Med, 211, pp.24~32 (1996)
- Kaneto H, "Reducing sugars trigger oxidative modification and apoptosis in pancreatic beta-cells by provoking oxidative stress through the glycation reaction", Biochem J, 320, pp.855~863 (1996)
- Tajiri Y, "Aminoguanidine exerts a beta-cell functionpreserving effect in high glucose-cultured beta-cells (INS-1)", Int J Exp Diabetes Res, 1, pp.111~119 (2000)
- Grankvist K, "Superoxide dismutase is a prophylactic against alloxan diabetes", Nature, 294, pp.158~160 (1981)
- Benhamou PY, "Adenovirusmediated catalase gene transfer reduces oxidant stress in human, porcine and rat pancreatic islets", Diabetologia, 41, pp.1093~1100 (1998)
- Krause MS, "L-arginine is essential for pancreatic beta-cell functional integrity, metabolism and defense from inflammatory challenge", J Endocrinol, 211, pp.87~97 (2011)
- Moriscot C, "Contribution of adenoviral-mediated superoxide dismutase gene transfer to the reduction in nitric oxide-induced cytotoxicity on human islets and INS-1 insulin-secreting cells", Diabetologia, 43, pp.625~631 (2000)
- Fariss MW, "Role of mitochondria in toxic oxidative stress", Mol Interv, 5, pp.94~111 (2005)
- Antonsson B, "The Bcl-2 protein family", Exp Cell Res, 256, pp.50~57 (2000)
- Porter AG, "Emerging roles of caspase-3 in apoptosis", Cell Death Differ, 6, pp.99~104 (1999)