Stevia rebaudiana의 항산화 효과
Anti-oxidant Effect on Stevia rebaudiana
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초록
Stevia rebaudiana is a traditional herb used as a sweetener in Brazil and Paraguay as well as Korea and China. This study investigated the efficacy of Stevia rebaudiana methanol extract (SRE) to protect cells against the mitochondrial dysfunction and apoptosis in hepatocyte. To determine the effects of SRE on oxidative stress, we used the human derived hepatocyte cell line, HepG2 cell. Treatment of arachidonic acid (AA)+iron in HepG2 cells synergistically amplified cytotoxicity, as indicated by the excess reactive oxygen species (ROS) and mitochondrial permeability transition by fluorescence activated cell sorter (FACS) and immunoblot analysis. Treatment with SRE protected hepatocytes from AA+iron-induced cellular toxicity, as shown by alterations in the protein levels related with cell viability such as procaspase-3. SRE also prevented the mitochondrial dysfunction induced by AA+iron, and showed anti-oxidant effects as inhibition of H2O2 production and GSH depletion. Moreover, we measured the effects of SRE on AMP-activated protein kinase (AMPK), a key regulator in determining cell survival or death. Acetyl-CoA Carboxylase (ACC), a direct downstream target of AMPK. SRE increased phosphorylation of ACC, and prevented the inhibition of ACC phosphorylation by AA+iron. These results indicated that SRE has the ability to protect cells against AA+iron-induced H2O2 production and mitochondrial impairment, which may be mediated with AMPK-ACC pathway.
Abstract
Stevia rebaudiana is a traditional herb used as a sweetener in Brazil and Paraguay as well as Korea and China. This study investigated the efficacy of Stevia rebaudiana methanol extract (SRE) to protect cells against the mitochondrial dysfunction and apoptosis in hepatocyte. To determine the effects of SRE on oxidative stress, we used the human derived hepatocyte cell line, HepG2 cell. Treatment of arachidonic acid (AA)+iron in HepG2 cells synergistically amplified cytotoxicity, as indicated by the excess reactive oxygen species (ROS) and mitochondrial permeability transition by fluorescence activated cell sorter (FACS) and immunoblot analysis. Treatment with SRE protected hepatocytes from AA+iron-induced cellular toxicity, as shown by alterations in the protein levels related with cell viability such as procaspase-3. SRE also prevented the mitochondrial dysfunction induced by AA+iron, and showed anti-oxidant effects as inhibition of H2O2 production and GSH depletion. Moreover, we measured the effects of SRE on AMP-activated protein kinase (AMPK), a key regulator in determining cell survival or death. Acetyl-CoA Carboxylase (ACC), a direct downstream target of AMPK. SRE increased phosphorylation of ACC, and prevented the inhibition of ACC phosphorylation by AA+iron. These results indicated that SRE has the ability to protect cells against AA+iron-induced H2O2 production and mitochondrial impairment, which may be mediated with AMPK-ACC pathway.
참고문헌 (39)
- Halliwell, B., "Free radicals, antioxidants and human disease: where are we now?", J Lab Clin Med, 119(6), pp.598~620 (1992)
- Kovacic, P., "Mechanisms of carcinogenesis : Focus on oxidative stress and electron transfer", Curr Med Chem, 8(7), pp.773~796 (2001)
- Valko, M., "Oxygen free radical generating mechanisms in the colon: Do the semiquinones of vitamin K play a role in the aetiology of colon cancer?", Biochim Biophys Acta, 1527(3), pp.161~166 (2001)
- Awasthi, Y. C., "Purification and properties of human erythrocyte glutathione peroxidase", J Biol Chem, 250(13), pp.5144~5149 (1975)
- 이소연, "산화스트레스가 유도된 인체 간암세포 (HepG2)에서 Sulforaphane과 Diallyl Sulfide, Capsaicin, Gingerol의 항산화효과 비교연구*", Journal of Nutrition and Health, 44(6), pp.488~497 (2011), 한국영양학회
- Gurpreet, K., "Evaluation of antioxidant activity of Cassia siamea flowers", J Ethnopharmacol, 108(3), pp.340~348 (2006)
- Scharffetter-Kochanek, K., "UV-induced reactive oxygen species in photo-carcinogenesis and photoaging", Bio Cehm, 378(11), pp.1247~1257 (1997)
- Horton, A. A., "Lipid peroxidation and mechanisms of toxicity", Crit Rev Toxicol, 18(1), pp.27~79 (1987)
- Ito N, "Carcinogenicity and modification of the carcinogenic respose by BHA, BHT, and other antioxidants", Crit Res Toxicol, 15(2), pp.109~150 (1985)
- 구강모, "고춧잎 품종별 항산화 활성과 항산화 성분", Journal of Applied Biological Chemistry, 52(2), pp.70~76 (2009), 한국응용생명화학회
- Balboa, M. A., "Oxidative stress and arachidonic acid mobilization", Biochim Biophys Acta, 1761(4), pp.385~391 (2006)
- Monjazeb, A. M., "Inhibitors of arachidonic acid metabolism act synergistically to signal apoptosis in neoplastic cells", Prostaglandins Leukot Essent Fatty Acids, 73(6), pp.463~474 (2005)
- Claria, J, "Regulation of cell proliferation and apoptosis by bioactive lipid mediators", Recent Pat Anticancer Drug Discov, 1(3), pp.369~382 (2006)
- Nakanishi, M., "Roles of cPLA2a and arachidonic acid in cancer", Biochim Biophys Acta, 1761(11), pp.1335~1343 (2006)
- Kim, K. K., "Characterization of ent-Kaurenoic Acid 13-Hydroxylase in Steviol Biosynthesis of stevia rebaudiana Bertoni", Agricultural Chemistry and Biotechnology, 40(6), pp.501~507 (1997)
- 김재훈, "다양한 추출방법으로 조제된 스테비아 잎 추출물의 항산화 활성", 한국식품영양과학회지, 39(2), pp.313~318 (2010), 한국식품영양과학회
- Nammi, S., "Protective effects of ethanolic extract of Zingiber officinale rhizome on the development of metabolic syndrome in high-fat diet-fed rats", Basic Clin Pharmacol Toxicol, 104(5), pp.366~373 (2009)
- Kim, Y. W., "Efficacy of sauchinone as a novel AMPK-activating lignan for preventing iron-induced oxidative stress and liver injury", Free Radic Biol Med, 47(7), pp.1082~1092 (2009)
- Kwon, Y. N., "Oxidized metabolites of oltipraz exert cytoprotective effects against arachidonic acid through AMP-activated protein kinase-dependent cellular antioxidant effect and mitochondrial protection", Drug Metab Dispos, 37(6), pp.1187~1197 (2009)
- Cocco, T., "Arachidonic acid interaction with the mitochondrial electron transport chain promotes reactive oxygen species generation", Free Radic Biol Med, 27(1-2), pp.51~59 (1999)
- Shin, S. M., "Inhibition of arachidonic acid and iron-induced mitochondrial dysfunction and apoptosis by oltipraz and novel 1, 2-dithiole-3-thione congeners", Mol Pharmacol, 75(1), pp.242~253 (2009)
- Zamzami, N., "Measurement of membrane permeability and the permeability transition of mitochondria", Methods Cell Biol, 65, pp.147~158 (2001)
- 하주헌, "생체 에너지 대사 조절에서 AMPK의 역할", Endocrinology and Metabolism, 25(1), pp.9~17 (2010), 대한내분비학회
- 변명우, "스테비아로부터 분리한 Austroinulin의 RAW264.7 세포에 대한 항염증 효과", 한국식품영양과학회지, 41(4), pp.456~461 (2012), 한국식품영양과학회
- Galaris, D., "Oxidative stress and iron homeostasis : mechanistic and health aspects", Crit Rev Clin Lab Sci, 45(1), pp.1~23 (2008)
- Neufeld, E. J, "Oral chelators deferasirox and deferiprone for transfusional iron overload in thalassemia major : new data, new questions", Blood, 107(9), pp.3436~3441 (2006)
- Takahashi, M., "Estimation of lipid peroxidation of live cells using fluorescent probe", Free Radic Biol Med, 31(2), pp.164~174 (2001)
- Melov, S. Mitochondrial oxidative stress, "Physiologic consequences and potential for a role in aging", Ann N Y Acad Sci, 908, pp.219~225 (2000)
- Minczuk, M., "Sequence-specific modification of mitochondrial DNA using a chimeric zinc finger methylase", Proc Natl Acad Sci USA, 103(52), pp.19689~19694 (2006)
- Shindo, Y., "Enzymic and non-enzymic antioxidants in epidermis and dermis of human skin", J Invest Dermatol, 102(1), pp.112~128 (1944)
- Zamzami, N., "The mitochondrion in apoptosis : how Pandora's box opens", Nat Rev Mol Cell Biol, 2(1), pp.67~71 (2001)
- Browning, J. D., "Molecular mediators of hepatic steatosis and liver injury", J Clin Invest, 114(2), pp.147~152 (2004)
- Yeom, J. B., "Mitochondrial Physiology", Korean Society of Medical Biochemistry and Molecular Biology, 15(1), pp.25~33 (2008)
- Towler, M. C., "AMP-activated protein kinase in metabolic control and insulin signaling", Circ Res, 100(3), pp.328~341 (2007)
- Carling, D, "The AMP-activated protein kinase cascade—a unifying system for energy control", Trends Biochem Sci, 29(1), pp.18~24 (2004)
- Hawley, S. A., "Haracterization of the AMP-activated protein kinase kinase from rat liver and identification of threonine 172 as the major site at which it phosphorylates AMP-activated protein kinase", J Biol Chem, 271(44), pp.27879~27887 (1996)
- Shwa, R. J., "The tumor suppressor LKB1 kinase directly activates AMP-activated kinase and regulates apoptosis in response to energy stress", Proc Natl Acad Sci USA, 101(10), pp.3329~3335 (2004)
- Cantó, C., "AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity", Nature, 458(7241), pp.1056~1060 (2009)
- Shin, S. M., "Resveratrol Protects Mitochondria against Oxidative Stress through AMP-Activated Protein Kinase-Mediated Glycogen Synthase Kinase-3βInhibition Downstream of Poly(ADPribose)polymerase-LKB1 Pathway", Mol Pharmacol, 76(4), pp.884~895 (2009)
