WANG Jing-ru1,2,YE Kai-he1,LÜ Yan-qing1,WEI Song-cheng1,ZHANG Xiao-qi3,YE Wen-cai3,YE Chun-ling1*
1.Department of Pharmacology, Pharmacy College of Jinan University, Guangzhou 510632, China; 2. The Karolinska Institute in Sweden, Stockholm 17177, Sweden; 3.Research Institute of Traditional Chinese Medicine and Natural Medicine of Jinan University, Guangzhou 510632, China
Abstract��
OBJECTIVE To investigate the effects of guavenoic acid(GA) on protection against oxidative damage in INS-1 cells. METHODS INS-1�� cells were cultured in vitro. Control group, medium group, model group, drug groups and positive group were set. INS-1 cells were treated with GA in 0.3, 1, 3, 10, 30 nmol��L-1 for 48 h. Using 650 ��mol��L-1H2O2 challenges the INS-1 cells for 2 h to establish the oxidative damage model(OD), the cell viability was tested by MTT assay, the nuclear was stained by Hoechst 33258, the ROS in OD-INS-1 was tested by ROS-kit and the apoptosis was tested via double staining-fcm. RESULTS Compared with model group, at low concentration GA(0.3, 1 nmol��L-1) could protect INS-1 cells exposed to H2O2 significantly(P<0.05 or P<0.01). Nuclear staining results showed that compared with the oxidative damage model group, cells treated with 0.3 nmol��L-1 GA, and emitting weaker cell fluorescence. 0.3, 1, 3 nmol��L-1 GA could reduce the DCF fluorescence intensity very significantly(P<0.01). At 0.3 nmol��L-1, GA can significantly resist the occurrence of apoptosis(P<0.05). CONCLUSION GA could protect INS-1 cells from oxidative damage, maybe achieved by inhibiting intracellular ROS generation and against apoptosis.
WANG Jing-Ru-,
,
YE Kai-He- etc
.Effects of Guavenoic Acid on Protection against Oxidative Damage in INS-1 Cells and Its Mechanisms[J] Chinese Pharmaceutical Journal, 2015,V50(3): 232-238
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[1]
GARDNER D G, SHOBACK D M. Greenspan��s Basic & Clinical Endocrinology. 9th ed. New York:McGraw-Hill Medical, 2011.[2] GEMBAL M, GILON P, HENQUIN J C. Evidence that glucose can control insulin release independently from its action on ATP-sensitive K+ channels in mouse B cells. J Clin Investigat, 1992, 89(4):1288-1295.[3] PI J, BAI Y, ZHANG Q, et al. Reactive oxygen species as a signal in glucose-stimulated insulin secretion. Diabetes, 2007, 56(7):1783-1791.[4] SINGH U, DEVARAG S, JIALAL I. Vitamin E, oxidative stress, and inflammation. Annu Rev Nut, 2005, 25:151-174.[5] PEREIRA E C, FERDERBAR S, BERTOLAMI M C, et al. Biomarkers of oxidative stress and endothelial dysfunction in glucose intolerance and diabetes mellitus.Clin Biochem, 2008, 41(18):1454-1460.[6] YOKOZAWA T, KIM Y, KIM H Y, et al. Protective effect of persimmon peel polyphenol against high glucose-induced oxidative stress in LLC-PK(1)cells. Food Chem Toxicol, 2007, 45(10):1979-1987.[7] LEE S H, HAN J S, HEO S J, et al. Protective effects of dieckol isolated from Ecklonia cava against high glucose-induced oxidative stress in human umbilical vein endothelial cells. Toxicol in Vitro, 2010, 24(2):375-381.[8] GUTIERREZ R M, MITCHELL S, SOLIS R V. Psidium guajava:A review of its traditional uses, phytochemistry and pharmacology. J Ethnopharmacol, 2008, 117(1):1-27.[9] MARTIN M A, FERNANDEZ-MILLAN E, RAMOS S, et al. Cocoa flavonoid epicatechin protects pancreatic beta cell viability and function against oxidative stress. Mol Nutr Food Res, 2014, 58(3):447-456. BRASNYO P, MOLNAR G A, MOHAS M, et al. Resveratrol improves insulin sensitivity, reduces oxidative stress and activates the Akt pathway in type 2 diabetic patients. Br J Nutr, 2011, 106(3):383-389. ROBERTSON R P, HARMON J, TRAN P O, et al. Glucose toxicity in ��-cells:Type 2 diabetes, good radicals gone bad, and the glutathione connection. Diabetes, 2003, 52(3):581-587. ROBERTSON R P, HARMON J S. Diabetes, glucose toxicity, and oxidative stress:A case of double jeopardy for the pancreatic islet �� cell. Free Rad Biol Med, 2006, 41(2):177-184. CUMAOGLU A, RACKOVA L, STEFEK M, et al. Effects of olive leaf polyphenols against H2O2 toxicity in insulin secreting ��-cells. Acta Biochim Pol, 2011,58(1):45-50. CHENG Q, DONG W, QIAN L, et al. Visfatin inhibits apoptosis of pancreatic ��-cell line, MIN6, via the mitogen-activated protein kinase/phosphoinositide 3-kinase pathway. J Mol Endocrinol, 2011, 47(1):13-21. LACRAZ G, FIGEAC F, MOVASSAT J, et al. Diabetic GK/Par rat ��-cells are spontaneously protected against H2O2-triggered apoptosis. A cAMP-dependent adaptive response. Am J Physiol-Endocrinol Metab, 2010, 298(1):17-27. RAALTE D H, KWA K A A, GENUGTEN R E, et al. Islet-cell dysfunction induced by glucocorticoid treatment:Potential role for altered sympathovagal balance. Metabolism, 2013, 62(4):568-577. WANG Y, LI Y, YIN J, et al. Autophagy regulates inflammation following oxidative injury in diabetes. Autophagy, 2013, 9(3):272-277. TAKAHASHI H, TRAN P O T, LEROY E, et al. D-Glyceraldehyde causes production of intracellular peroxide in pancreatic islets, oxidative stress, and defective beta cell function via non-mitochondrial pathways. J Biol Chem, 2004, 279(36):37316-37323. BONEKAMP N A, VOLKL A, FAHIMI H D, et al. Reactive oxygen species and peroxisomes:Struggling for balance. Biofactors, 2009, 35(4):346-355. SCHRADER M, FAHIMI H D. Peroxisomes and oxidative stress. Biochim Biophys Acta(BBA)-Mol Cell Res, 2006, 1763(12):1755-1766. DEL GUEEEA S, LUPI R, MARSELLI L, et al. Functional and molecular defects of pancreatic islets in human type 2 diabetes. Diabetes, 2005, 54(3):727-735. LEE S H. Bioactive compounds extracted from ecklonia cava by using enzymatic hydrolysis protects high glucose-induced damage in INS-1 pancreatic beta-cells. Appl Biochem Biotechnol, 2012, 167(7):1973-1985.
2015, Vol. 50 
