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PLOS ONE  2011 

EGb761, a Ginkgo Biloba Extract, Is Effective Against Atherosclerosis In Vitro, and in a Rat Model of Type 2 Diabetes

DOI: 10.1371/journal.pone.0020301

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Background EGb761, a standardized Ginkgo biloba extract, has antioxidant and antiplatelet aggregation and thus might protect against atherosclerosis. However, molecular and functional properties of EGb761 and its major subcomponents have not been well characterized. We investigated the effect of EGb761 and its major subcomponents (bilobalide, kaemferol, and quercetin) on preventing atherosclerosis in vitro, and in a rat model of type 2 diabetes. Methods and Results EGb761 (100 and 200 mg/kg) or normal saline (control) were administered to Otsuka Long-Evans Tokushima Fatty rats, an obese insulin-resistant rat model, for 6 weeks (from 3 weeks before to 3 weeks after carotid artery injury). Immunohistochemical staining was performed to investigate cell proliferation and apoptosis in the injured arteries. Cell migration, caspase-3 activity and DNA fragmentation, monocyte adhesion, and ICAM-1/VCAM-1 levels were explored in vitro. Treatment with EGb761 dose-dependently reduced intima-media ratio, proliferation of vascular smooth muscle cells (VSMCs) and induced greater apoptosis than the controls. Proliferation and migration of VSMCs in vitro were also decreased by the treatment of EGb761. Glucose homeostasis and circulating adiponectin levels were improved, and plasma hsCRP concentrations were decreased in the treatment groups. Caspase-3 activity and DNA fragmentation increased while monocyte adhesion and ICAM-1/VCAM-1 levels decreased significantly. Among subcomponents of EGb761, kaemferol and quercetin reduced VSMC migration and increased caspase activity. Conclusions EGb761 has a protective role in the development of atherosclerosis and is a potential therapeutic agent for preventing atherosclerosis.


[1]  Valli G, Giardina EG (2002) Benefits, adverse effects and drug interactions of herbal therapies with cardiovascular effects. J Am Coll Cardiol 39: 1083–95.
[2]  Zimmermann M, Colciaghi F, Cattabeni F, Di LM (2002) Ginkgo biloba extract: from molecular mechanisms to the treatment of Alzheimer's disease. Cell Mol Biol 48: 613–23.
[3]  Smith JV, Luo Y (2003) Elevation of oxidative free radicals in Alzheimer's disease models can be attenuated by Ginkgo biloba extract EGb 761. J Alzheimers Dis 5: 287–300.
[4]  Gohil K, Packer L (2002) Global gene expression analysis identifies cell and tissue specific actions of Ginkgo biloba extract, EGb 761. Cell Mol Biol 48: 625–31.
[5]  Oken BS, Storzbach DM, Kaye JA (1998) The efficacy of Ginkgo biloba on cognitive function in Alzheimer disease. Arch Neurol 55: 1409–15.
[6]  Smith JV, Luo Y (2004) Studies on molecular mechanisms of Ginkgo biloba extract. Appl Microbiol Biotechnol 64: 465–72.
[7]  Smith PF, Maclennan K, Darlington CL (1996) The neuroprotective properties of the Ginkgo biloba leaf: a review of the possible relationship to platelet-activating factor (PAF). J Ethnopharmacol 50: 131–9.
[8]  Oyama Y, Chikahisa L, Ueha T, Kanemaru K, Noda K (1996) Ginkgo biloba extract protects brain neurons against oxidative stress induced by hydrogen peroxide. Brain Res 712: 349–52.
[9]  Defeudis FV (2002) Bilobalide and neuroprotection. Pharmacol Res 46: 565–8.
[10]  Luo Y, Smith JV, Paramasivam V, Burdick A, Curry KJ, et al. (2002) Inhibition of amyloid-beta aggregation and caspase-3 activation by the Ginkgo biloba extract EGb761. Proc Natl Acad Sci U S A 99: 12197–202.
[11]  Rapin JR, Yoa RG, Bouvier C, Drieu K (1997) Effects of repeated treatments with an extract of Ginkgo biloba (EGb 761) and bilobalide on liver and muscle glycogen contents in the non-insulin-dependent diabetic rat. Drug Dev Res 40: 68–74.
[12]  Tanaka S, Han LK, Zheng YN, Okuda H (2004) Effects of the flavonoid fraction from Ginkgo biloba extract on the postprandial blood glucose elevation in rats. Yakugaku Zasshi 124: 605–11.
[13]  Clowes AW, Reidy MA, Clowes MM (1983) Kinetics of cellular proliferation after arterial injury. I. Smooth muscle growth in the absence of endothelium. Lab Invest 49: 327–33.
[14]  Lim S, Ahn BY, Chung SS, Park HS, Cho BJ, et al. (2009) Effect of a peroxisome proliferator-activated receptor gamma sumoylation mutant on neointimal formation after balloon injury in rats. Atherosclerosis 206: 411–7.
[15]  Ansari B, Coates PJ, Greenstein BD, Hall PA (1993) In situ end-labelling detects DNA strand breaks in apoptosis and other physiological and pathological states. J Pathol 170: 1–8.
[16]  Henke N, Schmidt-Ullrich R, Dechend R, Park JK, Qadri F, et al. (2007) Vascular endothelial cell-specific NF-kappaB suppression attenuates hypertension-induced renal damage. Circ Res 101: 268–76.
[17]  Schwartz RS, Murphy JG, Edwards WD, Camrud AR, Vliestra RE, et al. (1990) Restenosis after balloon angioplasty. A practical proliferative model in porcine coronary arteries. Circulation 82: 2190–200.
[18]  Ferns GA, Avades TY (2000) The mechanisms of coronary restenosis: insights from experimental models. Int J Exp Pathol 81: 63–88.
[19]  Walsh K, Smith RC, Kim HS (2000) Vascular cell apoptosis in remodeling, restenosis, and plaque rupture. Circ Res 87: 184–8.
[20]  Lim S, Jin CJ, Kim M, Chung SS, Park HS, et al. (2006) PPARgamma gene transfer sustains apoptosis, inhibits vascular smooth muscle cell proliferation, and reduces neointima formation after balloon injury in rats. Arterioscler Thromb Vasc Biol 26: 808–13.
[21]  Mahadevan S, Park Y (2008) Multifaceted therapeutic benefits of Ginkgo biloba L.: chemistry, efficacy, safety, and uses. J Food Sci 73: R14–R19.
[22]  Kim KS, Rhee KH, Yoon JH, Lee JG, Lee JH, et al. (2005) Ginkgo biloba extract (EGb 761) induces apoptosis by the activation of caspase-3 in oral cavity cancer cells. Oral Oncol 41: 383–9.
[23]  Chao JC, Chu CC (2004) Effects of Ginkgo biloba extract on cell proliferation and cytotoxicity in human hepatocellular carcinoma cells. World J Gastroenterol 10: 37–41.
[24]  Welt FG, Edelman ER, Simon DI, Rogers C (2000) Neutrophil, not macrophage, infiltration precedes neointimal thickening in balloon-injured arteries. Arterioscler Thromb Vasc Biol 20: 2553–8.
[25]  Moreno PR, Bernardi VH, Lopez-Cuellar J, Newell JB, McMellon C, et al. (1996) Macrophage infiltration predicts restenosis after coronary intervention in patients with unstable angina. Circulation 94: 3098–102.
[26]  Moreno PR, Fallon JT, Murcia AM, Leon MN, Simosa H, et al. (1999) Tissue characteristics of restenosis after percutaneous transluminal coronary angioplasty in diabetic patients. J Am Coll Cardiol 34: 1045–9.
[27]  Rogers C, Welt FG, Karnovsky MJ, Edelman ER (1996) Monocyte recruitment and neointimal hyperplasia in rabbits. Coupled inhibitory effects of heparin. Arterioscler Thromb Vasc Biol 16: 1312–8.
[28]  Mori E, Komori K, Yamaoka T, Tanii M, Kataoka C, et al. (2002) Essential role of monocyte chemoattractant protein-1 in development of restenotic changes (neointimal hyperplasia and constrictive remodeling) after balloon angioplasty in hypercholesterolemic rabbits. Circulation 105: 2905–10.
[29]  Pasceri V, Willerson JT, Yeh ET (2000) Direct proinflammatory effect of C-reactive protein on human endothelial cells. Circulation 102: 2165–8.
[30]  Choi SE, Shin HC, Kim HE, Lee SJ, Jang HJ, et al. (2007) Involvement of Ca2+, CaMK II and PKA in EGb 761-induced insulin secretion in INS-1 cells. J Ethnopharmacol 110: 49–55.
[31]  Kudolo GB (2001) The effect of 3-month ingestion of Ginkgo biloba extract (EGb 761) on pancreatic beta-cell function in response to glucose loading in individuals with non-insulin-dependent diabetes mellitus. J Clin Pharmacol 41: 600–11.
[32]  Hu E, Liang P, Spiegelman BM (1996) AdipoQ is a novel adipose-specific gene dysregulated in obesity. J Biol Chem 271: 10697–703.
[33]  Lim S, Koo BK, Cho SW, Kihara S, Funahashi T, et al. (2008) Association of adiponectin and resistin with cardiovascular events in Korean patients with type 2 diabetes: the Korean atherosclerosis study (KAS): a 42-month prospective study. Atherosclerosis 196: 398–404.
[34]  Pischon T, Girman CJ, Hotamisligil GS, Rifai N, Hu FB, et al. (2004) Plasma adiponectin levels and risk of myocardial infarction in men. JAMA 291: 1730–7.
[35]  Schulze MB, Shai I, Rimm EB, Li T, Rifai N, et al. (2005) Adiponectin and future coronary heart disease events among men with type 2 diabetes. Diabetes 54: 534–9.
[36]  Kumada M, Kihara S, Sumitsuji S, Kawamoto T, Matsumoto S, et al. (2003) Association of hypoadiponectinemia with coronary artery disease in men. Arterioscler Thromb Vasc Biol 23: 85–9.
[37]  Zietz B, Herfarth H, Paul G, Ehling A, Muller-Ladner U, et al. (2003) Adiponectin represents an independent cardiovascular risk factor predicting serum HDL-cholesterol levels in type 2 diabetes. FEBS Lett 545: 103–4.
[38]  Chen H, Montagnani M, Funahashi T, Shimomura I, Quon MJ (2003) Adiponectin stimulates production of nitric oxide in vascular endothelial cells. J Biol Chem 278: 45021–6.
[39]  Han SH, Quon MJ, Kim JA, Koh KK (2007) Adiponectin and cardiovascular disease: response to therapeutic interventions. J Am Coll Cardiol 49: 531–8.
[40]  Bhagat K, Vallance P (1997) Inflammatory cytokines impair endothelium-dependent dilatation in human veins in vivo. Circulation 96: 3042–7.
[41]  Pepys MB, Rowe IF, Baltz ML (1985) C-reactive protein: binding to lipids and lipoproteins. Int Rev Exp Pathol 27: 83–111.
[42]  Cermak J, Key NS, Bach RR, Balla J, Jacob HS, et al. (1993) C-reactive protein induces human peripheral blood monocytes to synthesize tissue factor. Blood 82: 513–20.
[43]  Wolbink GJ, Brouwer MC, Buysmann S, ten Berge IJ, Hack CE (1996) CRP-mediated activation of complement in vivo: assessment by measuring circulating complement-C-reactive protein complexes. J Immunol 157: 473–9.
[44]  Lagrand WK, Visser CA, Hermens WT, Niessen HW, Verheugt FW, et al. (1999) C-reactive protein as a cardiovascular risk factor: more than an epiphenomenon? Circulation 100: 96–102.
[45]  Mohamed-Ali V, Goodrick S, Rawesh A, Katz DR, Miles JM, et al. (1997) Subcutaneous adipose tissue releases interleukin-6, but not tumor necrosis factor-alpha, in vivo. J Clin Endocrinol Metab 82: 4196–200.
[46]  Orban Z, Remaley AT, Sampson M, Trajanoski Z, Chrousos GP (1999) The differential effect of food intake and beta-adrenergic stimulation on adipose-derived hormones and cytokines in man. J Clin Endocrinol Metab 84: 2126–33.
[47]  Rodriguez M, Ringstad L, Schafer P, Just S, Hofer HW, et al. (2007) Reduction of atherosclerotic nanoplaque formation and size by Ginkgo biloba (EGb 761) in cardiovascular high-risk patients. Atherosclerosis 192: 438–44.
[48]  Kudolo GB, Delaney D, Blodgett J (2005) Short-term oral ingestion of Ginkgo biloba extract (EGb 761) reduces malondialdehyde levels in washed platelets of type 2 diabetic subjects. Diabetes Res Clin Pract 68: 29–38.
[49]  Zhang Q, Wang GJ, JY A, Wu D, Zhu LL, et al. (2009) Application of GC/MS-based metabonomic profiling in studying the lipid-regulating effects of Ginkgo biloba extract on diet-induced hyperlipidemia in rats. Acta Pharmacol Sin 30: 1674–87.
[50]  Boveris AD, Galleano M, Puntarulo S (2007) In vivo supplementation with Ginkgo biloba protects membranes against lipid peroxidation. Phytother Res 21: 735–40.
[51]  Liu F, Zhang J, Yu S, Wang R, Wang B, et al. (2008) Inhibitory effect of Ginkgo biloba extract on hyperhomocysteinemia-induced intimal thickening in rabbit abdominal aorta after balloon injury. Phytother Res 22: 506–10.


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