Some aspects of the in vivo neuroprotective capacity of flavonoids: Bioavailability and structure-activity relationship Authors
Received: 09 July 2004 Revised: 08 October 2004 DOI:
Cite this article as: Rivera, F., Urbanavicius, J., Gervaz, E. et al. neurotox res (2004) 6: 543. doi:10.1007/BF03033450 Abstract
On the basis of previous work showing that flavonoids structurally related to quercetin are neuroprotective for cells in culture, this work was directed towards determining if several flavonoids (quercetin, fisetin and catechin) could acutely and by an intraperitoneal (IP) route reach significant cerebral concentrations and either prevent or facilitate recovery from a brain lesion induced by focal ischemia in rats.
Aqueous and liposomal preparations of quercetin, fisetin and catechin were administered IP in a single dose and assessed in the brain by HPLC at 30 min, 1 h, 2 h and 4 h. Ischemic damage from focal middle cerebral artery occlusion was assessed spectrophotometrically with 2,3,5,-triphenylltetrazolium chloride (TTC). Infarct volume was assessed by an image analysis system following perfusion with TTC. The status of the cerebral tissue was evaluated by hematoxylin-eosin.
Flavonoids administered in aqueous preparations were undetected in the brain. Cerebral concentrations of catechin (10.5 ng/g), fisetin (8.23 ng/g) and quercetin (509 ng/g) were detected in the brain only after IP injection of the liposomal preparations. Spectrophotometric analysis of brain tissue with the TTC-technique showed that liposomal quercetin reduced ischemic damage and infarct volume after permanent occlusion of the middle cerebral artery (ischemic: 41.3 mm
3 vs liposomal quercetin: 17 mm 3). In liposomal quercetin-treated animals there was also recovery of the cytoarchitecture in ischemic areas of striatum and cortex. Although a liposomal preparation of fisetin had similar effects, catechin failed to protect brain tissue.
In conclusion, early administration of liposomal preparations of quercetin and structurally related flavonoids are beneficial and neuroprotective in experimental focal ischemia.
Keywords Quercetin Fisetin Catechin Neuroprotection Permanent focal ischemia References
Agullo G, L Gamet-Payrastre, S Manenti, C Viala C Rémésy H Chap and B Payrastre (1997) Relationship between flavonoid structure and inhibition of phosphatidylinositol 3-kinase: a com parison with tyrosine kinase and protein kinase C inhibition.
Aherne SA and NM O’Brien (1999) Protection by the flavonoids myricetin, quercetin, and rutin against hydrogen peroxideinduced DNA damage in Caco-2 and Hep G2 cells.
Ahlemeyer B, V Junker, R Huhne and J Kriegelstein (2001) Neuroprotective effects of NV-31, a bilobalide-derived compound: evidence for an antioxidative mechanism.
Aoki Y, M Tamura and Y Itoh (2001) Cerebroprotective action of a Na
channel blocker NS-7. I. Effect on the cerebral infarction and edema at the acute stage of permanent middle cerebral artery occlusion in rats.
Azuma K, K Ippoushi, H Ito, H Higashio and J Terao (2002) Combination of lipids and emulsifiers enhances the absorption of orally administered quercetin in rats.
J. Agr. Food Chemical.
Bastianetto S and R Quirino (2002) Natural extracts as possible protective agents of brain aging.
Belayev L, OF Alonso, R Busto, W Zhao and MD Ginsberg (1996) Middle cerebral artery occlusion in the rat by intraluminal suture: neurological and pathological evaluation of an improved model.
Belayev L, L Yitao, Z Weizhao, R Busto and MD Ginsberg (2001) Human albumin therapy of acute ischemic stroke.
Bertorelli R, M Adami, E Di Santo and P Ghezzi (1998) MK 801 and dexamethasone reduce both tumor necrosis factor levels and infarct volume after focal cerebral ischemia in the rat brain.
Cao X.H. and JW Phillis (1994) ?-Phenyl-ter-butyl-nitrone reduces cortical infarct and edema in rats subjected to focal ischemia.
Chan P, JT Cheng, JC Tsai, GS Lien, FC Chen, PF Kao, JC Liu, YJ Chen and MH Hsieh (2002) Effect of catechin on the activity and gene expression of superoxide dismutase in cultured rat brain astrocytes.
Chang WS, YJ Lee, FJ Lu and HC Chiang (1993) Inhibitory effects of flavonoids on xanthine oxidase.
Cho EJ, T Yokozawa, DY Rhyu, SC Kim, N Shibahara and JC Park (2003) Study on the inhibitory effects of Korean medicinal plants and their main compounds on the 1,1-diphenyl 2-picrylhydrazyl radical.
Chung SY, MF Wang, JY Lin, HM Liu and FC Cheng (2003) Effect of one week treatment with Ginkgo biloba extract (Egb761) on ischemia-induced infarct volume in gerbils.
Am. J. Chin. Med.
Clark WM, LG Rinker, NS Lessov, SL Lowery and MJ Cipolla (2001) Efficacy of antioxidant therapies in transient focal ischemia in mice.
Clemens JA and JA Panetta (1994) Neuroprotection by antioxidants in models of global and focal ischemia.
Ann. NYAcad. Sci.
Corbett D and S Nurse (1998) The problem of assessing effective neuroprotection in experimental cerebral ischemia.
Dajas F, F Rivera, F Blasina, F Arredondo, JA Abín-Carriquiry, G Costa, C Echeverry, L Lafón, H Heinzen, M Ferreira and A Morquio (2003a) Neuroprotection by flavonoids.
Braz. J. Med. Biol. Res.
Dajas F, F Rivera, F Blasina, F Arredondo, C Echeverry, L Lafon, A Morquio and H Heizen (2003b) Cell culture protection and
neuroprotective capacity of flavonoids.
Davies CA, SA Loddick, RP Stroemer, J Hunt and NJ Rothwell (1998) An integrated analysis of the progression of cell responses induced by permanent focal middle cerebral artery occlusion in the rat.
Denizot F and R Lang (1986) Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and realibility.
J. Immunol. Methods
Emmert DH and JT Kirchner (1999) The role of vitamin E in the prevention of heart disease.
Arch. Fam. Med.
Erden Inal M, A Kahraman and T Koken (2001) Beneficial effects of quercetin on oxidative stress induced by ultraviolet A.
Clin. Exp. Dermatol.
Esposito E, D Rotilio, V Di Matteo, C Di Giulio, M Cacchio and S Algeri (2002) A review of specific dietary antioxidants and the effects on biochemical mechanisms related to neurodegenerative processes.
Ferriola PC, V Cody and E Middleton (1989) Protein kinase C inhibition by plant flavonoids. Kinetic mechanisms and structureactivity relationships.
Fiorani M, R De Sanctis, P Menghinello, B Cellini and M Dacha (2001) Quercetin prevents glutathione depletion induced by dehydroascorbic acid in rabbit red blood cells.
Free Radical Res.
Gregoriadis G (1995) Engineering liposomes for drug delivery: progress and problems.
Gupta R, M Singh and A Sharma (2003) Neuroprotective effect of antioxidants on ischaemia and reperfusion-induced cerebral injury.
Hertog MGL (1996) Epidemiological evidence on potential health properties of flavonoids.
Proc. Nutr. Soc.
Huh PW, L Belayev, W Zhao, JA Clemens, JA Panetta, R Busto and MD Ginsberg (2000) Neuroprotection by LY341122, a novel inhibitor of lipid peroxidation, against focal ischemic brain damage in rats.
Eur. J. Pharmacol.
Huk I, V Brovkovych, VJ Nanobash, G Weigel, C Neumayer, L Partyka, S Patton and T Malinski (1998) Bioflavonoid quercetin scavenges superoxide and increases nitric oxide concentration in ischaemia-reperfusion injury: an experimental study.
Br. J. Surgery
Hussain RF, AM Nouri and RT Oliver (1993) A new approach for measurement of cytotoxicity using colorimetric assay.
J. Immunol. Methods
Inanami O, Y Watanabe, B Syuto, M Nakano, M Tsuji and M Kuwabara (1998) Oral administration of (-)catechin protects against ischemia-reperfusion-induced neuronal death in the gerbil.
Free Radical Res.
Ishige K, D Schbert and Y Sagara (2001) Flavonoids protect neuronal cells from oxidative stress by three distinct mechanisms.
Free Radical Biol. Med.
Kannno A, Y Terada, K Tsuzaki, Y Matsumoto and R Ueoka (1999) Therapeutic effects of hybrid liposomes on mice model of lung carcinoma.
Drug Delivery System
Kinouchi H, CJ Epstein, T Mizui, E Carlson, SF Chen and PH Chan (1991) Attenuation of focal cerebral ischemic injury in transgenic mice overexpressing Cu/Zn superoxide dismutase.
Proc. Natl. Acad. Sci. USA
Kolominsky-Rabas PL C Sarti, PU Heuschmann, C Graf, S Siemonsen, B Neundoerfer, A Katalinic, KG Gassmann and TR von Strockert (1998) A prospective community-based study of stroke in Germany. The Erlangen Stroke Project (Es Pro) incidence and case fatality at 1, 3 and 12 months.
Legos J, JA Erhardt, R White, S Lenhard, S Chandra, A Parsons, R Tuma and F Barone (2001) SB 239063, a novel p38 inhibitor, attenuates early neuronal injury following ischemia.
Leppala J, J Virtamon, R Fogelholm, D Albanes and O Heinonen (1999) Different risk factors for different stroke subtypes: association of blood pressure, colesterol and antioxidants.
Lipton P (1999) Ischemic cell death in brain neurons.
Liu TH, JS Beckman, BA Freeman, EL Hogan and CY Hsu (1989) Polyethylene glycol-conjugated superoxide dismutase and catalase reduce ischemic brain injury.
Am. J. Physiol.
Mandel S, L Reznichenko, T Amit and MB Youdim (2003) Green tea polyphenol (-)-epigallocatechin-3-gallate protects rat PC 12 cells from apoptosis induced by serum withdrawal independent of P13-Akt pathway.
Margaill I, S Parmentier, J Callebert, M Allix, R Boulu and M Plotkine (1996). Short therapeutic window for MK801 in transient focal cerebral ischemia in normotensive rats.
J. Cereb. Blood Flow Metab.
Miura T, S Muraoka and Y Fujimoto (2003) Inactivation of creatine kinase induced by quercetin with horseradish peroxidase and hydrogen peroxide pro-oxidative and anti-oxidative actions of quercetin.
Food Chem. Toxicol.
Nagao A, M Seki and H Kobayashi (1999) Inhibition of xanthine oxidase by flavonoids.
Biosci. Biotechnol. Biochem.
Nagata H, S Takekoshi, T Honma and K Watanabe (1999) Antioxidative action of flavonoids, quercetin and catechin, mediated by the activation of glutathione peroxidase.
Tokai J. Exp. Clin. Med.
Naidu PS, A Singh and SK Kulkarni (2003) Quercetin, a bioflavonoid, attenuates haloperidol-induced orofacial dyskinesia.
Naidu PS, A Singh, and SK Kulkarni (2004) Reversal of reserpineinduced orofacial dyskinesia and cognitive dysfunction by quercetin.
Nardi GM, R Felippi, S DalBo, JM Siqueira-Jr, DC Arruda, F Delle Monache, AK Timbola, MG Pizzolatti, K Ckless and RM Ribeiro-do-valle (2003) Anti-inflammatory and antioxidant effects of Croton celtidifolius bark.
Nicotera P and S Lipton (1999) Excitotoxins in neuronal apoptosis and necrosis.
J. Cereb. Blood Flow Metab.
Park CK, AD Mendelow, DI Graham, J McCulloch and GM Teasdale (1988) Correlation of triphenyltetrazolium chloride perfusion staining with conventional neurohistology in the detection of early brain ischemia.
Neuropathol. Appl. Neurobiol.
Park JB (1999) Flavonoids are potential inhibitors of glucose uptake in U937 cells.
Biochem. Biophys. Res. Comm.
Picq M, M Dubois, Y Munari-Silem, AF Prigent and H Pacheco (1989) Flavonoid modulation of protein kinase C activation.
Prajda N, RL Singhai, YA Yeh, E Olah, KY Look and G Weber (1995) Linkage of reduction in 1-phosphatidylinositol 4-kinase activity and inositol 1,4,5-biphosphate concentration in human ovarian carcinoma cells treated with quercetin.
Preston E and J Webster (2000) Spectrophotometric measurement of experimental brain injury.
J. Neurosci. Methods
Ratty AK and NP Das (1988) Effects of flavonoids on nonenzymatic lipid peroxidation: structure-activity relationship.
Biochem. Med. Metab. Biol.
Rice-Evans C (2001) Flavonoid antioxidants.
Curr. Med. Chem.
Sakas DE, RM Crowell and NT Zervas (1994) Effects of lecithinemulsified perfluorochemical compounds in ischemic brain injury. Artif. Cells Blood Substit. Immobil. Biotechnol.
Sanderson J, W Lauchlan and G Williamson (1999) Quercetin inhibits hydrogen peroxide-induced oxidation of the rat lens.
Free. Radic. Biol. Med.
Sanhueza J, J Valdes, R Campos, A Garrido and A Valenzuela (1992) Changes in the xanthine dehydrogenase/xanthine oxidase ratio in the rat kidney subjected to ischemia-reperfusion stress: preventive effect of some flavonoids.
Res. Commun. Chem. Pathol. Pharmacol.
Shichinohe H, S Kuroda, T Abumiya, J Ikeda, T Kobayashi, T Yoshimoto and Y Iwasaki (2004) FK506 reduces infarct volume due to permanent focal cerebral ischemia by maintaining BAD turnover and inhibiting cytochrome c release.
Singh A, PS Naidu and SK Kulkarni (2003) Reversal of aging and chronic ethanol-induced cognitive dysfunction by quercetin, a bioflavonoid.
Free Radic. Res.
Soliman KF and EA Mazzio (1998)
attenuation of nitric oxide production in C6 astrocyte cell cultures by various dietary compounds.
Proc. Soc. Exp. Biol. Med.
Steinberg G, N Panahian, M Perez-Pinzon, G Sun, M Modi and J Sepinwal (1995) Narrow temporal therapeutic window for NMDA antagonist protection against focal cerebral ischemia.
Suzuki S, M Furushiro, M Takahashi, M Sakai and S Kudo (1999) Oral administration of soybean lecithin transphosphatidylated phosphatidylase (SB-tPS) reduces ischemic damage in the gerbil hippocampus.
Jpn. J. Pharmacol.
Swanson RA, MT Morton, G Tsao-Wu, RA Savalos, C Davidson and FC Sharp (1990) A semiautomated method of measurement of brain infarct volume.
J. Cereb. Blood Flow Metab.
Sydserff SG, AJ Cross and AR Green (1995) The neuroprotective effect of chlormethiazole on ischaemic neuronal damage following permanent middle cerebral artery ischaemia in the rat.
Terao J, M Piskula and Q Yao (1994) Protective effect of epicatechin, epicatechin gallate, and quercetin lipid peroxidation in phospholipid bilayers.
Arch. Biochem. Biophys.
Ueoka R, Y Matsumoto, A Kanno, K Tsuzaki and H Ichihara (2000) Marked therapeutic effects of dimyristoylphosphatidylcholine liposomes on carcinoma mice model
Biol. Pharm. Bull.
Varming T, J Drejer, A Frandsen and A Schousboe (1996) Characterization of a chemical anoxia model in cerebellar granule neurons using sodium azide.
J. Neurosci. Res.
Welton AF, LD Tobias and C Fiedler-Nagy (1986) Effect of flavonoids on arachidonic acid metabolism.
Prog. Clin. Biol. Res.
Wen TC, M Rogido, P Gressens and A Sola (2004) A reproducible experimental model of focal cerebral ischemia in the neonatal rat.
Brain Res. Brain Res. Protoc.
Williams AJ, JR Dave, JB Phillips, Y Lin, T Mcaber and FC Tortella (2000) Neuroprotective efficacy and therapeutic window of the high-affinity N-methyl-D-aspartate antagonist conantokin-G:
(primary cerebellar neurons) and
(rat model of transient focal brain ischemia) studies.
J. Pharmacol. Exp. Ther.
Youdim KA, MS Dobbie, G Kuhnle, AR Protegente, NJ Abbott and C Rice-Evans (2003) Interaction between flavonoids and the blood-brain barrier:
Yu Z, AJ Bruce-Keller, Y Goodman and MP Mattson (1998) Uric acid protects neurons against excitotoxic and metabolic insults in cell culture, and against focal ischemic brain injury
J. Neurosci. Res.
Yulug B, U Kilic, E Kilic and M Bahr (2004) Rifampicin attenuates brain damage in focal ischemia.
Zausinger S, E Hungerhuber, A Baethmann, H Reulen and R Schmid-Elsaesser (2000) Neurological imparirment in rats after transient middle cerebral artery occlusion: a comparative study under various treatment paradigms.
Zhang DL, YT Zhang, JJ Yin and BL Zhao (2004) Oral administration of Crataegus flavonoids protects against ischemia/reperfusion brain damage in gerbils.