Pinocembrin protects against β-amyloid-induced toxicity in neurons through inhibiting receptor for advanced glycation end products (RAGE)-independent signaling pathways and regulating mitochondrion-mediated apoptosis

BMC Medicine

Guan-hua Du¹  Tian-tai Zhang¹  Li Zhang¹  Shuo Tian¹  Fan Yang¹  Dan Zhou¹  Cai-xia Wu²  Rui Liu¹ 

[1] State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, P R China;Shenyang Pharmaceutical University, Shenyang, 110016, P R China

关键词: receptor for advanced glycation end products;    pinocembrin;    apoptosis;    amyloid-β peptide;    Alzheimer's disease;   

Others  :  857352
DOI  :  10.1186/1741-7015-10-105

 received in 2012-07-13, accepted in 2012-09-18,  发布年份 2012

【 摘 要 】

Background

It is known that amyloid-β peptide (Aβ) plays a pivotal role in the pathogenesis of Alzheimer's disease (AD). Interaction between Aβ and the receptor for advanced glycation end products (RAGE) has been implicated in neuronal degeneration associated with this disease. Pinocembrin, a flavonoid abundant in propolis, has been reported to possess numerous biological activities beneficial to health. Our previous studies have demonstrated that pinocembrin has neuroprotective effects on ischemic and vascular dementia in animal models. It has been approved by the State Food and Drug Administration of China for clinical use in stroke patients. Against this background, we investigated the effects of pinocembrin on cognitive function and neuronal protection against Aβ-induced toxicity and explored its potential mechanism.

Methods

Mice received an intracerebroventricular fusion of Aβ_25-35. Pinocembrin was administrated orally at 20 mg/kg/day and 40 mg/kg/day for 8 days. Behavioral performance, cerebral cortex neuropil ultrastructure, neuronal degeneration and RAGE expression were assessed. Further, a RAGE-overexpressing cell model and an AD cell model were used for investigating the mechanisms of pinocembrin. The mechanisms underlying the efficacy of pinocembrin were conducted on target action, mitochondrial function and potential signal transduction using fluorescence-based multiparametric technologies on a high-content analysis platform.

Results

Our results showed that oral administration of pinocembrin improved cognitive function, preserved the ultrastructural neuropil and decreased neurodegeneration of the cerebral cortex in Aβ_25-35-treated mice. Pinocembrin did not have a significant effect on inhibiting Aβ_1-42 production and scavenging intracellular reactive oxygen species (ROS). However, pinocembrin significantly inhibited the upregulation of RAGE transcripts and protein expression both in vivo and in vitro, and also markedly depressed the activation of p38 mitogen-activated protein kinase (MAPK)-MAPKAP kinase-2 (MK2)-heat shock protein 27 (HSP27) and stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK)-c-Jun pathways and the downstream nuclear factor κB (NFκB) inflammatory response subsequent to Aβ-RAGE interaction. In addition, pinocembrin significantly alleviated mitochondrial dysfunction through improving mitochondrial membrane potential and inhibiting mitochondrial oxidative stress, and regulated mitochondrion-mediated apoptosis by restoration of B cell lymphoma 2 (Bcl-2) and cytochrome c and inactivation of caspase 3 and caspase 9.

Conclusions

Pinocembrin was shown to infer cognitive improvement and neuronal protection in AD models. The mechanisms of action of the compound were illustrated on RAGE-dependent transduction inhibition and mitochondrion protection. It appears to be a promising candidate for the prevention and therapy of AD.

【 授权许可】

   
2012 Liu et al; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Selkoe DJ: Normal and abnormal biology of the beta-amyloid precursor protein. Annu Rev Neurosci 1994, 17:489-517.
• [2]Rohn TT, Head E, Su JH, Anderson AJ, Bahr BA, Cotman CW, Cribbs DH: Correlation between caspase activation and neurofibrillary tangle formation in Alzheimer's disease. Am J Pathol 2001, 158:189-198.
• [3]Su JH, Nichol KE, Sitch T, Sheu P, Chubb C, Miller BL, Tomaselli KJ, Kim RC, Cotman CW: DNA damage and activated caspase-3 expression in neurons and astrocytes: evidence for apoptosis in frontotemporal dementia. Exp Neurol 2000, 163:9-19.
• [4]Zilka N, Ferencik M, Hulin I: Neuroinflammation in Alzheimer's disease: protector or promoter? Bratisl Lek Listy 2006, 107:374-383.
• [5]Eckert A, Keil U, Marques CA, Bonert A, Frey C, Schussel K, Muller WE: Mitochondrial dysfunction, apoptotic cell death, and Alzheimer's disease. Biochem Pharmacol 2003, 66:1627-1634.
• [6]Parks JK, Smith TS, Trimmer PA, Bennett JP Jr, Parker WD Jr: Neurotoxic Aβ peptides increase oxidative stress in vivo through NMDA-receptor and nitric-oxide-synthase mechanisms, and inhibit complex IV activity and induce a mitochondrial permeability transition in vitro. J Neurochem 2001, 76:1050-1056.
• [7]Yan SD, Chen X, Fu J, Chen M, Zhu H, Roher A, Slattery T, Zhao L, Nagashima M, Morser J, Migheli A, Nawroth P, Stern D, Schmidt AM: RAGE and amyloid-beta peptide neurotoxicity in Alzheimer's disease. Nature 1996, 382:685-691.
• [8]Auld DS, Kornecook TJ, Bastianetto S, Quirion R: Alzheimer's disease and the basal forebrain cholinergic system: relations to β-amyloid peptides, cognition, and treatment strategies. Prog Neurobiol 2002, 68:209-245.
• [9]Bartus RT, Dean RL: Pharmaceutical treatment for cognitive deficits in Alzheimer's disease and other neurodegenerative conditions: exploring new territory using traditional tools and established maps. Psychopharmacology (Berl) 2009, 202:15-36.
• [10]Selkoe DJ: Alzheimer's disease is a synaptic failure. Science 2002, 298:789-791.
• [11]Yan SD, Yan SF, Chen X, Fu J, Chen M, Kuppusamy P, Smith MA, Perry G, Godman GC, Nawroth P, Zweier JL, Stern D: Nonenzymatically glycated tau in Alzheimer's disease induces neuronal oxidant stress resulting in cytokine gene expression and release of Aβ. Nat Med 1995, 1:693-699.
• [12]Lue LF, Walker DG, Brachova L, Beach TG, Rogers J, Schmidt AM, Stern DM, Yan SD: Involvement of microglial receptor for advanced glycation endproducts (RAGE) in Alzheimer's disease: identification of a cellular activation mechanism. Exp Neurol 2001, 171:29-45.
• [13]Yan SD, Bierhaus A, Nawroth PP, Stern DM: RAGE and Alzheimer's disease: a progression factor for amyloid-β-induced cellular perturbation? J Alzheimers Dis 2009, 16:833-843.
• [14]Schmidt AM, Yan SD, Yan SF, Stern DM: The biology of the receptor for advanced glycation end products and its ligands. Biochim Biophys Acta 2000, 1498:99-111.
• [15]Onyango IG, Tuttle JB, Bennett JP Jr: Altered intracellular signaling and reduced viability of Alzheimer's disease neuronal cybrids is reproduced by beta-amyloid peptide acting through receptor for advanced glycation end products (RAGE). Mol Cell Neurosci 2005, 29:333-343.
• [16]Mucke L, Yu GQ, McConlogue L, Rockenstein EM, Abraham CR, Masliah E: Astroglial expression of human α(1)-antichymotrypsin enhances Alzheimer-like pathology in amyloid protein precursor transgenic mice. Am J Pathol 2000, 157:2003-2010.
• [17]Arancio O, Zhang HP, Chen X, Lin C, Trinchese F, Puzzo D, Liu S, Hegde A, Yan SF, Stern A, Luddy JS, Lue LF, Walker DG, Roher A, Buttini M, Mucke L, Li W, Schmidt AM, Kindy M, Hyslop PA, Stern DM, Du Yan SS: RAGE potentiates Aβ-induced perturbation of neuronal function in transgenic mice. EMBO J 2004, 23:4096-4105.
• [18]Ma L, Carter RJ, Morton AJ, Nicholson LF: RAGE is expressed in pyramidal cells of the hippocampus following moderate hypoxic-ischemic brain injury in rats. Brain Res 2003, 966:167-174.
• [19]Rong LL, Gooch C, Szabolcs M, Herold KC, Lalla E, Hays AP, Yan SF, Yan SS, Schmidt AM: RAGE: a journey from the complications of diabetes to disorders of the nervous system - striking a fine balance between injury and repair. Restor Neurol Neurosci 2005, 23:355-365.
• [20]Kumazawa S, Shimoi K, Hayashi K, Ishii T, Hamasaka T, Nakayama T: Identification of metabolites in plasma and urine of Uruguayan propolis-treated rats. J Agric Food Chem 2004, 52:3083-3088.
• [21]Yang ZH, Sun X, Qi Y, Mei C, Sun XB, Du GH: Uptake characteristics of pinocembrin and its effect on p-glycoprotein at the blood-brain barrier in in vitro cell experiments. J Asian Nat Prod Res 2012, 14:14-21.
• [22]Liu R, Gao M, Yang ZH, Du GH: Pinocembrin protects rat brain against oxidation and apoptosis induced by ischemia-reperfusion both in vivo and in vitro. Brain Res 2008, 1216:104-115.
• [23]Gao M, Zhang WC, Liu QS, Hu JJ, Liu GT, Du GH: Pinocembrin prevents glutamate-induced apoptosis in SH-SY5Y neuronal cells via decrease of bax/bcl-2 ratio. Eur J Pharmacol 2008, 591:73-79.
• [24]Gao M, Liu R, Zhu SY, Du GH: Acute neurovascular unit protective action of pinocembrin against permanent cerebral ischemia in rats. J Asian Nat Prod Res 2008, 10:551-558.
• [25]Shi LL, Chen BN, Gao M, Zhang HA, Li YJ, Wang L, Du GH: The characteristics of therapeutic effect of pinocembrin in transient global brain ischemia/reperfusion rats. Life Sci 2011, 88:521-528.
• [26]Meng F, Liu R, Gao M, Wang Y, Yu X, Xuan Z, Sun J, Yang F, Wu C, Du G: Pinocembrin attenuates blood-brain barrier injury induced by global cerebral ischemia-reperfusion in rats. Brain Res 2011, 1391:93-101.
• [27]Guang HM, Du GH: Protections of pinocembrin on brain mitochondria contribute to cognitive improvement in chronic cerebral hypoperfused rats. Eur J Pharmacol 2006, 542:77-83.
• [28]Morris R: Developments of a water-maze procedure for studying spatial learning in the rat. J Neurosci Methods 1984, 11:47-60.
• [29]Schmued LC, Hopkins KJ: Fluoro-Jade B: a high affinity fluorescent marker for the localization of neuronal degeneration. Brain Res 2000, 874:123-130.
• [30]Candelario-Jalil E, Alvarez D, Merino N, Leon OS: Delayed treatment with nimesulide reduces measures of oxidative stress following global ischemic brain injury in gerbils. Neurosci Res 2003, 47:245-253.
• [31]Rüster C, Bondeva T, Franke S, Tanaka N, Yamamoto H, Wolf G: Angiotensin II upregulates RAGE expression on podocytes: role of AT2 receptors. Am J Nephrol 2009, 29:538-550.
• [32]Liu R, Meng F, Zhang L, Liu A, Qin H, Lan X, Li L, Du G: Luteolin isolated from the medicinal plant Elsholtzia rugulosa (Labiatae) prevents copper-mediated toxicity in β-amyloid precursor protein Swedish mutation overexpressing SH-SY5Y cells. Molecules 2011, 16:2084-2096.
• [33]Satoh T, Enokido Y, Aoshima H, Uchiyama Y: Changes in mitochondrialmembrane potential during oxidative stress-Induced apoptosis in PC12 cells. Neurosci Res 1997, 50:413-420.
• [34]Hu D, Cao P, Thiels E, Chu CT, Wu GY, Oury TD, Klann E: Hippocampal long-term potentiation, memory, and longevity in mice that overexpress mitochondrial superoxide dismutase. Neurobiol Learn Mem 2007, 87:372-384.
• [35]Trask OJ, Nickischer D, Burton A, Williams RG, Kandasamy RA, Johnston PA, Johnston PA: High-throughput automated confocal microscopy imaging screen of a kinase-focused library to identify p38 mitogen-activated protein kinase inhibitors using the GE InCell 3000 analyzer. Methods Mol Biol 2009, 565:159-186.
• [36]Bertelsen M: Multiplex analysis of inflammatory signaling pathways using a high-content imaging system. Methods Enzymol 2006, 414:348-363.
• [37]Butterfield DA, Reed T, Newman SF, Sultan R: Roles of amyloid beta-peptide-associated oxidative stress and brain protein modifications in the pathogenesis of Alzheimer's disease and mild cognitive impairment. Free Radic Biol Med 2007, 43:658-677.
• [38]Nitta A, Itoh A, Hasegawa T, Nabeshima T: β-Amyloid protein-induced Alzheimer's disease animal model. Neurosci Lett 1994, 170:63-66.
• [39]Hoshi M, Takashima A, Murayama M, Yasutake K, Yoshida N, Ishiguro K, Hoshino T, Imahori K: Nontoxic amyloid β peptide 1-42 suppresses acetylcholine synthesis. J Biol Chem 1997, 272:2038-2041.
• [40]Kar S, Issa AM, Seto D, Auld DS, Collier B, Quirion R: Amyloid β-peptide inhibits high-affinity choline uptake and acetylcholine release in rat hippocampal slices. J Neurochem 1998, 70:2179-2187.
• [41]Sigurdsson EM, Lee JM, Dong XW, Hejna MJ, Lorens SA: Bilateral injections of amyloid-β 25-35 into the amygdala of young Fischer rats: behavioral, neurochemical, and time dependent histopathological effects. Neurobiol Aging 1997, 18:591-608.
• [42]Pavia J, Alberch J, Alverez I, Toledano A, de Ceballos ML: Repeated intracerebroventricular administration of β-amyloid 25-35 to rats decreases muscarinic receptors in cerebral cortex. Neurosci Lett 2000, 278:69-72.
• [43]Nakdook W, Khongsombat O, Taepavarapruk P, Taepavarapruk N, Ingkaninan K: The effects of Tabernaemontana divaricata root extract on amyloid beta-peptide 25-35 peptides induced cognitive deficits in mice. J Ethnopharmacol 2010, 130:122-126.
• [44]Rovira C, Arbez N, Mariani J: Aβ(25-35) and Aβ(1-40) act on different calcium channels in CA1 hippocampal neurons. Biochem Biophys Res Commun 2002, 296:1317-1321.
• [45]Gruden MA, Davidova TB, Malisauskas M, Sewell RD, Voskresenskaya NI, Wilhelm K, Elistratova EI, Sherstnev VV, Morozova-Roche LA: Differential neuroimmune markers to the onset of Alzheimer's disease neuro-degeneration and dementia: autoantibodies to Aβ(25-35) oligomers. S100b and neurotransmitters. J Neuroimmunol 2007, 186:181-192.
• [46]Zhang L, Yu H, Zhao X, Lin X, Tan C, Cao G, Wang Z: Neuroprotective effects of salidroside against beta-amyloid-induced oxidative stress in SH-SY5Y human neuroblastoma cells. Neurochem Int 2010, 57:547-555.
• [47]Quintana C, Lancin M, Marhic C, Pérez M, Martin-Benito J, Avila J, Carrascosa JL: Initial studies with high resolution TEM and electron energy loss spectroscopy studies of ferritin cores extracted from brains of patients with progressive supranuclear palsy and Alzheimer disease. Cell Mol Biol 2000, 46:807-820.
• [48]Multhaup G, Scheuermann S, Schlicksupp A, Simons A, Strauss M, Kemmling A, Oehler C, Cappai R, Pipkorn R, Bayer TA: Possible mechanisms of APP-mediated oxidative stress in Alzheimer's disease. Free Radic Biol Med 2002, 33:45-51.
• [49]Sayre LM, Perry G, Harris PL, Liu Y, Schubert KA, Smith MA: In situ oxidative catalysis by neurofibrillary tangles and senile plaques in Alzheimer's disease: a central role for bound transition metals. J Neurochem 2000, 74:270-279.
• [50]Juurlink BH, Paterson PG: Review of oxidative stress in brain and spinal cord injury: suggestions for pharmacological and nutritional management strategies. J Spinal Cord Med 1998, 21:309-334.
• [51]Anandatheerthavarada HK, Biswas G, Robin MA, Avadhani NG: Mitochondrial targeting and a novel transmembrane arrest of Alzheimer's amyloid precursor protein impairs mitochondrial function in neuronal cells. J Cell Biol 2003, 161:41-54.
• [52]LaFerla FM, Green KN, Oddo S: Intracellular amyloid-β in Alzheimer's disease. Nat Rev Neurosci 2007, 8:499-509.
• [53]Lin MT, Beal MF: Alzheimer's APP mangles mitochondria. Nat Med 2006, 12:1241-1243.
• [54]Takuma K, Fang F, Zhang W, Yan S, Fukuzaki E, Du H, Sosunov A, McKhann G, Funatsu Y, Nakamichi N, Nagai T, Mizoguchi H, Ibi D, Hori O, Ogawa S, Stern DM, Yamada K, Yan SS: RAGE-mediated signaling contributes to intraneuronal transport of amyloid-β and neuronal dysfunction. Proc Natl Acad Sci USA 2009, 106:20021-20026.
• [55]Chen X, Walker DG, Schmidt AM, Arancio O, Lue LF, Yan SD: RAGE: A potential target for Aβ-mediated cellular perturbation in Alzheimer's disease. Curr Mol Med 2007, 7:735-742.
• [56]Ding Q, Keller JN: Evaluation of rage isoforms, ligands, and signaling in the brain. Biochim Biophys Acta 2005, 1746:18-27.
• [57]Origlia N, Righi M, Capsoni S, Cattaneo A, Fang F, Stern DM, Chen JX, Schmidt AM, Arancio O, Yan SD, Domenici L: Receptor for advanced glycation end product-dependent activation of p38 mitogen-activated protein kinase contributes to amyloid-β-mediated cortical synaptic dysfunction. J Neurosci 2008, 28:3521-3530.
• [58]Origlia N, Capsoni S, Cattaneo A, Fang F, Arancio O, Yan SD, Domenici L: Aβ-dependent Inhibition of LTP in different intracortical circuits of the visual cortex: The role of RAGE. J Alzheimers Dis 2009, 17:59-68.
• [59]Onyango IG, Tuttle JB, Bennett JP Jr: Altered intracellular signaling and reduced viability of Alzheimer's disease neuronal cybrids is reproduced by beta-amyloid peptide acting through receptor for advanced glycation end products (RAGE). Mol Cell Neurosci 2005, 29:333-343.
• [60]Habtemariam S: Flavonoids as inhibitors or enhancers of the cytotoxicity of tumor necrosis factor in L-929 tumor cells. J Nat Prod 1997, 60:775-778.
• [61]Sala A, Recio MC, Schinella GR, Máñez S, Giner RM, Cerdá-Nicolás M, Rosí JL: Assessment of the anti-inflammatory activity and free radical scavenger activity of tiliroside. Eur J Pharmacol 2003, 461:53-61.
• [62]Christophe M, Nicolas S: Mitochondria: a target for neuroprotective interventions in cerebral ischemia-reperfusion. Curr Pharm Des 2006, 12:739-757.
• [63]Chang L, Karin M: Mammalian MAP kinase signalling cascades. Nature 2001, 410:37-40.
• [64]Lao Y, Chang DC: Study of the functional role of Bcl-2 family proteins in regulating Ca(2+) signals in apoptotic cells. Biochem Soc Trans 2007, 35:1038-1039.
• [65]Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES, Wang X: Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 1997, 91:479-489.
• [66]Eminel S, Klettner A, Roemer L, Herdegen T, Waetzig V: JNK2 translocates to the mitochondria and mediates cytochrome c release in PC 12 cells in response to 6-hydroxydopamine. J Biol Chem 2004, 279:55385-55392.
• [67]Tournier C, Hess P, Yang DD, Xu J, Turner TK, Nimnual A, Bar-Sagi D, Jones SN, Flavell RA, Davis RJ: Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. Science 2000, 288:870-874.
• [68]King M, Nafar F, Clarke J, Mearow K: The small heat shock protein Hsp27 protects cortical neurons against the toxic effects of beta-amyloid peptide. J Neurosci Res 2009, 87:3161-3175.