Fermented Citrus reticulata (ponkan) fruit squeezed draff that contains a large amount of 4′-demethylnobiletin prevents MK801-induced memory impairment
A previous study reported biotransformation of a citrus peel polymethoxyflavone, nobiletin, by Aspergillus enabling production of 4′-demethylnobiletin, and the product’s antimutagenic activity. However, the effects of fermented citrus peel on the basal forebrain–hippocampal system remain unidentified. Citrus reticulata (ponkan) fruit squeezed draffs are generated as mass waste in beverage factories. In this study using PC12D cells and cultured central nervous system neurons, we therefore examined whether Aspergillus kawachii-fermented citrus fruit squeezed draff could affect cAMP response element (CRE)- and choline acetyltransferase gene (ChAT) promoter region-mediated transcriptional activities relevant to memory formation and cholinergic function. Our current fermentation yielded approximately 80% nobiletin bioconversion, and a sample of hot-water extract of the fermented fruit squeezed draff was stronger than that of the unfermented one in facilitating CRE-mediated transcription in cultured hippocampal neurons as well as in PC12D cells. A sample of 0–80% ethanol-eluted fraction of Diaion HP-20 column-adsorbed components of the preparation obtained by the fermentation concentration-dependently and more strongly facilitated CRE-mediated transcription than did the fraction of the unfermented one in both cell culture systems. In a separate study, this polymethoxyflavone-rich fraction of the fermented fruit squeezed draff showed a potent ability to facilitate CRE-mediated and ChAT transcription in a co-culture of hippocampal neurons and basal forebrain neurons. Repeated oral gavage of mice with the fermented fraction sample prevented MK801-impaired memory formation in mice. These findings suggest that the 4′-demethylnobiletin-rich fraction prepared from the Aspergillus-fermented ponkan squeezed draff has a potential anti-dementia effect.
KeywordsAspergillus kawachii-fermented Citrus reticulata fruit squeezed draff 4′-demethylnobiletin CRE-mediated transcription ChAT transcription CNS neurons Dementia
We thank Dr. Brygida Berse of Boston University, School of Medicine, for providing the plasmid pGL3-ChAT.
Compliance with ethical standards
Conflict of interest
Tohru Yamakuni has received a research Grant from Fuji Sangyo Co., Ltd.
- 6.Nakajima A, Yamakuni T, Matsuzaki K, Nakata N, Onozuka H, Yokosuka A, Sashida Y, Mimaki Y, Ohizumi Y (2007) Nobiletin, a citrus flavonoid, reverses learning impairment associated with N-methyl-D-aspartate receptor antagonism by activation of extracellular signal-regulated kinase signaling. J Pharmacol Exp Ther 321(2):784–790. doi: 10.1124/jpet.106.117010 CrossRefPubMedGoogle Scholar
- 7.Onozuka H, Nakajima A, Matsuzaki K, Shin RW, Ogino K, Saigusa D, Tetsu N, Yokosuka A, Sashida Y, Mimaki Y, Yamakuni T, Ohizumi Y (2008) Nobiletin, a citrus flavonoid, improves memory impairment and Abeta pathology in a transgenic mouse model of Alzheimer’s disease. J Pharmacol Exp Ther 326(3):739–744. doi: 10.1124/jpet.108.140293 CrossRefPubMedGoogle Scholar
- 9.Al Rahim M, Nakajima A, Saigusa D, Tetsu N, Maruyama Y, Shibuya M, Yamakoshi H, Tomioka Y, Iwabuchi Y, Ohizumi Y, Yamakuni T (2009) 4′-Demethylnobiletin, a bioactive metabolite of nobiletin enhancing PKA/ERK/CREB signaling, rescues learning impairment associated with NMDA receptor antagonism via stimulation of the ERK cascade. Biochemistry 48(32):7713–7721. doi: 10.1021/bi901088w CrossRefPubMedGoogle Scholar
- 23.Kawahata I, Yoshida M, Sun W, Nakajima A, Lai Y, Osaka N, Matsuzaki K, Yokosuka A, Mimaki Y, Naganuma A, Tomioka Y, Yamakuni T (2013) Potent activity of nobiletin-rich Citrus reticulata peel extract to facilitate cAMP/PKA/ERK/CREB signaling associated with learning and memory in cultured hippocampal neurons: identification of the substances responsible for the pharmacological action. J Neural Transm 120(10):1397–1409. doi: 10.1007/s00702-013-1025-x CrossRefPubMedGoogle Scholar
- 24.Matsuzaki K, Miyazaki K, Sakai S, Yawo H, Nakata N, Moriguchi S, Fukunaga K, Yokosuka A, Sashida Y, Mimaki Y, Yamakuni T, Ohizumi Y (2008) Nobiletin, a citrus flavonoid with neurotrophic action, augments protein kinase A-mediated phosphorylation of the AMPA receptor subunit, GluR1, and the postsynaptic receptor response to glutamate in murine hippocampus. Eur J Pharmacol 578(2–3):194–200. doi: 10.1016/j.ejphar.2007.09.028 CrossRefPubMedGoogle Scholar
- 25.Nakatani K, Yamakuni T, Kondo N, Arakawa T, Oosawa K, Shimura S, Inoue H, Ohizumi Y (2004) Gamma-Mangostin inhibits inhibitor-kappaB kinase activity and decreases lipopolysaccharide-induced cyclooxygenase-2 gene expression in C6 rat glioma cells. Mol Pharmacol 66(3):667–674. doi: 10.1124/mol.104.002626 CrossRefPubMedGoogle Scholar
- 38.Madziar B, Shah S, Brock M, Burke R, Lopez-Coviella I, Nickel AC, Cakal EB, Blusztajn JK, Berse B (2008) Nerve growth factor regulates the expression of the cholinergic locus and the high-affinity choline transporter via the Akt/PKB signaling pathway. J Neurochem 107(5):1284–1293. doi: 10.1111/j.1471-4159.2008.05681.x CrossRefPubMedGoogle Scholar
- 40.Berse B, Blusztajn JK (1995) Coordinated up-regulation of choline acetyltransferase and vesicular acetylcholine transporter gene expression by the retinoic acid receptor alpha, cAMP, and leukemia inhibitory factor/ciliary neurotrophic factor signaling pathways in a murine septal cell line. J Biol Chem 270(38):22101–22104CrossRefPubMedGoogle Scholar
- 44.Shankar GM, Li S, Mehta TH, Garcia-Munoz A, Shepardson NE, Smith I, Brett FM, Farrell MA, Rowan MJ, Lemere CA, Regan CM, Walsh DM, Sabatini BL, Selkoe DJ (2008) Amyloid-beta protein dimers isolated directly from Alzheimer’s brains impair synaptic plasticity and memory. Nat Med 14(8):837–842. doi: 10.1038/nm1782 CrossRefPubMedPubMedCentralGoogle Scholar
- 46.Nakajima A, Yamakuni T, Haraguchi M, Omae N, Song SY, Kato C, Nakagawasai O, Tadano T, Yokosuka A, Mimaki Y, Sashida Y, Ohizumi Y (2007) Nobiletin, a citrus flavonoid that improves memory impairment, rescues bulbectomy-induced cholinergic neurodegeneration in mice. J Pharmacol Sci 105(1):122–126CrossRefPubMedGoogle Scholar