Protective effect of donepezil against Abeta(1-40) neurotoxicity in rat septal neurons.
Donepezil, a potent acetylcholinesterase (AChE) inhibitor used for the treatment of Alzheimer's disease (AD), is thought to have a neuroprotective effect in AD patients. Because a deficit in cholinergic neurotransmission is a major feature in AD, and amyloid-beta (Abeta) accumulation has been proposed as a possible causative phenomenon, we were interested to examine the effect of donepezil on Abeta(1-40) induced neurotoxicity in primary cultures of rat septal neurons. Using immunohistochemical staining, almost all the neurons were found to be positive for vesicular acetylcholine transporter (VAChT) in these septal cultures. Septal neuronal cells were cultured for 7 days and then 15 micromol/L of Abeta(1-40) was added to the cell medium for 48 h. The cultured septal neurons were highly susceptible to Abeta toxicity, as shown by morphological examination and lactate dehydrogenase (LDH) assay. Donepezil concentration-dependently reduced the LDH efflux induced by Abeta(1-40), and the effect was significant at 100 nmol/L and above. Donepezil decreased both the negative peak at around 215 nm in the circular dichroism (CD) spectrum and the fluorescence intensity of thioflavin T in the presence of Abeta(1-40). These results suggest that donepezil exerts a neuroprotective effect by reducing the amount of the toxic form of Abeta fibrils in septal neuron cultures. These findings support the idea that the clinical efficacy of donepezil in AD is due to not only activation of cholinergic transmission, but also attenuation of neuronal damage.
Kimura
Manami
M
Department of Drug Discovery, Eisai Tsukuba Research Laboratories, 5-1-3 Tokodai, Tsukuba, Ibaraki 300-2635, Japan. m4-kimura@hhc.eisai.co.jp
Akasofu
Shigeru
S
Ogura
Hiroo
H
Sawada
Kohei
K
D000109:Acetylcholine / Q000378:metabolism; D000544:Alzheimer Disease / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D016229:Amyloid beta-Peptides / Q000037:antagonists & inhibitors* / Q000633:toxicity; D000818:Animals; D052160:Benzothiazoles; D002478:Cells, Cultured; D002799:Cholinergic Fibers / Q000187:drug effects* / Q000378:metabolism; D002800:Cholinesterase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D000077265:Donepezil; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology; D005456:Fluorescent Dyes / Q000737:chemistry; D007150:Immunohistochemistry; D007189:Indans / Q000737:chemistry / Q000494:pharmacology*; D007770:L-Lactate Dehydrogenase / Q000187:drug effects / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000378:metabolism; D015394:Molecular Structure; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D018696:Neuroprotective Agents / Q000737:chemistry / Q000494:pharmacology*; D010446:Peptide Fragments / Q000037:antagonists & inhibitors* / Q000633:toxicity; D010880:Piperidines / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D012686:Septal Nuclei / Q000187:drug effects* / Q000378:metabolism / Q000503:physiopathology; D013844:Thiazoles / Q000737:chemistry; D050494:Vesicular Acetylcholine Transport Proteins
D016229:Amyloid beta-Peptides; D052160:Benzothiazoles; D002800:Cholinesterase Inhibitors; D005456:Fluorescent Dyes; D007189:Indans; D026901:Membrane Transport Proteins; D018696:Neuroprotective Agents; D010446:Peptide Fragments; D010880:Piperidines; C493461:Slc18a3 protein, rat; D013844:Thiazoles; D050494:Vesicular Acetylcholine Transport Proteins; C078592:amyloid beta-protein (1-40); C009462:thioflavin T; D000077265:Donepezil; D007770:L-Lactate Dehydrogenase; D000109:Acetylcholine