title: In vitro acetylcholinesterase inhibitory activity and the antioxidant properties of Aegle marmelos leaf extract: implications for the treatment of Alzheimer's disease.
abstract: BACKGROUND
  Alzheimer's disease (AD) is a progressive neurodegenerative disorder clinically characterized by loss of memory and cognition. The effective therapeutic options for AD are limited and thus there is a demand for new drugs. Aegle marmelos (Linn.) (A. marmelos) leaves have been used in traditional medicine to promote intellect and enhance memory. In this study, we evaluated A. marmelos for its acetylcholinesterase (AChE) inhibitory activity and antioxidant property in vitro in the treatment of AD.
  
  
  METHODS
  A crude methanol extract and four fractions (petroleum ether, chloroform, ethyl acetate and aqueous) were prepared from the leaves of A. marmelos. The preparations were assessed for AChE inhibitory activity by the Ellman method, and their antioxidant properties were assessed by several assays: reducing power, scavenging of 1,1-diphenyl-2-picrylhydrazyl free radical and hydroxyl radical, and inhibition of lipid peroxidation. Qualitative and quantitative analyses of endogenous substances in A. marmelos were performed by the standard phytochemical methods.
  
  
  RESULTS
  Among the different extracts tested, the ethyl acetate fraction exhibited the highest inhibition of AChE activity. In the same way, ethyl acetate fraction showed the highest reducing activity and radical scavenging ability towards the 1,1-diphenyl-2-picrylhydrazyl (half maximal inhibitory concentration = 3.84 μg/mL) and hydroxyl free radicals (half maximal inhibitory concentration = 5.68 μg/mL). The antiradical activity of the ethyl acetate fraction appeared to be similar to that of the reference standard butylated hydroxytoluene and catechin used in this study. In addition, the ethyl acetate fraction displayed higher inhibition of brain lipid peroxidation. Phytochemical screening of different extractives of A. marmelos showed the presence of phenols and flavonoids, alkaloid, saponin, glycoside, tannin and steroids. Quantitative analysis revealed higher contents of phenolics (58.79-mg gallic acid equivalent/g dried extract) and flavonoids (375.73-mg gallic acid equivalent/g dried extract) in the ethyl acetate fraction.
  
  
  CONCLUSIONS
  The results suggest that the ethyl acetate fraction of A. marmelos is a significant source of polyphenolic compounds with potential AChE inhibitory property and antioxidant activity and, thus, may be useful in the treatment of AD.
authors.affiliation: Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh.
authors.forename: Md
authors.initials: M
authors.lastname: Asaduzzaman
authors.forename: Md Josim
authors.initials: MJ
authors.lastname: Uddin
authors.forename: M A
authors.initials: MA
authors.lastname: Kader
authors.forename: A H M K
authors.initials: AH
authors.lastname: Alam
authors.forename: Aziz Abdur
authors.initials: AA
authors.lastname: Rahman
authors.forename: Mamunur
authors.initials: M
authors.lastname: Rashid
authors.forename: Kiyoko
authors.initials: K
authors.lastname: Kato
authors.forename: Toshihisa
authors.initials: T
authors.lastname: Tanaka
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Takeda
authors.forename: Golam
authors.initials: G
authors.lastname: Sadik
mesh_terms: D000110:Acetylcholinesterase / Q000187:drug effects*; D045471:Aegle / Q000737:chemistry*; D000544:Alzheimer Disease; D000975:Antioxidants / Q000737:chemistry / Q000494:pharmacology*; D005419:Flavonoids / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D010636:Phenols / Q000737:chemistry / Q000494:pharmacology; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D018515:Plant Leaves / Q000737:chemistry
chemical_list: D000975:Antioxidants; D005419:Flavonoids; D010636:Phenols; D010936:Plant Extracts; D000110:Acetylcholinesterase
keywords: Aegle marmelos; Alzheimer's disease; acetylcholinesterase inhibition; antioxidant activity; phytochemical screening