Investigations of anticholinestrase and antioxidant potentials of methanolic extract, subsequent fractions, crude saponins and flavonoids isolated from Isodon rugosus.
BACKGROUND
Based on the ethnomedicinal uses and the effective outcomes of natural products in various diseases, this study was designed to evaluate Isodon rugosus as possible remedy in oxidative stress, alzheimer's and other neurodegenerative diseases. Acetylecholinestrase (AChE) and butyrylcholinesterase (BChE) inhibitory activities of crude methanolic extract (Ir.Cr), resultant fractions (n-hexane (Ir.Hex), chloroform (Ir.Cf), ethyl acetate (Ir.EtAc), aqueous (Ir.Aq)), flavonoids (Ir.Flv) and crude saponins (Ir.Sp) of I. rugosus were investigated using Ellman's spectrophotometric method. Antioxidant potential of I. rugosus was determined using DPPH, H2O2 and ABTS free radicals scavenging assays. Total phenolic and flavonoids contents of plant extracts were determined and expressed in mg GAE/g dry weight and mg RTE/g of dry sample respectively.


RESULTS
Among different fractions Ir.Flv and Ir.Cf exhibited highest inhibitory activity against AChE (87.44 ± 0.51, 83.73 ± 0.64%) and BChE (82.53 ± 0.71, 88.55 ± 0.77%) enzymes at 1 mg/ml with IC50 values of 45, 50 for AChE and 40, 70 μg/ml for BChE respectively. Activity of these fractions were comparable to galanthamine causing 96.00 ± 0.30 and 88.61 ± 0.43% inhibition of AChE and BChE at 1 mg/ml concentration with IC50 values of 20 and 47 μg/ml respectively. In antioxidant assays, Ir.Flv, Ir.Cf, and Ir.EtAc demonstrated highest radicals scavenging activities in DPPH and H2O2 assays which were comparable to ascorbic acid. Ir.Flv was found most potent with IC50 of 19 and 24 μg/ml against DPPH and H2O2 radicals respectively. Whereas antioxidant activates of plant samples against ABTS free radicals was moderate. Ir.Cf, Ir.EtAc and Ir.Cr showed high phenolic and flavonoid contents and concentrations of these compounds in different fractions correlated well to their antioxidant and anticholinestrase activities.


CONCLUSIONS
It may be inferred from the current investigations that the Ir.Sp, Ir.Flv and various fractions of I. rugosus are good sources of anticholinesterase and antioxidant compounds. Different fractions can be subjected to activity guided isolation of bioactive compounds effective in neurological disorders.
Department of Pharmacy, University of Malakand, Chakdara, Dir, Pakistan. zeb.takor@yahoo.com.
Anwar
A
Zeb
Department of Pharmacy, University of Malakand, Chakdara, Dir, Pakistan. sadiquom@yahoo.com.
Abdul
A
Sadiq
Department of Pharmacy, University of Malakand, Chakdara, Dir, Pakistan. farhataziz80@hotmail.com.
Farhat
F
Ullah
Department of Pharmacy, University of Malakand, Chakdara, Dir, Pakistan. sajad4u2000@yahoo.com.
Sajjad
S
Ahmad
Department of Pharmacy, University of Malakand, Chakdara, Dir, Pakistan. ayazuop@gmail.com.
Muhammad
M
Ayaz
D000085:Acetates; D000110:Acetylcholinesterase / Q000187:drug effects; D000975:Antioxidants / Q000032:analysis*; D052160:Benzothiazoles / Q000378:metabolism; D001713:Biphenyl Compounds / Q000378:metabolism; D002091:Butyrylcholinesterase / Q000187:drug effects; D002725:Chloroform; D002800:Cholinesterase Inhibitors / Q000032:analysis*; D045424:Complex Mixtures; D005419:Flavonoids / Q000032:analysis* / Q000302:isolation & purification; D016166:Free Radical Scavengers / Q000378:metabolism; D005609:Free Radicals / Q000032:analysis; D006586:Hexanes; D006861:Hydrogen Peroxide / Q000378:metabolism; D020128:Inhibitory Concentration 50; D037602:Isodon / Q000737:chemistry* / Q000145:classification / Q000201:enzymology; D008519:Medicine, Traditional; D000432:Methanol; D018384:Oxidative Stress / Q000187:drug effects; D010851:Picrates / Q000378:metabolism; D035261:Plant Components, Aerial / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry*; D012503:Saponins / Q000032:analysis* / Q000302:isolation & purification; D013053:Spectrophotometry / Q000379:methods; D013451:Sulfonic Acids / Q000378:metabolism
D000085:Acetates; D000975:Antioxidants; D052160:Benzothiazoles; D001713:Biphenyl Compounds; D002800:Cholinesterase Inhibitors; D045424:Complex Mixtures; D005419:Flavonoids; D016166:Free Radical Scavengers; D005609:Free Radicals; D006586:Hexanes; D010851:Picrates; D010936:Plant Extracts; D012503:Saponins; D013451:Sulfonic Acids; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; C026385:n-hexane; C007650:ethyl acetate; D002725:Chloroform; D006861:Hydrogen Peroxide; C004931:1,1-diphenyl-2-picrylhydrazyl; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase; D000432:Methanol