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In vitro screening on amyloid precursor protein modulation of plants used in Ayurvedic and traditional Chinese medicine for memory improvement
Authors:Liu Liang-Feng  Durairajan Siva Sundara Kumar  Lu Jia-Hong  Koo Irene  Li Min
Institution:School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong.
Abstract:

Ethnopharmacological relevance

The 15 herbs for the screening have been traditionally used in Ayurvedic medicine or in Traditional Chinese medicine (TCM) for the treatment of cognitive disorders clinically.

Aim of the study

Fifteen plant species were investigated for the inhibition of amyloid peptide (Aβ) production and modulation of amyloid precursor protein (APP) processing.

Materials and methods

The selected plants were extracted successively with 70% ethyl alcohol and absolute alcohol concentrated with rotary evaporation then lyophilized. Using a mouse neuroblastoma cells expressing Swedish APP (N2a-SweAPP), MTT assay was performed to determine the toxicity concentration of each herbal extract. In order to evaluate the activity of ethanol extracts on Aβ inhibition, the N2a-SweAPP cells were treated with a high and low dosage of different extracts for 24 h, Aβs levels in the supernatant of conditioned media were assessed by ELISA. The most active extracts were then subjected to test the effect on APP modulation in N2a-SweAPP cells by determining their effect on sAPPα and sAPPβ through western blot analysis.

Results

Among the screened herbal extracts, only Polygonum multiflorum Thunb. (root) and Convolvulus pluricaulis Choisy. (leaves) showed profound inhibition of Aβ production. MTT assay demonstrated that the anti-Aβ effect of these extracts was not a sequential consequence of their cytotoxicity. The extract of Polygonum multiflorum Thunb. (root) could reduce Aβ production only through APP modulation, which was exhibited together with the up-regulation of sAPPα and down-regulation of sAPPβ.

Conclusion

The results show that extract of Polygonum multiflorum Thunb. (root) can lower Aβ generation by modulating APP processing in the N2a-SwedAPP cell line. These results corroborate the traditional use of Polygonum multiflorum Thunb. (root) for the treatment of cognitive disorders including Alzheimer's disease (AD).
Keywords:AD  Alzheimer's disease    β-amyloid peptide  APP  amyloid-β precursor protein  ADAMs  a disintegrin and metalloproteinases  sAPP  α soluble APP  AICD  APP intracellular domain  BACE1  beta-site APP cleaving enzyme1  sAPPβ  β soluble APP  BBB  blood–brain barrier  CNS  central nervous system  DMEM  Dulbecco's modified Eagle's medium  FBS  fetal bovine serum  DMSO  dimethyl sulfoxide  MTT  3-(4  5-dimethylthiazol-2-yl)-2  5-diphenyltetrazolium bromide  DMF  N  N-dimethylformamide  ELISA  enzyme-linked immunosorbent assay  PBS  phosphate-buffered saline  PBST  PBS with 0  05% Tween 20  SDS  sodium dodecyl sulfate  SDS–PAGE  sodium dodecyl sulfate polyacrylamide gel electrophoresis  BCA  bicinchoninic acid  PVDF  polyvinylidene fluoride  FL-APP  full length APP
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