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Int J Clin Exp Pathol 2012;5(9):900-913

Original Article
The protection of acetylcholinesterase inhibitor on β-amyloid-induced injury of neurite
outgrowth via regulating axon guidance related genes expression in neuronal cells

Jiao-Ning Shen, Deng-Shun Wang, Rui Wang

Department of Pharmaceutical Sciences, School of Pharmacy, Shanghai Key Laboratory of New Drug Design, East China University of Science
and Technology, Shanghai 200237, China; 2Department of Pathology and Laboratory Medicine, School of Medicine and Public Health,
University of Wisconsin, Madison, Wisconsin, USA.

Received August 21, 2012; Accepted September 27, 2012; Epub October 20, 2012; Published October 30, 2012

Abstract: Cognitive deficits in AD correlate with progressive synaptic dysfunction and loss. The Rho family of small GTPases, including Rho,
Rac, and Cdc42, has a central role in cellular motility and cytokinesis. Acetylcholinesterase inhibitor has been found to protect cells against a
broad range of reagents-induced injuries. Present studies examined if the effect of HupA on neurite outgrowth in Aβ-treated neuroblastoma
cells executed via regulating Rho- GTPase mediated axon guidance relative gene expression. Affymetrix cDNA microarray assay followed by
real-time RT-PCR and Western Blotting analysis were used to elucidate and analyze the signaling pathway involved in Aβ and HupA’s effects.
The effects of Aβ and HupA on the neurite outgrowth were further confirmed via immunofluorescence staining. Aβ up-regulated the mRNA
expressions of NFAT5, LIMK1, EPHA1, NTN4 and RAC2 markedly in SH-SY5Y cells. Co-incubation of Aβ and HupA reversed or decreased the
changes of NFAT5, NTN4, RAC2, CDC42 and SEMA4F. HupA treated alone increased NFAT5, LIMK1, NTN4 significantly. Following qRT-PCR
validation showed that the correlation of the gene expression ratio between microarray and qRT-PCR is significant. Western blot result showed
that the change of CDC42 protein is consistent with the mRNA level while RAC2 is not. The morphological results confirmed that HupA
improved, or partly reversed, the Aβ-induced damage of neurite outgrowth. The protective effect of HupA from Aβ induced morphological injury
might be correlative to, at least partially, regulating the network of neurite outgrowth related genes. (IJCEP1208019).

Keywords: β-amyloid, axon guidance, neurite outgrowth, acetylcholinesterase inhibitor, huperzine A

Address all correspondence to:
Dr. Rui Wang
130 Meilong Road
East China University of Science and Technology
Shanghai 200237, China.
Tel/Fax: +86-21-64250823
E-mail: ruiwang@ecust.edu.cn