Preventing Expression of the Nicotinic Receptor Subunit α7 in SH-SY5Y Cells with Interference RNA Indicates that this Receptor may Protect Against the Neurotoxicity of Aβ

• Published in: Neurochemical research Vol. 38; no. 5; pp. 943 - 950
• Main Authors: Qi, Xiao-Lan, Ou-Yang, Kai, Ren, Jia-Mou, Wu, Chang-Xue, Xiao, Yan, Li, Yi, Guan, Zhi-Zhong
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.05.2013
• Subjects: alpha7 Nicotinic Acetylcholine Receptor; Amyloid beta-Peptides - genetics; Amyloid beta-Peptides - physiology; Base Sequence; Biochemistry; Biomedical and Life Sciences; Biomedicine; Cell Biology; Cell Line, Tumor; DNA Primers; Humans; Neurochemistry; Neurology; Neurosciences; Original Paper; Real-Time Polymerase Chain Reaction; Receptors, Nicotinic - metabolism; RNA Interference; RNA, Small Interfering - genetics; Oxidative stress; β-Amyloid peptide; SH-SY5Y cells; Secretase; α7 Nicotinic acetylcholine receptor
• ISSN: 0364-3190; 1573-6903
• DOI: 10.1007/s11064-013-1001-8

The present aim was to characterize the influence of the α7 nicotinic acetylcholine receptor (nAChR) on BACE, the enzyme that cleaves the amyloid precursor protein (APP) at the β-site, as well as on the oxidative stress induced by amyloid-β peptide (Aβ). To this end, human neuroblastoma SH-SY5Y cells were transfected with siRNAs targeting the α7 nAChR subunit and/or exposed to Aβ 1–42 . For α7 nAChR, BACE1 (cleaving at the β-site of APP) and BACE2 (cleaving within the Aβ domain), α-secretase (ADAM10), and the two components of γ-secretase, PS and NCT, the mRNA and protein levels were determined by real-time PCR and Western blotting, respectively. The level of Aβ 1–42 in the cell culture medium was determined by an ELISA procedure. The extent of lipid peroxidation and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were assayed spectrophotometrically. In the transfected SH-SY5Y cells, expression of α7 nAChR was reduced; the level of BACE1 increased and that of BACE2 decreased; the amount of ADAM10 lowered; and the level of PS raised. Moreover, the level of Aβ 1–42 in the culture medium was elevated. Treatment of non-transfected cells with Aβ elevated the level of malondialdehyde (MDA) and lowered the activities of SOD and GSH-Px and these changes were potentiated by inhibiting expression of α7 nAChR. These results indicate that α7 nAChR plays a significant role in amyloidogenic metabolism of APP and the oxidative stress evoked by Aβ, suggesting that this receptor might help protect against the neurotoxicity of Aβ.