[Influence of inhibited gene expression of alpha3 nicotinic acetylcholine receptor by RNA interference on anti-oxidation in SH-SY5Y cells.]
 

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12-20-08 08:00 AM
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[Influence of inhibited gene expression of alpha3 nicotinic acetylcholine receptor by RNA interference on anti-oxidation in SH-SY5Y cells.]
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[Influence of inhibited gene expression of alpha3 nicotinic acetylcholine receptor by RNA interference on anti-oxidation in SH-SY5Y cells.]

Zhonghua Bing Li Xue Za Zhi. 2008 Sep;37(9):620-4

Authors: Tang Z, An Y, Qi XL, Xiao Y, Shan KR, Guan ZZ

OBJECTIVE:s To investigate the neuroprotective function of alpha3 nicotinic acetylcholine receptor (nAChR) by inhibiting the gene expression in human neuroblastoma (SH-SY5Y) cells using small interference RNA (siRNA). METHODS: The siRNA coding oligonucleotide sequences targeting alpha3 nAChR were designed and synthesized. The annealed product was cloned into pSilencer 3.1-H1 neo vector. The recombinant alpha3 nAChR pSilencer 3.1-H1 neo vector was transfected into the SH-SY5Y cells. The stable clones were screened by G418 medium, and the levels of alpha3 nAChR mRNA and protein were monitored by using real-time PCR and Western blotting, respectively. After the SH-SY5Y cells with siRNA treatment were exposed to 1 micromol/L Abeta(1 - 42), MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide], SOD, GSH-px and the lipid peroxidation were measured by spectrophotometry. RESULTS: Compared with the controls, the expression levels of mRNA and protein in the stable SH-SY5Y clone cells transfected with the recombinant alpha3 nAChR pSilencer 3.1-H1 neo vector were decreased with inhibitory efficiency of 98% and 66%, respectively, the MTT reduction decreased; the product of lipid peroxidation was increased and the activities of SOD and GSH-px were decreased. Biologically, the gene expression inhibition of alpha3 nAChR enhanced the toxicity induced by Abeta in SH-SY5Y cells. CONCLUSIONS: The expression inhibition of alpha3 nAChR as a result of recombinant alpha3 nAChR siRNA can induce oxidative stress and improve the toxicity of Abeta on SH-SY5Y cells, indicating that alpha3 nAChR may play a significant neuroprotective role in the pathogenesis of Alzheimer disease.

PMID: 19094587 [PubMed - in process]