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[Effects of α3 neuronal nicotinic acetylcholine receptor on cell apoptosis and p38 MAPK signal transduction pathway in SH-SY5Y cells]. | LitMetric

AI Article Synopsis

  • The study aims to explore how the α3 neuronal nicotinic acetylcholine receptor influences cell death (apoptosis) and the p38 signaling pathway in SH-SY5Y cells, with a focus on its relevance to Alzheimer's disease (AD).
  • Researchers used techniques like real-time PCR and Western blot to assess the expression of α3 nAChR and related proteins in cells treated with a specific peptide (Aβ25-35).
  • Results indicated that silencing α3 nAChR led to increased apoptosis and heightened p38 activation when exposed to Aβ25-35, suggesting that α3 nAChR may contribute to AD progression by altering apoptosis-related gene expression.

Article Abstract

Objective: To investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD).

Methods: The levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aβ25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again.

Results: Compared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aβ25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aβ25-35. Compared to controls, the Aβ25-35-treated SH-SY5Y cells and the Aβ25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former.

Conclusion: The siRNA silencing of α3 nAChR mRNA may enhance the effect of Aβ25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.

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Source
http://dx.doi.org/10.3760/cma.j.issn.0529-5807.2013.02.010DOI Listing

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