Di-san junyi daxue xuebao (Jun 2020)

Insulin promotes amyloid precursor protein metabolism in an Alzheimer's disease cell model

  • HU Rong,
  • WANG Dan,
  • LOU Dandan,
  • LI Tingyu,
  • ZHOU Weihui,
  • SONG Weihong

DOI
https://doi.org/10.16016/j.1000-5404.202001195
Journal volume & issue
Vol. 42, no. 12
pp. 1201 – 1208

Abstract

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Objective To investigate how insulin affects the metabolism of amyloid precursor protein (APP) in a cell model of Alzheimer's disease (AD). Methods Human neuroblastoma SH-SY5Y cells and the nonneural cell lines HEK293 and 2EB2 were treated with different concentrations of insulin for 48 h, and the changes in cell viability were evaluated with CCK-8 assay. The expression levels of APP, C-terminal fragment of APP (CTF), β-site APP cleaving enzyme 1 (BACE1), the α-secretase ADAM10 (a disintegrin and metalloproteinase 10) and γ-secretase presenilin 1 (PS1) in the cells were detected using Western blotting. ELISA was used to determine the cellular expression levels of Aβ40 and Aβ42. The key signaling pathways mediated by insulin were screened using a protein chip. The effect of insulin on protein phosphorylation levels of AKT and GSK3β pathways was examined by detecting p-AKT and p-GSK3β expression in the cells with Western blotting. Results Treatment with insulin below 10 μmol/L did not obviously affect the viability of the cells (P>0.05). Western blotting showed that treatment with 1.00 and 10.00 μmol/L insulin significantly increased the protein expression of APP and CTF (P 0.05). Conclusion Insulin promotes the metabolism of APP into CTF and Aβ42, which play certain rolesin pathogenesis of AD.

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