[Chronic high glucose inhibits AKT phosphorylation and promotes M1 polarization of mouse RAW264.7 macrophages]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2019 Oct;35(10):910-917.
[Article in Chinese]

Abstract

Objective To explore the effect of chronic high glucose on mouse macrophages in vitro and its mechanism. Methods RAW264.7 macrophages were cultured in normal growth medium and glucose-containing 40 or 60 mmol/L growth medium for 1, 3, 5 and 7 days. Cell morphology was observed by phase contrast microscopy; cell proliferation was measured by CCK-8 assay; interleukin-1β (IL-1β) and IL-10 protein levels were detected by Western blot analysis; IL-1β and IL-10 secretion were detected by ELISA; the levels of AKT mRNA in the cells were tested by real-time quantitative PCR; the levels of AKT and phosphorylated AKT (p-AKT) were examined by Western blot analysis. Results After 60 mmol/L glucose treatment for 1 day, the longer spindle cells appeared, the extended pseudopod was visible, and the number and volume increased over time. After treatment for 1 day, cell proliferation was significantly inhibited, and the inhibitory effect was ameliorated with time. After treatment for 3, 5, and 7 days, IL-1β protein level increased and IL-10 protein level decreased. After treatment for 1 day, the secretion of IL-1β increased significantly, but after treatment for 3 days, the secretion of IL-10 decreased significantly and decreased further over time. AKT protein phosphorylation was inhibited after treatment for 3, 5, and 7 days, but AKT mRNA level did not change obviously. Glucose (40 mmol/L) treatment played a role to some extent, but it was not as significant and stable as 60 mmol/L glucose. Conclusion Chronic high glucose induces macrophages to transform into pro-inflammatory M1 type and the continuous chronic inflammatory response, which may be related to the inhibition of AKT protein phosphorylation.

MeSH terms

  • Animals
  • Glucose / pharmacology*
  • Inflammation
  • Interleukin-10 / metabolism
  • Interleukin-1beta / metabolism
  • Macrophages / drug effects
  • Macrophages / metabolism*
  • Mice
  • Phosphorylation
  • Proto-Oncogene Proteins c-akt / metabolism*
  • RAW 264.7 Cells

Substances

  • IL10 protein, mouse
  • IL1B protein, mouse
  • Interleukin-1beta
  • Interleukin-10
  • Proto-Oncogene Proteins c-akt
  • Glucose