High-glucose-induced changes in macrophage secretome: regulation of immune response

Molecular and Cellular Biochemistry
Taniya Kaewarpai, Visith Thongboonkerd

Abstract

Secretory products from infiltrating macrophages have been thought to play crucial roles in development and progression of diabetic complications in various tissues/organs. Nevertheless, diabetes-induced changes in macrophage secretory products remained largely unknown. We thus analyzed high-glucose (HG)-induced changes in secretome of human macrophages derived from U937 human monocytic cell line after phorbol 12-myristate 13-acetate (PMA) activation. Serum-free culture supernatants were collected from macrophages exposed to 5.5 mM glucose (NG-M-sup) (normal control), 25 mM glucose (HG-M-sup), or 5.5 mM glucose + 19.5 mM mannitol (MN-M-sup) (osmotic control) for 16 h. After dialysis and lyophilization, secreted proteins were subjected to 2-DE analysis (n = 5 gels derived from 5 independent cultures per group). Quantitative analysis and statistics revealed 23 protein spots whose secretory levels significantly differed among the three conditions. These proteins were successfully identified by nanoLC-ESI-MS/MS analyses and changes in levels of heat shock protein 90 (HSP90), HSP70, HSP60, and β-actin were confirmed by Western blotting. Global protein network and functional enrichment analyses revealed that the altered proteins in H...Continue Reading

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Citations

Feb 9, 2020·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Shelton W WrightT Eoin West

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Methods Mentioned

BETA
protein assay
Electrophoresis
PMA
ubiquitination
protein folding

Software Mentioned

SPSS
Mascot
ImageMaster 2D Platinum
STRING
FunRich

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