High glucose promotes macrophage switching to the M1 phenotype via the downregulation of STAT-3 mediated autophagy

PLoS One. 2024 Dec 31;19(12):e0314974. doi: 10.1371/journal.pone.0314974. eCollection 2024.

Abstract

Aim: Imbalanced M1/M2 macrophage phenotype activation is a key point in diabetic kidney disease (DKD). Macrophages mainly exhibit the M1 phenotype, which contributes to inflammation and fibrosis in DKD. Studies have indicated that autophagy plays an important role in M1/M2 activation. However, the mechanism by which autophagy regulates the macrophage M1/M2 phenotype in DKD is unknown. Thus, the aim of the present study was to explore whether high glucose-induced macrophages switch to the M1 phenotype via the downregulation of STAT-3-mediated autophagy.

Methods: DKD model rats were established in vivo via the intraperitoneal injection of streptozocin (STZ). The rats were sacrificed at 18 weeks for histological and molecular analysis. RAW264.7 cells were cultured in vitro with 30 mM glucose in the presence or absence of a STAT-3 activator (colivelin) and an autophagy activator (rapamycin). Moreover, M1 and M2 macrophage activation models were established as a control group. Immunofluorescence and Western blot analyses were used to detect the expression of autophagy-related proteins (LC3 and Beclin-1), M1 markers (iNOS and CD11c) and M2 markers (MR and CD206).

Results: In DKD, macrophages exhibit an M1 phenotype. Under high-glucose conditions, RAW264.7 macrophages switched to the M1 phenotype. Autophagy was downregulated in high glucose-induced M1 macrophages. Both the STAT-3 activator and the autophagy activator promoted the transition of glucose-induced M1 macrophages to M2 macrophages. Moreover, STAT-3 activation increased the expression of autophagy markers (LC3 and Beclin-1). However, the autophagy activator had no effect on STAT-3 phosphorylation.

Conclusion: High glucose promotes macrophage switching to the M1 phenotype via the downregulation of STAT-3-mediated autophagy.

MeSH terms

  • Animals
  • Autophagy* / drug effects
  • Beclin-1 / genetics
  • Beclin-1 / metabolism
  • Diabetes Mellitus, Experimental / metabolism
  • Diabetes Mellitus, Experimental / pathology
  • Diabetic Nephropathies / metabolism
  • Diabetic Nephropathies / pathology
  • Down-Regulation* / drug effects
  • Glucose* / metabolism
  • Glucose* / pharmacology
  • Macrophage Activation / drug effects
  • Macrophages* / drug effects
  • Macrophages* / metabolism
  • Male
  • Mice
  • Phenotype
  • RAW 264.7 Cells
  • Rats
  • Rats, Sprague-Dawley
  • STAT3 Transcription Factor* / metabolism

Substances

  • STAT3 Transcription Factor
  • Glucose
  • Beclin-1

Grants and funding

This work was supported by grants from the National Natural Science Foundation Youth Fund (NO.82000650); Jiangsu Province Key R&D Program-Social Development (BE2021737). Zhongda Hospital Affiliated to Southeast University, Jiangsu Province High-Level Hospital Pairing Assistance Construction Funds (zdyyxy14, zdyyxy02, zdlyg02).