Proteomics identifies hypothermia induced adiponectin protects corneal endothelial cells via AMPK mediated autophagy in phacoemulsification

Graefes Arch Clin Exp Ophthalmol. 2024 Oct;262(10):3207-3217. doi: 10.1007/s00417-024-06542-6. Epub 2024 Jun 8.

Abstract

Aim: To explore the molecular mechanism underlying the protective effect of hypothermic perfusion on the corneal endothelium during phacoemulsification.

Methods: Phacoemulsification was performed on New Zealand white rabbits. Perfusate at different temperatures was used during the operation, and the aqueous humor was collected for proteomic sequencing after the operation. Corneal endothelial cell injury was simulated by a corneal endothelial cell oxygen-glucose deprivation/reoxygenation (OGD/R) model in vitro. Flow cytometry and evaluation of fluorescent LC3B puncta were used to detect apoptosis and autophagy, and western blotting was used to detect protein expression.

Results: A total of 381 differentially expressed proteins were identified between the two groups. In vitro, 4 ℃ hypothermia significantly reduced apoptosis and promoted autophagy. Apoptosis increased after autophagy was inhibited by 3-Methyladenine (3-MA). Furthermore, adiponectin (ADIPOQ) knockdown inhibited phospho-AMPK and blocked the protective effect of hypothermia on corneal endothelial cells.

Conclusions: We investigated the differential expression of proteins between the hypothermia group and normothermia group by proteomics. Moreover, hypothermia-induced ADIPOQ can reduce apoptosis by promoting AMPK-mediated autophagy.

Keywords: Apoptosis; Autophagy; Hypothermia; Phacoemulsification.

MeSH terms

  • AMP-Activated Protein Kinases / metabolism
  • Adiponectin* / metabolism
  • Animals
  • Apoptosis*
  • Autophagy*
  • Blotting, Western
  • Cells, Cultured
  • Disease Models, Animal*
  • Endothelium, Corneal* / metabolism
  • Endothelium, Corneal* / pathology
  • Flow Cytometry
  • Hypothermia, Induced* / methods
  • Male
  • Phacoemulsification*
  • Proteomics* / methods
  • Rabbits

Substances

  • Adiponectin
  • AMP-Activated Protein Kinases