Loss of pleckstrin homology domain and leucine-rich repeat protein phosphatase 2 has protective effects on high glucose-injured retinal ganglion cells via the effect on the Akt-GSK-3β-Nrf2 pathway

Inflamm Res. 2023 Mar;72(3):373-385. doi: 10.1007/s00011-022-01680-1. Epub 2022 Dec 23.

Abstract

Objective: Pleckstrin homology domain and leucine-rich repeat protein phosphatase 2 (PHLPP2) is linked to various pathological states. However, whether PHLPP2 mediates diabetic retinopathy is unaddressed. This work explored the biological function of PHLPP2 in modulating high glucose (HG)-elicited damage of retinal ganglion cells (RGCs), an in vitro model for studying diabetic retinopathy.

Methods: Mouse RGCs were treated with HG to establish a cell model. PHLPP2 was silenced by transfecting specific shRNAs targeting PHLPP2. RT-qPCR, immunoblotting, CCK-8 assay, flow cytometry, TUNEL assay, and ELISA were carried out.

Results: Significant increases in PHLPP2 levels were observed in cultured RGCs exposed to HG. The severe damages evoked by HG to RGCs were remarkably weakened in PHLPP2-silenced RGCs, including improved cell survival, attenuated cell apoptosis, repressed oxidative stress, and prohibited proinflammatory response. The silencing of PHLPP2 strengthened the activation of Nrf2 in HG-treated RGCs via modulation of the Akt-GSK-3β axis. Interruption of the Akt-GSK-3β axis reversed PHLPP2-silencing-elicited Nrf2 activation. The protective effects of PHLPP2 silencing on HG-induced injury of RGCs were diminished by Nrf2 inhibition.

Conclusions: The loss of PHLPP2 was beneficial for HG-injured RGCs through the effect on the Akt-GSK-3β-Nrf2 pathway. This work suggests a possible role of PHLPP2 in diabetic retinopathy.

Keywords: Diabetic retinopathy; Inflammation; Nrf2; Oxidative stress; PHLPP2; Retinal ganglion cells.

MeSH terms

  • Animals
  • Apoptosis
  • Diabetic Retinopathy* / genetics
  • Glucose / pharmacology
  • Glycogen Synthase Kinase 3 beta / genetics
  • Glycogen Synthase Kinase 3 beta / metabolism
  • Leucine-Rich Repeat Proteins
  • Mice
  • NF-E2-Related Factor 2 / genetics
  • NF-E2-Related Factor 2 / metabolism
  • Oxidative Stress
  • Pleckstrin Homology Domains
  • Protein Phosphatase 2 / metabolism
  • Protein Phosphatase 2 / pharmacology
  • Proto-Oncogene Proteins c-akt* / metabolism
  • Retinal Ganglion Cells / metabolism
  • Signal Transduction

Substances

  • Proto-Oncogene Proteins c-akt
  • NF-E2-Related Factor 2
  • Protein Phosphatase 2
  • Glycogen Synthase Kinase 3 beta
  • Leucine-Rich Repeat Proteins
  • Glucose