Overproduction of phosphoprotein enriched in diabetes (PED) induces mesangial expansion and upregulates protein kinase C-beta activity and TGF-beta1 expression

Diabetologia. 2009 Dec;52(12):2642-52. doi: 10.1007/s00125-009-1528-z. Epub 2009 Sep 30.

Abstract

Aims/hypothesis: Overproduction of phosphoprotein enriched in diabetes (PED, also known as phosphoprotein enriched in astrocytes-15 [PEA-15]) is a common feature of type 2 diabetes and impairs insulin action in cultured cells and in mice. Nevertheless, the potential role of PED in diabetic complications is still unknown.

Methods: We studied the effect of PED overproduction and depletion on kidney function in animal and cellular models.

Results: Transgenic mice overexpressing PED (PEDTg) featured age-dependent increases of plasma creatinine levels and urinary volume, accompanied by expansion of the mesangial area, compared with wild-type littermates. Serum and kidney levels of TGF-beta1 were also higher in 6- and 9-month-old PEDTg. Overexpression of PED in human kidney 2 cells significantly increased TGF-beta1 levels, SMAD family members (SMAD)2/3 phosphorylation and fibronectin production. Opposite results were obtained following genetic silencing of PED in human kidney 2 cells by antisense oligonucleotides. Inhibition of phospholipase D and protein kinase C-beta by 2-butanol and LY373196 respectively reduced TGF-beta1, SMAD2/3 phosphorylation and fibronectin production. Moreover, inhibition of TGF-beta1 receptor activity and SMAD2/3 production by SB431542 and antisense oligonucleotides respectively reduced fibronectin secretion by about 50%. TGF-beta1 circulating levels were significantly reduced in Ped knockout mice and positively correlated with PED content in peripheral blood leucocytes of type 2 diabetic patients.

Conclusions/interpretation: These data indicate that PED regulates fibronectin production via phospholipase D/protein kinase C-beta and TGF-beta1/SMAD pathways in kidney cells. Raised PED levels may therefore contribute to the abnormal accumulation of extracellular matrix and renal dysfunction in diabetes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics
  • Animals
  • Astrocytes / metabolism
  • Blood Pressure
  • DNA Primers
  • Diabetes Mellitus, Type 1 / complications
  • Diabetes Mellitus, Type 1 / genetics
  • Diabetes Mellitus, Type 1 / physiopathology
  • Diabetes Mellitus, Type 2 / complications
  • Diabetes Mellitus, Type 2 / genetics
  • Diabetes Mellitus, Type 2 / physiopathology
  • Diabetic Nephropathies / epidemiology
  • Fatty Acids, Nonesterified / blood
  • Fibronectins / genetics
  • Gene Expression Regulation
  • Heart Rate
  • Humans
  • Insulin / blood
  • Kidney / physiology
  • Kidney Failure, Chronic / etiology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Transgenic
  • Phenotype
  • Phosphoproteins / biosynthesis
  • Phosphoproteins / genetics
  • Protein Kinase C / genetics*
  • Protein Kinase C beta
  • Reverse Transcriptase Polymerase Chain Reaction
  • Smad2 Protein / genetics
  • Transforming Growth Factor beta1 / genetics*
  • Up-Regulation

Substances

  • Actins
  • DNA Primers
  • Fatty Acids, Nonesterified
  • Fibronectins
  • Insulin
  • Phosphoproteins
  • SMAD2 protein, human
  • Smad2 Protein
  • Transforming Growth Factor beta1
  • Protein Kinase C
  • Protein Kinase C beta