Antisense to protein kinase C-alpha and p47phox attenuates the pro-inflammatory effects of human C-reactive protein in macrophages of biobreeding diabetic rats

Diab Vasc Dis Res. 2012 Oct;9(4):315-9. doi: 10.1177/1479164112452165. Epub 2012 Jul 16.

Abstract

Objective: Type 1 diabetes mellitus (T1DM) is a pro-inflammatory state characterized by high C-reactive protein (CRP) levels. Previously, we showed that CRP accentuated a macrophage (MO) activity in spontaneously diabetic biobreeding (BB) rats and increased the MO activity of protein kinase C-alpha (PKC-α) and p47phox. In this report, we tested the effects of molecular inhibition of CRP effects on MO activity using antisense oligonucleotide (ASO) to both PKC-α and p47phox.

Methods: Prior to administration of human C-reactive protein (hCRP) daily for 3 days, ASO or scrambled ASO to either PKC-α or p47phox was also delivered for 3 days and after killing on day 4, peritoneal MOs were isolated.

Results: The increase in the levels of superoxide anion, interleukin (IL)-1, monocyte chemoattractant protein-1 (MCP-1), tumour necrosis factor-alpha (TNF-α) and IL-6 release in MOs with hCRP compared to human albumin was significantly attenuated by antisense to either PKC-α and p47phox (p < 0.01 vs. scrambled ASO; n = 5 per group).

Conclusion: Our novel data suggest that antisense to either PKC-α or p47phox attenuates the pro-inflammatory effects of human CRP on MOs in diabetic rats.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • C-Reactive Protein / administration & dosage
  • C-Reactive Protein / metabolism*
  • Chemokine CCL2 / metabolism
  • Diabetes Mellitus, Type 1 / enzymology
  • Diabetes Mellitus, Type 1 / genetics
  • Diabetes Mellitus, Type 1 / immunology
  • Diabetes Mellitus, Type 1 / therapy*
  • Disease Models, Animal
  • Genetic Therapy / methods*
  • Humans
  • Inflammation Mediators / administration & dosage
  • Inflammation Mediators / metabolism*
  • Injections, Intraperitoneal
  • Interleukin-1 / metabolism
  • Interleukin-6 / metabolism
  • Macrophages, Peritoneal / enzymology*
  • Macrophages, Peritoneal / immunology
  • Male
  • NADPH Oxidases / genetics
  • NADPH Oxidases / metabolism*
  • Oligoribonucleotides, Antisense / administration & dosage*
  • Protein Kinase C-alpha / genetics
  • Protein Kinase C-alpha / metabolism*
  • Rats
  • Superoxides / metabolism
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Ccl2 protein, rat
  • Chemokine CCL2
  • Inflammation Mediators
  • Interleukin-1
  • Interleukin-6
  • Oligoribonucleotides, Antisense
  • Tumor Necrosis Factor-alpha
  • Superoxides
  • C-Reactive Protein
  • NADPH Oxidases
  • neutrophil cytosolic factor 1
  • Protein Kinase C-alpha