September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Blockade of NLRP3 inflammasome activation in diabetic retina by the ketone metabolite beta-hydroxybutyrate is mediated by GPR109A
Author Affiliations & Notes
  • Ollya Fromal
    Biochemistry and Molecular Biology, Medical College of Georgia at Georgia Regents University, Augusta, Georgia, United States
  • Folami Lamoke
    Biochemistry and Molecular Biology, Medical College of Georgia at Georgia Regents University, Augusta, Georgia, United States
  • Matthew Kaufman
    Biochemistry and Molecular Biology, Medical College of Georgia at Georgia Regents University, Augusta, Georgia, United States
  • Manuela Bartoli
    Ophthalmology & Culver Vision Discovery Institute, Medical College of Georgia at Georgia Regents University, Augusta, Georgia, United States
  • Pamela M Martin
    Biochemistry and Molecular Biology, Medical College of Georgia at Georgia Regents University, Augusta, Georgia, United States
    Ophthalmology & Culver Vision Discovery Institute, Medical College of Georgia at Georgia Regents University, Augusta, Georgia, United States
  • Footnotes
    Commercial Relationships   Ollya Fromal, None; Folami Lamoke, None; Matthew Kaufman, None; Manuela Bartoli, None; Pamela Martin, None
  • Footnotes
    Support  5R01EY022704-02
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 5451. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Ollya Fromal, Folami Lamoke, Matthew Kaufman, Manuela Bartoli, Pamela M Martin; Blockade of NLRP3 inflammasome activation in diabetic retina by the ketone metabolite beta-hydroxybutyrate is mediated by GPR109A. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5451.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : NLRP3 inflammasome pathway activation has emerged as a key source of subclinical inflammation in diabetic retinopathy (DR). The ketone body beta-hydroxybutyrate (β-HB) has been shown to block the NLRP3 inflammasome pathway in chronic inflammatory states, but its utility has not been investigated in diabetic retina. Here, we evaluated whether β-HB negatively regulates NLRP3 activation through GPR109A, a Gi-protein coupled receptor for β-HB, by assessing the effects of β-HB in streptozotocin (STZ)-induced diabetic Gpr109a+/+ (wildtype, WT) and Gpr109a-/- (knockout, KO) mouse retina.

Methods : Diabetes was induced in male, WT and KO mice using STZ (75 mg/kg in 0.01M sodium-citrate buffer, pH 4.5). The GPR109A cognate ligand β-HB was administered daily (40 mg/mouse, i.p.) to diabetic (DB) and age-matched, control non-diabetic (non-DB) WT and KO mice for a period of 2 weeks beginning 16-wks post-diabetes onset. WT and KO, non-DB and DB, animals injected with PBS only served as controls. Multiplex cytokine arrays (R&D systems) and immunofluorescence techniques were used to assess retinal inflammation. Western blotting was used to confirm changes in the expression of key inflammasome-associated cytokines (IL-1 beta and IL-18) and, pro-caspase 1 and cleaved caspase 1, proteins integral to NLRP3 inflammasome activation. Blood-retinal barrier breakdown and increased vascular permeability was assessed by albumin extravasation and FITC-dextran staining and, FITC-ConA perfusion to evaluate leukostasis.

Results : Basal levels of pro-inflammatory proteins and pro-inflammatory cytokines were higher in WT DB and KO non-DB compared to WT non-DB retinas. The increases were exacerbated even further in KO DB eyes. β-HB treatment significantly reduced pro-inflammatory protein expression, pro-inflammatory cytokine levels, and cleaved caspase 1 expression in WT DB mice, but had little to no effect in KO control and KO DB animals. Notably, we also observed accelerated and worsened pathology in KO DB animals, evidenced by increased vascular permeability and leukocyte adhesion.

Conclusions : These data show that β-HB reduces NLRP3-derived inflammation in the diabetic retina through GPR109A, identifying this metabolite and its receptor as a new potential therapeutic drug and target for combating inflammation in DR.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×