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Diabetologia

, Volume 62, Issue 12, pp 2365–2374 | Cite as

Neutrophil elastase contributes to the pathological vascular permeability characteristic of diabetic retinopathy

  • Haitao Liu
  • Emma M. Lessieur
  • Aicha Saadane
  • Sarah I. Lindstrom
  • Patricia R. Taylor
  • Timothy S. KernEmail author
Article

Abstract

Aims/hypothesis

Levels of neutrophil elastase, a serine protease secreted by neutrophils, are elevated in diabetes. The purpose of this study was to determine whether neutrophil elastase (NE) contributes to the diabetes-induced increase in retinal vascular permeability in mice with streptozotocin-induced diabetes, and, if so, to investigate the potential role of IL-17 in this process.

Methods

In vivo, diabetes was induced in neutrophil elastase-deficient (Elane−/−), Il-17a−/− and wild-type mice. After 8 months of diabetes, Elane−/− mice and wild-type age-matched control mice were injected with FITC-BSA. Fluorescence microscopy was used to assess leakage of FITC-BSA from the retinal vasculature into the neural retina. The level of NE in Il-17a−/− diabetic retina and sera were determined by ELISA. In vitro, the effect of NE on the permeability and viability of human retinal endothelial cells and the expression of junction proteins and adhesion molecules were studied.

Results

Eight months of diabetes resulted in increased retinal vascular permeability and levels of NE in retina and plasma of wild-type animals. All of these abnormalities were significantly inhibited in mice lacking the elastase. The diabetes-induced increase in NE was inhibited in mice lacking IL-17. In vitro, NE increased retinal endothelial cell permeability, which was partially inhibited by a myeloid differentiation primary response 88 (MyD88) inhibitor, NF-κB inhibitor, and protease-activated receptor (PAR)2 inhibitor. NE degraded vascular endothelial-cadherin (VE-cadherin) in a concentration-dependent manner.

Conclusions/interpretation

IL-17 regulates NE expression in diabetes. NE contributes to vascular leakage in diabetic retinopathy, partially through activation of MyD88, NF-κB and PAR2 and degradation of VE-cadherin.

Keywords

Diabetic retinopathy Elane IL-17 Neutrophil elastase Vascular permeability 

Abbreviations

hREC

Human retinal endothelial cell

ICAM-1

Intercellular adhesion molecule 1

INL

Inner nuclear layer

IPL

Inner plexiform layer

MyD88

Myeloid differentiation primary response 88

NE

Neutrophil elastase

ONL

Outer nuclear layer

OPL

Outer plexiform layer

PAR

Protease-activated receptor

SD-OCT

Spectral-domain optical coherence tomography

TLR4

Toll-like receptor 4

VE-cadherin

Vascular endothelial cadherin

WT

Wild-type

ZO-1

Zona occludens 1

Notes

Acknowledgements

The authors thank D. A. Antonetti and X. Liu (University of Michigan, Ann Arbor, MI, USA) for the retinal whole mount immunostaining. Chieh Allen Lee, Katie Franke and Heather Butler (Case Western Reserve University, Cleveland, OH, USA) who maintained the mouse colonies. Dawn Smith (Case Western Reserve University) who maintained the hRECs.

Contribution statement

HL performed molecular analyses and wrote the manuscript. EML analysed data and wrote the manuscript. AS and PRT acquired data, were involved in the analysis and interpretation of data and reviewed the manuscript. SIL performed ELISA, was involved in the analysis and interpretation of data and edited the manuscript. TSK designed experiments, acquired data and reviewed/edited the manuscript. All the authors approved the final version of the manuscript to be published. TSK is the guarantor of this work.

Funding

This work was supported by NIH grants RO1 EY022938 and R24 EY024864 (to TSK), and grants BX003604 (to TSK) and BX003403 (to PRT) from the Department of Veterans Affairs, and core grant P30 EY011373 to Case Western Reserve University. 81900884 (to HL) from The First Affiliated Hospital of Dalian Medical University. This research received no specific grant from any funding agency in commercial or not-for-profit sectors.

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2019_4998_MOESM1_ESM.pdf (121 kb)
ESM Figure (PDF 121 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Haitao Liu
    • 1
    • 2
  • Emma M. Lessieur
    • 3
  • Aicha Saadane
    • 3
  • Sarah I. Lindstrom
    • 4
  • Patricia R. Taylor
    • 4
    • 5
  • Timothy S. Kern
    • 3
    • 5
    • 6
    Email author
  1. 1.Department of Biology, School of MedicineCase Western Reserve UniversityClevelandUSA
  2. 2.Department of OphthalmologyThe First Affiliated Hospital of Dalian Medical UniversityDalianPeople’s Republic of China
  3. 3.Center for Translational Vision Research, Department of Ophthalmology, Gavin Herbert Eye Institute, School of MedicineUniversity of California-IrvineIrvineUSA
  4. 4.Department of Ophthalmology and Visual SciencesCase Western Reserve UniversityClevelandUSA
  5. 5.Veterans Administration Medical Center Research Service 151ClevelandUSA
  6. 6.Veterans Administration Medical Center Research ServiceLong BeachUSA

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