June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Exposure to a diabetic-like milieu in vitro alters metabolism in human microvascular endothelial cells
Author Affiliations & Notes
  • David Hughes
    SCHOOL OF MEDICINE, DENTISTRY AND BIOMEDICAL SCIENCES, Queen's University Belfast, Belfast, Belfast, United Kingdom
  • Pietro Bertelli
    SCHOOL OF MEDICINE, DENTISTRY AND BIOMEDICAL SCIENCES, Queen's University Belfast, Belfast, Belfast, United Kingdom
  • Edoardo Pedrini
    SCHOOL OF MEDICINE, DENTISTRY AND BIOMEDICAL SCIENCES, Queen's University Belfast, Belfast, Belfast, United Kingdom
  • Cristina Branco
    SCHOOL OF MEDICINE, DENTISTRY AND BIOMEDICAL SCIENCES, Queen's University Belfast, Belfast, Belfast, United Kingdom
  • Tim Curtis
    SCHOOL OF MEDICINE, DENTISTRY AND BIOMEDICAL SCIENCES, Queen's University Belfast, Belfast, Belfast, United Kingdom
  • Reinhold J. Medina
    SCHOOL OF MEDICINE, DENTISTRY AND BIOMEDICAL SCIENCES, Queen's University Belfast, Belfast, Belfast, United Kingdom
  • Alan Stitt
    SCHOOL OF MEDICINE, DENTISTRY AND BIOMEDICAL SCIENCES, Queen's University Belfast, Belfast, Belfast, United Kingdom
  • Footnotes
    Commercial Relationships   David Hughes, None; Pietro Bertelli, None; Edoardo Pedrini, None; Cristina Branco, None; Tim Curtis, None; Reinhold Medina, None; Alan Stitt, None
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 2252. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      David Hughes, Pietro Bertelli, Edoardo Pedrini, Cristina Branco, Tim Curtis, Reinhold J. Medina, Alan Stitt; Exposure to a diabetic-like milieu in vitro alters metabolism in human microvascular endothelial cells. Invest. Ophthalmol. Vis. Sci. 2021;62(8):2252.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : The retinal capillary endothelium experiences marked fluctuations in hyperglycaemia and ischemia as diabetic retinopathy (DR) progresses. This study has assessed the changes of metabolism and related function in human retinal microvascular endothelial cells (HRMEC) exposed to long term high glucose and hypoxia conditions in vitro

Methods : HRMEC obtained from human donors were grown in to 25 mM L-glucose or 25 mM D-glucose (HLG, HDG) for four weeks and then also exposed to hypoxia (1% O2, 5% CO2 ) for 24 hours. The metabolic flux of both the extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) was assessed using the Seahorse XF analyser. To assess in vitro functionality assays such as tubulogenesis, proliferation, and migration, along with barrier analysis using the xCELLigence system were utilised. Pharmaceutical compounds were also utilised to modulate the responses of key metabolic genes

Results : Compared to controls HDG cells demonstrated reduced ECAR from 24.4 to 16.4 (mpH/min) in normoxia and 44.8 and 35 (mpH/min) in hypoxia (P ≤ 0.01). Furthermore, OCR was reduced in normoxia from 61.4 to 52.9 (pmol/min) (P ≤ 0.05), however in hypoxia HDG showed increased levels in relation to control 25.8 to 17.3 (pmol/min) (P ≤ 0.01) assessed by the Seahorse XF analyser. HDG treated HRMECs also displayed decreased tubulogenic capacity in normoxia which further decreased in hypoxia (P ≤ 0.001). HDG reduced proliferative potential (P ≤ 0.001) and was shown to increase median cellular size (P ≤ 0.001). Migration of cells was also reduced in scratch wound assays and migration and barrier formation were shown to be hindered (P ≤ 0.01). HDG cells demonstrated reduced ECAR from 24.4 to 16.4 (mpH/min) in normoxia and 44.8 and 35 (mpH/min) in hypoxia (P ≤ 0.01). Furthermore, OCR was reduced in normoxia from 61.4 to 52.9 (pmol/min) (P ≤ 0.05), however in hypoxia HDG showed increased levels in relation to control 25.8 to 17.3 (pmol/min) (P ≤ 0.01) assessed by the Seahorse XF analyser.

Conclusions : HRMEC exposed to long term exposure of HDG and then hypoxia appear to have severely affected functionality and altered metabolism. As endothelial cells normally have a remarkable dependency on glycolytic metabolism which allows for growth and repair, these alterations are relevant to vascular pathology occurring in DR.

This is a 2021 ARVO Annual Meeting abstract.

×
×

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.

×