The effect of circulating iron on barrier integrity of primary human endothelial cells
Abstract Iron is hypothesized to be one of the contributors to cardiovascular disease and its levels in the circulation may correlate with cardiovascular risk. The aim of this study is to investigate the mechanisms that underlie the effects of iron on the barrier function of primary human endotheliu...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-10-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-44122-6 |
| _version_ | 1797453305569345536 |
|---|---|
| author | M. C. Madsen F. Podieh M. C. Overboom A. Thijs M. den Heijer P. L. Hordijk |
| author_facet | M. C. Madsen F. Podieh M. C. Overboom A. Thijs M. den Heijer P. L. Hordijk |
| author_sort | M. C. Madsen |
| collection | DOAJ |
| description | Abstract Iron is hypothesized to be one of the contributors to cardiovascular disease and its levels in the circulation may correlate with cardiovascular risk. The aim of this study is to investigate the mechanisms that underlie the effects of iron on the barrier function of primary human endothelium. We used Human Umbilical Vein Endothelial Cells (HUVEC) to investigate the effects of Fe3+ using electric cell-substrate impedance sensing, microscopy, western blot and immunofluorescence microscopy. Exposure to Fe3+ caused EC elongation and upregulation of stress-induced proteins. Analysis of barrier function showed a dose-dependent drop in endothelial integrity, which was accompanied by Reactive Oxygen Species (ROS) production and could partly be prevented by ROS scavengers. Inhibition of contractility by the ROCK inhibitor Y27632, showed even more effective rescue of barrier integrity. Using western blot, we detected an increase in expression of the small GTPase RhoB, an inducer of EC contraction, and a small decrease in VE-cadherin, suggestive for an iron-induced stress response. Co-stimulation by TNFα and iron, used to investigate the role of low-grade inflammation, revealed an additive, negative effect on barrier integrity, concomitant with an upregulation of pro-inflammatory markers ICAM-1 and RhoB. Iron induces a response in HUVEC that leads to endothelial activation and a pro-inflammatory state measured by loss of barrier integrity which can be reversed by ROS scavengers, combined with inhibition of contractility. These data suggest that ROS-mediated damage of the vascular endothelium could contribute to the increased cardiovascular risk which is associated with elevated levels of circulating iron. |
| first_indexed | 2024-03-09T15:20:54Z |
| format | Article |
| id | doaj.art-d9c946d39f454f2086e1d48f86aceb0e |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-09T15:20:54Z |
| publishDate | 2023-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-d9c946d39f454f2086e1d48f86aceb0e2023-11-26T12:50:58ZengNature PortfolioScientific Reports2045-23222023-10-0113111110.1038/s41598-023-44122-6The effect of circulating iron on barrier integrity of primary human endothelial cellsM. C. Madsen0F. Podieh1M. C. Overboom2A. Thijs3M. den Heijer4P. L. Hordijk5Department of Physiology, Amsterdam UMCDepartment of Physiology, Amsterdam UMCDepartment of Physiology, Amsterdam UMCDepartment of Internal Medicine, Amsterdam UMCDepartment of Internal Medicine, Amsterdam UMCDepartment of Physiology, Amsterdam UMCAbstract Iron is hypothesized to be one of the contributors to cardiovascular disease and its levels in the circulation may correlate with cardiovascular risk. The aim of this study is to investigate the mechanisms that underlie the effects of iron on the barrier function of primary human endothelium. We used Human Umbilical Vein Endothelial Cells (HUVEC) to investigate the effects of Fe3+ using electric cell-substrate impedance sensing, microscopy, western blot and immunofluorescence microscopy. Exposure to Fe3+ caused EC elongation and upregulation of stress-induced proteins. Analysis of barrier function showed a dose-dependent drop in endothelial integrity, which was accompanied by Reactive Oxygen Species (ROS) production and could partly be prevented by ROS scavengers. Inhibition of contractility by the ROCK inhibitor Y27632, showed even more effective rescue of barrier integrity. Using western blot, we detected an increase in expression of the small GTPase RhoB, an inducer of EC contraction, and a small decrease in VE-cadherin, suggestive for an iron-induced stress response. Co-stimulation by TNFα and iron, used to investigate the role of low-grade inflammation, revealed an additive, negative effect on barrier integrity, concomitant with an upregulation of pro-inflammatory markers ICAM-1 and RhoB. Iron induces a response in HUVEC that leads to endothelial activation and a pro-inflammatory state measured by loss of barrier integrity which can be reversed by ROS scavengers, combined with inhibition of contractility. These data suggest that ROS-mediated damage of the vascular endothelium could contribute to the increased cardiovascular risk which is associated with elevated levels of circulating iron.https://doi.org/10.1038/s41598-023-44122-6 |
| spellingShingle | M. C. Madsen F. Podieh M. C. Overboom A. Thijs M. den Heijer P. L. Hordijk The effect of circulating iron on barrier integrity of primary human endothelial cells Scientific Reports |
| title | The effect of circulating iron on barrier integrity of primary human endothelial cells |
| title_full | The effect of circulating iron on barrier integrity of primary human endothelial cells |
| title_fullStr | The effect of circulating iron on barrier integrity of primary human endothelial cells |
| title_full_unstemmed | The effect of circulating iron on barrier integrity of primary human endothelial cells |
| title_short | The effect of circulating iron on barrier integrity of primary human endothelial cells |
| title_sort | effect of circulating iron on barrier integrity of primary human endothelial cells |
| url | https://doi.org/10.1038/s41598-023-44122-6 |
| work_keys_str_mv | AT mcmadsen theeffectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT fpodieh theeffectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT mcoverboom theeffectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT athijs theeffectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT mdenheijer theeffectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT plhordijk theeffectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT mcmadsen effectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT fpodieh effectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT mcoverboom effectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT athijs effectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT mdenheijer effectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells AT plhordijk effectofcirculatingirononbarrierintegrityofprimaryhumanendothelialcells |