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Pigment epithelium-derived factor is an interleukin-6 antagonist in the RPE: Insight of structure-function relationships
Retinal and choroidal inflammatory lesions increase the levels of the pro-inflammatory cytokine interleukin-6 (IL-6). Pigment epithelium-derived factor (PEDF) has anti-inflammatory properties, but it is not known if it can prevent the production of IL-6 by the retinal pigment epithelium. To investig...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9709256/ https://www.ncbi.nlm.nih.gov/pubmed/36467689 http://dx.doi.org/10.3389/fphys.2022.1045613 |
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author | Bernardo-Colón, Alexandra Lerner, Miriam Becerra, S. Patricia |
author_facet | Bernardo-Colón, Alexandra Lerner, Miriam Becerra, S. Patricia |
author_sort | Bernardo-Colón, Alexandra |
collection | PubMed |
description | Retinal and choroidal inflammatory lesions increase the levels of the pro-inflammatory cytokine interleukin-6 (IL-6). Pigment epithelium-derived factor (PEDF) has anti-inflammatory properties, but it is not known if it can prevent the production of IL-6 by the retinal pigment epithelium. To investigate the anti-inflammatory effects of PEDF in the RPE, we used human ARPE-19 cells stimulated with human recombinant tumor necrosis factor-alpha (TNF-α) to induce overexpression of the IL6 gene. We found that the viability of ARPE-19 cells decreased by 22% with TNF-α at 10 ng/ml, being drastically decreased at ≥50 ng/ml. TNF-α at 5–100 ng/ml elevated the production and secretion of IL-6 protein, as measured by ELISA. To challenge the TNF-α-mediated stimulation of IL-6, we used recombinant human PEDF protein. PEDF at 100 nM recovered the TNF-α-mediated loss of cell viability and repressed IL-6 gene expression as determined by RT-PCR. PEDF at 10–100 nM attenuated the IL-6 protein secretion in a dose dependent fashion (IC50 = 65 nM), being abolished with 100 nM PEDF. To map the region that confers the IL-6 blocking effect to the PEDF polypeptide, we used chemically synthesized peptides designed from its biologically active domains, pro-death 34-mer, and pro-survival 44-mer and 17-mer (H105A), to challenge the IL-6 overproduction. The pro-survival peptides recovered the TNF-α-mediated cell viability loss, and inhibited IL-6 secretion, while the 34-mer did not have an effect, suggesting a role for the pro-survival domain in blocking TNF-α-mediated cell death and IL-6 stimulation. Our findings position PEDF as a novel antagonistic agent of IL-6 production in RPE cells, underscoring its use for the management of retinal disease-related inflammation. |
format | Online Article Text |
id | pubmed-9709256 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-97092562022-12-01 Pigment epithelium-derived factor is an interleukin-6 antagonist in the RPE: Insight of structure-function relationships Bernardo-Colón, Alexandra Lerner, Miriam Becerra, S. Patricia Front Physiol Physiology Retinal and choroidal inflammatory lesions increase the levels of the pro-inflammatory cytokine interleukin-6 (IL-6). Pigment epithelium-derived factor (PEDF) has anti-inflammatory properties, but it is not known if it can prevent the production of IL-6 by the retinal pigment epithelium. To investigate the anti-inflammatory effects of PEDF in the RPE, we used human ARPE-19 cells stimulated with human recombinant tumor necrosis factor-alpha (TNF-α) to induce overexpression of the IL6 gene. We found that the viability of ARPE-19 cells decreased by 22% with TNF-α at 10 ng/ml, being drastically decreased at ≥50 ng/ml. TNF-α at 5–100 ng/ml elevated the production and secretion of IL-6 protein, as measured by ELISA. To challenge the TNF-α-mediated stimulation of IL-6, we used recombinant human PEDF protein. PEDF at 100 nM recovered the TNF-α-mediated loss of cell viability and repressed IL-6 gene expression as determined by RT-PCR. PEDF at 10–100 nM attenuated the IL-6 protein secretion in a dose dependent fashion (IC50 = 65 nM), being abolished with 100 nM PEDF. To map the region that confers the IL-6 blocking effect to the PEDF polypeptide, we used chemically synthesized peptides designed from its biologically active domains, pro-death 34-mer, and pro-survival 44-mer and 17-mer (H105A), to challenge the IL-6 overproduction. The pro-survival peptides recovered the TNF-α-mediated cell viability loss, and inhibited IL-6 secretion, while the 34-mer did not have an effect, suggesting a role for the pro-survival domain in blocking TNF-α-mediated cell death and IL-6 stimulation. Our findings position PEDF as a novel antagonistic agent of IL-6 production in RPE cells, underscoring its use for the management of retinal disease-related inflammation. Frontiers Media S.A. 2022-11-16 /pmc/articles/PMC9709256/ /pubmed/36467689 http://dx.doi.org/10.3389/fphys.2022.1045613 Text en Copyright © 2022 Bernardo-Colón, Lerner and Becerra. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Physiology Bernardo-Colón, Alexandra Lerner, Miriam Becerra, S. Patricia Pigment epithelium-derived factor is an interleukin-6 antagonist in the RPE: Insight of structure-function relationships |
title | Pigment epithelium-derived factor is an interleukin-6 antagonist in the RPE: Insight of structure-function relationships |
title_full | Pigment epithelium-derived factor is an interleukin-6 antagonist in the RPE: Insight of structure-function relationships |
title_fullStr | Pigment epithelium-derived factor is an interleukin-6 antagonist in the RPE: Insight of structure-function relationships |
title_full_unstemmed | Pigment epithelium-derived factor is an interleukin-6 antagonist in the RPE: Insight of structure-function relationships |
title_short | Pigment epithelium-derived factor is an interleukin-6 antagonist in the RPE: Insight of structure-function relationships |
title_sort | pigment epithelium-derived factor is an interleukin-6 antagonist in the rpe: insight of structure-function relationships |
topic | Physiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9709256/ https://www.ncbi.nlm.nih.gov/pubmed/36467689 http://dx.doi.org/10.3389/fphys.2022.1045613 |
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