Hyperosmolar stress induces monocyte chemoattractant protein 1 expression in retinal pigmented epithelial arising retinal pigmented epithelial 19 cells.
| Title: | Hyperosmolar stress induces monocyte chemoattractant protein 1 expression in retinal pigmented epithelial arising retinal pigmented epithelial 19 cells. |
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| Authors: | Hamou MO; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Department of Ophthalmology, CHU St Pierre and Brugmann, Université Libre de Bruxelles, Brussels, Belgium.; Masset M; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Department of Ophthalmology, CHU St Pierre and Brugmann, Université Libre de Bruxelles, Brussels, Belgium.; Weber C; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Department of Ophthalmology, CHU St Pierre and Brugmann, Université Libre de Bruxelles, Brussels, Belgium.; Scifo L; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Department of Ophthalmology, CHU St Pierre and Brugmann, Université Libre de Bruxelles, Brussels, Belgium.; Libert S; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Department of Ophthalmology, Erasme Hospital, Hôpital Universitaire de Bruxelles, Université Libre de Bruxelles, Brussels, Belgium.; Bryla A; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Gregoire F; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Delforge V; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Bolaky N; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Datlibagi A; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Department of Ophthalmology, Erasme Hospital, Hôpital Universitaire de Bruxelles, Université Libre de Bruxelles, Brussels, Belgium.; Springael JY; I.R.I.B.H.M., Université Libre de Bruxelles, Brussels, Belgium.; Perret J; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium.; Willermain F; Department of Ophthalmology, CHU St Pierre and Brugmann, Université Libre de Bruxelles, Brussels, Belgium.; I.R.I.B.H.M., Université Libre de Bruxelles, Brussels, Belgium.; Delporte C; Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium. |
| Source: | Molecular vision [Mol Vis] 2025 Oct 05; Vol. 31, pp. 380-394. Date of Electronic Publication: 2025 Oct 05 (Print Publication: 2025). |
| Publication Type: | Journal Article; Research Support, Non-U.S. Gov't |
| Language: | English |
| Journal Info: | Publisher: Molecular Vision Country of Publication: United States NLM ID: 9605351 Publication Model: eCollection Cited Medium: Internet ISSN: 1090-0535 (Electronic) Linking ISSN: 10900535 NLM ISO Abbreviation: Mol Vis Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: Atlanta Ga : Molecular Vision, 1995- |
| MeSH Terms: | Chemokine CCL2*/genetics ; Chemokine CCL2*/metabolism ; Retinal Pigment Epithelium*/metabolism ; Retinal Pigment Epithelium*/cytology ; Epithelial Cells*/metabolism ; Epithelial Cells*/drug effects ; Osmotic Pressure*; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Transcription Factors/metabolism ; Transcription Factors/genetics ; NFATC Transcription Factors/metabolism ; NFATC Transcription Factors/genetics ; p38 Mitogen-Activated Protein Kinases/metabolism ; p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors ; Protein Kinase C/metabolism ; Protein Kinase C/antagonists & inhibitors ; Humans ; Cell Line ; Gene Expression Regulation ; Animals ; Signal Transduction |
| Abstract: | Purpose: Diabetes is a chronic inflammatory disease that may damage the blood-retinal barrier, leading to diabetic retinopathy (DR). Blood-retinal barrier rupture may subject the retinal pigmented epithelial cells to a hyperosmolar stress (HOS), activating the transcription factor nuclear factor of activated T cells 5 (NFAT5). In addition, inflammatory cytokines, such as monocyte chemoattractant protein 1 (MCP-1/CCL2), play a crucial role in DR. The aims of our study were to determine whether HOS induces MCP-1 levels in arising retinal pigmented epithelial 19 (ARPE-19) cells and to decipher the responsible intracellular cascade involved in such stimulation.; Methods: ARPE-19 cells or ARPE-19 cells transfected with dominant negative NFAT5 plasmid or NFAT5 short hairpin RNA plasmids were preincubated or not for 1 h in the absence or presence of a protein kinase or transcription factor inhibitor and then incubated for 8 h with iso-osmolar or hyperosmolar medium in the absence or presence of inhibitor. NFAT5 reporter gene activity was quantified by luminescence. MCP-1 messenger RNA (mRNA) and protein levels were determined by quantitative real-time PCR and enzyme-linked immunosorbent assay, respectively. Biologically active MCP-1 was assessed by a calcium mobilization assay performed using Chinese hamster ovary cells expressing or not the MCP-1 receptor and apoaequorin.; Results: In response to HOS, ARPE-19 cells showed a significant increase in MCP-1 mRNA levels independent of NFAT5 activation. Moreover, the MCP-1 protein secreted by ARPE-19 in response to HOS is biologically active. The use of various inhibitors of protein kinase and transcription factors suggest that the HOS-induced increase in MCP-1 mRNA levels is dependent on a protein kinase C (PKC) and/or a MEK1/2-p38 pathway activating p53, as well as a PKC-p38-PI3K-PDK1-AKT activating hypoxia-inducible factor 1 alpha (HIF1α).; Conclusion: HOS increases the expression of MCP-1 mRNA and protein levels in ARPE-19 cells, and the secreted MCP-1 is biologically active. The HOS-induced increase of MCP-1 mRNA appears to be independent of NFAT5 activation. Despite the activation of NFAT5 upon HOS and the presence of NFAT5 binding sites in the MCP-1 gene promoter, activated NFAT5 may not be sufficient to induce MCP-1 gene transactivation in response to HOS in ARPE-19 cells. The intracellular cascade involved in the HOS-induced increase of MCP-1 mRNA in ARPE-19 cells may consist of a PKC-p38-PI3K-PDK1-AKT-HIF1α axis and/or a MEK1/2-p38-p53 axis.; (Copyright © 2025 Molecular Vision.) |
| Substance Nomenclature: | 0 (Chemokine CCL2); 0 (NFAT5 protein, human); 0 (CCL2 protein, human); 0 (RNA, Messenger); 0 (Transcription Factors); 0 (NFATC Transcription Factors); EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases); EC 2.7.11.13 (Protein Kinase C) |
| Entry Date(s): | Date Created: 20260323 Date Completed: 20260323 Latest Revision: 20260513 |
| Update Code: | 20260513 |
| PubMed Central ID: | PMC13002342 |
| PMID: | 41867363 |
| Database: | MEDLINE |
Journal Article; Research Support, Non-U.S. Gov't