Artikel
Synovial fibroblasts from patients with rheumatoid arthritis influence the cell network of endothelial cells via CXCL2
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Autoren
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Frederik Lötfering
- Justus Liebig University Giessen, Department of Rheumatology and Clinical Immunology, Campus Kerckhoff, Bad Nauheim
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Corinna Heck
- Justus Liebig University Giessen, Department of Rheumatology and Clinical Immunology, Campus Kerckhoff, Bad Nauheim
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Daria Kürsammer
- Justus Liebig University Giessen, Department of Rheumatology and Clinical Immunology, Campus Kerckhoff, Bad Nauheim
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Klaus Frommer
- Justus Liebig University Giessen, Department of Rheumatology and Clinical Immunology, Campus Kerckhoff, Bad Nauheim
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Mona Arnold
- Justus Liebig University Giessen, Department of Rheumatology and Clinical Immunology, Campus Kerckhoff, Bad Nauheim
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Stefan Rehart
- Agaplesion Markus Hospital, Department of Orthopedics and Trauma Surgery, Frankfurt
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Ulf Müller-Ladner
- Justus Liebig University Giessen, Department of Rheumatology and Clinical Immunology, Campus Kerckhoff, Bad Nauheim
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Elena Neumann
- Justus Liebig University Giessen, Department of Rheumatology and Clinical Immunology, Campus Kerckhoff, Bad Nauheim
| Veröffentlicht: | 18. September 2024 |
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Gliederung
Text
Introduction: In rheumatoid arthritis (RA), increased vascularization contributes to synovial inflammation. RA synovial fibroblasts (RASF), on the other hand, have an enhancing effect on angiogenesis. The pro-inflammatory chemokine CXCL2 is secreted by various immune cells as well as RASF. CXCL2 promotes the migration of leukocytes to sites of inflammation. In addition, CXCL2 has an enhancing effect on angiogenesis, e.g. in neoplastic diseases. Therefore, the aim was to investigate CXCL2 in the context of RASF-altered angiogenesis.
Methods: 2D tube formation assays were performed by plating Human Umbilical Vein Endothelial Cells (HUVEC) on Matrigel®. First, HUVEC were treated with 25/50/100 ng/ml CXCL2. Then, HUVEC were treated with 50 ng/ml CXCL2 plus 0.015/0.03/0.06 μM of the CXCR2-antagonist SB225002. In addition, RASF were stimulated once or three times with 1 ng/ml IL-1β, then the supernatants were harvested. Subsequently, RASF supernatants ±0.015 µM SB225002 were added to HUVEC. After 4 h of incubation, standardized images of the HUVEC cell network were taken and the cell network area was measured using the Keyence software tool.
Results: All CXCL2 concentrations reduced the HUVEC cell network area compared to control (control=100%). The CXCL2 concentration of 50 ng/ml showed the strongest effect (71%, p=0.02), followed by 25 ng/ml (73%, p=0.04) and 100ng/ml CXCL2 (75%, p=0.08). The addition of SB225002 partially abolished the CXCL2 effect (0.015 µM 93%; 0.03 µM 87%; 0.06 µM 92%). Addition of supernatants of single and triple IL-1β-stimulated RASF reduced the HUVEC cell network area to 78% (p=0.01) and 77% (p=0.04), respectively. The addition of SB225002 partially counteracted the observed effect of the supernatants, resulting in a less prominent reduction of the HUVEC cell network area to 83% (single) and 81% (triple). The supernatants of unstimulated RASF had slightly weaker effects on the HUVEC cell network compared to supernatants of stimulated RASF.
Conclusion: CXCL2 and RASF supernatants containing CXCL2 reduced the HUVEC cell network area in a similar manner, whereas the CXCR2-antagonist SB225002 attenuated the observed effects. Thus, CXCL2 secreted by RASF may have a key influence on the altered neovascularization observed in RA.
