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Analysis of <t>cytokines,</t> interleukins and endothelial mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including <t>GM-CSF,</t> <t>MCP-1</t> and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).
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Analysis of <t>cytokines,</t> interleukins and endothelial mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including <t>GM-CSF,</t> <t>MCP-1</t> and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).
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Analysis of <t>cytokines,</t> interleukins and endothelial mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including <t>GM-CSF,</t> <t>MCP-1</t> and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).
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Analysis of <t>cytokines,</t> interleukins and endothelial mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including <t>GM-CSF,</t> <t>MCP-1</t> and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).
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Analysis of <t>cytokines,</t> interleukins and endothelial mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including <t>GM-CSF,</t> <t>MCP-1</t> and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).
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Analysis of cytokines, interleukins and endothelial mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including GM-CSF, MCP-1 and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).

Journal: Redox Biology

Article Title: Fenofibrate attenuates the adverse effects of radiation on endothelial cells through modulation of ROS-NO signalling and inflammation

doi: 10.1016/j.redox.2025.103994

Figure Lengend Snippet: Analysis of cytokines, interleukins and endothelial mesenchymal transition markers. qPCR analysis of inflammation markers 2d (A) and 7d (B), inflammatory regulators 2d (E) and 7d (F) and EndMT markers 2d (G) and 7d (H) were performed in irradiated HCAECs with and without fenofibrate (Feno). The release of different cytokines including GM-CSF, MCP-1 and IL-6 which were quantified using flow cytometry after 2d (C) and 7d (D). The error bars represent the standard deviation (±SD) (Two-way ANOVA, Tukey's multiple comparisons test; ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗ p ≤ 0.0001; n = 3).

Article Snippet: Extracellular release of inflammatory cytokines (e.g., M-CSF, Granzyme B, IFN-γ, IL-2, IL-4, IL-6, IL-10, IL-17A, IL-21, MCP-1 (CCL2), Perforin and TNF-α) in cell culture supernatants were quantitatively measured based on a fluorescent bead-based system using MACSPlex Cytokine Kits (#130-125-800; Miltenyi Biotec, Germany) according to the manufacturer's protocol.

Techniques: Irradiation, Flow Cytometry, Standard Deviation

Proposed mechanism of action of fenofibrate in irradiated HCAECs in-vitro . Irradiation reduced NO signalling via inactivation of the PI3K–AKT–eNOS pathway, whereas fenofibrate reactivated this pathway and restored NO production (A). Consistent with this, irradiation increased ROS generation, NOX activity, MDA levels, and 3-NT levels, while fenofibrate mitigated this effect (B). Irradiation also triggered an inflammatory response, which was counteracted by fenofibrate (C). The released cytokines contribute to inflammation, changes in cytoskeleton organisation and initiation of EndMT, and fenofibrate effectively attenuated the processes (D). Alterations in the pathways described above play a crucial role in the remodelling of vascular endothelial cells involved in the initiation and progression of atherosclerosis. Fenofibrate acts to reduce or restore the effects of irradiation on these pathways (E–F). Solid lines indicate correlations validated in this study, while dashed lines indicate unvalidated correlations.

Journal: Redox Biology

Article Title: Fenofibrate attenuates the adverse effects of radiation on endothelial cells through modulation of ROS-NO signalling and inflammation

doi: 10.1016/j.redox.2025.103994

Figure Lengend Snippet: Proposed mechanism of action of fenofibrate in irradiated HCAECs in-vitro . Irradiation reduced NO signalling via inactivation of the PI3K–AKT–eNOS pathway, whereas fenofibrate reactivated this pathway and restored NO production (A). Consistent with this, irradiation increased ROS generation, NOX activity, MDA levels, and 3-NT levels, while fenofibrate mitigated this effect (B). Irradiation also triggered an inflammatory response, which was counteracted by fenofibrate (C). The released cytokines contribute to inflammation, changes in cytoskeleton organisation and initiation of EndMT, and fenofibrate effectively attenuated the processes (D). Alterations in the pathways described above play a crucial role in the remodelling of vascular endothelial cells involved in the initiation and progression of atherosclerosis. Fenofibrate acts to reduce or restore the effects of irradiation on these pathways (E–F). Solid lines indicate correlations validated in this study, while dashed lines indicate unvalidated correlations.

Article Snippet: Extracellular release of inflammatory cytokines (e.g., M-CSF, Granzyme B, IFN-γ, IL-2, IL-4, IL-6, IL-10, IL-17A, IL-21, MCP-1 (CCL2), Perforin and TNF-α) in cell culture supernatants were quantitatively measured based on a fluorescent bead-based system using MACSPlex Cytokine Kits (#130-125-800; Miltenyi Biotec, Germany) according to the manufacturer's protocol.

Techniques: Irradiation, In Vitro, Activity Assay