Limits...
Prednisolone as preservation additive prevents from ischemia reperfusion injury in a rat model of orthotopic lung transplantation.

Paulus P, Holfeld J, Urbschat A, Mutlak H, Ockelmann PA, Tacke S, Zacharowski K, Reissig C, Stay D, Scheller B - PLoS ONE (2013)

Bottom Line: Hypoxia induced vasoactive cytokines such as VEGF were reduced.Together with this, prednisolone treated animals displayed significantly reduced lung protein levels of neutrophil chemoattractants like CINC-1, CINC-2α/β and LIX and upregulated tissue inhibitor of matrix metalloproteinase (TIMP)-1.Additionally, prednisolone preconditioning might also lead to macrophage polarization as a beneficial long-term effect.

View Article: PubMed Central - PubMed

Affiliation: Clinic of Anesthesiology, Intensive Care Medicine and Pain Therapy, Goethe-University Hospital Frankfurt, Frankfurt am Main, Germany. Patrick.Paulus@kgu.de

ABSTRACT
The lung is, more than other solid organs, susceptible for ischemia reperfusion injury after orthotopic transplantation. Corticosteroids are known to potently suppress pro-inflammatory processes when given in the post-operative setting or during rejection episodes. Whereas their use has been approved for these clinical indications, there is no study investigating its potential as a preservation additive in preventing vascular damage already in the phase of ischemia. To investigate these effects we performed orthotopic lung transplantations (LTX) in the rat. Prednisolone was either added to the perfusion solution for lung preservation or omitted and rats were followed for 48 hours after LTX. Prednisolone preconditioning significantly increased survival and diminished reperfusion edema. Hypoxia induced vasoactive cytokines such as VEGF were reduced. Markers of leukocyte invasiveness like matrix metalloprotease (MMP)-2, or common pro-inflammatory molecules like the CXCR4 receptor or the chemokine (C-C motif) ligand (CCL)-2 were downregulated by prednisolone. Neutrophil recruitment to the grafts was only increased in Perfadex treated lungs. Together with this, prednisolone treated animals displayed significantly reduced lung protein levels of neutrophil chemoattractants like CINC-1, CINC-2α/β and LIX and upregulated tissue inhibitor of matrix metalloproteinase (TIMP)-1. Interestingly, lung macrophage invasion was increased in both, Perfadex and prednisolone treated grafts, as measured by MMP-12 or RM4. Markers of anti-inflammatory macrophage transdifferentiation like MRC-1, IL-13, IL-4 and CD163, significantly correlated with prednisolone treatment. These observations lead to the conclusion that prednisolone as an additive to the perfusion solution protects from hypoxia triggered danger signals already in the phase of ischemia and thus reduces graft edema in the phase of reperfusion. Additionally, prednisolone preconditioning might also lead to macrophage polarization as a beneficial long-term effect.

Show MeSH

Related in: MedlinePlus

Prednisolone induces M2 macrophage polarization.(A) The gene expressions of the anti-inflammatory cytokine IL-13 and of a marker for M2 polarization, MRC-1, were analyzed by RT-PCR. (B) Selective immunohistochemical CD163 staining (brown cells) and counting of the cells (graph). N=3 fields per rat have been evaluated. Significance level was set to P<0.05. P<0.01: **, § § and P<0.001: § § §. * = significantly different from shams and § = significantly different from Perfadex-group. RT-PCR experiments were performed in triplicates and means are expressed as % of Perfadex group (Perfadex group has been set at 100%). ANOVA followed by Bonferroni’s post-hoc multiple comparisons test.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3756949&req=5

pone-0073298-g007: Prednisolone induces M2 macrophage polarization.(A) The gene expressions of the anti-inflammatory cytokine IL-13 and of a marker for M2 polarization, MRC-1, were analyzed by RT-PCR. (B) Selective immunohistochemical CD163 staining (brown cells) and counting of the cells (graph). N=3 fields per rat have been evaluated. Significance level was set to P<0.05. P<0.01: **, § § and P<0.001: § § §. * = significantly different from shams and § = significantly different from Perfadex-group. RT-PCR experiments were performed in triplicates and means are expressed as % of Perfadex group (Perfadex group has been set at 100%). ANOVA followed by Bonferroni’s post-hoc multiple comparisons test.

Mentions: Tissue protein expression of tumor necrosis factor (TNF)-α was significantly upregulated in Perfadex-treated lungs (362.0 ± 24.42 mean pixel density) when compared to prednisolone-treated animals (202.5 ± 15.75 mean pixel density) or shams (192.2 ± 14.94 mean pixel density; P<0.05). Tissue protein expression of chemokine (C-X-C motif) ligand (CXCL) 7 was significantly upregulated in Perfadex-treated lungs (4025 ± 163.0 mean pixel density) when compared to prednisolone-treated animals (2794 ± 178.5 mean pixel density; P<0.05) or shams (2192 ± 63.88 mean pixel density; P<0.01). Tissue protein expression of macrophage inflammatory protein (MIP)-1α was significantly upregulated in Perfadex-treated lungs (898.0 ± 49.28 mean pixel density) when compared to prednisolone-treated animals (528.0 ± 16.41 mean pixel density; P<0.05) or shams (321 ± 19.35 mean pixel density; P<0.01). Tissue protein expression of interleukin-1 receptor antagonist (IL-1ra) was significantly downregulated in Perfadex-treated lungs (993.6 ± 0.44 mean pixel density) when compared to prednisolone-treated animals (3582 ± 108.4 mean pixel density; P<0.01) or shams (2972 ± 161.1 mean pixel density; P<0.01). Tissue protein expression of regulated on activation, normal T cell expressed and secreted (RANTES) was significantly downregulated in Perfadex-treated lungs (1527 ± 149.0 mean pixel density) compared to prednisolone-treated animals (2950 ± 139.0 mean pixel density; P<0.05) but not to shams (2248 ± 234.8 mean pixel density; n.s.). Finally, tissue protein expression of the anti-inflammatory IL-4 was significantly upregulated in prednisolone-treated lungs (2672 ± 96.09 mean pixel density) when compared to Perfadex-treated animals (923.1 ± 50.30 mean pixel density; P<0.001) or shams (662.1 ± 11.89 mean pixel density; P<0.001) (Figure 7A).


Prednisolone as preservation additive prevents from ischemia reperfusion injury in a rat model of orthotopic lung transplantation.

Paulus P, Holfeld J, Urbschat A, Mutlak H, Ockelmann PA, Tacke S, Zacharowski K, Reissig C, Stay D, Scheller B - PLoS ONE (2013)

Prednisolone induces M2 macrophage polarization.(A) The gene expressions of the anti-inflammatory cytokine IL-13 and of a marker for M2 polarization, MRC-1, were analyzed by RT-PCR. (B) Selective immunohistochemical CD163 staining (brown cells) and counting of the cells (graph). N=3 fields per rat have been evaluated. Significance level was set to P<0.05. P<0.01: **, § § and P<0.001: § § §. * = significantly different from shams and § = significantly different from Perfadex-group. RT-PCR experiments were performed in triplicates and means are expressed as % of Perfadex group (Perfadex group has been set at 100%). ANOVA followed by Bonferroni’s post-hoc multiple comparisons test.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3756949&req=5

pone-0073298-g007: Prednisolone induces M2 macrophage polarization.(A) The gene expressions of the anti-inflammatory cytokine IL-13 and of a marker for M2 polarization, MRC-1, were analyzed by RT-PCR. (B) Selective immunohistochemical CD163 staining (brown cells) and counting of the cells (graph). N=3 fields per rat have been evaluated. Significance level was set to P<0.05. P<0.01: **, § § and P<0.001: § § §. * = significantly different from shams and § = significantly different from Perfadex-group. RT-PCR experiments were performed in triplicates and means are expressed as % of Perfadex group (Perfadex group has been set at 100%). ANOVA followed by Bonferroni’s post-hoc multiple comparisons test.
Mentions: Tissue protein expression of tumor necrosis factor (TNF)-α was significantly upregulated in Perfadex-treated lungs (362.0 ± 24.42 mean pixel density) when compared to prednisolone-treated animals (202.5 ± 15.75 mean pixel density) or shams (192.2 ± 14.94 mean pixel density; P<0.05). Tissue protein expression of chemokine (C-X-C motif) ligand (CXCL) 7 was significantly upregulated in Perfadex-treated lungs (4025 ± 163.0 mean pixel density) when compared to prednisolone-treated animals (2794 ± 178.5 mean pixel density; P<0.05) or shams (2192 ± 63.88 mean pixel density; P<0.01). Tissue protein expression of macrophage inflammatory protein (MIP)-1α was significantly upregulated in Perfadex-treated lungs (898.0 ± 49.28 mean pixel density) when compared to prednisolone-treated animals (528.0 ± 16.41 mean pixel density; P<0.05) or shams (321 ± 19.35 mean pixel density; P<0.01). Tissue protein expression of interleukin-1 receptor antagonist (IL-1ra) was significantly downregulated in Perfadex-treated lungs (993.6 ± 0.44 mean pixel density) when compared to prednisolone-treated animals (3582 ± 108.4 mean pixel density; P<0.01) or shams (2972 ± 161.1 mean pixel density; P<0.01). Tissue protein expression of regulated on activation, normal T cell expressed and secreted (RANTES) was significantly downregulated in Perfadex-treated lungs (1527 ± 149.0 mean pixel density) compared to prednisolone-treated animals (2950 ± 139.0 mean pixel density; P<0.05) but not to shams (2248 ± 234.8 mean pixel density; n.s.). Finally, tissue protein expression of the anti-inflammatory IL-4 was significantly upregulated in prednisolone-treated lungs (2672 ± 96.09 mean pixel density) when compared to Perfadex-treated animals (923.1 ± 50.30 mean pixel density; P<0.001) or shams (662.1 ± 11.89 mean pixel density; P<0.001) (Figure 7A).

Bottom Line: Hypoxia induced vasoactive cytokines such as VEGF were reduced.Together with this, prednisolone treated animals displayed significantly reduced lung protein levels of neutrophil chemoattractants like CINC-1, CINC-2α/β and LIX and upregulated tissue inhibitor of matrix metalloproteinase (TIMP)-1.Additionally, prednisolone preconditioning might also lead to macrophage polarization as a beneficial long-term effect.

View Article: PubMed Central - PubMed

Affiliation: Clinic of Anesthesiology, Intensive Care Medicine and Pain Therapy, Goethe-University Hospital Frankfurt, Frankfurt am Main, Germany. Patrick.Paulus@kgu.de

ABSTRACT
The lung is, more than other solid organs, susceptible for ischemia reperfusion injury after orthotopic transplantation. Corticosteroids are known to potently suppress pro-inflammatory processes when given in the post-operative setting or during rejection episodes. Whereas their use has been approved for these clinical indications, there is no study investigating its potential as a preservation additive in preventing vascular damage already in the phase of ischemia. To investigate these effects we performed orthotopic lung transplantations (LTX) in the rat. Prednisolone was either added to the perfusion solution for lung preservation or omitted and rats were followed for 48 hours after LTX. Prednisolone preconditioning significantly increased survival and diminished reperfusion edema. Hypoxia induced vasoactive cytokines such as VEGF were reduced. Markers of leukocyte invasiveness like matrix metalloprotease (MMP)-2, or common pro-inflammatory molecules like the CXCR4 receptor or the chemokine (C-C motif) ligand (CCL)-2 were downregulated by prednisolone. Neutrophil recruitment to the grafts was only increased in Perfadex treated lungs. Together with this, prednisolone treated animals displayed significantly reduced lung protein levels of neutrophil chemoattractants like CINC-1, CINC-2α/β and LIX and upregulated tissue inhibitor of matrix metalloproteinase (TIMP)-1. Interestingly, lung macrophage invasion was increased in both, Perfadex and prednisolone treated grafts, as measured by MMP-12 or RM4. Markers of anti-inflammatory macrophage transdifferentiation like MRC-1, IL-13, IL-4 and CD163, significantly correlated with prednisolone treatment. These observations lead to the conclusion that prednisolone as an additive to the perfusion solution protects from hypoxia triggered danger signals already in the phase of ischemia and thus reduces graft edema in the phase of reperfusion. Additionally, prednisolone preconditioning might also lead to macrophage polarization as a beneficial long-term effect.

Show MeSH
Related in: MedlinePlus