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Phosphodiesterase-4 inhibition with rolipram attenuates hepatocellular injury in hyperinflammation in vivo and in vitro without influencing inflammation and HO-1 expression.

Wollborn J, Wunder C, Stix J, Neuhaus W, Bruno RR, Baar W, Flemming S, Roewer N, Schlegel N, Schick MA - J Pharmacol Pharmacother (2015 Jan-Mar)

Bottom Line: Untreated LPS-induced rats showed significantly decreased liver microcirculation and increased hepatic cell death, whereas LPS + PD-4-I treatment could improve hepatic volumetric flow and cell death to control level whithout influencing the inflammatory impact.The heme oxygenase 1 (HO-1) pathway did not induce the protective effect of PD-4-I.Intravenous PD-4-I treatment was effective in improving hepatic microcirculation and hepatic integrity, while it had a direct protective effect on HepG2 viability during inflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Anaesthesia and Critical Care, University Hospital Würzburg, Germany.

ABSTRACT

Objective: To investigate the impact of the phophodiesterase-4 inhibition (PD-4-I) with rolipram on hepatic integrity in lipopolysaccharide (LPS) induced hyperinflammation.

Materials and methods: Liver microcirculation in rats was obtained using intravital microscopy. Macrohemodynamic parameters, blood assays, and organs were harvested to determine organ function and injury. Hyperinflammation was induced by LPS and PD-4-I rolipram was administered intravenously one hour after LPS application. Cell viability of HepG2 cells was measured by EZ4U-kit based on the dye XTT. Experiments were carried out assessing the influence of different concentrations of tumor necrosis factor alpha (TNF-α) and LPS with or without PD-4-I.

Results: Untreated LPS-induced rats showed significantly decreased liver microcirculation and increased hepatic cell death, whereas LPS + PD-4-I treatment could improve hepatic volumetric flow and cell death to control level whithout influencing the inflammatory impact. In HepG2 cells TNF-α and LPS significantly reduced cell viability. Coincubation with PD-4-I increased HepG2 viability to control levels. The heme oxygenase 1 (HO-1) pathway did not induce the protective effect of PD-4-I.

Conclusion: Intravenous PD-4-I treatment was effective in improving hepatic microcirculation and hepatic integrity, while it had a direct protective effect on HepG2 viability during inflammation.

No MeSH data available.


Related in: MedlinePlus

HepG2 cell culture: Cell viability of PD-4-I-incubated cells (A) and in presence of 50 μg/ml LPS, showing a compromised viability with 50 ng/ml LPS and a cytoprotective effect of 2.5 μM and 10 μM PD-4-I. (C) Cell viability of different concentrations of TNF-α with and without 5μM PD-4-I, revealing cytoprotective effects of PD-4-I incubation at 50-400 ng/ml. (*vs. 0%, #vs. 50 μg/ml LPS; §vs. corresponding TNF-α-only incubated cells, n = 32-64, P≤ 0.05, mean ± SEM)
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Figure 7: HepG2 cell culture: Cell viability of PD-4-I-incubated cells (A) and in presence of 50 μg/ml LPS, showing a compromised viability with 50 ng/ml LPS and a cytoprotective effect of 2.5 μM and 10 μM PD-4-I. (C) Cell viability of different concentrations of TNF-α with and without 5μM PD-4-I, revealing cytoprotective effects of PD-4-I incubation at 50-400 ng/ml. (*vs. 0%, #vs. 50 μg/ml LPS; §vs. corresponding TNF-α-only incubated cells, n = 32-64, P≤ 0.05, mean ± SEM)

Mentions: Hepatoma cell line, HepG2, cell culture was used as an in vitro model of hepatocytes to test our hypothesis of direct effects of PD-4-I on hepatocytes. First we tested the influence of PD-4-I at different doses on cell viability using the EZ4U-tests. Doses of rolipram at 2.5-10 μM did not affect cell viability while higher doses of 20-40 μM rolipram significantly reduced cell viability [Figure 6]. Application of 50μg LPS significantly reduced cell viablility and coincubation with PD-4-I resulted in significantly augmented cell viability. Interestingly, this was also observed when higher doses of rolipram (20-40 μM) were used. Proinflammatory cytokine TNF-α (25-400 ng/ml) led to significantly decreased cell viability in a concentration-dependent manner. Because 5 μM rolipram showed no derangements to HepG2 cells, the coincubation with TNF-α + PD-4-I 5μM was investigated and revealed improved cell viability to control levels.


Phosphodiesterase-4 inhibition with rolipram attenuates hepatocellular injury in hyperinflammation in vivo and in vitro without influencing inflammation and HO-1 expression.

Wollborn J, Wunder C, Stix J, Neuhaus W, Bruno RR, Baar W, Flemming S, Roewer N, Schlegel N, Schick MA - J Pharmacol Pharmacother (2015 Jan-Mar)

HepG2 cell culture: Cell viability of PD-4-I-incubated cells (A) and in presence of 50 μg/ml LPS, showing a compromised viability with 50 ng/ml LPS and a cytoprotective effect of 2.5 μM and 10 μM PD-4-I. (C) Cell viability of different concentrations of TNF-α with and without 5μM PD-4-I, revealing cytoprotective effects of PD-4-I incubation at 50-400 ng/ml. (*vs. 0%, #vs. 50 μg/ml LPS; §vs. corresponding TNF-α-only incubated cells, n = 32-64, P≤ 0.05, mean ± SEM)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4319242&req=5

Figure 7: HepG2 cell culture: Cell viability of PD-4-I-incubated cells (A) and in presence of 50 μg/ml LPS, showing a compromised viability with 50 ng/ml LPS and a cytoprotective effect of 2.5 μM and 10 μM PD-4-I. (C) Cell viability of different concentrations of TNF-α with and without 5μM PD-4-I, revealing cytoprotective effects of PD-4-I incubation at 50-400 ng/ml. (*vs. 0%, #vs. 50 μg/ml LPS; §vs. corresponding TNF-α-only incubated cells, n = 32-64, P≤ 0.05, mean ± SEM)
Mentions: Hepatoma cell line, HepG2, cell culture was used as an in vitro model of hepatocytes to test our hypothesis of direct effects of PD-4-I on hepatocytes. First we tested the influence of PD-4-I at different doses on cell viability using the EZ4U-tests. Doses of rolipram at 2.5-10 μM did not affect cell viability while higher doses of 20-40 μM rolipram significantly reduced cell viability [Figure 6]. Application of 50μg LPS significantly reduced cell viablility and coincubation with PD-4-I resulted in significantly augmented cell viability. Interestingly, this was also observed when higher doses of rolipram (20-40 μM) were used. Proinflammatory cytokine TNF-α (25-400 ng/ml) led to significantly decreased cell viability in a concentration-dependent manner. Because 5 μM rolipram showed no derangements to HepG2 cells, the coincubation with TNF-α + PD-4-I 5μM was investigated and revealed improved cell viability to control levels.

Bottom Line: Untreated LPS-induced rats showed significantly decreased liver microcirculation and increased hepatic cell death, whereas LPS + PD-4-I treatment could improve hepatic volumetric flow and cell death to control level whithout influencing the inflammatory impact.The heme oxygenase 1 (HO-1) pathway did not induce the protective effect of PD-4-I.Intravenous PD-4-I treatment was effective in improving hepatic microcirculation and hepatic integrity, while it had a direct protective effect on HepG2 viability during inflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Anaesthesia and Critical Care, University Hospital Würzburg, Germany.

ABSTRACT

Objective: To investigate the impact of the phophodiesterase-4 inhibition (PD-4-I) with rolipram on hepatic integrity in lipopolysaccharide (LPS) induced hyperinflammation.

Materials and methods: Liver microcirculation in rats was obtained using intravital microscopy. Macrohemodynamic parameters, blood assays, and organs were harvested to determine organ function and injury. Hyperinflammation was induced by LPS and PD-4-I rolipram was administered intravenously one hour after LPS application. Cell viability of HepG2 cells was measured by EZ4U-kit based on the dye XTT. Experiments were carried out assessing the influence of different concentrations of tumor necrosis factor alpha (TNF-α) and LPS with or without PD-4-I.

Results: Untreated LPS-induced rats showed significantly decreased liver microcirculation and increased hepatic cell death, whereas LPS + PD-4-I treatment could improve hepatic volumetric flow and cell death to control level whithout influencing the inflammatory impact. In HepG2 cells TNF-α and LPS significantly reduced cell viability. Coincubation with PD-4-I increased HepG2 viability to control levels. The heme oxygenase 1 (HO-1) pathway did not induce the protective effect of PD-4-I.

Conclusion: Intravenous PD-4-I treatment was effective in improving hepatic microcirculation and hepatic integrity, while it had a direct protective effect on HepG2 viability during inflammation.

No MeSH data available.


Related in: MedlinePlus