Limits...
17-oxo-DHA displays additive anti-inflammatory effects with fluticasone propionate and inhibits the NLRP3 inflammasome

View Article: PubMed Central - PubMed

ABSTRACT

Chronic obstructive pulmonary disease (COPD) is characterized by reduced lung function associated with increased local and systemic inflammatory markers, such as TNFα and IL-1β. Glucocorticoids are used to treat this chronic disease, however their efficacy is low and new drugs are very much required. 17-oxo-DHA is a cyclooxygenase-2-dependent, electrophilic, α,β-unsaturated keto-derivative of docosahexaenoic acid with anti-inflammatory properties. We evaluated the action of 17-oxo-DHA alone or in combination with the steroid fluticasone propionate (FP) in peripheral blood mononuclear cells (PBMCs) from COPD patients and healthy individuals exposed to lipopolysaccharide. We show that PBMCs from COPD patients released higher levels of TNFα and IL-1β compared to controls. 17-oxo-DHA displayed strong anti-inflammatory effects. The addition of 17-oxo-DHA in combination with FP showed enhanced anti-inflammatory effects through the modulation of transcriptional and post-transcriptional mechanisms. 17-oxo-DHA, but not FP, was able to suppress the release of mature IL-1β through inhibition of the NLRP3 inflammasome. Furthermore, 17-oxo-DHA inhibited inflammasome-dependent degradation of the glucocorticoid receptor (GR). Our findings suggest that 17-oxo-DHA in combination with FP or other steroids might achieve higher therapeutic efficacy than steroids alone. Combined treatment might be particularly relevant in those conditions where increased inflammasome activation may lead to GR degradation and steroid-unresponsive inflammation.

No MeSH data available.


Related in: MedlinePlus

Modulation of LPS-induced TNFα and IL-1β release by 17-oxo-DHA alone or in combination with FP in PBMCs from COPD subjects and healthy individuals.Freshly isolated PBMCs from COPD patients (grey dots, N = 10) and healthy individuals (black dots, N = 11) were stimulated with 1 μg/ml LPS for 18 h. Where indicated, 17-oxo-DHA (5 μM) or FP (10 nM) were added, alone or in combination, 1 h before LPS. TNFα (a) and IL-1β (b) were measured in supernatants by ELISA. Data are presented in Box and whiskers plots with individual data points. The box extends from the 25th to 75th percentiles and the line in the middle of the box is the median. The whiskers represent the minimum and the maximum values. *p-value < 0.05; **p-value < 0.005; ns, p-value > 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5121625&req=5

f1: Modulation of LPS-induced TNFα and IL-1β release by 17-oxo-DHA alone or in combination with FP in PBMCs from COPD subjects and healthy individuals.Freshly isolated PBMCs from COPD patients (grey dots, N = 10) and healthy individuals (black dots, N = 11) were stimulated with 1 μg/ml LPS for 18 h. Where indicated, 17-oxo-DHA (5 μM) or FP (10 nM) were added, alone or in combination, 1 h before LPS. TNFα (a) and IL-1β (b) were measured in supernatants by ELISA. Data are presented in Box and whiskers plots with individual data points. The box extends from the 25th to 75th percentiles and the line in the middle of the box is the median. The whiskers represent the minimum and the maximum values. *p-value < 0.05; **p-value < 0.005; ns, p-value > 0.05.

Mentions: Freshly isolated PBMCs from moderate to severe COPD patients (N = 10) and healthy individuals (N = 11, control group) were treated for 1 h with 17-oxo-DHA, FP or 17-oxo-DHA + FP, then stimulated with LPS for 18 hours. Afterwards, TNFα and IL-1β concentrations were measured in the supernatants. The induction of TNFα and IL-1β release in response to LPS stimulation was significantly higher in PBMCs from COPD patients (median (range): 17.88 ng/ml (9.003; 23.91) for TNFα and 1.528 ng/ml (1.173; 3.901) for IL-1β) compared to PBMCs from healthy individuals (median (range): 8.92 ng/ml (4.96; 11.5) for TNFα and 1.096 ng/ml (0.8440; 1.528) for IL-1β). 17-oxo-DHA and FP, when administered alone, displayed a similar effect in suppressing TNFα release, while 17-oxo-DHA was more potent than FP in inhibiting IL-1β release (Fig. 1). The combination of 17-oxo-DHA and FP resulted in even greater suppression of TNFα and IL-1β release, showing additive effects in both study groups. To further investigate the impact of 17-oxo-DHA on FP actions, dose-response experiments were performed and the effect of 17-oxo-DHA on half maximal inhibitory concentration (IC50) and maximum inhibition (Imax) of FP was evaluated (Fig. 2, Supplementary Figures S1, S2). No differences of IC50 and Imax were observed between the two study groups. When administered in combination with FP, 17-oxo-DHA significantly increased FP-Imax for both cytokines and in both study groups, without modifying FP-IC50.


17-oxo-DHA displays additive anti-inflammatory effects with fluticasone propionate and inhibits the NLRP3 inflammasome
Modulation of LPS-induced TNFα and IL-1β release by 17-oxo-DHA alone or in combination with FP in PBMCs from COPD subjects and healthy individuals.Freshly isolated PBMCs from COPD patients (grey dots, N = 10) and healthy individuals (black dots, N = 11) were stimulated with 1 μg/ml LPS for 18 h. Where indicated, 17-oxo-DHA (5 μM) or FP (10 nM) were added, alone or in combination, 1 h before LPS. TNFα (a) and IL-1β (b) were measured in supernatants by ELISA. Data are presented in Box and whiskers plots with individual data points. The box extends from the 25th to 75th percentiles and the line in the middle of the box is the median. The whiskers represent the minimum and the maximum values. *p-value < 0.05; **p-value < 0.005; ns, p-value > 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5121625&req=5

f1: Modulation of LPS-induced TNFα and IL-1β release by 17-oxo-DHA alone or in combination with FP in PBMCs from COPD subjects and healthy individuals.Freshly isolated PBMCs from COPD patients (grey dots, N = 10) and healthy individuals (black dots, N = 11) were stimulated with 1 μg/ml LPS for 18 h. Where indicated, 17-oxo-DHA (5 μM) or FP (10 nM) were added, alone or in combination, 1 h before LPS. TNFα (a) and IL-1β (b) were measured in supernatants by ELISA. Data are presented in Box and whiskers plots with individual data points. The box extends from the 25th to 75th percentiles and the line in the middle of the box is the median. The whiskers represent the minimum and the maximum values. *p-value < 0.05; **p-value < 0.005; ns, p-value > 0.05.
Mentions: Freshly isolated PBMCs from moderate to severe COPD patients (N = 10) and healthy individuals (N = 11, control group) were treated for 1 h with 17-oxo-DHA, FP or 17-oxo-DHA + FP, then stimulated with LPS for 18 hours. Afterwards, TNFα and IL-1β concentrations were measured in the supernatants. The induction of TNFα and IL-1β release in response to LPS stimulation was significantly higher in PBMCs from COPD patients (median (range): 17.88 ng/ml (9.003; 23.91) for TNFα and 1.528 ng/ml (1.173; 3.901) for IL-1β) compared to PBMCs from healthy individuals (median (range): 8.92 ng/ml (4.96; 11.5) for TNFα and 1.096 ng/ml (0.8440; 1.528) for IL-1β). 17-oxo-DHA and FP, when administered alone, displayed a similar effect in suppressing TNFα release, while 17-oxo-DHA was more potent than FP in inhibiting IL-1β release (Fig. 1). The combination of 17-oxo-DHA and FP resulted in even greater suppression of TNFα and IL-1β release, showing additive effects in both study groups. To further investigate the impact of 17-oxo-DHA on FP actions, dose-response experiments were performed and the effect of 17-oxo-DHA on half maximal inhibitory concentration (IC50) and maximum inhibition (Imax) of FP was evaluated (Fig. 2, Supplementary Figures S1, S2). No differences of IC50 and Imax were observed between the two study groups. When administered in combination with FP, 17-oxo-DHA significantly increased FP-Imax for both cytokines and in both study groups, without modifying FP-IC50.

View Article: PubMed Central - PubMed

ABSTRACT

Chronic obstructive pulmonary disease (COPD) is characterized by reduced lung function associated with increased local and systemic inflammatory markers, such as TNF&alpha; and IL-1&beta;. Glucocorticoids are used to treat this chronic disease, however their efficacy is low and new drugs are very much required. 17-oxo-DHA is a cyclooxygenase-2-dependent, electrophilic, &alpha;,&beta;-unsaturated keto-derivative of docosahexaenoic acid with anti-inflammatory properties. We evaluated the action of 17-oxo-DHA alone or in combination with the steroid fluticasone propionate (FP) in peripheral blood mononuclear cells (PBMCs) from COPD patients and healthy individuals exposed to lipopolysaccharide. We show that PBMCs from COPD patients released higher levels of TNF&alpha; and IL-1&beta; compared to controls. 17-oxo-DHA displayed strong anti-inflammatory effects. The addition of 17-oxo-DHA in combination with FP showed enhanced anti-inflammatory effects through the modulation of transcriptional and post-transcriptional mechanisms. 17-oxo-DHA, but not FP, was able to suppress the release of mature IL-1&beta; through inhibition of the NLRP3 inflammasome. Furthermore, 17-oxo-DHA inhibited inflammasome-dependent degradation of the glucocorticoid receptor (GR). Our findings suggest that 17-oxo-DHA in combination with FP or other steroids might achieve higher therapeutic efficacy than steroids alone. Combined treatment might be particularly relevant in those conditions where increased inflammasome activation may lead to GR degradation and steroid-unresponsive inflammation.

No MeSH data available.


Related in: MedlinePlus