Relationship between platelet and urinary 8-Iso-PGF2α levels in subjects with different degrees of NOX2 regulation

Roberto Carnevale, Luigi Iuliano, Cristina Nocella, Simona Bartimoccia, Stefano Trapè, Roberta Russo, Maria Cristina Gentile, Roberto Cangemi, Lorenzo Loffredo, Pasquale Pignatelli, Francesco Violi, IPINET group, Roberto Carnevale, Luigi Iuliano, Cristina Nocella, Simona Bartimoccia, Stefano Trapè, Roberta Russo, Maria Cristina Gentile, Roberto Cangemi, Lorenzo Loffredo, Pasquale Pignatelli, Francesco Violi, IPINET group

Abstract

Background: Urinary 8-iso-PGF2α, a marker of oxidative stress, is influenced by the activation of NOX2. It is unclear if platelets 8-iso-PGF2α contribute to urinary 8-iso-PGF2α.

Methods and results: In a cross-sectional study, platelet, urinary, and serum 8-iso-PGF2α were determined in subjects with downregulation (X-linked chronic granulomatous disease [X-CGD], n=25) and upregulation (type II diabetic patients [T2D], n=121) of NOX2 and 153 controls matched for sex and age. In diabetic patients (n=18), the above variables were repeated before and after 7 days treatment with 100 mg/day aspirin or 100 mg/day aspirin plus 40 mg/day atorvastatin. In vitro study was performed to see the contribution of blood cells to serum 8-iso-PGF2α. Compared with controls, X-CGD patients had lower platelet, serum, and urinary 8-iso-PGF2α values; conversely, diabetic patients had higher values of 8-iso-PGF2α compared with controls. Urinary 8-iso-PGF2α significantly correlated with both platelet and serum 8-iso-PGF2α in the 2 cohorts. A parallel increase of platelet, serum, and urinary 8-iso-PGF2α by aspirin and a parallel decrease by aspirin plus atorvastatin were detected in the interventional study. In vitro study demonstrated that platelets contribute to 37% of serum 8-iso-PGF2α and that only 13% of it is of extravascular origin.

Conclusions: The study suggests that NOX2 contributes to the formation of 8-iso-PGF2α in both platelets and urine. The direct correlation between platelet and urinary 8-iso-PGF2α suggests that, at least partly, urinary 8-iso-PGF2α reflects platelet 8-iso-PGF2α production. Analysis of serum 8-iso-PGF2α may represent a novel tool to investigate the production of 8-iso-PGF2α by blood cells including platelets.

Clinical trial registration: URL: ClinicalTrials.gov. Unique Identifier: NCT01250340.

Keywords: 8‐iso‐PGF2α; NOX2; oxidative stress; platelets.

Figures

Figure 1.
Figure 1.
Urinary (A), platelet (B), and serum (C) 8‐iso‐PGF2α levels in X‐CGD patients (n=25), controls (n=153), and diabetic patients (n=121); P<0.001. The error bar represents the standard error. X‐CGD indicates X‐linked chronic granulomatous disease; PGF2a, 8‐iso‐Prostaglandin F2‐alpha.
Figure 2.
Figure 2.
Serum (A) and urinary (B) 8‐iso‐PGF2α formation in diabetic patients treated or not treated with aspirin (100 mg/day). The error bar represented the standard error. ASA indicates aspirin; PGF2a, 8‐iso‐Prostaglandin F2‐alpha.
Figure 3.
Figure 3.
A, Serum sNOX‐2‐dp levels in X‐CGD patients, controls, and diabetic patients (the error bar represented the standard error). B, Urinary (on the x axis) and platelet (on the y axis) 8‐iso‐PGF2α levels in both hereditary deficiency of NOX2, controls, and diabetic patients (highlighted with different symbols; data are represented as a scatter plot). X‐CGD indicates X‐linked chronic granulomatous disease; NOX, NADPH oxidase; sNOX‐2‐dp, soluble NADPH oxidase 2‐derived peptide.
Figure 4.
Figure 4.
Platelet (A), serum (B), and urinary (C) 8‐iso‐PGF2α formation in diabetic patients at baseline and after 7 days of aspirin (100 mg/day) or aspirin (100 mg/day) plus atorvastatin (10 mg/day) treatment. The error bar represented the standard error. ASA indicates aspirin; ATO, atorvastatin.
Figure 5.
Figure 5.
8‐Iso‐PGF2α levels in serum kept for 60 minutes at 37°C (n=5) or at room temperature (n=5) in platelets stimulated with AA in the presence or absence of NOX2‐tat inhibitor (n=5), in PMNs stimulated with PMA in the presence or absence of NOX2‐tat inhibitor (NADPH oxidase inhibitor; n=5), and in LYN/MON stimulated with LPS in the presence or absence of NOX2‐tat inhibitor (n=5); *P<0.001. The error bar represented the standard error. AA indicates arachidonic acid; PMNs, polymorphonuclear leukocytes; PMA, phorbol 12‐myristate 13‐acetate; PGF2a, 8‐iso‐Prostaglandin F2‐alpha; NOX, NADPH oxidase; LYM/MON, lymphocytes/monocyte; LPS, lipopolysaccharide.

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Source: PubMed

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