Integrated Haematological Profiles of Redox Status, Lipid, and Inflammatory Protein Biomarkers in Benign Obesity and Unhealthy Obesity with Metabolic Syndrome

Abstract

The pathogenesis of obesity (OB) and metabolic syndrome (MetS) implies free radical-, oxidized lipid- (LOOH-), and inflammatory cytokine-mediated altered pathways in target organs. Key elements of the transition from benign OB to unhealthy OB+MetS remain unclear. Here, we measured a panel of redox, antioxidant, and inflammation markers in the groups of OB patients (67 with, 45 without MetS) and 90 controls. Both OB groups displayed elevated levels of adipokines and heavy oxidative stress (OS) evidenced by reduced levels of glutathione, downregulated glutathione-S-transferase, increased 4-hydroxynonenal-protein adducts, reactive oxygen species, and membrane-bound monounsaturated fatty acids (MUFA). Exclusively in OB+MetS, higher-than-normal glutathione peroxidase activity, tumor necrosis factor-α, and other proinflammatory cytokines/chemokines/growth factors were observed; a combination of high adipokine plasminogen activator inhibitor-1 and MUFA was consistent with increased cardiovascular risk. The uncomplicated OB group showed features of adaptation to OS such as decreased levels of vitamin E, activated superoxide dismutase, and inhibited catalase, suggesting H2O2 hyperproduction. Proinflammatory cytokine pattern was normal, except few markers like RANTES, a suitable candidate for therapeutic approaches to prevent a setting of MetS by inhibition of LOOH-primed leukocyte chemotaxis/recruitment to target tissues.

Figure 1

Metabolic redox parameters: erythrocyte levels of antioxidant/detoxifying enzymatic activities of glutathione peroxidase (a), CuZn-superoxide dismutase (b), catalase (c), glutathione-S-transferase (d), and reduced, oxidized glutathione (e-f), in the groups of obese patients with metabolic syndrome (OB w.MetS, n = 67) and without metabolic syndrome (OB w.o.MetS, n = 45), and of control healthy subjects (CTR, n = 90). Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), and 1.96 × standard error (upper and lower whiskers). Intergroup significant differences (p) are indicated in each panel. Abbreviations: GPX (glutathione peroxidase), CuZnSOD (CuZn-superoxide dismutase); CAT (catalase); GST (glutathione S-transferase); GSH (reduced glutathione); GSSG (oxidized glutathione); RBC (red blood cells); U (units); prot. (proteins); Hb (haemoglobin). Techniques: spectrophotometric methods (a–d), HPLC with array photodiode detection (e).

Figure 2

Metabolic redox parameters: levels of plasma 4-hydroxynonenal protein adducts (a) and a representative blot of the relative gel bands of samples and of loading control (ponceau staining) (b), of whole blood luminol-dependent chemiluminescence (c), of plasma α-tocopherol (d), in the groups of obese patients with metabolic syndrome (OB w.MetS, n = 67) and without metabolic syndrome (OB w.o.MetS, n = 45), and of control healthy subjects (CTR, n = 90). Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), and 1.96 × standard error (upper and lower whiskers). Intergroup significant differences (p) are indicated in each panel. Abbreviations: 4-HNE PA (4-hydroxynonenal protein adducts); kDa (kiloDalton); CL (luminol-dependent chemiluminescence); cps (counts per second); ALPHA-TOC (α-tocopherol). Techniques: Western blot (a-b), chemiluminescence (c), and HPLC with array photodiode detection (d).

Figure 3

Selected representative parameters describing fatty acid profile of erythrocyte membrane phospholipids: ratio saturated/monounsaturated fatty acids (a), ratio ω6/ω3 polyunsaturated fatty acids (b), ω6-arachidonic acid percent of total FA content of phospholipids (c), and ω-3 eicosapentaenoic acid % of total FA content of phospholipids (d), in the groups of obese patients with metabolic syndrome (OB w.MetS, n = 45) and without metabolic syndrome (OB w.o.MetS, n = 33) and of control healthy subjects (CTR, n = 49). Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), and 1.96 × standard error (upper and lower whiskers). Intergroup significant differences (p) are indicated in each panel. Abbreviations: SAT (saturated fatty acids); MUFA (monounsaturated fatty acids); PUFA (polyunsaturated fatty acids); C20:4 (arachidonic acid); C20:5 (eicosapentaenoic acid); and RBC (red blood cells). Techniques: gas chromatography-mass spectrometry with selected ion monitoring (a–d).

Figure 4

Plasma levels of selected adipokines: C-peptide (a), plasminogen activator inhibitor-1 (b), leptin (d), glucose-dependent insulinotropic polypeptide (e), ghrelin (f), and ATP levels in platelet-enriched plasma (c), in the groups of obese patients with metabolic syndrome (OB w.MetS, n = 67) and without metabolic syndrome (OB w.o.MetS, n = 45), and control healthy subjects (CTR, n = 90). Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), and 1.96 × standard error (upper and lower whiskers). Intergroup significant differences (p) are indicated in each panel. Abbreviations: PAI-1 (plasminogen activator inhibitor-1); PRP (platelet-rich plasma); and GIP (glucose-dependent insulinotropic polypeptide). Techniques: multiplexed Bio-Plex Suspension Array System (a-b, d–f), bioluminescence (c).

Figure 5

(a) Plasma levels of selected cytokines/chemokines/growth factors: platelet-derived growth factor (A), interleukin 6 (B), interleukin 7 (C), interleukin 10 (D), interleukin 4 (E), and interleukin 8 (F), in the groups of obese patients with metabolic syndrome (OB w.MetS, n = 67) and without metabolic syndrome (OB w.o.MetS, n = 45) and of control healthy subjects (CTR, n = 90). Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), and 1.96 × standard error (upper and lower whiskers). Intergroup significant differences (p) are indicated in each panel. Abbreviations: PDGF-bb (platelet-derived growth factor); IL-6 (interleukin 6); IL-7 (interleukin 7); IL-10 (interleukin 10); IL-4 (interleukin 4); and IL-8 (interleukin 8). Techniques: multiplexed Bio-Plex Suspension Array System (A–F). (b) Plasma levels of selected cytokines/chemokines/growth factors: interleukin 9 (A), granulocyte colony stimulating factor (B), tumor necrosis factor-α (C), vascular endothelial growth factor (D), granulocyte-macrophage colony stimulating factor (E), regulated on activation, normal T cell expressed and secreted (F), in the groups of obese patients with metabolic syndrome (OB w.MetS, n = 67) and without metabolic syndrome (OB w.o.MetS, n = 45), and control healthy subjects (CTR, n = 90). Values are represented as mean (□), standard error of the mean (upper and lower limits of the box), and 1.96 × standard error (upper and lower whiskers). Intergroup significant differences (p) are indicated in each panel. Abbreviations: IL-9 (interleukin 9); G-CSF (granulocyte colony stimulating factor); TNF-alpha (tumor necrosis factor-α); VEGF (vascular endothelial growth factor); GM-CSF (granulocyte-macrophage colony stimulating factor); and RANTES (regulated on activation, normal T cell expressed and secreted). Techniques: multiplexed Bio-Plex Suspension Array System (A–F).