Concentration Of Pro-inflammatory Factors Research Articles

MiR-26a-5p Attenuates Oxidative Stress and Inflammation in Diabetic Retinopathy through the USP14/NF-κB Signaling Pathway.

Diabetic retinopathy (DR) is an ocular disease caused by diabetes and may lead to vision impairment and even blindness. Oxidative stress and inflammation are two key pathogenic factors of DR. Recently, regulatory roles of different microRNAs (miRNAs) in DR have been widely verified. miR-26a-5p has been confirmed to be a potential biomarker of DR. Nevertheless, the specific functions of miR-26a-5p in DR are still unclear. Primary cultured mouse retinal Müller cells in exposure to high glucose (HG) were used to establish an in vitro DR model. Müller cells were identified via morphology observation under phase contrast microscope and fluorescence staining for glutamine synthetase. The in vivo animal models for DR were constructed using streptozotocin-induced diabetic C57BL/6 mice. Western blotting was performed to quantify cytochrome c protein level in the cytoplasm and mitochondria of Müller cells and to measure protein levels of glial fibrillary acidic protein (GFAP), ubiquitin-specific peptidase 14 (USP14), as well as factors associated with NF-κB signaling (p-IκBα, IκBα, p-p65, and p65) in Müller cells or murine retinal tissues. ROS production was detected by CM-H2DCFDA staining, and the concentration of oxidative stress markers (MDA, SOD, and CAT) was estimated by using corresponding commercial kits. Quantification of mRNA expression was conducted by RT-qPCR analysis. The concentration of proinflammatory factors (TNF-α, IL-1β, and IL-6) was evaluated by ELISA. Hematoxylin-eosin staining for murine retinal tissues was performed for histopathological analysis. Immunofluorescence staining was conducted to determine NF-κB p65 nuclear translocation in Müller cells. Furthermore, the interaction between miR-26a-5p and USP14 was verified via the luciferase reporter assays. HG stimulation contributed to Müller cell dysfunction by inducing inflammation, oxidative injury, and mitochondrial damage to Müller cells. miR-26a-5p was downregulated in Müller cells under HG condition, and overexpression of miR-26a-5p relieved HG-induced Müller cell dysfunction. Moreover, miR-26a-5p targeted USP14 and inversely regulated USP14 expression. Additionally, HG-evoked activation of NF-κB signaling was suppressed by USP14 knockdown or miR-26a-5p upregulation. Rescue assays showed that the protective impact of miR-26a-5p upregulation against HG-induced Müller cell dysfunction was reversed by USP14 overexpression. Furthermore, USP14 upregulation and activation of NF-κB signaling in the retinas of DR mice were detected in animal experiments. Injection with miR-26a-5p agomir improved retinal histopathological injury and weakened the concentration of proinflammatory cytokines and oxidative stress markers in the retinas of DR mice. miR-26a-5p inhibits oxidative stress and inflammation in DR progression by targeting USP14 and inactivating the NF-κB signaling pathway.

Application of Comfort Therapy under eCASH Concept in Acute and Chronic Wound Treatment.

Given the new ideas on wound care offered by the eCASH (early Comfort using Analgesia, minimal Sedatives, and maximal Humane care) and the substantial differences in clinical treatment between acute and chronic wounds, we aimed to investigate the effect of comfort therapy under the eCASH concept on analgesic sedation and accelerated wound healing in patients with acute or chronic wounds. This randomized clinical study was conducted in two parts: acute wounds and chronic wounds. Patients with acute wounds were allocated into the acute wound control group (AWCG) and the acute wound experimental group (AWEG). Patients with chronic wounds were allocated into the chronic wound control group (CWCG) and two experimental groups, in which they received intermittent negative pressure therapy (IPTEG) and continuous negative pressure therapy (CPTEG). On the basis of the standard treatment for patients in the control group, eCASH therapy was used in the experimental groups. In addition, pain intensity and procedural anxiety were evaluated using the visual analogue score (VAS) and the Hamilton Anxiety Scale (HAM-A). In addition, clinical effects were assessed on the basis of the size of the surface area, rate of healing, and concentration of pro-inflammatory factors (IL-1, IL-6, TNF-α) and growth factors (VEGF, bFGF, TGF-β1). Compared with the control group, the VAS score and HAM-A score in the experimental groups were significantly decreased after intervention (P < 0.05). After intervention, the levels of IL-1β, IL-6, and TNF-α in AWEG, IPTEG, and CPTEG were significantly lower than those in AWCG. In addition, the levels of VEGF, bFGF, and TGF-β1 in IPTEG and CPTEG were significantly higher than those in CWCG (P < 0.05). These results indicated that comfort therapy under the eCASH concept has a significant effect on ameliorating the pain and anxiety of patients, reducing the inflammatory reaction during the period of wound healing in the treatment of acute and chronic wounds. The trial has been registered in the Chinese Clinical Trial Registry (ChiCTR2200057981).

Cholecalciferol supplementation lowers leptin and TMAO but increases NO and VEGF-A levels in obese vitamin D deficient patients: Is it one of the potential cardioprotective mechanisms of vitamin D?

BackgroundVitamin D deficiency is one of the most common health issues in developed countries. Obese patients are most at risk of having serum 25-hydroxyvitamin D3 (25(OH)D3) levels that are too low due to the accumulation of vitamin D in adipose tissue. While the effects of a deficiency on the skeletal or immune system are known, the effects on the cardiovascular system are not yet clear. Our study investigates the effect of cholecalciferol supplementation in obese patients on selected biomarkers associated with cardiovascular diseases (CVDs).MethodsThe study enrolled 33 obese patients with insufficient 25(OH)D3 levels. For three months, the subjects supplemented with cholecalciferol at a dose of 2000 IU/day. Concentrations of nitric oxide (NO), vascular endothelial growth factor A (VEGF-A), leptin, trimethylamine N-oxide (TMAO) and soluble suppression of tumorigenicity 2 (sST2) were measured in baseline samples using ELISA (BioTek EPOCH). 25(OH)D3 levels measured on Beckman Coulter DXI 800 by chemiluminescence method.ResultsAfter supplementation, 25(OH)D3 levels increased significantly. Normal levels were achieved in most patients. A statistically significant reduction leptin and TMAO levels was observed. At the same time, NO and VEGF-A levels increased statistically significantly.ConclusionThis study indicates that restoring normal 25(OH)D3 levels in obese people reduces the concentration of pro-inflammatory factors associated with cardiovascular diseases. Reducing inflammation and the potential impact on vascular reactivity leads to the conclusion that cholecalciferol supplementation in obese patients may benefit the cardiovascular system.

Spinal cord injury target-immunotherapy with TNF-α autoregulated and feedback-controlled human umbilical cord mesenchymal stem cell derived exosomes remodelled by CRISPR/Cas9 plasmid

Human umbilical cord mesenchymal stem cell (hucMSC) derived exosomes (EXOs) have been investigated as a new treatment for spinal cord injury (SCI) because of their anti-inflammatory, anti-apoptotic, angiogenesis-promoting, and axonal regeneration properties. The CAQK peptide found in the brains of mice and humans after trauma has recently been found to specifically bind to the injured site after SCI. Thus, we developed a nanocarrier system called EXO-C@P based on hucMSC exosomes remodelled by the CRISPR/Cas9 plasmid to control inflammation and modified by the CAQK peptide. EXO-C@P was shown to effectively accumulate at the injury site and saturate the macrophages to significantly reduce the expression of inflammatory cytokines in a mouse model of SCI. Moreover, EXO-C@P treatment improved the performance of mice in behavioural assessments and upregulated soluble tumour necrosis factor receptor-1 (sTNFR1) in serum and at the trauma site after SCI surgery, but lowered the proportion of iNOS+ cells and the concentration of proinflammatory factors. In conclusion, EXO-C@P provides an effective alternative to multiple topical administration and drug delivery approaches for the treatment of SCI. Statement of significanceSCI is a serious disease characterised by a high incidence, high disability rate, and high medical costs, and has become a global medical problem. Several studies have shown that the inflammatory response is the critical inducer of secondary injury after SCI. The inflammatory cytokine TNF-α is considered to be one of the most significant therapeutic targets for autoimmune diseases. Antibodies targeting TNF-α and sTNFR1 are capable of neutralising free TNF-α. In this study, exosomes in the CRISPR/Cas9 system were used to establish stem cells with an autoregulated and feedback-controlled TNF-α response, with these cells secreting sTNFR1, which neutralised TNF-α and antagonised the inflammation stimulated by TNF-α. Moreover, the plasmid was combined with CAQK, which targeted the injury site and promoted the recovery of SCI function.

Mesenchymal Stem Cell-Derived Exosomes Improved Cerebral Infarction via Transferring miR-23a-3p to Activate Microglia.

Mesenchymal stem cells-derived exosome (MSCs-exo) is a potential method for cerebral infarction (CI) treatment. Here, western blot and qRT-PCR were carried out to measure the expression of proteins and genes, respectively. Modified neurological severity score and TTC staining were used to evaluate the brain injury of middle cerebral artery occlusion (MCAO) rats. Immunohistochemistry was performed to detect the expression of Iba-1, iNOS, and Arg-1 in tissues. Moreover, the rate of M1/M2 microglia was ensured by flow cytometry, and the concentration of pro-inflammatory factors in medium was measured using ELISA. Here, we found that miR-23a-3p is increased in human umbilical cord blood MSCs-exo. Bone marrow MSCs-exo (BMSCs-exo) could improve the injury in neuronal function and reduce the infarct size in vivo. However, the improvement of BMSCs-exo to CI was reversed by miR-23a-3p knockdown. The inhibition of BMSCs-exo to MCAO-induced microglia activation and M1 polarization and the upregulation of pro-inflammatory factors were limited by miR-23a-3p knockdown, which also confirmed in lipopolysaccharide-induced microglia. Overall, our data indicated that MSCs-exo improves CI via transferring miR-23a-3p, thus to induce the deactivation of microglia and M2 polarization. Our study revealed a new regulatory mechanism of CI.

Cannabinoid receptor agonist WIN55212-2 reduces unpredictable mild stress-induced depressive behavior of rats.

BackgroundDepression is a neurological disorder characterized by persistent low mood. A number of studies have suggested that the use of type 1 cannabinoid receptor (CB1R) agonists can reduce depressive behavior, but its effect on the depressive behavior and nerve damage of rats exposed to chronic unpredictable mild stress (CUMS) has not been reported.MethodsRats were exposed to CUMS for 4 weeks to induce depressive behavior. Male Sprague-Dawley (SD) rats aged 6–8 weeks were randomly divided into six groups: control group (control), depression group (CUMS), depression + fluoxetine group (Flu), depression + WIN55212-2 group (WIN), depression + NF-κB inhibitor group (PDTC), and depression + WIN + PDTC group (WIN + PDTC). We performed four behavioral experiments test to evaluate the depressive behaviors of rats. Hematoxylin and eosin (HE) and Nissl staining were performed to observe the neuron structures of the hippocampus. Enzyme-linked immunosorbent assay (ELISA) was used to measure the concentrations of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and cyclooxygenase-2 (COX-2). Biochemical experiments were performed to evaluate the concentrations of nitric oxide (NO), malondialdehyde (MDA), reactive oxygen species (ROS), and superoxide dismutase (SOD). Fluorescence quantitative PCR was used to detect the mRNA expression of brain-derived neurotrophic factor (BDNF), tyrosine kinase receptor B (TrkB), and inducible nitric oxide synthase (iNOS) in the hippocampus, and western blot was performed to detect protein expression levels related to the NF-κB signaling pathway in the hippocampus.ResultsCompared with the normal control group, CUMS significantly induced abnormal behaviors in stressed rats. The concentrations of pro-inflammatory factors and oxidative stress injury factors in the hippocampus of the CUMS group increased significantly. The interventions of Flu, WIN, and PDTC significantly reduced neuroinflammation and oxidative stress injury. Compared with the WIN group, the WIN + PDTC intervention group had better results. In addition, WIN could significantly inhibit the activation of the NF-κB signaling pathway.ConclusionsThis study showed that cannabinoid receptor agonists can reduce the CUMS-induced depressive behaviors of rats by blocking the NF-κB signaling pathway to alleviate neuroinflammation and oxidative stress injury.

Determination of Polyphenols in Ilex kudingcha and Insect Tea (Leaves Altered by Animals) by Ultra-high-performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry (UHPLC-QqQ-MS) and Comparison of Their Anti-Aging Effects.

Ilex kudingcha C.J. Tseng tea and insect tea, as traditional Chinese teas, are favored for their original craftsmanship, unique flavor, and biological functionality. In this study, ultra high-performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-QqQ-MS) was used to analyze the bioactive components of the extracts of Ilex kudingcha and insect tea, and D-galactose-induced aging mice were used to compare the in vivo anti-aging effects of Ilex kudingcha and insect tea extracts. The results were remarkable, UHPLC-QqQ-MS analysis showed that ITP contains 29 ingredients, while IKDCP contains 26 ingredients. However, due to the large differences in the content of the main chemical components in IKDCP and ITP, the effects are equally different. At the same time, the in vivo research results suggesting that the anti-aging effects of IKDCP and ITP (500 mg/kg) include the regulation of viscera indices of major organs; improvement in liver, skin, and spleen tissue morphology; decreased production of inflammatory cytokines; up regulation of SOD, CAT, GSH, GSH-PX, and T-AOC and down regulation of NO and MDA levels in serum and liver tissue; reductions in the concentration of pro-inflammatory factors, and increases in the concentration of anti-inflammatory factor. RT-qPCR and western blot assay also showed that IKDCP and ITP affect anti-aging by regulating the gene and protein expression of GSH-PX, GSH1, SOD1, SOD2, and CAT. The overall results indicate that ITP is more effective in treating oxidative damage in aging mice induced by D-galactose. Thus, ITP appears to be an effective functional drink owing to its rich nutritional components and anti-aging activities.

Gold nanoparticles alleviates the lipopolysaccharide-induced intestinal epithelial barrier dysfunction

ABSTRACT Nanotechnology is used in the immune response manipulation to treat various human diseases. In the present study, we explored the effects of Au nanoparticles (AuNPs) on the lipopolysaccharide (LPS)-induced epithelial barrier dysfunction and inflammatory response of colonic epithelial NCM460 cells. According to the results of cell counting kit-8 and flow cytometry analysis, the viability of NCM460 cells was inhibited, and the apoptosis was increased after LPS treatment, and AuNPs reversed these changes in a dose-dependent way. The permeability was evaluated by detecting the flux of fluorescein isothiocyanate-dextran and transepithelial electrical resistance. LPS enhanced the permeability and promoted barrier dysfunction of NCM460 cells. Enzyme-linked immunosorbent sorbent assay results revealed that the concentrations of pro-inflammatory factors and nitric oxide were elevated by LPS treatment and decreased by the AuNPs. LPS aggravated the inflammatory response, which was rescued by the AuNPs. Moreover, LPS promoted the activation of the nuclear factor kappa-B and extracellular signal-regulated kinase/c-Jun NH-terminal kinase signaling pathways, which were inhibited by AuNPs.

P0526MIR-181D-5P TARGETS KLF6 TO IMPROVE ISCHEMIC/REPERFUSION AKI THROUGH EFFECTS ON RENAL FUNCTION,APOPTOSIS AND INFLAMMATION

Abstract Background and Aims Inflammation and renal tubular cell death is the most crucial pathophysiological changes in acute kidney ischemic/reperfusion injury(IRI). The miR-181d family plays diverse roles in cell proliferation, apoptosis and inflammation, but its renal target and potential role in IRI are unknown. Method We built hypoxia/reoxygenation (H/R) renal cell model (HK-2 cells) and renal ischemia/reperfusion (I/R) injury mice model. We used observe the expression location of miR-181d-5p and KLF6 by in situ hybridization and immunohistochemistry, the blood creatinine and urea nitrogen levels through fully automatic biochemical detector, the severity of kidney injury by HE staining, the apoptosis and necrosis of renal tubular epithelial cells by TUNEL and Annexin V-FITC/PI flow cytometry, the concentrations of pro-inflammatory factors in the cell supernatant by ELISA and the mRNA and protein levels of related factors and predicted and verified the relationship between miR-181d-5p and KLF6 via bioinformatics systems and luciferase reporter genes. Results We showed that the expression of miR-181d-5p decreased in HK-2 cells of H/R and renal mouse model of I/R injury. Mechanistically, KLF6 was predicted as a new potential target gene of miR-181d-5p through bioinformatics and the Luciferase reporter verification. After renal ischemia/ reperfusion, overexpression of microRNA-181d significantly attenuated the level of KLF6 and inhibited inflammatory mediators such as IL-6, TNF-α, reducing apoptosis and further improving renal function. Critical NF-κB signaling pathways mediated this inflammatory response. Conclusion In conclusion, upregulation of microRNA-181d inhibits KLF6 expression, which have protective anti-apoptosis and anti-inflammatory effects against I/R kidney in vivo, and H/R HK-2 cells in vitro, thereby providing novel insights into the molecular mechanisms associated with ischemia/reperfusion injury and a potential novel therapeutic target.

Protective effects of isorhynchophylline against silicon-dioxide-induced lung injury in mice

Inhalation of silicon dioxide (SD) results in pulmonary inflammatory responses and fibrosis. Isorhynchophylline (Isorhy) is the main alkaloid in the traditional Chinese herb Tripterygium wilfordii, which is reported to have anti-inflammatory activities in the nervous system. However, the effects of Isorhy on SD-induced pulmonary inflammation and fibrosis in mice are unknown. Male mice were exposed to a single dose of SD (2.5 mg/kg, intranasal inoculation) to induce pulmonary fibrosis (PF). The mice were woken up and immediately treated with Isorhy (20 mg/kg, intraperitoneal injection) for 14 or 42 days. The effects of Isorhy on inflammatory responses and lung fibrosis induced by SD were then investigated. After the 14-day treatment, there was a significant reduction in inflammatory cell infiltration in the lungs of mice, with reduced recruitment of inflammatory cells to the lungs. The concentration of pro-inflammatory factors in the bronchoalveolar lavage fluid was reduced, which alleviated inflammatory injury in the lung tissue. After the 42-day treatment, Isorhy alleviated inflammation and inhibited the release of fibrogenic factors in mice with PF. Isorhy also significantly reduced collagen deposition in the lung tissues of mice. Isorhy has the ability to reduce inflammatory responses and fibrosis associated with SD-induced acute lung injury.

Rubber seed oil and flaxseed oil supplementation on serum fatty acid profile, oxidation stability of serum and milk, and immune function of dairy cows

ObjectiveThis study was designed to investigate the effect of diet supplementation with rubber seed oil and flaxseed oil on serum fatty acids profile, oxidation stability of serum and milk, and immune function of dairy cows.MethodsForty-eight mid-lactation Holstein dairy cows were randomly assigned to one of four treatments for 8 wk, including basal diet (CON) or the basal diet supplemented with 4% rubber seed oil (RO), 4% flaxseed oil (FO) or 2% rubber seed oil plus 2% flaxseed oil (RFO) on a dry matter basis.ResultsCompared with CON, all the oil groups increased the levels of trans-11 C18:1 (vaccenic acid), cis-9, trans-11 C18:2 (conjugated linoleic acid, CLA) and C18:3 (α-linolenic acid, ALA) in serum. Both the activity of glutathione peroxidase and catalase in serum and milk in oil groups were decreased, which were negatively correlated with the levels of cis-9, trans-11 CLA and ALA. The concentrations of proinflammatory factors (tumor necrosis factor α and interferon γ) in serum of oil groups were lower than that from the CON cows.ConclusionThese results indicate that diet supplementation with RO or FO could alter serum fatty acid profile and enhance the immune function of dairy cows. However, the negative effect on milk oxidation stability should be considered when feeding these n-3 polyunsaturated fatty acid-enriched oils in dairy production.

Protective effects of astragaloside IV on IL-8-treated diaphragmatic muscle cells.

The diaphragmatic fatigue that results from airflow obstruction is associated with the severe morbidity of patients with chronic obstructive pulmonary disease. Astragaloside IV (AS-IV) has antioxidant, anti-apoptotic and anti-inflammatory activities in various cell types. The present study aimed to evaluate the protective effects of AS-IV in diaphragmatic muscle cells. Diaphragmatic muscle cells extracted from neonatal rats were treated with a series of AS-IV concentrations (5, 10 or 20 mg/l) and the AKT inhibitor GSK690693 in the presence of interleukin-8 (IL-8). Cell proliferation and AKT phosphorylation were measured using Cell Counting Kit-8 and western blot assays, respectively. Cell apoptosis and reactive oxygen species (ROS) production were evaluated using flow cytometric analysis. Caspase activity and concentrations of proinflammatory factors (tumor necrosis factor-α, IL-6 and IL-8) were assessed using a caspase colorimetric assay and ELISA, respectively. IL-8 treatment resulted in decreased rates of cell proliferation and increased rates of AKT phosphorylation, cell apoptosis, caspase 3/9 activity, ROS production and proinflammatory factor production. AS-IV and GSK690693 treatment reversed the effects of IL-8. The effects of AS-IV were dose-dependent. The present results suggested that AS-IV is a candidate for the treatment of diaphragmatic fatigue due to its antioxidant, anti-apoptotic and anti-inflammatory activity.

Protecting neurons from cerebral ischemia/reperfusion injury via nanoparticle-mediated delivery of an siRNA to inhibit microglial neurotoxicity

Complement component C3 (C3) plays a central role in microglial neurotoxicity following cerebral ischemia/reperfusion (I/R) injury. In this study, we focused on the role of nanoparticles loaded with C3 siRNA (NPsiC3) in inhibiting microglial neurotoxicity after brain (I/R) injury. NPsiC3 inhibited the hypoxia/re-oxygenation-induced increase in C3 expression in microglia in vitro. Importantly, treatment with NPsiC3 decreased C3b deposition on neurons and reduced microglia-mediated neuronal damage under hypoxia/re-oxygen conditions. Nanoparticles could effectively deliver C3-siRNA from the blood into ischemic penumbra across the blood-brain barrier (BBB) and significantly decrease C3 expression in microglia and ischemic brain tissue, while reducing the number of infiltrating inflammatory cells and the concentration of pro-inflammatory factors in the penumbra. Furthermore, NPsiC3 also prevented neuronal apoptosis, reduced the volume of the ischemic zone, and substantially improved functional recovery after I/R injury. Therefore, the NPsiC3-induced inhibition of microglial neurotoxicity represents a novel therapeutic strategy for treating brain I/R injury.

Inflammatory biomarkers associated with obesity and insulin resistance: a focus on lipocalin-2 and adipocyte fatty acid-binding protein

Obesity is an important risk factor for a cluster of metabolic and cardiovascular diseases, including insulin resistance, Type 2 diabetes, nonalcoholic fatty liver disease and atherosclerosis. Systemic low-grade inflammation, characterized by elevated circulating concentrations of proinflammatory factors, has recently been proposed to be a key mediator that links obesity with its medical complications. Adipose tissue is now recognized as the major contributor to systemic inflammation associated with obesity. As obesity develops, adipose tissue is infiltrated with activated macrophages. The ‘inflamed’ adipose tissue secretes a large number of proinflammatory adipokines and/or cytokines, which can act either in an autocrine manner to perpetuate local inflammation or in an endocrine manner to induce insulin resistance and endothelial dysfunction. In this review, we summarize recent advances in several newly identified adipose tissue-derived inflammatory factors, with the focus on lipocalin-2 and adipocyte fatty acid-binding protein (A-FABP). Both lipocalin-2 and A-FABP possess lipid-binding properties and are important integrators of metabolic and inflammatory pathways. A growing body of evidence from experimental, epidemiological and genetic studies suggests that both lipocalin-2 and A-FABP represent a novel class of serum biomarkers for risk prediction and therapeutic intervention of obesity-related medical complications.

Proinflammatory Cytokines in Obesity: Impact of Type 2 Diabetes Mellitus and Gastric Bypass

Obesity and obesity-associated type 2 diabetes mellitus (T2DM) are frequently related to a low-grade chronic inflammatory state, which increases the risk of developing cardiovascular diseases. The aim of the present work was to evaluate the effect of obesity and T2DM on the concentrations of pro-inflammatory factors and to study the effect of weight loss after Roux-en-Y gastric bypass (RYGBP). Plasma concentrations of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-alpha), serum amyloid A (SAA) and sialic acid (SA) were measured in 25 female volunteers. The concentrations of these cytokines were determined in 14 female obese patients before and after weight loss following RYGBP. Additionally, visceral adipose tissue (VAT) obtained from 15 females was used to quantify expression levels of MCP-1 and CD68 by Real-Time PCR. Both obese normoglycemic (NG) and T2DM groups exhibited significantly higher MCP-1 (P < 0.05), TNF-alpha (P < 0.01), SAA (P < 0.05) and SA (P < 0.05) concentrations, compared to the lean group. No differences were found between obese NG and obese T2DM subjects. A significant positive correlation was found between body fat percentage (BF) and all inflammatory markers (P < 0.05) studied. MCP-1 expression levels in VAT were upregulated in obese NG (P = 0.008) and obese T2DM (P = 0.032) patients compared to lean subjects, but no additional detrimental effect of T2DM was observed between both obese groups. After weight loss, SAA (P < 0.001) and SA (P < 0.05) concentrations diminished, whereas circulating levels of MCP-1 showed a tendency to decrease (P = 0.093) and TNF-alpha did not change. The present findings suggest that elevated pro-inflammatory cytokine levels found in obese patients relate mainly to obesity rather than to T2DM. Moreover, surgery-induced weight loss reduces circulating concentrations of key pro-inflammatory factors, which contribute to the improvement in the cardiovascular co-morbidity following excess weight loss.