interleukin-8 and Diabetic-Retinopathy

Increasing evidence shows that systemic inflammation is an embedded mechanism of proliferative diabetic retinopathy (PDR). However, the specific systemic inflammatory factors involved in this process remained obscure. The study aimed to identify the upstream and downstream systemic regulators of PDR by using Mendelian randomization (MR) analyses.. We performed a bidirectional two-sample MR analysis implementing the results from genome-wide association studies for 41 serum cytokines from 8,293 Finnish individuals, and PDR from FinnGen consortium (2,025 cases vs. 284,826 controls) and eight cohorts of European ancestry (398 cases vs. 2,848 controls), respectively. The inverse-variance-weighted method was adopted as the main MR method, and four additional MR methods (MR-Egger, weighted-median, MR-pleiotropy residual sum and outlier (MR-PRESSO), and MR-Steiger filtering methods) were used for the sensitivity analyses. Results from FinnGen and eight cohorts were pooled into a meta-analysis.. Our results showed that genetically predicted higher stem cell growth factor-β (SCGFb) and interleukin-8 were positively associated with an elevated risk of PDR, with a combined effect of one standard deviation (SD) increase in SCGFb and interleukin-8 causing 11.8% [95% confidence interval (CI): 0.6%, 24.2%]) and 21.4% [95% CI: 3.8%, 41.9%]) higher risk of PDR, respectively. In contrast, genetically predisposition to PDR showed a positive association with the increased levels of growth-regulated oncogene-α (GROa), stromal cell-derived factor-1 alpha (SDF1a), monocyte chemotactic protein-3 (MCP3), granulocyte colony-stimulating factor (GCSF), interleukin-12p70, and interleukin-2 receptor subunit alpha (IL-2ra).. Our MR study identified two upstream regulators and six downstream effectors of PDR, providing opportunities for new therapeutic exploitation of PDR onset. Nonetheless, these nominal associations of systemic inflammatory regulators and PDR require validation in larger cohorts.

Topics: Diabetes Mellitus; Diabetic Retinopathy; Genome-Wide Association Study; Humans; Interleukin-12; Interleukin-8; Mendelian Randomization Analysis

PurposeThe aim of the present study is to investigate the association of the polymorphism of two genes in CXC chemokine family, interleukin-8 (IL-8) and interferon-inducible protein 10 (IP-10), with both susceptibility and progression of DR in T2D population of northern China.Patients and methodsA total of 1043 eligible type 2 diabetic patients from Heilongjiang of northern China were recruited for this study. They were grouped into: with diabetic retinopathy (DR, 528 cases) and without diabetic retinopathy (DNR, 515 cases). Single nucleotide polymorphism (SNP) genotyping of IL-8(-251T/A) and IP-10(-1596C/T) was performed by polymerase chain reaction. Multivariate analysis and stepwise multiple logistic progression analysis were conducted to evaluate the association between gene SNP and DR susceptibility and progression. Pooled odds ratio (OR) with 95% confidence interval (CI) was applied to assess the strength of the association among study groups.ResultsThe occurring of IL-8(-251) AA genotype was correlated with susceptibility (OR: 2.286, 95% CI: 1.382-3.782, P=0.001) and progression of high-risk proliferative diabetic retinopathy (PDR) (OR: 0.354, 95% CI: 0.162-0.770, P=0.009). Reversely, T allele of IP-10 (-1596) C/T was correlated with a reduced risk of DR (OR: 0.341, 95% CI: 0.249-0.466, P<0.001). However, gene polymorphisms of IL-8-251T/A and IP-10-1596C/T were not associated with diabetic macular edema (DME)(P>0.05).ConclusionsAA genotype of IL-8-251T/A was closely correlated to DR and high-risk proliferative diabetic retinopathy (PDR). -1596T allele of the IP-10 is a beneficial genotype for DR.

Topics: Adult; Aged; Aged, 80 and over; Asian People; Case-Control Studies; Chemokine CXCL10; China; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Disease Progression; Female; Genetic Predisposition to Disease; Genotype; Humans; Interleukin-8; Male; Middle Aged; Polymorphism, Single Nucleotide

Matrix metalloproteinases (MMPs) comprise a family of over 20 structurally related proteins which are zinc-dependent and calcium-activated endopeptidases. The members of this family are able to degrade most extracellular matrix (ECM) proteins and are thus involved in tissue remodeling and contribute to cell migration by eliminating extracellular matrix and basement membrane barriers. Of the MMPs, MMP-2 and MMP-9 are especially active in the degradation of type IV collagen, the main constituent of the basement membrane. MMPs also cleave a variety of non-ECM proteins, including cytokines, chemokines, and growth factors. MMPs and their inhibitors (TIMPs) play important roles in physiological processes such as embryogenesis and wound healing; however, these enzymes are also involved in the pathogeneses of many diseases, such as cancer and atherosclerosis. In these pathological conditions the balance between MMPs and TIMPs shifts in favor of MMPs, resulting in excessive degradation of ECM. Research results published recently show that these enzymes can also be involved in the pathogenesis of diabetes mellitus and diabetic complications such as diabetic retinopathy. MMP-9 has the ability to degrade insulin and is able to activate IL-8, the main chemoattractant factor for neutrophils and monocytes. In addition, MMP-9 enables infl ammatory cell migration and pancreas colonization by eliminating the basement membrane barriers. Type IV collagenases are also important for endothelial cell invasion occurring during neovascularization (diabetic retinopathy), as angiogenesis needs extracellular matrix degradation; what is more, these enzymes are able to degrade pigment epithelium-derived factor, which is the principal antiangiogenic protein of the eye.

Topics: Basement Membrane; Cell Movement; Diabetes Mellitus; Diabetic Retinopathy; Disease Progression; Humans; Interleukin-8; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinases; Neovascularization, Pathologic; Retinal Vessels

Inhibition of inflammatory cytokines may have therapeutic effects in diabetic retinopathy (DR).. To compare aqueous and vitreous levels of 17 inflammatory cytokines in patients treated preoperatively with topical ketorolac tromethamine, 0.45%, or placebo before pars plana vitrectomy for complications related to proliferative DR (PDR).. A prospective, randomized, placebo-controlled, patient-masked interventional study performed in a university academic hospital included 20 eyes from 20 patients undergoing pars plana vitrectomy for complications of PDR.. Eyes were randomized to ketorolac tromethamine, 0.45% (Acuvail), or placebo 4 times daily for 3 days before pars plana vitrectomy. Undiluted aqueous and vitreous samples were taken at the time of surgery and immediately frozen at -80°C.. Aqueous and vitreous levels of prostaglandin E2 and 16 other inflammatory cytokines implicated in the pathogenesis of DR.. Prostaglandin E2, platelet-derived growth factor (PDGF) AA, eotaxin, vascular endothelial growth factor, interferon γ-inducible protein of 10 kDa, monocyte chemoattractant protein 1, growth-related oncogene, interleukin 6, interleukin 8 (IL-8), and tumor necrosis factor were detectable in the aqueous and vitreous of at least half of the eyes, and these cytokines were analyzed further. Aqueous levels were lower in the ketorolac group for all cytokines detected, but only the difference in IL-8 was statistically significant (52% reduction; P = .04). Levels of IL-8 (41% reduction; P = .002) and PDGF-AA (21% reduction; P = .009) were significantly lower in the vitreous of patients treated with ketorolac.. Topical ketorolac tromethamine, 0.45%, significantly lowered aqueous IL-8 levels and vitreous IL-8 and PDGF-AA levels in this series of eyes, suggesting that it may cause meaningful inhibition of inflammatory cytokines implicated in the pathogenesis of DR.. clinicaltrials.gov Identifier: NCT01609881.

Topics: Administration, Topical; Anti-Inflammatory Agents, Non-Steroidal; Aqueous Humor; Diabetic Retinopathy; Dinoprostone; Double-Blind Method; Female; Humans; Immunoenzyme Techniques; Interleukin-8; Ketorolac Tromethamine; Male; Middle Aged; Ophthalmic Solutions; Platelet-Derived Growth Factor; Prospective Studies; Vitrectomy; Vitreous Body

The purpose of this study was to determine the concentrations of angiogenic and inflammatory markers in human eyes with diffuse diabetic macular edema before and during therapy with intravitreal bevacizumab and their association with disease activity.. In a prospective clinical trial, 10 eyes of 10 consecutive patients with vision loss because of diabetic macular edema were compared with 10 eyes of 10 age-matched controls. Bevacizumab was administered at baseline; retreatments were given monthly according to disease activity. During a follow-up of 6 months, aqueous humor samples were taken each time intravitreal therapy was administered. A multiplex assay was used for measurement of 12 different growth factors and cytokines.. Aqueous humor of eyes with diabetic macular edema demonstrated a significantly increased expression of monocyte chemoattractant protein-1 and interleukin-8 and higher, but not significant, levels of interleukin-6 and vascular endothelial growth factor. Intravitreal therapy with bevacizumab resulted in a significant decrease of vascular endothelial growth factor below physiologic levels. This change was not associated with clinical disease activity as measured by visual acuity and central retinal thickness.. Eyes with diabetic macular edema showed a different profile of monocyte chemoattractant protein-1 and interleukin-8 as compared with controls. The intraocular vascular endothelial growth factor expression decreased significantly after the first intravitreal injection of bevacizumab; this reduction was prolonged by consecutive monthly retreatment.

Topics: Aged; Angiogenesis Inhibitors; Angiogenic Proteins; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Aqueous Humor; Bevacizumab; Biomarkers; Chemokine CCL2; Diabetic Retinopathy; Humans; Inflammation Mediators; Interleukin-8; Intravitreal Injections; Macular Edema; Prospective Studies; Retreatment; Vascular Endothelial Growth Factor A

Diabetic retinopathy is a frequent complication of diabetes mellitus and a leading cause of blindness in adults. The objective of this study was to elucidate the diabetic retinopathy pathophysiology in more detail by comparing protein alterations in human vitreous of different diabetic retinopathy stages.. Vitreous samples were obtained from 116 patients undergoing pars plana vitrectomy. Quantitative immunoassays were performed of angiogenic factors (VEGF-A, PIGF, Angiopoietin-1, Angiopoietin-2, Galectin-1) as well as cytokines (IL-1β, IL-8, IFN-γ, TNF-α, CCL3) in samples from control patients (patients who don't suffer from diabetes; n = 58) as well as diabetes mellitus patients without retinopathy (n = 25), non-proliferative diabetic retinopathy (n = 12), and proliferative diabetic retinopathy patients (n = 21). In addition, correlation analysis of protein levels in vitreous samples and fasting glucose values of these patients as well as correlation analyses of protein levels and VEGF-A were performed.. We detected up-regulated levels of VEGF-A (p = 0.001), PIGF (p<0.001), Angiopoietin-1 (p = 0.005), Angiopoietin-2 (p<0.001), IL-1β (p = 0.012), and IL-8 (p = 0.018) in proliferative diabetic retinopathy samples. Interestingly, we found a strong positive correlation between Angiopoietin-2 and VEGF-A levels as well as a positive correlation between Angiopoietin-1 and VEGF-A.. This indicated that further angiogenic factors, besides VEGF, but also pro-inflammatory cytokines are involved in disease progression and development of proliferative diabetic retinopathy. In contrast, factors other than angiogenic factors seem to play a crucial role in non-proliferative diabetic retinopathy development. A detailed breakdown of the pathophysiology contributes to future detection and treatment of the disease.

Topics: Adult; Angiopoietin-1; Angiopoietin-2; Cytokines; Diabetes Mellitus; Diabetic Retinopathy; Female; Humans; Interleukin-8; Placenta Growth Factor; Vascular Endothelial Growth Factor A; Vitrectomy; Vitreous Body

The aim was to study aqueous humour inflammatory mediators' levels in a cohort of Egyptian patients with diabetic macular oedema (DMO).. This was a case-control prospective study conducted on 2 groups: 25 eyes of 22 (11 females) patients seeking treatment for DMO as patients group, and 10 eyes of 10 (4 females) cataract patients as a control group. Aqueous humour was aspirated before intravitreal injection (patients' group) or cataract surgery (control group). Inflammatory mediators in aqueous humour were measured using a multiplex bead immunoassay kit of 27 pre-mixed cytokines.. Eotaxin, interferon gamma-induced protein 10 (IP-10), monocyte chemoattractant protein-1 (MCP-1/CCL2) and interleukin-8 (IL-8/CXCL8) were found significantly higher in patients' group compared to control group (p = 0.043, 0.037, 0.001, 0.015 respectively). On the contrary, interferon-gamma (IFN-gamma) and granulocyte colony-stimulating factor (G-CSF) were found significantly higher in control group than patients' group (p = 0.003, 0.019 respectively). Basic fibroblast growth factor (Basic-FGF/FGF-2) and interleukin-1 receptor antagonist (IL-1ra) were found higher (but not statistically significant) in controls (p = 0.100 and 0.070 respectively). Additionally, a negative and significant correlation was found between Eotaxin level in aqueous humour and central macular thickness.. Some mediators might be implicated in the pathogenesis of DMO either augmenting or suppressing role. Eotaxin, IP-10, MCP-1 and IL-8 might have a role in cases not responding to standard anti-vascular endothelial growth factor (VEGF) therapy. IL-1ra might have a protective role; therefore, the effectiveness of intravitreal injection of IL-1ra homologue needs to be studied in future clinical trials.

Topics: Aqueous Humor; Cataract; Chemokine CXCL10; Cytokines; Diabetes Mellitus; Diabetic Retinopathy; Egypt; Female; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-8; Macular Edema; Prospective Studies

Diabetic retinopathy (DR) is a microvascular inflammatory and neurodegenerative disease. The purpose of this study was to analyze the relationship between DR severity and the levels of potential biomarkers in the serum and/or vitreous.. A prospective, consecutive, controlled, observational study was performed between June 2018 and January 2020. Blood and vitreous samples were collected on the day of vitrectomy in patients without diabetes and in patients with diabetes with epiretinal membrane, macular edema, and indication for vitrectomy.. Transthyretin (TTR) was the only blood biomarker with levels statistically higher in patients with diabetes (p = 0.037). However, no correlation with DR severity was observed. Erythropoietin (EPO) was the only blood biomarker whose levels were associated with DR severity (p = 0.036). In vitreous samples, levels of EPO (p = 0.011), interleukin (IL)-6 (p < 0.001), IL-8 (p < 0.001), IL-17 (p = 0.022), monokine induced by interferon-γ (MIG) (p < 0.001), and interferon gamma-induced protein 10 (IP-10) (p = 0.005) were significantly higher in patients with diabetes. Additionally, in vitreous, IL-6, IL-8, MIG, and IPL-10 levels were also higher in more severe DR cases (p < 0.05).. Among the studied biomarkers, vitreous IL-6, IL-8, MIG, and IP-10 were the ones whose levels had the strongest coherent relationship with DR severity prediction and, thus, have the best potential post-vitrectomy prognostic value.

Topics: Biomarkers; Chemokine CXCL10; Diabetes Mellitus; Diabetic Retinopathy; Humans; Interleukin-6; Interleukin-8; Neurodegenerative Diseases; Prospective Studies; Vitrectomy; Vitreous Body

We sought to investigate the association between inflammatory cytokine levels and retinal capillary nonperfusion area in eyes with quiescent proliferative diabetic retinopathy (PDR).. Samples of aqueous humor were collected from 67 eyes (n = 42 patients) with treatment-naïve PDR. Levels of interleukin (IL)-10, IL-1β, IL-6, IL-8, monocyte chemoattractant protein 1 (MCP-1), and tumor necrosis factor-α (TNF-α) were obtained using multiplex bead assay. Areas of capillary nonperfusion at the posterior pole and peripheral retina were measured via ultra-widefield fluorescein angiography and correlated with cytokine levels.. The levels of IL-10, IL-6, IL-8, MCP-1, and TNF-α were positively correlated with the nonperfusion area of the peripheral retina (r = 0.298, 0.401, 0.265, 0.435, and 0.393; all P ≤ 0.030). There were positive correlations between IL and 10, IL-6, IL-8, MCP-1, and TNF-α (all R ≥ 0.247; all P ≤ 0.043). IL-1β did not show a significant correlation with the nonperfusion area (P = 0.972 for posterior pole and 0.392 for periphery) but was positively correlated with TNF-α (r = 0.334; P = 0.006).. An increased level of inflammation was observed in PDR eyes with larger nonperfusion areas, which suggests inflammation as a possible target for suppressing PDR progression associated with nonperfusion.

Topics: Cytokines; Diabetes Mellitus; Diabetic Retinopathy; Humans; Inflammation; Interleukin-6; Interleukin-8; Retina; Tumor Necrosis Factor-alpha

Diabetic retinopathy (DR) is characterized by microvascular changes including ischemia. Degradation and metabolic changes of various retinal cells occur during ischemia. Ischemic region containing more cells will lead to greater metabolic impairment. We analyzed the non-perfusion region (NPR) by integrating histologic mapping with ultra-widefield fluorescein angiography (UWF FA) images. We also investigated the correlations of the weighted ischemic index (ISI) considering the regional distribution of retinal cells with cytokines, macular edema (ME), and neovascularization (NV). In this study, 32 patients with treatment-naïve DR and 21 age-matched control participants were included. The difference between the non-weighted and weighted ISI of NPR with leakage was greatest at the posterior region. The weighted ISI of NPR with leakage was correlated with MCP-1, IL-8, IL-6, PlGF, and VEGF-A levels, while the non-weighted ISI of NPR with leakage was correlated with IL-8 and IL-6 levels. The presence of baseline ME or NV in patients with DR was associated with the weighted ISI, with a stronger association when cones and rods were weighted. The weighted ISI reflecting both metabolic activity and cell distribution demonstrated a better correlation with clinical features and was more valuable in NPR with leakage than non-weighted ISI, which previous studies conventionally used.

Topics: Diabetes Mellitus; Diabetic Retinopathy; Fluorescein Angiography; Humans; Interleukin-6; Interleukin-8; Ischemia; Macular Edema; Retinal Vessels; Tomography, Optical Coherence; Vascular Endothelial Growth Factor A; Visual Acuity

To determine the associations between aqueous humor cytokine levels and the severity of diabetic retinopathy and the prior panretinal photocoagulation (PRP) status of patients with diabetic macular edema (DME).. We divided 98 DME patients into those with nonproliferative diabetic retinopathy (NPDR), proliferative diabetic retinopathy (PDR), and PRP patients. We compared the concentrations of interleukin- (IL-) 1. The aqueous humor levels of IL-6, IL-8, VEGF, and PlGF were significantly higher in the PDR group than in the NPDR group. The PlGF and VEGF levels in the PDR group were significantly higher than those in the PRP group. On PRP subgroup analyses, patients who had undergone PRP within 6 months prior exhibited higher levels of VEGF, PlGF, and TNF-. PDR patients exhibited higher concentrations of VEGF and certain inflammatory cytokines than did NPDR and PRP patients. In the latter patients, the intraocular VEGF and inflammatory cytokine levels fell gradually over time.

Topics: Aqueous Humor; Cytokines; Diabetes Mellitus; Diabetic Retinopathy; Female; Humans; Interleukin-6; Interleukin-8; Light Coagulation; Macular Edema; Placenta Growth Factor; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Background and Objectives: Cytokines are cell-signaling proteins whose identification may serve as inflammatory markers or early indicators for progressive disease. The aim of our study was to quantify several cytokines in aqueous humor (AH) and their correlations with biochemical parameters in diabetic eyes with non-proliferative diabetic retinopathy (NPDR). Materials and Methods: A total of 62 eyes from 62 patients were included in the study: 37 eyes from nondiabetic patients (group 1), 13 diabetic eyes with no retinopathy changes (group 2) and 12 diabetic eyes with early and moderate NPDR (group 3). AH samples were collected during uneventful cataract surgery. The cytokines IL-1β, IL-6, IL-8, IL-10, IL-12, IP-10, MCP-1, TNF-α and VEGF were quantified using multiplex bead-based immunoassay. Due to unreliable results, IL-1β, TNF-α, IL-10 and IL-12 were excluded. Concentrations were compared between groups. Biochemical parameters (fasting blood sugar, glycated hemoglobin, C-reactive protein) and the duration of diabetes were recorded. Results: VEGF levels were significantly different between groups (p = 0.001), while levels of IL-6, IL-8, IP-10 and MCP-1 were comparable across all groups (p > 0.05). IL-6 concentration correlated with VEGF in group 1 (rho = 0.651, p = 0.003) and group 3 (rho = 0.857, p = 0.007); no correlation could be proved between IL-6, IL-8, IP-10, MCP-1 or VEGF and biochemical parameters. Duration of diabetes was not correlated with the cytokine levels in groups 2 and 3. The receiver operating characteristic (ROC) curve revealed that VEGF concentrations could discriminate early and moderate NPDR from diabetes, with an area under the curve (AUC) of 0.897 (p = 0.001, 95% CI = 0.74−1.0). Conclusions: Diabetes mellitus induces significant intraocular changes in the VEGF expression in diabetic patients vs. normal subjects, even before proliferative complications appear. VEGF was increasingly expressed once the diabetes progressed from no retinopathy to early or moderate retinopathy.

Topics: Aqueous Humor; Chemokine CXCL10; Cytokines; Diabetes Mellitus; Diabetic Retinopathy; Humans; Interleukin-10; Interleukin-12; Interleukin-6; Interleukin-8; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Diabetic retinopathy (DR), as one of the most important causes of blindness in Western societies, is a common micro-vasculopathy associated with diabetes. There is growing evidence of the role of inflammation in its development. This study was designed to measure cytokines in patients with diabetes with different stages of retinopathy .In this study, tear concentrations of three types of cytokines with different angiogenic properties including IL-1RA, IL-8, and TNF-α were measured in patients with diabetes without retinopathy, with non-proliferative retinopathy, with proliferative retinopathy, and in a healthy control group. The results showed that concentrations of TNF-α and IL-8 were higher in the tear sample of diabetics than in the control group and the concentrations of these cytokines were higher in patients with more advanced stages of diabetes, while the tear level of IL-1RA was significantly lower in diabetics. Based on these findings, it can be concluded that diabetes and its progression of severity affects the tear levels of IL-1RA, IL-8, and TNF-α inflammatory cytokines.

Topics: Cytokines; Diabetes Mellitus; Diabetic Retinopathy; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-8; Tears; Tumor Necrosis Factor-alpha

Quantitative analysis of vitreous inflammatory and angiogenic factors from patients with proliferative diabetic retinopathy (PDR) or diabetic macular edema (DME).. Collection of undiluted vitreous samples from 20 diabetic patients: 13 with proliferative diabetic retinopathy (PDR) and 7 with diabetic macular edema (DME). DME patients had suboptimal response to anti-VEGF treatment. Samples from 11 control patients, with vitreomacular interface pathology such as idiopathic epiretinal membrane (iERM) (n = 4), vitreomacular traction syndrome (VMT) (n = 3) and full thickness macular hole (FTMH) (n = 3), were also collected. The levels of IL1b, IL6, IL8, IL27, TNFα, ICAM-1, VCAM, MCP-1, VEGFA and LCN2 were measured using cytometry flow analysis. Median values were compared with Mann-Whitney test since the distributions were skewed. Statistical analysis was performed with the Statistical Package for Social Sciences software (IBM Corp. Released 2012. IBM SPSS Statistics for Windows, Version 21.0. Armonk, NY: IBM Corp.).. The median concentration of LCN2, IL6, IL8, IL1b, IL27, ICAM, VCAM-1, MCP-1, TNFa and VEGFA was higher in PDR patients than in controls. Similarly, the median concentration of LCN2, IL6, IL8, IL27, ICAM, VCAM-1, TNFa and VEGFA was higher in DME patients than in controls. In particular, median LCN2 concentration in diabetic patients was 5,711 pg/ml (interquartile range [IR] = 2,534), while in controls was 2,586 pg/ml (IR = 2,345). Moreover, median LCN2 was 6,534 pg/ml in the DME group (IR = 6,850) and 4,785 pg/ml in the PDR group (IR = 2,608), (p = 0.025).. Various inflammatory and angiogenic factors are involved in the pathophysiology of PDR and DME. Elevated vitreous levels of LCN2 in PDR and especially in DME patients reveal a potential pathogenic association. More extended studies could verify LCN2 as an alternative therapeutic target.

Topics: Angiogenesis Inducing Agents; Diabetes Mellitus; Diabetic Retinopathy; Humans; Interleukin-27; Interleukin-6; Interleukin-8; Lipocalin-2; Macular Edema; Vascular Cell Adhesion Molecule-1; Vitreous Body

Topics: Apoptosis; bcl-2-Associated X Protein; Diabetic Retinopathy; Glucose; Glutathione Peroxidase; Humans; Interleukin-8; NF-E2-Related Factor 2; Oxidative Stress; Polygonatum; Polysaccharides; Reactive Oxygen Species; Retinal Pigment Epithelium; Superoxide Dismutase; Tumor Necrosis Factor-alpha

In eyes with diabetic macular oedema (DME), aqueous humour (AH) cytokine levels before and after anti-vascular endothelial growth factor (VEGF) treatment were compared and correlated with optical coherence tomography structural parameters.. This prospective study included 56 control patients with cataracts and 83 patients with DME manifesting as diffuse retinal thickening (DRT), cystoid macular oedema and serous retinal detachment (SRD). AH samples were obtained before intravitreal injection of anti-VEGF or cataract surgery. VEGF, interleukin (IL)-6, IL-8, IL-10, interferon-inducible protein 10 (IP-10) and monocyte chemotactic protein 1 (MCP-1) levels were measured by multiplex bead assay. AH cytokine levels, central macular thickness (CMT), number of hyper-reflective foci (HF), continuity of external limiting membrane and ellipsoid zone (EZ) and best-corrected visual acuity were evaluated.. In SRD, IL-6 and MCP-1 levels and HF were increased (all p < 0.05) compared to DRT. At baseline, the number of HF was correlated with VEGF, IL-6, IL-8, IP-10 and MCP-1 (all p < 0.05). Eyes sensitive to anti-VEGF treatment had high baseline levels of VEGF, MCP-1, HF and many EZ disruptions (all p < 0.05). DME patients with normal VEGF levels but with high levels of IL-8, IP-10 and MCP-1 (all p < 0.05) had little change in CMT after anti-VEGF treatment (p = 0.678).. AH concentrations of some inflammatory cytokines in DME were differentially expressed among the three DME morphologies. HF was associated with VEGF and other inflammatory cytokine levels. Multiple HF at baseline predicted a significant decrease in CMT, and eyes with normal VEGF but increased inflammatory cytokines may be insensitive to anti-VEGF treatment.

Topics: Angiogenesis Inhibitors; Chemokine CXCL10; Cytokines; Diabetes Mellitus; Diabetic Retinopathy; Humans; Interleukin-6; Interleukin-8; Intravitreal Injections; Macular Edema; Prospective Studies; Tomography, Optical Coherence; Vascular Endothelial Growth Factor A; Visual Acuity

Identification of nonresponders prior to anti-vascular endothelial growth factor (anti-VEGF) therapy would help in the judicious clinical management of diabetic macular edema (DME) patients. Thus, a systematic study was initiated to identify nonresponding DME patient population undergoing ranibizumab treatment to figure out additional inflammatory components that may contribute to their nonresponsiveness to anti-VEGF therapy.. A total of 40 patients recruited to this investigator-initiated trial received intravitreal ranibizumab monthly for 3 months. The fourth- and fifth-month injections were according to PRN protocol and the sixth-month injection was mandatory. Best-corrected visual acuity (BCVA), central macular thickness (CMT), and VEGF in aqueous humor were measured for all the patients. Patients were grouped into responders/nonresponders on the formulated criteria and the levels of key pro-inflammatory cytokines were also measured between the two groups at baseline, 2 month and 5 months using cytometric bead array (CBA).. Eleven patients were categorized (29.72%) as responders and 10 patients (27.02%) as nonresponders. Nonresponders showed poorer BCVA (P = 0.024, 0.045, and 0.048 for 4, 5, and 6 months) and higher CMT (P = 0.021, 0.0008 and <0.0001 for baseline, 1, 2, 3, 4, 5, and 6 months) compared to responders. The cytokines IL-8, MCP-1 were significantly up regulated (P = 0.0048 and 0.029 for MCP-1 and IL-8) in nonresponders.. Elevated MCP-1 and IL-8 levels found in the nonresponders could be used as a prognostic marker to identify these groups of patients and can help in developing alternative treatment options along with anti-VEGF therapy.

Topics: Angiogenesis Inhibitors; Chemokine CCL2; Diabetes Mellitus; Diabetic Retinopathy; Humans; Interleukin-8; Intravitreal Injections; Macular Edema; Ranibizumab; Tomography, Optical Coherence; Vascular Endothelial Growth Factor A; Visual Acuity

To determine the associations between the levels of certain cytokines in the aqueous humor and the severity of diabetic retinopathy.. A total of 103 patients (one eye per patient) who received intravitreal injection with ranibizumab for diabetic retinopathy were enrolled and divided into 3 groups: nonproliferative diabetic retinopathy (NPDR) with macular edema group (42 eyes), proliferative diabetic retinopathy (PDR) group (40 eyes) and neovascular glaucoma due to PDR (NVG-PDR) group (21 eyes). The concentrations of interleukin (IL)-6, IL-8, IL-10, vascular endothelial growth factor (VEGF), transforming growth factor-β (TGF-β), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and monocyte chemoattractant protein-1 (MCP-1) in the aqueous humor were measured.. In this study, 42, 40 and 21 patients (one eye per patient) were included in the NPDR, PDR and NVG-PDR groups, respectively. The median concentrations of IL-6, IL-8, IL-10, VEGF, TGF-β, VCAM-1, ICAM-1 and MCP-1 in the groups were measured. The levels of these 8 cytokines increased with the severity of diabetic retinopathy, especially in the NVG-PDR group. Compared with those in the NPDR group, the aqueous concentrations of these 8 cytokines were higher in the PDR group and were the highest in the NVG-PDR group. There were significant differences in all cytokines among the three groups (P < 0.05). Multivariate analysis showed that in the NPDR and PDR groups, the risk of PDR associated with elevated levels of TGF-β (P = 0.0004, OR 1.11, 95% CI [1.05-1.18]) and ICAM-1 (P = 0.0408, OR 10.75, 95% CI [1.10-104.61]). In the PDR and NVG groups, the risk of NVG associated with elevated levels of IL-10 (P = 0.0486, OR 0.7040, 95% CI [0.4966, 0.9979]), VEGF (P = 0.0279, OR 0.9963, 95% CI [0.9931, 0.9996]), and VCAM-1 (P = 0.0316, OR 0.9998, 95% CI [0.9996, 0.99998]). In the three groups, the risk of developing NVG associated with elevated levels of TGF-β (P < 0.001, OR 1.04, 95% CI [1.02, 1.05]).. The levels of these eight cytokines in the aqueous humor increased with the severity of diabetic retinopathy, especially in NVG-PDR. This study suggests that TGF-β, ICAM-1, IL-10, VEGF, and VCAM-1 may play a role in the progression of diabetic retinopathy, especially TGF-β, which may plays a significant role in NVG-PDR. These cytokines potentially may be used as biomarkers to predict the progress of diabetic retinopathy, contribute to the choice of treatment options and/or monitor treatment responses.

Topics: Aqueous Humor; Chemokine CCL2; Cytokines; Diabetes Mellitus; Diabetic Retinopathy; Glaucoma, Neovascular; Humans; Intercellular Adhesion Molecule-1; Interleukin-10; Interleukin-6; Interleukin-8; Macular Edema; Ranibizumab; Transforming Growth Factor beta; Vascular Cell Adhesion Molecule-1; Vascular Endothelial Growth Factor A

The purpose of this study was to investigate the levels of cytokines in the vitreous, and their correlation with the density of inflammatory cells in fibrovascular membranes (FVMs) in patients with proliferative diabetic retinopathy (PDR) to evaluate intraocular inflammatory conditions with regard to disease activity.. Thirty-three patients (33 eyes) with PDR requiring vitreoretinal surgery because of FVMs and tractional detachment were enrolled in the study, and compared with 20 patients (20 eyes) with macular hole (MH; control group). All patients underwent complete ophthalmological examinations before surgery. The activity of the disease was noted in patients with PDR. Samples of vitreous and blood were taken, and cytokine (MCP-1, IL-8, IL-6, VEGF, IL-1β, TNF-α, MIP-1α, MIP-1β, IL-10, and IL-12) levels were measured using cytometric bead array (CBA). Samples of FVMs were analyzed with immunohistochemical methods for the presence of inflammatory cells (CD45+, CD14+, CD3+, CD4+, CD8+, and CD19+ cells), and the numerical areal density was calculated (N. Patients with active PDR had statistically significantly higher levels of MCP-1 (p = 0.003), VEGF (p = 0.009), and IL-8 (p = 0.02) in the vitreous in comparison with those with inactive PDR. CD45+, CD14+, CD3+, CD4+, CD8+, and CD19+ cells were identified in FVMs for patients with PDR. Statistically significantly higher numerical areal density of T lymphocytes (CD3+, CD4+, and CD8+) was demonstrated in patients with active PDR in comparison with patients with inactive PDR. Moderate to strong correlations were found between either MCP-1 or IL-8 in the vitreous, and the numerical areal density of cells (CD45+, CD3+, CD4+, and CD8+) in the FVMs, and weaker between either MCP-1 or IL-8 in the vitreous and the numerical areal density of CD14+ cells in the FVMs.. The correlation of cytokine (MCP-1 and IL-8) vitreous levels with the density of inflammatory cells in FVMs, and differences in cytokine levels in the vitreous between patients with active and inactive PDR, and between the vitreous and serum in PDR indicate the importance of local intraocular inflammation in patients with PDR.

Topics: Adaptor Proteins, Signal Transducing; Aged; Aged, 80 and over; Antigens, CD; Case-Control Studies; Chemokine CCL2; Diabetic Retinopathy; Female; Gene Expression; Humans; Inflammation; Interleukin-10; Interleukin-12; Interleukin-1beta; Interleukin-6; Interleukin-8; Lymphocyte Count; Male; Middle Aged; Retina; Retinal Perforations; T-Lymphocytes; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Vitreoretinal Surgery; Vitreous Body

We designed this study to determine the association between the duration of action of intravitreal dexamethasone implants and aqueous humor biomarkers or optical coherence tomography (OCT) findings of diabetic macular edema (DME) patients. We measured the concentrations of interleukin (IL)-1β, -8, -10, -17; placental growth factor; and vascular endothelial growth factor in the aqueous humor, and identified the number of hyperreflective foci (HF), grades of ellipsoid zone disruptions, and baseline central subfield thicknesses (CSTs) using OCT of patients with DME. The average duration of action of dexamethasone implants was 4.32 ± 1.18 months. In multivariate linear regression analyses, the duration of action was associated with aqueous IL-8 levels and the number of HF (β = -0.016, p = 0.037 and β = -0.073, p = 0.035, respectively). Multivariate logistic regression showed that the number of HF (>10) was significantly associated with a shorter duration (<4 months) of action (odds ratio: 17.17, p = 0.010). The duration of action of intravitreal dexamethasone implants in DME patients was associated with the level of aqueous IL-8 and the number of HF using OCT. Specifically, higher number of HF in the OCT was associated with a shorter duration of action.

Topics: Dexamethasone; Diabetic Retinopathy; Female; Glycated Hemoglobin; Humans; Interleukin-10; Interleukin-17; Interleukin-1beta; Interleukin-8; Intravitreal Injections; Macular Edema; Male; Middle Aged; Multivariate Analysis; Placenta Growth Factor; Prospective Studies; Tomography, Optical Coherence; Vascular Endothelial Growth Factor A

To determine the cytokine levels in vitreous samples of diabetic macular edema (DME) patients in comparison with nondiabetic patients, and to evaluate the effect of subretinal fluid on the cytokine levels of vitreous samples.. In this prospective case-control study, 11 eyes of 11 patients with DME and subretinal fluid, 11 eyes of 11 patients with DME without subretinal fluid, and 14 eyes of 14 patients who had undergone vitreoretinal surgery for the epiretinal membrane or a macular hole (control group) were evaluated. The blood glycated hemoglobin (HbA1c) level, vitreous vascular endothelial growth factor (VEGF), and interleukin-8 (IL-8) levels were determined.. The vitreous VEGF level of patients in DME groups was significantly higher than the control group (p < 0.001) without significant difference between DME patients with and without subretinal fluid (p = 0.796). The vitreous IL-8 level of DME patients with subretinal fluid was significantly higher than both control (p = 0.002) and DME without subretinal fluid groups (p = 0.019). The blood HbA1c level was significantly higher in DME group with subretinal fluid than those without subretinal fluid (8.7 ± 1.32 and 7.1 ± 1.13%, respectively, p = 0.010). The only significant correlation was between vitreous VEGF level and blood HbA1c level in DME patients without subretinal fluid (r = 0.813, p = 0.002).. IL-8 level in vitreous samples was higher in DME patients with subretinal fluid than those without subretinal fluid, suggesting that inflammation is an important factor in the progression of DME leading to the subretinal fluid formation in diabetic patients.

Topics: Adult; Aged; Analysis of Variance; Case-Control Studies; Diabetic Retinopathy; Female; Glycated Hemoglobin; Humans; Interleukin-8; Macular Edema; Male; Middle Aged; Prospective Studies; Retrospective Studies; Subretinal Fluid; Vascular Endothelial Growth Factor A

Chronic hyperglycemia, oxidative stress and inflammation are key players in the pathogenesis of diabetic retinopathy (DR). In this work we study the role of phospholipase D (PLD) pathway in an in vitro model of high glucose (HG)-induced damage. To this end, we exposed human retinal pigment epithelium (RPE) cell lines (ARPE-19 and D407) to HG concentrations (16.5 or 33 mM) or to normal glucose concentration (NG, 5.5 mM) for 4, 24 or 72 h. Exposure to HG increased reactive oxygen species levels and caspase-3 cleavage and reduced cell viability after 72 h of incubation. In addition, short term HG exposure (4 h) induced the activation of early events, that involve PLD and ERK1/2 signaling, nuclear factor kappa B (NFκB) nuclear translocation and IκB phosphorylation. The increment in pro-inflammatory interleukins (IL-6 and IL-8) and cyclooxygenase-2 (COX-2) mRNA levels was observed after 24 h of HG exposure. The effect of selective pharmacological PLD1 (VU0359595) and PLD2 (VU0285655-1) inhibitors demonstrated that ERK1/2 and NFκB activation were downstream events of both PLD isoforms. The increment in IL-6 and COX-2 mRNA levels induced by HG was reduced to control levels in cells pre-incubated with both PLD inhibitors. Furthermore, the inhibition of PLD1, PLD2 and MEK/ERK pathway prevented the loss of cell viability and the activation of caspase-3 induced by HG. In conclusion, our findings demonstrate that PLD1 and PLD2 mediate the inflammatory response triggered by HG in RPE cells, pointing to their potential use as a therapeutic target for DR treatment.

Topics: Blotting, Western; Caspase 3; Cell Line; Cyclooxygenase 2; Diabetic Retinopathy; Extracellular Signal-Regulated MAP Kinases; Glucose; Humans; Interleukin-6; Interleukin-8; Microscopy, Confocal; Microscopy, Fluorescence; NF-kappaB-Inducing Kinase; Oxidative Stress; Phospholipase D; Phosphorylation; Protein Serine-Threonine Kinases; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Retinal Pigment Epithelium; RNA, Messenger

This study evaluates diabetic macular-edema (DME) patients for the effect of intravitreal bevacizumab (IVB) \ injection on macular thickness and proangiogenic biomarkers in serum and vitreous.. Forty DME patients were analyzed for macular thickness (MT). Twelve proangiogenic biomarkers in serum \ and vitreous were analyzed before and after IVB.. Significant decrease in MT with vitreal vascular endothelial growth factor-A (VEGF-A) was observed as expected after IVB, \ while serum VEGF-A did not follow a decreasing trend in contrast to VEGF-C, which decreased both in serum and vitreous. Other \ vitreal factors like bone morphogenetic protein-9 (BMP9) and fibroblast growth factor (FGF) were also significantly decreased, while \ endothelial growth factor (EGF) increased following IVB. Before IVB, significant negative correlations were vitreous BMP9 with serum \ FGF, vitreous human growth factor (HGF) and interleukin-8 (IL-8) with serum endothelin, and vitreous and serum FGF and serum \ placental growth factor (PLGF) with EGF. After IVB, negative correlations in serum vs. vitreous were found for both HGF and PLGF \ with BMP9, and angiopoietin with FGF. Cube average thickness was negatively correlated with serum FGF and positively correlated \ with vitreous PLGF and endothelin.. Vascular endothelial growth factors are not the only factors that cause macular edema in diabetic patients. The effect of IVB \ on different proangiogenic biomarkers indicated a complex interplay of other factors in DME.

Topics: Adult; Aged; Angiogenesis Inhibitors; Angiopoietins; Bevacizumab; Biomarkers; Diabetic Retinopathy; Endothelins; Epidermal Growth Factor; Female; Fibroblast Growth Factors; Hepatocyte Growth Factor; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Intravitreal Injections; Macula Lutea; Macular Edema; Male; Middle Aged; Neovascularization, Pathologic; Placenta Growth Factor; Vascular Endothelial Growth Factor A; Vitreous Body

To investigate the immunopathogenesis of endophthalmitis, and determine if cytokine profiles could serve as biomarkers of disease severity in infectious endophthalmitis.. Vitreous samples of 46 patients clinically diagnosed as endophthalmitis (of which 25 were culture positive) and 20 non-infectious controls from patients with Retinal Detachment (RD) or diabetic retinopathy were included in the study. The cytokine and chemokine expression patterns of 40 immune mediators including 6 antiinflammatory cytokines, 15 proinflammatory cytokines, 9 Growth factors and 10 proinflammatory chemokines in the vitreous were were analyzed by multiplex cytokine immunoassay. In addition, significant immune mediators were correlated with initial and final visual acuity (VA).. Our results demonstrated elevated expression of 16 mediators such as GCSF, GRO, IFN-γ, IL-1α, IL-1β, IL-1 RA, IL-6, IL-8, IP-10, MCP-1, MCP-3, MIP-1α, IL-1β, TGF-α, TNF-α in patients with culture positive endophthalmitis. Cytokine profile expression significantly differed between patients with proven endophthalmitis and the non-infectious controls in heat map analysis. PCoA plot indicated five mediators (IL-1RA, IL-6, IL-8, GRO, G-CSF) as biomarkers that could be Independent Predictors of Disease especially in culture negative cases. Correlation of cytokines with VA revealed strong association between the initial VA and intraocular levels of TGF-α, IL-1β and IL-8 but there was no correlation with the severity or visual outcome of infection.. In comparison to non-infectious ocular conditions, the pathogenesis of infectious endophthalmitis correlates with increased expression levels of IL-1RA, IL-6, IL-8, GRO, G-CSF. Understanding cytokine profiles in culture negative endophthalmitis patients could aid in therapy in non-responders to empirical antibiotic therapy.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Bacterial Infections; Biomarkers; Chemokine CXCL1; Child; Child, Preschool; Diabetic Retinopathy; Endophthalmitis; Female; Gene Expression; Gram-Negative Bacteria; Gram-Positive Bacteria; Granulocyte Colony-Stimulating Factor; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-6; Interleukin-8; Male; Middle Aged; Prospective Studies; Retinal Detachment; Severity of Illness Index; Visual Acuity; Vitreous Body

To compare aqueous angiogenic and inflammatory cytokine concentrations in different patterns of diabetic macular edema (DME) based on optical coherence tomography (OCT).. This prospective study was conducted between July 1, 2015, and March 31, 2016, for 9 months. Aqueous samples were obtained from 52 consecutive DME patients and 16 controls. DME patients were divided according to OCT patterns into diffuse retinal thickening (DRT; n = 17), cystoid macular edema (CME; n = 20) and serous retinal detachment (SRD; n = 15) groups. Interleukin (IL)-6, IL-8, vascular endothelial growth factor (VEGF) and tumor necrosis factor alpha (TNF-α) levels were measured by RayBio. IL-6, IL-8 and VEGF levels differed significantly between three DME groups (p < 0.001 in all cases), but the differences in TNF-α levels were not significant (p = 0.226). VEGF and IL-6 levels correlated with central foveal thickness in DRT and SRD groups, respectively.. Aqueous cytokine levels vary with different morphological patterns of DME though the role of TNF-α needs to be studied further, and both anti-angiogenic and anti-inflammatory agents are required simultaneously for treatment of DME.

Topics: Aqueous Humor; Biomarkers; Cytokines; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; Fovea Centralis; Humans; Interleukin-6; Interleukin-8; Macular Edema; Male; Middle Aged; Prospective Studies; Severity of Illness Index; Tomography, Optical Coherence; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Visual Acuity

To evaluate whether NETosis is involved in cytokine-induced ocular inflammation and to track neutrophil extracellular traps (NET) complexes in patients with proliferative diabetic retinopathy (PDR).. For the animal model, the eyes of C57BL/6J mice were intravitreally injected with interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-α), or saline. Histology and immunofluorescence staining for CD11b, neutrophil elastase (NE), myeloperoxidase (MPO), citrullinated histone 3 (H3Cit), and net-like structure were performed. Vitreous samples were collected from patients with PDR; the PDR1 group had no need for repeated surgical intervention, and the PDR2 group had repeated vitreous bleeding or other complication and controls. Levels of MPO, H3Cit-MPO, and NE-MPO complex were measured with enzyme-linked immunosorbent assay (ELISA).. Massive influx of CD11+ inflammatory cells, involving the anterior and posterior chambers, was observed in the murine eyes 24 h after the IL-8 or TNF-α injections. Cells excreted to their surroundings an extracellular net-like structure positive for NE, MPO, and H3Cit. H3Cit staining was abolished with the DNase I treatment, indicating the presence of extracellular DNA in the net-like structures. The vitreous samples of the patients with PDR2 contained statistically significantly higher levels of MPO (173±230) compared to those of the patients with PDR1 (12.0±33.0, p<0.05) or the controls (0.00, p<0.01). The levels of H3Cit-MPO and NE-MPO complexes were also statistically significantly higher in the patients with PDR2 (776.0±1274, 573.0±911.0, respectively) compared to those in the patients with PDR1 (0, p<0.05) and the controls (0, p<0.05).. This study showed the existence of NETosis in cytokine-induced ocular inflammation in a mouse model and human samples. Furthermore, the extent of NET complex formation was higher in a subset of patients who exhibited more complicated PDR.

Topics: Adult; Aged; Animals; Diabetic Retinopathy; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Extracellular Traps; Female; Histones; Humans; Inflammation; Interleukin-8; Leukocyte Elastase; Male; Mice; Mice, Inbred C57BL; Middle Aged; Peroxidase; Tumor Necrosis Factor-alpha; Uveitis, Anterior; Uveitis, Posterior; Vitreous Body

Clinical trials suggest that fenofibrate reduces the progression of retinopathies in patients with type 2 diabetes. Furthermore, patients with retinopathies have elevated levels of inflammatory chemokines and dysfunctional retinal angiogenesis. Therefore, we investigated the effects of fenofibrate on the production of inflammatory chemokines and genes associated with angiogenesis. Retinal pigment epithelial cells (RPECs) were cultured with IL-1β and fenofibrate ranging from 1-50 μM. ENA-78, IL-8, and RANTES were measured in cell culture by ELISA. ENA-78, ABCA1, and ABCG1 gene expression were tested by RT-PCR. IL-1β significantly induced the production of ENA-78, IL-8, and RANTES. Fenofibrate at concentrations of 25-50 uM blunted the IL-1β induced production of ENA-78 (p < 0.05) with no significant effects on RANTES and IL-8. Fenofibrate also reduced the expression of the ENA-78 gene as well as ABCA1 and ABCG1, which are genes involved in angiogenesis. Fenofibrate decreases ENA-78 production and ABCA1/ABCG1 gene expression in RPECs.

Topics: ATP Binding Cassette Transporter 1; ATP Binding Cassette Transporter, Subfamily G, Member 1; Biomarkers; Cells, Cultured; Chemokine CCL5; Chemokine CXCL5; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Fenofibrate; Humans; Hypolipidemic Agents; Interleukin-1beta; Interleukin-8; Neovascularization, Pathologic; Real-Time Polymerase Chain Reaction; Retinal Pigment Epithelium

Diabetic retinopathy (DR), a major complication of diabetes mellitus, is the leading cause of adult blindness. The Toll-like receptor (TLR) family is believed to be involved in the pathogenesis and progression of DR. Here, we investigated the expression profiles of TLR-2 and TLR-4 in retinal ganglion cells (RGCs), in an attempt to elucidate the role of these molecules in the etiology of DR. In vitro cultured RGCs were divided into control and high-glucose groups. The mRNA and protein levels of TLR-2, TLR-4, and nuclear factor (NF)-κB were detected by real-time PCR and western blotting. RGCs were further transfected with specific siRNA targeting TLR2/TLR4; the proliferation of transfected RGCs and their tumor necrosis factor (TNF)-α and interleukin (IL)-8 secretory capacity were analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and enzyme-linked immunosorbent assays (ELISA), respectively. In a high-glucose environment, TLR-2/4 expression was significantly upregulated in RGCs (while their viability decreased); additionally, NF-κB expression and secretion of TNF-α and IL-8 were significantly increased. Co-silencing of the TLR-2 and TLR-4 genes inhibited NF- κB expression and TNF-α/IL-8 secretion, while increasing the survival rate of RGCs. Therefore, a high-glucose environment can potentiate the expression of TLR-2 and TLR-4 in RGCs, activate the downstream signaling pathway, and increase the secretion of pro-inflammatory factors, thereby aggravating DR.

Topics: Cell Line; Cell Survival; Diabetes Mellitus; Diabetic Retinopathy; Gene Expression Regulation; Glucose; Humans; Interleukin-8; NF-kappa B; Retinal Ganglion Cells; Signal Transduction; Toll-Like Receptor 2; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Limited mechanistic understanding of diabetic retinopathy (DR) has hindered therapeutic advances. Berberine, an isoquinolone alkaloid, has shown favorable effects on glucose and lipid metabolism in animal and human studies, but effects on DR are unknown. We previously demonstrated intraretinal extravasation and modification of LDL in human diabetes, and toxicity of modified LDL to human retinal Müller cells. We now explore pathogenic effects of modified LDL on Müller cells, and the efficacy of berberine in mitigating this cytotoxicity.. Confluent human Müller cells were exposed to in vitro-modified 'highly oxidized, glycated (HOG-) LDL versus native-LDL (N-LDL; 200 mg protein/L) for 6 or 24 hours, with/without pretreatment with berberine (5 μM, 1 hour) and/or the adenosine monophosphate (AMP)-activated protein kinase (AMPK) inhibitor, Compound C (5 μM, 1 hour). Using techniques including Western blots, reactive oxygen species (ROS) detection assay, and quantitative real-time PCR, the following outcomes were assessed: cell viability (CCK-8 assay), autophagy (LC3, Beclin-1, ATG-5), apoptosis (cleaved caspase 3, cleaved poly-ADP ribose polymerase), oxidative stress (ROS, nuclear factor erythroid 2-related factor 2, glutathione peroxidase 1, NADPH oxidase 4), angiogenesis (VEGF, pigment epithelium-derived factor), inflammation (inducible nitric oxide synthase, intercellular adhesion molecule 1, IL-6, IL-8, TNF-α), and glial cell activation (glial fibrillary acidic protein).. Native-LDL had no effect on cultured human Müller cells, but HOG-LDL exhibited marked toxicity, significantly decreasing viability and inducing autophagy, apoptosis, oxidative stress, expression of angiogenic factors, inflammation, and glial cell activation. Berberine attenuated all the effects of HOG-LDL (all P < 0.05), and its effects were mitigated by AMPK inhibition (P < 0.05).. Berberine inhibits modified LDL-induced Müller cell injury by activating the AMPK pathway, and merits further study as an agent for preventing and/or treating DR.

Topics: Analysis of Variance; Antioxidants; Apoptosis; Autophagy; Berberine; Biomarkers; Cell Survival; Cells, Cultured; Diabetic Retinopathy; Ependymoglial Cells; Eye Proteins; Humans; Interleukin-6; Interleukin-8; Lipoproteins, LDL; Nerve Growth Factors; Nitric Oxide Synthase Type II; Oxidative Stress; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Serpins; Tumor Necrosis Factor-alpha

To determine whether vitreal concentrations of MCP-1, IL-6 and IL-8 are altered after vitrectomy in patients with proliferative diabetic retinopathy (PDR) and to investigate whether the altered levels of these cytokines are associated with postoperative macular oedema.. Vitreous samples were collected from 36 eyes of 33 patients with PDR before pars plana vitrectomy without intraocular lens (IOL) implantation, and also from the same 36 eyes during IOL implantation surgery approximately 7 months after the initial vitrectomy. Levels of MCP-1, IL-6, IL-8 and vascular endothelial growth factor were measured by flow cytometry using cytometric bead array (CBA) technology.. The mean vitreous levels of MCP-1, IL-6 and IL-8 in the samples collected before vitrectomy were significantly higher in patients with PDR than in control patients (p<0.0001). The levels of MCP-1 and IL-6 in the samples collected at the time of IOL implantation were significantly higher than those collected before vitrectomy (p<0.05). In contrast, the level of IL-8 was significantly lower after vitrectomy (p<0.05). The levels of IL-6 and IL-8, but not MCP-1, in the vitreous from eyes with PDR were inversely correlated with the interval between the initial vitrectomy and the time of implantation surgery. Among the vitrectomised patients, the mean vitreous level of MCP-1 in eyes with diabetic macular oedema (DME) was significantly higher than in those without DME (p=0.028).. The elevated levels of MCP-1 and IL-6 may indicate prolonged inflammation even after successful vitrectomy, which can cause postoperative DME.

Topics: Chemokine CCL2; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Humans; Interleukin-6; Interleukin-8; Lens Implantation, Intraocular; Macular Edema; Male; Middle Aged; Phacoemulsification; Postoperative Complications; Vascular Endothelial Growth Factor A; Visual Acuity; Vitrectomy; Vitreous Body

To determine the relationship between the retinal morphologic changes and concentrations of intravitreal cytokines in eyes with diabetic macular edema.. A retrospective comparative study was performed. The preoperative optical coherence tomography images were evaluated to determine the presence of serous retinal detachments (SRDs), retinal cystic changes, and retinal swelling. The concentrations of vascular endothelial growth factor, interleukin (IL)-6, and IL-8 in vitreous samples collected during vitrectomy were determined. The correlations between optical coherence tomography parameters, other clinical factors, and the concentration of cytokines were calculated.. Fifty-two eyes (52 patients) were investigated. An SRD was found in 19 of the 52 eyes (36.5%). Multivariate regression analysis showed that IL-6 was the only factor significantly associated with the presence of an SRD (P = 0.001; odds ratio, 1.268; 95% confidence interval, 1.105-1.452). The other morphologic changes, such as retinal cystic changes and retinal swellings, were not significantly associated with the concentrations of intravitreal cytokines.. The significant association of SRD with intravitreal IL-6 indicates that inflammation may play an important role in the development of SRD in diabetic macular edema.

Topics: Adult; Aged; Cytokines; Diabetic Retinopathy; Female; Humans; Interleukin-6; Interleukin-8; Macular Edema; Male; Middle Aged; Retina; Retinal Detachment; Retrospective Studies; Tomography, Optical Coherence; Vascular Endothelial Growth Factor A; Vitrectomy; Vitreous Body

Inflammatory markers have been observed in proliferative diabetic retinopathy (PDR). We assessed vitreous concentrations of adhesion molecules and cytokines in PDR and non-diabetic controls and plasma concentrations to differentiate local inflammation from the breakdown of the blood-retina barrier.. 38 patients with PDR and 16 controls with macular hole or epiretinal membrane underwent vitrectomy. Vitreous and plasma soluble adhesion molecules [sE-selectin, intercellular adhesion molecule (sICAM)-1 and -3, platelet-endothelial cell adhesion molecule (sPECAM)-1, sP-selectin, vascular cell adhesion molecule (sVCAM)-1] and cytokines [interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12 (p70), tumour necrosis factor-α and -β, γ-interferon] were detected by the multiplex assay.. Levels of IL-6 and IL-8 were 26-fold (p = 0.001) and 6-fold higher (p = 0.001) in vitreous than in plasma in PDR. Vitreous IL-10 (p = 0.004), sPECAM-1, sE-selectin, sICAM-1 and sVCAM-1 were higher in PDR than controls (p = 0.001 for all). Adhesion molecule concentrations in vitreous in PDR were less than 10% of those in plasma. IL-10 was lower in vitreous than plasma (3.0 vs. 12.8 pg/ml, p = 0.007), and the vitreous IL-10/IL-8 ratio was significantly lower in PDR than in controls (0.10 vs. 0.55 pg/ml, p = 0.003).. The elevated IL-6 and IL-8 levels in vitreous, but not in plasma, are evidence favouring local over systemic inflammation in PDR. Furthermore, there was imbalance between inflammatory and anti-inflammatory cytokines in the vitreous.

Topics: Aged; Cell Adhesion Molecules; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Female; Humans; Interleukin-6; Interleukin-8; Macular Edema; Male; Middle Aged; Retinal Perforations; Vitrectomy; Vitreous Body

To investigate whether concurrent hypertension affects vitreous cytokine levels in diabetic retinopathy.. Vitreous samples from 41 patients with diabetic retinopathy with or without concurrent hypertension, who underwent vitrectomy, were collected. Vitreous cytokine concentrations were simultaneously measured using flow cytometry. Patients were stratified according to hypertension or other clinical conditions, and the differences in vitreous levels of monocyte chemotactic protein 1, interleukin 8, vascular endothelial growth factor, interferon-inducible protein 10, and monokine induced by interferon gamma were examined.. Vitreous levels of monocyte chemotactic protein 1 and interleukin 8 were significantly (P < 0.05) higher in hypertensive patients than in nonhypertensive patients and were significantly (P < 0.05) higher in active diabetic retinopathy than in inactive diabetic retinopathy. Vitreous levels of vascular endothelial growth factor, interferon-inducible protein 10, and monokine induced by interferon gamma were not affected by the coexistence of hypertension. In multivariate models, active diabetic retinopathy (P = 0.004 and P = 0.007), systolic blood pressure (P = 0.039 and P = 0.041), and hypertension (P = 0.032 and P = 0.035) were significant and independent predictors for increased vitreous monocyte chemotactic protein 1 and interleukin 8 levels.. Both monocyte chemotactic protein 1 and interleukin 8 levels were elevated in the vitreous of patients with diabetic retinopathy and concurrent hypertension. These findings may help to explain the epidemiologic and clinical evidence that systemic hypertension exacerbates diabetic retinopathy.

Topics: Adult; Aged; Chemokine CCL2; Chemokine CXCL10; Diabetic Retinopathy; Female; Fluorescein Angiography; Humans; Hypertension; Immunoassay; Interleukin-8; Male; Middle Aged; Monokines; Vascular Endothelial Growth Factor A; Vitrectomy; Vitreous Body

The purpose of this study was to simultaneously measure the concentrations of multiple cytokines, including vascular endothelial growth factor, in the vitreous of patients with diabetic retinopathy and to examine their relation with clinical findings.. Vitreous samples from 46 eyes with diabetic retinopathy and 19 eyes with nondiabetic macular disease (controls) were used. Nine cytokines were simultaneously measured using a FACSCalibur flow cytometer.. Vascular endothelial growth factor, interleukin-8, monocyte chemotactic protein-1, interferon-inducible protein-10, and monokine induced by interferon-gamma were detected in the vitreous samples, and the concentrations were significantly (P < 0.001) higher in patients with diabetic retinopathy compared with control subjects. Vascular endothelial growth factor, interleukin-8, monocyte chemotactic protein-1, and monokine induced by interferon-gamma concentrations were significantly (P < 0.05) higher in active retinopathy than in inactive retinopathy. Furthermore, a significant (P < 0.01) positive correlation was observed between vascular endothelial growth factor concentration and interleukin-8, monocyte chemotactic protein-1, interferon-inducible protein-10, or monokine induced by interferon-gamma concentration in the vitreous.. Vascular endothelial growth factor, interleukin-8, monocyte chemotactic protein-1, interferon-inducible protein-10, and monokine induced by interferon-gamma were expressed at high levels locally in ocular tissues in diabetic retinopathy, and these cytokines may form a network and interact to impact the pathogenesis of the disease.

Topics: Aged; Chemokine CCL2; Chemokine CXCL10; Chemokine CXCL9; Chemokines; Diabetic Retinopathy; Female; Flow Cytometry; Humans; Immunoassay; Interleukin-8; Male; Middle Aged; Vascular Endothelial Growth Factor A; Vitrectomy; Vitreous Body

The purpose of this study was to compare intravitreous levels of lipopolysaccharide-binding protein and soluble CD14 (sCD14) between patients with proliferative diabetic retinopathy (PDR) and nondiabetic subjects.. This study included 19 consecutive Type 2 diabetic patients with PDR in whom a vitrectomy was performed. Sixteen vitreous humors from nondiabetic patients matched by age, with idiopathic macular holes, were selected from our vitreous bank and used as a control group. Lipopolysaccharide-binding protein was assessed by enzyme-linked immunosorbent assay and sCD14 by a solid-phase enzyme-amplified sensitive immunoassay.. Lipopolysaccharide-binding protein and sCD14 levels were significantly higher in patients with PDR than in the control group (lipopolysaccharide-binding protein, P < 0.001; sCD14, P < 0.01). After correcting for vitreal proteins, the results remained significantly higher in patients with PDR. No differences in serum levels were observed, and we did not find any correlation between serum and vitreous levels. A direct correlation between lipopolysaccharide-binding protein and sCD14 was detected in the vitreous fluid (r = 0.57; P < 0.001) but not in the plasma. Finally, a significant correlation between intravitreal levels of both lipopolysaccharide-binding protein and sCD14 and interleukin-8 and monocyte chemotactic protein-1 was also detected.. Lipopolysaccharide-binding protein and sCD14 are elevated in the vitreous fluid of patients with PDR and thus may play a role in the innate immune response triggered by the inflammatory injury characteristic of PDR.

Topics: Acute-Phase Proteins; Adult; Aged; Carrier Proteins; Case-Control Studies; Chemokine CCL2; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Lipopolysaccharide Receptors; Male; Membrane Glycoproteins; Middle Aged; Retinal Perforations; Vitrectomy; Vitreous Body

The pathophysiology of the early events leading to diabetic retinopathy is not fully understood. It has been suggested that Inflammatory processes are involved in the development of the disease; however, the concentrations of tissue retinal inflammatory mediators and their possible alteration in diabetic retinopathy have not been described. The aim of this work was to study T-helper cell cytokine and chemokine profiles, and tyrosine nitration in retinal tissue of diabetic rats.. Cytokines (interleukin IL-1a, IL-1b, IL-2, IL-4, IL-6, IL-10, TNFa, GM-CSF, IFN-g), chemokines (MIP-1a, MIP-2, MIP-3a, MCP-1, GRO/KC, RANTES, Fractalkine), and tyrosine nitration were measured in retinal homogenate obtained from Long-Evans rats after 5 months of experimental diabetes.. The T-helper type 1 cytokines IL-2 and INF-gamma, in addition to NO production (measured as nitrotyrosine), were found to be significantly elevated in diabetic rat retina homogenates. None of the other cytokines and chemokines studied were affected by the diabetic condition.. Immunoregulatory cytokines belonging to the Th-1 group (IL-2 and IFN-gamma) were increased in the retina of experimental diabetic rats. Moreover, the nitrotyrosine formation (as an expression of increased NO production) was significantly elevated in the diabetic retina, supporting the concept of an inflammatory element in the development of diabetic retinopathy.

Topics: Animals; Cataract; Chemokines; Diabetes Mellitus, Experimental; Diabetic Retinopathy; Disease Models, Animal; Interferon-gamma; Interleukin-10; Interleukin-1alpha; Interleukin-1beta; Interleukin-2; Interleukin-4; Interleukin-6; Interleukin-8; Male; Nitric Oxide; Rats; Rats, Long-Evans; Retina; T-Lymphocytes, Helper-Inducer; Tumor Necrosis Factor-alpha; Tyrosine

The purpose of this study was to determine the inhibitory activity of trans-resveratrol against hyperglycemia-induced inflammation and degradation of gap junction intercellular communication in retinal pigment epithelial cells. Retinal (ARPE-19) cells were incubated with 5.5 mM glucose, 5.5 mM glucose and 10 microM resveratrol, 33 mM glucose, or 33 mM glucose and 0-10 microM trans-resveratrol at 37 degrees C and 5% CO(2) for 9 days. Cell viability was determined by the crystal violet assay. The levels of low-grade inflammation biomarkers interleukin-6 and interleukin-8 (IL-6 and IL-8), angiogenic factors, and vascular endothelial growth factor (VEGF) were determined by the enzyme-linked immunosorbent assay (ELISA). Gap junction intercellular communication (GJIC) was determined by the scrape-load/dye transfer method. The expression levels of protein kinase Cbeta (PKCbeta), connexin 43 (Cx43), transforming growth factor-beta1 (TGF-beta1), and cyclooxygenase-2 (COX-2) were determined by Western blot. Incubation of retinal cells with 10 microM trans-resveratrol in the presence of 5.5 mM glucose did not affect any of the biomarkers investigated. Incubation of ARPE-19 cells with 33 mM glucose for 9 days significantly induced the accumulation of VEGF, IL-6, IL-8, TGF-beta, and COX-2, activation of PKCbeta, and reduction of Cx43 and GJIC. Incubation of ARPE-19 cells with 33 mM glucose in the presence of 0-10 microM trans-resveratrol dose-dependently inhibited VEGF, TGF-beta1, COX-2, IL-6, and IL-8 accumulation, PKCbeta activation, and Cx43 degradation and enhanced GJIC. These data suggest that trans-resveratrol can protect the retinal pigment epithelial cells against hyperglycemia-induced low-grade inflammation and GJIC degradation.

Topics: Cell Line; Connexin 43; Diabetic Retinopathy; Down-Regulation; Gap Junctions; Humans; Hyperglycemia; Interleukin-6; Interleukin-8; Resveratrol; Retinal Pigment Epithelium; Stilbenes; Transforming Growth Factor beta1

To determine whether the vitreous levels of interleukin 8 (IL-8) and vascular endothelial growth factor (VEGF) of patients with proliferative diabetic retinopathy (PDR) were associated with poor visual acuity after vitrectomy.. Observational cross-sectional study. Patient clinical characteristics and preoperative eye characteristics (63 eyes): visual acuity, iris neovascularization, vitreous haemorrhage, macular detachment, macular oedema, active retinal neovascularization, neovascularization of the disc, burned out PDR (defined as natural end stage of PDR with inactive membranes without previously performed laser photocoagulation) and panretinal photocoagulation were registered prior to vitrectomy for each patient. Vitreous VEGF and IL-8 levels were measured using the cytometric bead array method. Poor postoperative visual acuity was defined as visual acuity of <20/200 and was checked 2 years after vitrectomy.. Twenty-one of the 63 eyes (33.3%) had poor visual acuity after vitrectomy. Univariate analysis showed that vitreous levels of IL-8, the absence of panretinal photocoagulation, preoperative macular detachment and poor preoperative visual acuity were significantly associated with poor final visual acuity after vitrectomy. A stepwise multiple logistic regression analysis showed that elevated vitreous levels of IL-8 (p < 0.0001), macular detachment (p = 0.011) and the absence of panretinal photocoagulation (p = 0.03) were independent predictors for poor visual outcome.. Elevated vitreous IL-8 level could either be a marker of ischaemic inflammatory reaction, or it could play a role in deteriorating visual acuity by DR progression or both. Further studies are needed to provide better understanding of IL-8 and inflammation involvement in visual prognosis in PDR.

Topics: Age of Onset; Aged; Aged, 80 and over; Cross-Sectional Studies; Diabetic Retinopathy; Female; Flow Cytometry; Glycated Hemoglobin; Humans; Interleukin-8; Male; Middle Aged; Postoperative Period; Preoperative Care; Prospective Studies; Vascular Endothelial Growth Factor A; Visual Acuity; Vitrectomy; Vitreous Body

To evaluate the usefulness of cytokine levels of peripheral blood in diabetic retinopathy (DR), demographic and biochemical parameters including low-density lipoprotein (LDL) diameter as well as cytokine profiles were analyzed in 74 patients with type 2 diabetes mellitus (DM), with DR (n=46) or without DR (n=28). DM duration was longer in the patients with DR than without (p<0.001). Serum glucose (p=0.005) and total cholesterol (p=0.029) levels were higher in DM patients with DR than DM patients without DR. Plasma LDL diameter, interleukin-6 (IL-6), and interleukin-8 (IL-8) showed significant differences among the different degrees of DR severity in analysis of variance (ANOVA) with no definite trend. The risk of DR in DM patients was decreased by an increase of interleukin-10 (IL-10) level [odds ratio (OR)=0.152; confidence interval (CI): 0.028-0.817]. Plasma LDL diameter was smaller and IL-6 and tumor necrosis factor-alpha (TNF-alpha) levels were higher in DM patients with proliferative diabetic retinopathy (PDR) compared to those with non-proliferative diabetic retinopathy (NPDR) (p<0.05). We found that higher IL-10 levels were related to lower risk of DR in DM patients. Levels of IL-6 and TNF-alpha as well as LDL diameter may be helpful in the prediction of PDR in DM patients with DR.

Topics: Adult; Aged; Cholesterol, HDL; Cholesterol, LDL; Cytokines; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Tumor Necrosis Factor-alpha

To determine the levels of interleukin 8 (IL-8) in the vitreous of patients with proliferative diabetic retinopathy (PDR).. Observational case-control study.. Vitreous fluid samples were obtained by vitreoretinal surgery from 71 eyes of patients with diabetes mellitus type 2 with PDR and from 17 eyes of nondiabetic patients with a macular hole. PDR was classified as active and inactive and subdivided according to the extent of large-vessel gliotic obliteration. The cytokine levels were measured by cytometric bead array method.. The vitreous levels of IL-8 were significantly higher in patients with PDR in comparison with the control subjects (P < .001) and in patients with higher extent of large vessel gliotic obliteration (P < .001). A vitreous level of IL-8 was not associated with the presence of active PDR.. Increased levels of IL-8 in PDR were associated with a higher extent of large-vessel gliotic obliteration.

Topics: Aged; Case-Control Studies; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Gliosis; Humans; Interleukin-8; Middle Aged; Retinal Neovascularization; Vitreous Body

We examined the levels of vitreous chemokines and Sho (Zheng in Chinese) of Chinese-Korean-Japanese medicine in diabetic patients. Patients undergoing vitrectomy were classified into Group 1 (no diabetic retinopathy), Group 2 (diabetic retinopathy with no or a few new vessels), and Group 3 (diabetic retinopathy with many new vessels). The levels of IL-8, MCP-1, MIP-1alpha, MIP-1beta, and RANTES in the vitreous fluid were measured using cytometric bead array method. Sho was determined by the standard diagnostic method of Chinese-Korean-Japanese medicine. Vitreous levels of IL-8 and MCP-1 in Groups 2 and 3 were higher than those in Group 1. MIP-1alpha, MIP-1beta, and RANTES levels in Groups 2 and 3 were almost the same as those in Group 1. The percentage of patients with Keishibukuryo-gan (Guizhifuling-wan in Chinese) sho in Group 3 was higher than that in Group 1. In conclusion, vitreous levels of IL-8 and MCP-1 were high in patients with diabetic vitreoretinopathy. Keishibukuryo-gan sho may be associated with diabetic vitreoretinopathy.

Topics: Aged; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CCL5; Chemokines; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Female; Humans; Interleukin-8; Macrophage Inflammatory Proteins; Male; Middle Aged; Prospective Studies; Vitrectomy; Vitreoretinopathy, Proliferative; Vitreous Body

To determine the intra-vitreous levels of two pro-inflammatory cytokines [interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1)] and the anti-inflammatory cytokine interleukin-10 (IL-10) in patients with proliferative diabetic retinopathy (PDR). In addition, the relationship between the profile of cytokines and PDR activity has also been evaluated.. The study included 22 consecutive diabetic patients with PDR (4 Type 1 and 18 Type 2) on whom a vitrectomy was performed. Sixteen age-matched non-diabetic patients with other conditions requiring vitrectomy, but in which the retina was not directly affected by neovascularization served as a control group. IL-8, MCP-1 and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA).. The vitreal levels of both IL-8 and MCP-1 were strikingly higher in diabetic patients with PDR in comparison with the control group [173.5 (64-1670) vs. 49 pg/ml (25-145), P < 0.001, and 2171 (388-6155) vs. 438 pg/ml (207-1344), P < 0.001, respectively]. In addition, the vitreous concentrations of IL-8 and MCP-1 were higher in patients with active PDR than in those patients with quiescent PDR [324.5 (80-1670) vs. 173.5 pg/ml (64-487), P = 0.06 and 3596 (1670-6155) vs. 1143 pg/ml (388-2500), P = 0.01, respectively]. However, vitreal levels of IL-10 in diabetic patients were similar to that obtained in the control group [2.89 (1.55-5.50) vs. 2.46 pg/ml (2.2-5.41), P = NS].. The pro-inflammatory cytokines IL-8 and MCP-1 are increased in the vitreous fluid of PDR patients without an increase in the anti-inflammatory cytokine IL-10. In addition, both IL-8 and MCP-1 intra-vitreous levels correlated with PDR activity, thus suggesting that these cytokines may be pathogenically important in PDR.

Topics: Aged; Chemokine CCL2; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-10; Interleukin-8; Male; Middle Aged; Vitreous Body

To investigate the role of chemokines in the pathogenesis of proliferative diabetic retinopathy (PDR).. In total, 41 eyes of 38 patients undergoing vitrectomy were divided into two groups; PDR and non-PDR. The PDR group was comprised of 30 eyes, and the non-PDR group of 11 eyes. Vitreous specimens obtained at vitrectomy were centrifuged and separated into supernatants and cellular components. Concentrations of vascular endothelial growth factor(VEGF), interleukin-8 (IL-8), monocyte chemotactic protein-1 (MCP-1), and regulated upon activation, normal T cell expressed and secreted(RANTES) in the supernatants were determined by enzyme-linked immunosorbent assay(ELISA) or chemiluminescence enzyme immunoassay(CLEIA). Expression of VEGF in the cellular components was determined by immunohistochemistry.. Vitreous levels of VEGF(p < 0.05), IL8 (p < 0.0001) and MCP-1 (p < 0.05) in the PDR group were significantly higher than in the non-PDR group. However, there was no significant difference in RANTES between the two groups. There was a significant correlation (p < 0.0001, r = 0.84) between vitreous IL-8 and MCP-1 levels in the PDR group. After immunohistochemical staining with antiVEGF monoclonal antibody, VEGF positivity was localized in polymorphonuclear leukocytes and monocytes of the cellular components of PDR vitreous specimens.. These results indicate that chemokines are possibly involved in the recruitment of neutrophils and monocytes into the vitreous and that they play a role in the intraocular neovascularization characteristic of PDR.

Topics: Chemokine CCL2; Chemokine CCL5; Chemokines; Diabetic Retinopathy; Endothelial Growth Factors; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Luminescent Measurements; Male; Retinal Neovascularization; Vitrectomy; Vitreous Body

Formation of epiretinal membranes (ERMs) after proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR) results in progressive deterioration of vision, but its pathogenic mechanisms are still unknown. This study was conducted to examine the role of nuclear factor kappa B (NF-kappaB) in the formation of ERMs after PDR and PVR.. ERM samples were obtained by vitrectomy from 10 patients with PDR (aged 53+/-12 years with 14+/-5 years of diabetes), 20 patients with PVR, and 17 patients with idiopathic ERMs. Ten PVR and 17 idiopathic ERM samples were processed for reverse transcription-polymerase chain reaction (RT-PCR) analysis. In addition, 10 PDR and 10 PVR membranes were processed for immunohistochemical analysis.. NF-kappaB mRNA expression levels were significantly higher (10 of 10 versus 9 of 17 subjects in idiopathic ERM, p=0.0119) in PVR subjects. Immunohistochemical analysis showed NF-kappaB protein expression in 8 of the 10 PDR samples as well as all 10 PVR samples, and NF-kappaB positive cells were partially double labeled with glial cell markers. Interestingly, NF-kappaB protein was also overlapped with angiogenic factor interleukin-8 (IL-8) in glial cells as well as vascular endothelial cells.. These results suggest that NF-kappaB is involved in the formation of both glial and vascular endothelial cell components, and that these two cell types might have functional interactions that lead to the enlargement of intraocular proliferative membranes.

Topics: Adult; Aged; Diabetic Retinopathy; Endothelium, Vascular; Epiretinal Membrane; Fluorescent Antibody Technique, Indirect; Humans; Interleukin-8; Middle Aged; Neuroglia; NF-kappa B; NF-kappa B p50 Subunit; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-ret; Receptor Protein-Tyrosine Kinases; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Vitrectomy; Vitreoretinopathy, Proliferative

Many cytokines are involved in the pathogenesis of retinal proliferative diseases, but none has been shown to be related to a specific disorder. The aim of this study was to provide a selective marker of diabetes induced proliferative retinopathies.. 10 vitreous samples from 10 subjects affected by quiescent proliferative diabetic retinopathy (PDR), 20 vitreous samples from 20 subjects affected by active PDR, and 15 samples from 15 patients with proliferative vitreoretinopathy (PVR) were studied. Samples from 18 patients with a macular hole (n = 8) or pucker (n = 10) served as controls. Vitreous samples were obtained via pars plana vitrectomy. The polyamines spermidine, putrescine, and spermine, vascular endothelial growth factor (VEGF), interleukin 8 (IL-8), and transforming growth factor 1beta (TGF-1beta) were measured by high performance liquid chromatography (HPLC) and enzyme linked immunosorbent assay (ELISA), and the correlation coefficients between the vitreous polyamine content and VEGF, IL-8, and TGF-1beta levels were determined.. Spermidine and putrescine were expressed in normal vitreous, but spermine was not detectable. In all the test groups spermidine was 3-4 times higher than in control vitreous and putrescine was similarly lower. The spermine content was up to 15 times higher only in vitreous from patients affected by PDR. Correlation coefficients showed that the spermidine and putrescine level variations correlated with the VEGF and IL-8 content in the active PDR and PVR groups, but not in those with quiescent PDR patients, while spermine was correlated to these cytokines in PDR, but not in PVR groups.. These data suggest a significant role for spermidine and putrescine as markers of proliferative diseases of the retina. The increase in spermine, restricted to diabetic states, may indicate that this polyamine is a unique and specific index of PDR.

Topics: Adult; Aged; Biomarkers; Chromatography, High Pressure Liquid; Diabetic Retinopathy; Endothelial Growth Factors; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Lymphokines; Middle Aged; Putrescine; Spermidine; Spermine; Transforming Growth Factor beta; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors; Vitreous Body

Vitreal interleukin (IL)-1beta (IL-1beta), IL-6, IL-8, tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) levels have previously been determined in patients with proliferative diabetic retinopathy (PDR). However, at present there is no cohort study linking serum levels of NO and many inflammatory cytokines such as TNF-alpha, IL-1beta, soluble IL-2 receptor (sIL-2R), IL-6 and IL-8 to the grade of the microvascular complications.. To determine the relation between the stages of DR and the levels of serum NO, TNF-alpha, IL-1beta, sIL-2R, IL-6 and chemokine IL-8 in patients with diabetes compared with healthy controls.. Fifty-three consecutive patients with diabetes (25 men, 28 women) with or without DR and 15 non-diabetic healthy subjects (seven men, eight women) as controls were included in this prospective study. As an indicator for NO, serum total nitrite (NO2- + NO3-) levels (end-product of NO) were measured by the Griess reaction. Serum TNF-alpha, IL-1beta, sIL-2R, IL-6 and IL-8 levels were determined by a spectrophotometric technique using an Immulite chemiluminescent immunometric assay. The patients with diabetes were classified into three groups according to the stage of DR: no DR (NDR; n = 16), non-proliferative DR (NPDR; n = 18) and PDR (n = 19). The data were analysed using a Mann-Whitney U-test and the results were expressed as mean +/- SE (range).. The levels of IL-1beta and IL-6 were below the detection limits of the assay (for each, <5.0 pg/ml) in all patients with diabetes and controls. Soluble IL-2R levels ranged from 260 to 958 U/ml, with the highest values observed in the patients with PDR. In 47 of the 53 samples (89%) tested for diabetic patients, IL-8 levels were above the detection limits of the assay (5.0 pg/ml). IL-8 levels ranged from <5.0 to 25.0 pg/ml, with the highest mean values observed in PDR patients. TNF-alpha was detectable in 46 of 53 patients with diabetes (87%), ranging from <4.0 to 26.4 pg/ml, with again the highest values obtained in the patients with PDR. Serum NO levels ranged from 80 to 188 micromol/l, with the highest values obtained in patients with PDR. Taken together, the mean serum NO, sIL-2R, IL-8 and TNF-alpha levels increased with the stage of DR and the highest levels were found in patients with PDR. The PDR patients had significantly (for each, P < 0.001) higher serum NO (166.8 +/- 3.2 micromol/l), sIL-2R (807.9 +/- 33.3 U/ml), IL-8 (17.9 +/- 0.4 pg/ml) and TNF-alpha (15.0 +/- 0.8 pg/ml) levels compared with NPDR patients (149.5 +/- 2.1, 659.4 +/- 23.4, 12.9 +/- 1.1, 11.5 +/- 0.6, respectively), NDR patients (115.9 +/- 5.8, 373.8 +/- 15.0, 8.3 +/- 1.0, 6.6 +/- 0.9, respectively) and controls (116.6 +/- 2.3, 392.4 +/- 16.6, 7.2 +/- 0.3, 7.3 +/- 0.5, respectively). Serum levels of these parameters for NPDR patients were also significantly (for each, P < 0.01) higher compared with those of NDR patients and controls. On the other hand, serum NO, sIL-2R, IL-8 and TNF-alpha levels of patients with NDR were comparable with those of controls (for each, P > 0.05).. The results of the present study suggest that NO, sIL-2R, IL-8 and TNF-alpha may play important roles in the pathophysiology and progression of DR. We think that these potentially inflammatory cytokines and NO with their endothelial implications may act together during the course and progression of DR. These molecules may serve as therapeutic targets for the treatment and/or prevention of diabetes with its systemic and ocular microvascular complications.

Topics: Adult; Aged; Diabetic Retinopathy; Female; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Interleukins; Male; Microcirculation; Middle Aged; Nitric Oxide; Prospective Studies; Receptors, Interleukin-2; Tumor Necrosis Factor-alpha

To determine vitreous levels of interleukin 8 (IL-8) and interferon-induced protein 10 (IP-10), which are members of the C-X-C chemokine family that promote and inhibit neovascularization, respectively.. We measured the levels of IL-8 and IP-10 by specific enzyme-linked immunosorbent assays in the vitreous from 30 patients with proliferative diabetic retinopathy (PDR) and 10 control patients undergoing vitrectomy for idiopathic macular holes or idiopathic macular puckers.. Detectable levels of IL-8 were found in 23 of 24 patients with active PDR, 4 of 6 patients with inactive PDR, and 6 of 10 controls. Levels of IL-8 were significantly increased in vitreous samples from the patients with active PDR (P = .02) when compared with vitreous samples from the controls. The IL-8 levels detected in vitreous samples from patients with inactive PDR were not significantly elevated over those found in the control samples. Interferon-induced protein 10 was detected in the vitreous samples from 23 of 24 patients with active PDR, all patients with inactive PDR, and 9 of 10 controls. Significant elevations of IP-10 were measured in samples from patients with active PDR (P = .004) and in those with inactive PDR (P = .00) over those from controls. In addition, levels of IP-10 were significantly elevated in vitreous samples from patients with inactive PDR compared with vitreous samples from patients with active PDR (P = .02).. Both IL-8 and IP-10 participate in the pathogenesis of PDR.

Topics: Adult; Aged; Chemokine CXCL10; Chemokines, CXC; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Humans; Interferon-gamma; Interleukin-8; Middle Aged; Vitrectomy; Vitreous Body

To investigate whether the chemokines monocyte chemotactic protein-1 (MCP-1) and interleukin-8 (IL-8) are involved in the pathogenesis of proliferative vitreoretinal disorders and to study their possible interaction with IL-6.. In a prospective study of 125 consecutive patients (125 eyes), vitreous and paired serum samples were obtained and were assayed for MCP-1 and IL-8. Levels of IL-6 were determined by proliferation of the IL-6-dependent hybridoma cell line 7TD1.. Monocyte chemotactic protein-1 was detected in 13 (48%) of 27 vitreous samples from patients with retinal detachment, in five (63%) of eight samples from patients with macular pucker, in 31 (72%) of 43 samples from patients with proliferative vitreoretinopathy, and in 32 (76%) of 42 samples from patients with proliferative diabetic retinopathy, but not in samples from five patients with idiopathic epiretinal membrane. There was a significant (P = .049) correlation between the incidence of MCP-1 detection in retinal detachment, macular pucker, and proliferative vitreoretinopathy groups and the severity of proliferation. Interleukin-8 was detected in two vitreous samples from eyes with retinal detachment, in two samples from eyes with proliferative vitreoretinopathy, and in three samples from eyes with proliferative diabetic retinopathy. Monocyte chemotactic protein-1 levels in the vitreous samples were positively correlated with IL-6 levels (r = .31, P = .01). Interleukin-6 levels were significantly (P = .0097) greater in vitreous samples with than without detectable levels of MCP-1.. Monocyte chemotactic protein-1 is present in a substantial percent of vitreous samples from eyes with proliferative vitreoretinal disorders and may help in stimulating the infiltration of monocytes and macrophages into eyes with these disorders.

Topics: Antibodies, Monoclonal; Chemokine CCL2; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-6; Interleukin-8; Prospective Studies; Retinal Detachment; Vitreoretinopathy, Proliferative; Vitreous Body

We determined whether interleukin-8, monocyte chemotactic protein-1, and macrophage-colony stimulating factor are present in the vitreous of patients with proliferative diabetic retinopathy (PDR) or proliferative vitreoretinopathy (PVR). The levels of these cytokines were measured by specific enzyme-linked immunoassays in vitreous from 30 patients with PDR, 13 patients with PVR, and 26 control individuals, including 10 cadaver eyes and 16 patients with idiopathic macular holes, idiopathic macular puckers, vitreous hemorrhages, or uncomplicated retinal detachments. Detectable levels of interleukin-8 were found in 90% of vitreous samples of patients with PDR, 85% with PVR, and 58% of control samples. IL-8 was significantly increased in PDR (mean +/- SEM; 25.0 +/- 5.3 ng/ml; p = 0.01), but not in PVR (11.9 +/- 3.9 ng/ml; p = 0.50) compared to control human vitreous (8.5 +/- 2.5 2.5 ng/ml). MCP-1 was detected in 90% of vitreous samples of patients with PDR, 92% with PVR, and 81% of control samples. MCP-1 was significantly increased in PDR (6.2 +/- 0.9 ng/ml, p = 0.001) and PVR (7.7 +/- 2.5 ng/ml, p = 0.001) over the levels in control vitreous (1.2 +/- 0.2 ng/ml). M-CSF was detected in 94% of vitreous samples of patients with PDR, 88% with PVR, and 92% from control vitreous. M-CSF was significantly elevated in PDR (32.3 +/- 8.3 ng/ml, p = 0.03), but not in PVR (23.6 +/- 12.8 ng/ml, p = 0.4) compared to control (10.7 +/- 3.5 ng/ml). Our results suggest that IL-8, MCP-1, and M-CSF participate in the pathogenesis of PDR and PVR.

Topics: Chemokine CCL2; Cytokines; Diabetic Retinopathy; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-8; Macrophage Colony-Stimulating Factor; Vitreoretinopathy, Proliferative; Vitreous Body