Nonsteroidal anti‐inflammatory drugs prevent early diabetic retinopathy via TNF‐α suppression

Leukocyte adhesion to the diabetic retinal vasculature results in blood‐retinal barrier breakdown, capillary nonperfusion, and endothelial cell injury and death. Intercellular adhesion molecule‐1 (ICAM‐1) and the leukocyte integrin CD18 are required for these processes. Diabetes was induced in Long Evans rats, resulting in a two‐ to threefold increase in retinal leukocyte adhesion. Following one week of diabetes, neutrophil CD11a, CD11b, and CD18 expression was increased significantly, as were retinal ICAM‐1 levels. Animals were treated with aspirin, a cyclooxygenase 2 (COX‐2) inhibitor (meloxicam), or a soluble tumor necrosis factor α (TNF‐α) receptor/Fc construct (TNFR‐Fc, etanercept). High‐dose aspirin, etanercept, and high‐dose meloxicam each reduced leukocyte adhesion and suppressed blood‐retinal barrier breakdown. High‐dose aspirin also reduced the expression of CD11a, CD11b, and CD18, whereas meloxicam and etanercept did not. High‐dose aspirin, etanercept, and high‐dose meloxicam each reduced retinal ICAM‐1 expression. Aspirin and meloxicam both lowered retinal TNF‐α levels. Notably, aspirin, meloxicam, and etanercept did not change retinal vascular endothelial growth factor levels. High‐dose aspirin, etanercept and high‐dose meloxicam, each suppressed the retinal expression of eNOS and the DNA‐binding capacity of retinal nuclear factor‐κΒ. High‐dose aspirin also suppressed Erk kinase activity, which is involved in CD18 up‐regulation. Taken together, these data identify COX‐2 and TNF‐α as operative in the early signature pathologies of diabetic retinopathy, a newly recognized inflammatory disease.

[1]  K Miyamoto,et al.  Integrin-mediated neutrophil adhesion and retinal leukostasis in diabetes. , 2000, Investigative ophthalmology & visual science.

[2]  G. Hitman,et al.  An association in non-insulin-dependent diabetes mellitus subjects between susceptibility to retinopathy and tumor necrosis factor polymorphism. , 1996, Human immunology.

[3]  D. Brenner,et al.  NF-κB stimulates inducible nitric oxide synthase to protect mouse hepatocytes from TNF-α– and Fas-mediated apoptosis , 2001 .

[4]  K Miyamoto,et al.  Vascular endothelial growth factor (VEGF)-induced retinal vascular permeability is mediated by intercellular adhesion molecule-1 (ICAM-1). , 2000, The American journal of pathology.

[5]  P. Hardy,et al.  Prolonged Hypercapnia-Evoked Cerebral Hyperemia via K+ Channel– and Prostaglandin E2–Dependent Endothelial Nitric Oxide Synthase Induction , 2000, Circulation research.

[6]  A. Malik,et al.  Inhibition of NF-kappaB activation by pyrrolidine dithiocarbamate prevents In vivo expression of proinflammatory genes. , 1999, Circulation.

[7]  K. Mccormack.  Roles of COX-1 and COX-2. , 1998, The Journal of rheumatology.

[8]  P. Rutgeerts,et al.  Anti-TNF agents in Crohn’s disease , 2000, Expert opinion on investigational drugs.

[9]  R. Medford,et al.  NF- kappa B independent suppression of endothelial vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 gene expression by inhibition of flavin binding proteins and superoxide production. , 2000, Journal of molecular and cellular cardiology.

[10]  G. Caughey,et al.  Differential Regulation of Prostaglandin E2 and Thromboxane A2 Production in Human Monocytes: Implications for the Use of Cyclooxygenase Inhibitors1 , 2000, The Journal of Immunology.

[11]  D. Furst,et al.  Etanercept therapy in rheumatoid arthritis. A randomized, controlled trial. , 1999, Annals of internal medicine.

[12]  J. Spranger,et al.  [TNF-alpha level in the vitreous body. Increase in neovascular eye diseases and proliferative diabetic retinopathy]. , 1995, Medizinische Klinik.

[13]  F. Luscinskas,et al.  Salicylates inhibit I kappa B-alpha phosphorylation, endothelial-leukocyte adhesion molecule expression, and neutrophil transmigration. , 1996, Journal of immunology.

[14]  S. Abramson,et al.  Nitric Oxide Synthase/COX Cross-Talk: Nitric Oxide Activates COX-1 But Inhibits COX-2-Derived Prostaglandin Production1 , 2000, The Journal of Immunology.

[15]  G. A. Limb,et al.  Distribution of TNF alpha and its reactive vascular adhesion molecules in fibrovascular membranes of proliferative diabetic retinopathy. , 1996, The British journal of ophthalmology.

[16]  G. Yancopoulos,et al.  VEGF-initiated blood-retinal barrier breakdown in early diabetes. , 2001, Investigative ophthalmology & visual science.

[17]  J. Folkman,et al.  Increased Vascular Endothelial Growth Factor Levels in the Vitreous of Eyes With Proliferative Diabetic Retinopathy , 1995 .

[18]  G. King,et al.  Characterization of protein kinase C beta isoform activation on the gene expression of transforming growth factor-beta, extracellular matrix components, and prostanoids in the glomeruli of diabetic rats. , 1997, The Journal of clinical investigation.

[19]  W. Erl,et al.  Aspirin inhibits nuclear factor-kappa B mobilization and monocyte adhesion in stimulated human endothelial cells. , 1995, Circulation.

[20]  Shao-Ling Zhang,et al.  Peroxynitrite induces integrin‐dependent adhesion of human neutrophils to endothelial cells via activation of the Raf‐1/MEK/Erk pathway , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[21]  S. Soker,et al.  Tumor Necrosis Factor-α Regulates Expression of Vascular Endothelial Growth Factor Receptor-2 and of Its Co-receptor Neuropilin-1 in Human Vascular Endothelial Cells* , 1998, The Journal of Biological Chemistry.

[22]  S. Harrison,et al.  Signal integration by transcription-factor assemblies: interactions of NF-AT1 and AP-1 on the IL-2 promoter. , 1999, Cold Spring Harbor symposia on quantitative biology.

[23]  D. Brenner,et al.  NF-kappaB stimulates inducible nitric oxide synthase to protect mouse hepatocytes from TNF-alpha- and Fas-mediated apoptosis. , 2001, Gastroenterology.

[24]  K Miyamoto,et al.  VEGF increases retinal vascular ICAM-1 expression in vivo. , 1999, Investigative ophthalmology & visual science.

[25]  G. Weissmann,et al.  Modes of action of aspirin-like drugs: salicylates inhibit erk activation and integrin-dependent neutrophil adhesion. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[26]  S. Jabbur,et al.  The role of cytokines and prostaglandin-E2 in thymulin induced hyperalgesia , 2000, Neuropharmacology.

[27]  Early Treatment Diabetic Retinopathy Study design and baseline patient characteristics. ETDRS report number 7. , 1991, Ophthalmology.

[28]  L. Aiello,et al.  Suppression of retinal neovascularization in vivo by inhibition of vascular endothelial growth factor (VEGF) using soluble VEGF-receptor chimeric proteins. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[29]  Hong-Hee Kim,et al.  Tumor Necrosis Factor-α Supports the Survival of Osteoclasts through the Activation of Akt and ERK* , 2001, The Journal of Biological Chemistry.

[30]  Z. Radisavljevic,et al.  Vascular Endothelial Growth Factor Up-regulates ICAM-1 Expression via the Phosphatidylinositol 3 OH-kinase/AKT/Nitric Oxide Pathway and Modulates Migration of Brain Microvascular Endothelial Cells* , 2000, The Journal of Biological Chemistry.

[31]  A. Adamis,et al.  Sensitive blood-retinal barrier breakdown quantitation using Evans blue. , 2001, Investigative ophthalmology & visual science.

[32]  T. Collins.  Endothelial nuclear factor-kappa B and the initiation of the atherosclerotic lesion. , 1993, Laboratory investigation; a journal of technical methods and pathology.

[33]  H. Ledebur,et al.  Transcriptional Regulation of the Intercellular Adhesion Molecule-1 Gene by Inflammatory Cytokines in Human Endothelial Cells , 1995, The Journal of Biological Chemistry.

[34]  S. Soker,et al.  Tumor necrosis factor-alpha regulates expression of vascular endothelial growth factor receptor-2 and of its co-receptor neuropilin-1 in human vascular endothelial cells. , 1998, The Journal of biological chemistry.

[35]  B Kirchhof,et al.  Leukocyte-mediated endothelial cell injury and death in the diabetic retina. , 2001, The American journal of pathology.

[36]  K. Nakagawa,et al.  The relation between expression of vascular endothelial growth factor and breakdown of the blood-retinal barrier in diabetic rat retinas. , 1996, Laboratory investigation; a journal of technical methods and pathology.

[37]  D. Wallach,et al.  Cell death induction by TNF: a matter of self control. , 1997, Trends in biochemical sciences.

[38]  B. Williams,et al.  Glucose-induced protein kinase C activity regulates arachidonic acid release and eicosanoid production by cultured glomerular mesangial cells. , 1993, The Journal of clinical investigation.

[39]  C. Long,et al.  Expression and regulation of adhesion molecules in cardiac cells by cytokines: response to acute hypoxia. , 1998, Circulation research.

[40]  R. Alexander,et al.  Vascular cell adhesion molecule-1 (VCAM-1) gene transcription and expression are regulated through an antioxidant-sensitive mechanism in human vascular endothelial cells. , 1993, The Journal of clinical investigation.

[41]  S. Schiekofer,et al.  The role of oxidative stress and NF‐κB activation in late diabetic complications , 1999, BioFactors.

[42]  R. Natarajan,et al.  Hyperglycemia-induced activation of nuclear transcription factor kappaB in vascular smooth muscle cells. , 1999, Diabetes.

[43]  P. Wright,et al.  Induction of vascular cell adhesion molecule‐1 expression by IL‐4 in human aortic smooth muscle cells is not associated with increased nuclear NF‐κB levels , 1999, Journal of cellular physiology.

[44]  S. Molotchnikoff,et al.  Developmental regulation of endothelial nitric oxide synthase in cerebral vessels of newborn pig by prostaglandin E(2). , 1999, The Journal of pharmacology and experimental therapeutics.

[45]  H. Kim,et al.  Nuclear factor-kappaB regulates cyclooxygenase-2 expression and cell proliferation in human gastric cancer cells. , 2001, Laboratory investigation; a journal of technical methods and pathology.

[46]  E. Chew,et al.  Effects of aspirin on vitreous/preretinal hemorrhage in patients with diabetes mellitus. Early Treatment Diabetic Retinopathy Study report no. 20. , 1995, Archives of ophthalmology.

[47]  G. Sonenshein,et al.  Expression of a constitutive NF-kappa B-like activity is essential for proliferation of cultured bovine vascular smooth muscle cells. , 1995, The Journal of clinical investigation.

[48]  Shuibang Wang,et al.  Superoxide Production and Reactive Oxygen Species Signaling by Endothelial Nitric-oxide Synthase* , 2000, The Journal of Biological Chemistry.

[49]  F. Ferris.  Aspirin effects on mortality and morbidity in patients with diabetes mellitus. Early Treatment Diabetic Retinopathy Study report 14. ETDRS Investigators. , 1992, JAMA.

[50]  D. Williams,et al.  Activation of NF-kappa B and induction of vascular cell adhesion molecule-1 and intracellular adhesion molecule-1 expression in human glial cells by IL-1. Modulation by antioxidants. , 1994, Journal of immunology.

[51]  K. Chadee,et al.  Proinflammatory cytokines regulate cyclooxygenase-2 mRNA expression in human macrophages. , 1995, Biochemical and biophysical research communications.

[52]  Hollifield,et al.  Evidence for control of tumour necrosis factor‐alpha (TNF‐α) activity by TNF receptors in patients with proliferative diabetic retinopathy , 1999, Clinical and experimental immunology.

[53]  Z. Dong,et al.  Antioxidant properties of aspirin: characterization of the ability of aspirin to inhibit silica-induced lipid peroxidation, DNA damage, NF-kappaB activation, and TNF-alpha production. , 1999, Molecular and cellular biochemistry.

[54]  A. Barner.  Review of clinical trials and benefit/risk ratio of meloxicam. , 1996, Scandinavian journal of rheumatology. Supplement.

[55]  M. Weisman,et al.  Phase I/II trial of recombinant methionyl human tumor necrosis factor binding protein PEGylated dimer in patients with active refractory rheumatoid arthritis. , 2000, The Journal of rheumatology.

[56]  B. T. Luong,et al.  Treatment Options for Rheumatoid Arthritis: Celecoxib, Leflunomide, Etanercept, and Infliximab , 2000, The Annals of pharmacotherapy.

[57]  G. Hansson,et al.  Unstable angina activates myocardial heat shock protein 72, endothelial nitric oxide synthase, and transcription factors NFkappaB and AP-1. , 2000, Cardiovascular research.

[58]  L. Aiello,et al.  Prevention of leukostasis and vascular leakage in streptozotocin-induced diabetic retinopathy via intercellular adhesion molecule-1 inhibition. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[59]  N L Carteron,et al.  Cytokines in rheumatoid arthritis: trials and tribulations. , 2000, Molecular medicine today.

[60]  A. Ianaro,et al.  Interaction between nitric oxide and cyclooxygenase pathways. , 1996, Prostaglandins, leukotrienes, and essential fatty acids.

[61]  D. Lefer,et al.  Enhanced expression of intracellular adhesion molecule-1 and P-selectin in the diabetic human retina and choroid. , 1995, The American journal of pathology.

[62]  G. Schmid-Schönbein,et al.  Activated monocytes and granulocytes, capillary nonperfusion, and neovascularization in diabetic retinopathy. , 1991, The American journal of pathology.

[63]  J. Cunha-Vaz,et al.  Nitric oxide synthase activity in retinas from non-insulin-dependent diabetic Goto-Kakizaki rats: correlation with blood-retinal barrier permeability. , 2000, Nitric oxide : biology and chemistry.

[64]  Z. Dong,et al.  Antioxidant properties of aspirin: Characterization of the ability of aspirin to inhibit silica-induced lipid peroxidation, DNA damage, NF-κB activation, and TNF-α production , 2004, Molecular and Cellular Biochemistry.

[65]  R. A. Field,et al.  DIABETIC RETINOPATHY AND RHEUMATOID ARTHRITIS. , 1964, Lancet.

[66]  G. Engelhardt,et al.  Distinct isoforms (COX‐1 and COX‐2) of cyclooxygenase: possible physiological and therapeutic implications , 1996, Fundamental & clinical pharmacology.

[67]  S. Grösch,et al.  Celecoxib loses its anti‐inflammatory efficacy at high doses through activation of NF‐κB , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[68]  R. Panettieri,et al.  Dexamethasone Pathway That Is Only Partially Sensitive to B-dependent Signaling Κ Involves an Nf- Human Tracheal Smooth Muscle Cells Up-regulation of Icam-1 by Cytokines In , 2013 .

[69]  T. Yue,et al.  Antisense oligonucleotides to the p65 subunit of NF-kB inhibit human vascular smooth muscle cell adherence and proliferation and prevent neointima formation in rat carotid arteries. , 1995, Biochemical and biophysical research communications.

[70]  Shilling,et al.  Platelet expression of tumour necrosis factor‐alpha (TNF‐α), TNF receptors and intercellular adhesion molecule‐1 (ICAM‐1) in patients with proliferative diabetic retinopathy , 1999, Clinical and experimental immunology.