title: Intraocular Delivery of miR-146 Inhibits Diabetes-Induced Retinal Functional Defects in Diabetic Rat Model.
abstract: Previously, we showed that microRNA-146 (miR-146) is a pivotal negative feedback regulator of multiple nuclear factor kappa-B (NF-κB) activation pathways in retinal endothelial cells (RECs). We hypothesized that miR-146 plays an important role in diabetic retinopathy (DR) by inhibiting diabetes-induced inflammatory response in the retina. The purpose of the current study is to test this hypothesis in vivo.
  
  
  
  Lentiviruses expressing rno-miR-146a, lenti-miR-146a, and negative control oligonucleotide with scrambled sequence, lenti-miR-neg ctl, were produced. Young male Sprague-Dawley rats were injected with a single dose of streptozotocin ([STZ] 65 mg/kg) to induce diabetes. One week after diabetes, animals were injected with lentivirus intravitreally (4 μl, ∼106 CFU/mL). Three months after diabetes, retinal microvascular leakage was tested by Evans blue assay; retinal function by electroretinogram (ERG). Total RNA and protein lysate were isolated from the retina for quantitative (q)RT-PCR and Western blot analyses.
  
  
  
  Lenti-miR-146a robustly transduced human retinal endothelial cells (HRECs) and increased the expression of miR-146a in vitro. In vivo, intravitreal injection of lenti-miR-146a increased the expression of miR-146a in the retina, while its key downstream target genes, including CARD10, IRAK1, and TRAF6, were downregulated. Intravitreal delivery of miR-146 inhibited diabetes-induced upregulation of NF-κB downstream gene, Intercellular Adhesion Molecule 1 (ICAM1), as well as microvascular leakage and retinal functional defects.
  
  
  
  Intravitreal delivery of miR-146 inhibited diabetes-induced NF-κB activation and retinal microvascular and neuronal functional defects in a diabetic rat model.
authors.affiliation: Department of Ophthalmology/Kresge Eye Institute, Wayne State University, Detroit, Michigan, United States 2Department of Anatomy and Cell Biology, School of Medicine, Wayne State University, Detroit, Michigan, United States.
authors.forename: Pei
authors.initials: P
authors.lastname: Zhuang
authors.affiliation: Department of Ophthalmology/Kresge Eye Institute, Wayne State University, Detroit, Michigan, United States 2Department of Anatomy and Cell Biology, School of Medicine, Wayne State University, Detroit, Michigan, United States.
authors.forename: Chithra K
authors.initials: CK
authors.lastname: Muraleedharan
authors.affiliation: Department of Ophthalmology/Kresge Eye Institute, Wayne State University, Detroit, Michigan, United States 2Department of Anatomy and Cell Biology, School of Medicine, Wayne State University, Detroit, Michigan, United States.
authors.forename: Shunbin
authors.initials: S
authors.lastname: Xu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003921:Diabetes Mellitus, Experimental; D003930:Diabetic Retinopathy / Q000175:diagnosis / Q000235:genetics / Q000517:prevention & control*; D004596:Electroretinography; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D058449:Intravitreal Injections; D008297:Male; D035683:MicroRNAs / Q000008:administration & dosage* / Q000096:biosynthesis / Q000235:genetics; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics; D012313:RNA / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D012160:Retina / Q000378:metabolism* / Q000473:pathology
chemical_list: C554180:MIRN146 microRNA, rat; D035683:MicroRNAs; D016328:NF-kappa B; D018799:Intercellular Adhesion Molecule-1; D012313:RNA