Inhibition of Ocular Aldose Reductase by a New Benzofuroxane Derivative Ameliorates Rat Endotoxic Uveitis

Abstract

The study investigated the effects of the aldose reductase (AR) inhibitor benzofuroxane derivative 5(6)-(benzo[d]thiazol-2-ylmethoxy) benzofuroxane (herein referred to as BF-5m) on the biochemical and tissue alterations induced by endotoxic uveitis in rats. BF-5m has been administered directly into the vitreous, in order to assess the expression and levels of (i) inflammatory markers such as the ocular ubiquitin-proteasome system, NF-κB, TNF-α, and MCP-1; (ii) prooxidant and antioxidant markers such as nitrotyrosine, manganese superoxide dismutase (MnSOD), and glutathione peroxidase (GPX); (iii) apoptotic/antiapoptotic factors caspases and Bcl-xl; (iv) markers of endothelial progenitor cells (EPCs) recruitment such as CD34 and CD117. 5 μL of BF-5m (0.01; 0.05; and 0.1 μM) into the right eye decreased in a dose-dependent manner the LPS-induced inflammation of the eye, reporting a clinical score 1. It reduced the ocular levels of ubiquitin, 20S and 26S proteasome subunits, NF-κB subunits, TNF-α, MCP-1, and nitrotyrosine. BF-5m ameliorated LPS-induced decrease in levels of MnSOD and GPX. Antiapoptotic effects were seen from BF-5m by monitoring the expression of Bcl-xl, an antiapoptotic protein. Similarly, BF-5m increased recruitment of the EPCs within the eye, as evidenced by CD34 and CD117 antibodies.

Activation of Melanocortin Receptors MC1and MC5Attenuates Retinal Damage in Experimental Diabetic Retinopathy

“…Ocular tissue sections (5 μ m) were serially cut from paraffin-embedded tissue and labelled for the detection of Ki-67 by immunohistochemistry, according to previous published protocol [ 31 ]. Briefly, sections were incubated with primary mouse monoclonal anti-ki67 (PP-67) antibody (dilution 1 : 250, Abcam, Cambridge, UK) for 30 min at room temperature.…”

Retinal and circulating miRNA expression patterns in diabetic retinopathy: An in silico and in vivo approach

“…Immunohistochemistry was performed in retinal section of the whole eye explanted from 5 to 10 weeks diabetic and control mice. Ocular tissue sections (5 μm) were serially cut from paraffin‐embedded tissue and labelled for the detection of BDNF were incubated with primary mouse monoclonal anti‐BDNF antibody (dilution 1:250, [EPR1292] (ab108319) Abcam, UK []) for 30 min at room temperature by immunohistochemistry, according to a previously published protocol (Di Filippo et al, ). Images from independent retinal samples ( n = 10 per experimental group) were used for quantitative analysis for BDNF nanoparticles by two investigators, without knowledge of the treatment groups.…”

Effects of the New Aldose Reductase Inhibitor Benzofuroxane Derivative BF-5m on High Glucose Induced Prolongation of Cardiac QT Interval and Increase of Coronary Perfusion Pressure

“…Western blotting analysis was performed from the homogenized frozen biopsy according to Di Filippo et al [ 19 ]. We used (i) primary anti-SIRT1 antibody (1 : 1000, Abcam, Cambridge, UK), (ii) specific monoclonal antibody directed against FOXO-1 (1 : 500, Millipore, California, USA), (iii) anti-MnSOD primary antibody (1 : 800, Millipore, California, USA), (iv) anti- β -actin monoclonal antibody (1 : 1000, Sigma-Aldrich), and (v) anti-eNOS sc-654 (1 : 500, Santa Cruz Biotech, USA) with an enhanced chemiluminescence detection reagent (ECL), quantified by densitometry using a BioRad ChemiDoc MP Imaging system.…”

“…Primers sequences were as follows: 5′-TGTTTCCTGTGGGATACCTGA-3′ (sense) and 5′-TGAAGAATGGTCTTGGGTCTTT-3′ (antisense) for SIRT1 and 5′-CGAGTACAACCTTCTTGCAG-3′ (sense) and 5′-TTCTGACCCATACCCACCAT-3′ (antisense) for β -actin. Relative quantification of gene expression was normalized to beta-actin using the 2 −ΔΔCt method [ 19 , 21 ].…”