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U. C. Yadav, S. K. Srivastava, K. V. Ramana; Amelioration of Endotoxin -Induced Uveitis by Aldose Reductase Inhibition in Rats. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2636.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the amelioration of inflammation in endotoxin-induced uveitis (EIU) in Lewis rats by aldose reductase (AR) inhibition and study the in-vitro anti-inflammatory effects of AR inhibition on lipopolysaccharide (LPS) -induced activation of NF-ΚB-dependent signals in human lens epithelial cells (HLEC).
EIU was induced by injection of E. coli LPS. The rats were sacrificed at 24 h after LPS injection, eyes were enucleated and aqueous humor was collected immediately. The number of infiltrating cells, protein concentration, and the levels of TNF-α, NO, and PGE2 were determined in the aqueous humor. Immunohistochemical staining with antibodies against iNOS, COX-2, TNF- α, activated NF-ΚB, and AR was performed in eye sections to evaluate the effect of AR inhibition during EIU. The level of ROS in rat eye was quantified by dihydroethidium fluorescence staining. For in-vitro studies HLEC were stimulated with LPS for 24 h, cell viability was assessed by cell counting and MTT assay and the apoptosis was measured by nucleosomal degradation. Electrophoretic mobility gel shift assays (EMSA) were carried out to determine the activation of NF-ΚB and AP1. The levels of TNF-α, nitric oxide, MMP2, MMP9, COX-2 and PGE2 were measured by specific ELISA kits.
Infiltration of inflammatory cells and increased protein concentration in the aqueous humor of EIU rats were significantly suppressed by AR inhibition. Similarly, the increase in TNF-α, NO and PGE2 in aqueous humor of EIU rats and levels of TNF-α in ciliary body and retina, and that of iNOS and COX-2 proteins in ciliary body, corneal epithelium and retina were inhibited by AR inhibition. The activation of NF-ΚB and increased level of ROS in ocular tissues of EIU rats were also prevented by AR inhibition. Also inhibition or siRNA ablation of AR in HLEC prevented the LPS-induced activation of NF-ΚB and AP1 and expression of iNOS, COX-2, MMP2, MMP9 and TNF-α proteins.
Our results provide evidence for an unanticipated role of AR in mediating NF-ΚB-dependent inflammatory response during acute inflammatory conditions and provide a novel concept that inhibition of AR could be therapeutically useful in preventing ocular inflammation such as commonly observed in uveitis.
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