Ganglion cells apoptosis in diabetic rats as early prediction of glaucoma:a study of Brn3b gene expression and association with change of quantity of NO, caspase-3, NF-κB, and TNF-α

AIM:To find a new concept to show whether or not apoptosis of retinal ganglion cells(RGCs)canbe determined in the histology of acute hyperglycemia in the role of expressed Brn3b gene related to nitric oxide(NO),caspase-3,nuclear factor kappa-B(NF-κB),and tumor necrosis factor-α(TNF-α)as an early predictor of primary open angle glaucoma(POAG)eyes and their associations.METHODS:Experimental in vivo study was carried out using adult male,white Sprague-Dawley rats aged≥2 mo,weighing 150-200 g.The animals were divided into two groups,one group receiving intraperitoneal injection of streptozotociz 50 mg/kg in 0.01 mol/L citricbuffer and p H 4.5 and a comparison made with the control group.Retinal tissue was divided into two parts(both experimental and control groups respectively):a)right retina for immunohistochemistry(IHC;caspase-3 and TNF-α);b)left retina was divided into two parts for the purpose of real-time polymerase chain reaction(PCR)test(RNA extraction for Brn3b gene expression analysis)and ELISA test(NO and NF-κB).RESULTS:The experimental group showed a decrease in Brn3b gene expression compared to the control group(1.3-fold lower in 2nd month;1.1-fold lower in 4th month and 2.5-fold lower in 6th month).However,there was a decrease of NO,caspase-3,and an increase of NF-κB and TNF-αquantity.CONCLUSION:The expression of m RNA Brn3b gene is inversely proportional to apoptosis in RGCs.The quantity of NO,caspase-3,NF-κB and TNF-αis influential in expression of Brn3b in RGCs caused by hyperglycemia in diabetic rats.