ABSTRACT
Objective To investigate the effect of advanced glycation end products(AGEs)on expression of vascular endothelial growth factor(VEGF)in rabbit retinal M?ller cells in vitro.Methods We successfully cultured rabbit retinal M?ller cells and made AGEs-BSA as well as its control in vitro.Study with M?ller cells were divided into AGEs-BSA group,AGEs-BSA control group and blank control group.AGEs-BSA group and AGEs-BSA control group were respectively treated with 5 different concentration series of AGEs-BSA and AGEs-BSA control and cultured for 3,6 and 9 days,while blank control group was incubated without any intervention.Then VEGF expression in M?ller cells was observed by immunocytochemistry(ICC).Results Compared with control group,VEGF expression in cultured retinal M?ller cells was significantly enhanced in AGEs-BSA group.And the effects were in the time-and concentration-dependent manners.Conclusions AGEs increases VEGF expression in rabbit retinal M?ller cells in vitro,which indicates that AGEs may promote the progression of diabetic retinopathy(DR)through the induction of VEGF expression.
ABSTRACT
Objective To observe the effects of static pressure on the number of cultured retinal M?ller glial cells(RMGC)and expression of glial fibrillary acidic protein(GFAP)and heat shock protein(HSP)70 by these cells.Design Experimental study. Participants Cultured rat RMGC.Methods Rat RMGCs were cultured and identified according to previous method described by Reichenbach.These cells were treated with different static pressures and divided into 4 groups:A(1.33kPa),B(2.67kPa),C(5.33kPa)and D(10.67 kPa)while the cells without treatment was as control group(NC).The morphologies of RMGC in these groups were observed under inverted phased contrast microscope,the number of RMGC counted with conservative method and the viability were studied with trypan blue staining.The expressions of GFAP and HSP70 in RMGCs were detected with the method of western blot.Main Outcome Measures The morphologies of RMGC,cell number,cell viability.Results There were pressure-dependent changes of RMGC number. The cell number of group C and D was less than that of group NC,A and B(P＜0.01).High static pressure resulted directly in the decreased ratio of unstained RMGCs(P＜0.01).The ratio of unstained RMGCs in group C and D was less than that in group NC,A and B(P＜0.01).Many cells in group C and D were injured and the higher the pressure elevated,the more the degree of injury became.The expressions of GFAP and HSP70 in group NC were less than other pressure treated groups and the expression of GFAP in group C and D was higher than that in group A and B.There was no obvious difference between these pressure treated groups.Conclusions High static pressure could cause the injuries of RMGCs.The increased expression of GFAP and HSP70 in RMGC might be regarded as a sign of retinal injury response to high intraocular pressure.