Objective To investigate the effect of pigment epitheliumderived factor (PEDF)on the expression of glutamine synthetase in retinal M uuml;ller cells of diabetic rats.Methods Diabetic rats were induced with streptozotocin injection.Before and after injection of 10 mu;l (0.1 mu;g/ mu;l) PEDF (experimental group) or 10 mu;l PBS (control group) into the vitreous cavities of diabetic rats respectively for 48 hours,the expressions of GS and IL-1 beta; in retina were analyzed by immunohistochemistry and real time RTPCR techniques. After being treated with 100 ng/ml PEDF for 24 hours in high glucose conditions,the expressions of GS and IL-1 beta; in cultured M uuml;ller cells were studied by western blot and real time RT-PCR techniques. Apoptosis was analyzed by flow cytometry after Annexin V fluorescein isothiocyanate/Propidium idoium (Annexin V-FITC/PI) staining.Results By immunohistochemistry (the protein level) and real time RT-PCR (the mRNA level),it was found that the expression of GS decreased and the expression of IL-1 beta; increased obviously (real time RT-PCR:GS:t=4.23,P<0.01;IL-1 beta;:t=16.73,P<0.01;immunohistochemistry:GS: t=5.13,P<0.01;IL-1 beta;:t=9.32,P<0.01) in diabetic rats. After injection of 10 mu;l (0.1 mu;g/ mu;l) PEDF into the vitreous cavities of diabetic rats for 48 hours,it was found that the expression of GS increased and the expression of IL-1 beta; decreased significantly(RT-PCR GS:t=3.87,P<0.01IL-1 beta;:t=3.61,P<0.05;immunohistochemistry:GS:t=3.32, P<0.05;IL-1 beta;: t=2.63,P<0.05). Under high glucose conditions, 100 ng/ml PEDF induced decreasing expression of IL-1 beta; and increasing expression of GS significantly (RT-PCR:GS: t=2.89, P<0.05;IL-1 beta;: t=3.37,P<0.05;Western blot:GS:t=2.66,P<0.05;IL-1 beta;:t=3.23,P<0.05).Apoptosis of M uuml;ller cells under high glucose conditions was inhibited significantly by the treatment with 100 nmol/ml PEDF (t=3.21,P<0.05). Conclusions In diabetic rats,PEDF may decrease expression of IL-1 beta; in rat retinal M uuml;ller cells, which may result in increasing expression of GS.To some degree,it inhibits possibly the death of retinal ganglion cells.
Citation: 沈玺,焦秦,钟一声,谢冰. The effect of pigment epithelium derived factor on expression of glutamine synthetase in diabetic rat retina. Chinese Journal of Ocular Fundus Diseases, 2010, 26(2): 155-159. doi: Copy