Effects of PEDF on rat retinal Müller cells apoptosis stimulated by high glucose

Objective To investigate the effects of pigment epithelium-derived factor( PEDF) on high glucose-stimulated rat retinal Muller cells apoptosis,and provide experimental basis for the treatment of diabetic retinopathy with PEDF. Methods T he cells were divided into normal control group,high glucose model( HG) group( containing 25 mmol·L- 1glucose in medium) and high glucose plus PEDF( HG + PED F) group( containing 25mmol·L- 1glucose and 25 mg·L- 1PED F in medium). Repeated pancreatic enzyme-degisting method was used to culture and purify the retinal M uller cells of SD rats. The purity was indentified by double-labeled immunofluorescence. The morphology was observed by confocalmicroscopy. The changes of Muller cells morphology and apoptosis were assayed with inverted phase contrast microscope,AnnexinV-FIT C ﹠ PI staining and flowcytometric,the expression of bcl-2 and bax mR N A were detected by R T-PC R. Results The purity of passage 3 retinal Muller cells was above 95%. The apoptotic rate of Muller cells in normal control group,HG group,HG + PEDF group were( 1. 223 ± 0. 106) %,( 2. 657 ± 0. 120) %,( 1. 907 ±0. 117) %,respectively,the relative gray values of bax mRNA were 0. 269 ± 0.041,0. 804 ± 0. 052,0. 528 ± 0. 018,respectively,the relative gray values of bcl-2 mR N A were 0. 904 ± 0. 021,0. 413 ± 0. 015,0. 521 ± 0. 010,respectively. Compared with normal control group,the morphology of retinal Muller cells in HG group were changed,and the apoptotic rate was obviously increased( P 0. 001); The expression of bax mR N A in Muller cells was obviously increased,while bcl-2 mR N A was obviously decreased( all P 0. 01);Compared with HG group,the apoptotic rate in HG + PED F group was obviously decreased( P 0. 001),the expression of bcl-2 mRNA was significantly decreased,while bax mRNA was significantly increased( all P 0. 01),but still had statistical difference compared with normal control group( all P 0. 05). Conclusion Giving exogenous PEDF can significant-ly reduce the high glucose-stimulated retinal Muller cells apoptosis,so PEDF has a significant protective effect on retinal Muller cells.