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Recent Advances in Ophthalmology. 2014, (06) Core

Effects of PEDF on rat retinal Müller cells apoptosis stimulated by high glucoseChinese Full Text

LI Meng-Yun;LI Yan;LIU Hong-Bo;BI Huan-Li;ZHANG Jie;the Department of Ophthalmology,Weifang Medical College;Linyi People’s Hospital;

Abstract: Objective To investigate the effects of pigment epithelium-derived factor（ PEDF） on high glucose-stimulated rat retinal Müller 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 üller cells of SD rats. The purity was indentified by double-labeled immunofluorescence. The morphology was observed by confocalmicroscopy. The changes of Müller 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 Müller cells was above 95%. The apoptotic rate of Müller 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 Müller cells in HG group were changed,and the apoptotic rate was obviously increased（ P ＜ 0. 001）; The expression of bax mR N A in Müller 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 Müller cells apoptosis,so PEDF has a significant protective effect on retinal Müller cells.
Keywords：

PEDF; high glucose stimulation; Müller cell; bcl-2; bax; cell apoptosis;

  • DOI：

    10.13389/j.cnki.rao.2014.0144

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  • Subject：

  • Classification Code：

    R774.1;R587.2

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