Müller细胞内色素上皮衍生因子与血管内皮生长因子的相互作用

Interaction between vascular endothelial growth factor and pigment epithelium derived factor in Müller cells

目的研究Müller细胞内色素上皮衍生因子(pigment epithelium derived factor,PEDF)与血管内皮生长因子(vascularendothelial growth factor,VEGF)的相互作用,为临床上治疗新生血管提供新的思路。方法大鼠Mller细胞原代培养,取第3代细胞用于实验。实验按以下操作进行:(1)观察VEGF对PEDF的作用:VEGF处理组细胞培养液中加入不同浓度VEGF(100nmol.L-1、500nmol.L-1、1000nmol.L-1、5000nmol.L-1),24h后收集细胞分别用RT-PCR和Western-blot法分析Mller细胞内PEDF mRNA和蛋白的表达。(2)观察PEDF对VEGF的作用:PEDF处理组细胞培养液中加入不同浓度PEDF(10nmol.L-1、40nmol.L-1、160nmol.L-1、640nmol.L-1),24h后收集细胞分别用RT-PCR和Western-blot法分析Mller细胞内VEGFmRNA和蛋白的表达。结果 VEGF处理组中,VEGF浓度为1000nmol.L-1、5000nmol.L-1时,PEDF mRNA分别为正常对照组的0.69倍(P<0.05)和0.44倍(P<0.01),PEDF蛋白分别为正常对照组的0.54倍(P<0.05)和0.52倍(P<0.05)。随VEGF浓度的升高,PEDF mRNA及蛋白表达均有逐渐减弱趋势。PEDF处理组中,PEDF浓度为40nmol.L-1和160nmol.L-1时,VEGF mRNA分别为正常对照组的0.61倍(P<0.05)和0.42倍(P<0.01),VEGF蛋白分别为对照组的0.70倍(P<0.05)、0.63倍(P<0.05)。随PEDF浓度的升高,VEGF mRNA及蛋白表达亦有逐渐减弱趋势。结论在Mller细胞内,VEGF对PEDF表达具有抑制作用,PEDF对VEGF也具有抑制作用,二者的负反馈调节作用可能在视网膜病理性新生血管中起重要作用。

Objective To investigate the interaction between vascular endothelial growth factor（VEGF）and pigment epithelium derived factor（PEDF）in Müller cells.Methods Müller cells of rats were primarily cultured and the third generation cells were used in the experiment.（1）Effects of VEGF on PEDF：Different concentrations of VEGF（100 nmol·L-1,500 nmol·L-1,1 000 nmol·L-1,5 000 nmol·L-1）were added into the cell culture medium,and cells were collected 24 hours later.The expression of PEDF mRNA and protein in Müller cells were detected with RT-PCR and Western blot.（2）Effects of PEDF on VEGF：Different concentrations of PEDF（10 nmol·L-1,40 nmol·L-1,160 nmol·L-1,640 nmol·L-1）were added into the cell culture medium,and cells were collected 24 hours later.The expression of VEGF mRNA and protein in Müller cells were detected with RT-PCR and Western blot.Results In VEGF treatment groups,when concentrations of VEGF were 1 000 nmol·L-1 and 5 000 nmol·L-1,the expression of PEDF mRNA were 0.69 times（P0.05）and 0.44 times（P0.01）to those of normal control group;The expression of PEDF protein were 0.54 times（P0.05）and 0.52 times（P0.05）to those of normal control group.The expression of PEDF mRNA and PEDF protein became lower with the increase of concentration of VEGF.In PEDF treatment groups,when concentrations of PEDF were 40 nmol·L-1 and 160 nmol·L-1,the expression of VEGF mRNA were 0.61 times（P0.05）and 0.42 times（P0.01）to those of normal control group;The expression of VEGF protein were 0.70 times（P0.05）and 0.63 times（P0.05）to those of normal control group.The expression of VEGF mRNA and VEGF protein became lower with the increase of concentration of PEDF.Conclusions In Müller cells,VEGF has inhibitive effect on PEDF,and same for PEDF on VEGF.The reciprocal interaction between VEGF and PEDF plays an important role in retinal pathologic neovascularization.