视网膜下瘢痕形成小鼠视网膜色素上皮-脉络膜中血管内皮生长因子、促纤维化因子及炎症因子的表达

Abstract

Objective To observe the expression of vascular endothelial growth factor (VEGF),fibrogenic mediators and inflammatory mediators in the retinal pigment epithelium (RPE)-choroid complex of mice with experimental subretinal fibrosis.Methods By subretinal injection of inflammatory macrophages after retinal photocoagulation,experimental subretinal fibrosis was induced in 64 adult C57BL/6(B6) female mice (7-8 weeks).Masson staining and glial fibrillary acidic protein (GFAP) staining of choroidal wholemont were performed to verify that the subretinal fibrosis at day 7 after subretinal injection.Before subretinal injection and at day 1,2,3,5 and 7 after subretinal injection,the mRNA expression level of VEGF,transforming growth factor (TGF)-β1,TGF-β2,TGF-β3,interleukin (IL)-6,IL-10 and IL-13 in RPE-choroid complex were evaluated by quantitative reverse transcription-polymerase chain reaction.Enzyme-linked immune sorbent assay was next used to detect protein expression of these factors.Results At seven days after subretinal injection of inflammatory macrophages,experimental subretinal fibrosis was detected by Masson staining and GFAP staining.The mRNA level of VEGF,TGF-β1 and TGF-β2 reached the peak at day 5 after modeling,while the mRNA expression of IL-6,IL-10,IL-13 reached the peak at day 2 after modeling.TGF-β3 mRNA was not detected either in naive mice or during the development of experimental subretinal fibrosis.At day 2,3,5 and 7 after modeling,compared with the pre-modeling,the mRNA expression of VEGF (t =2.38,3.65,4.03,2.26),TGF-β1 (t =2.58,2.30,3.89,4.15) and TGF-β2 (t=4.37,4.20,3.77,3.98) were significantly increased (P＜0.05).At day 1,2 and 3 after modeling,compared with the pre-modeling,the mRNA expression of IL-6 were significantly increased (t=2.36,4.54,4.01; P＜0.05).At day 2 and 3 after modeling,compared with the pre-modeling,the mRNA expression of IL-10 and IL-13 were significantly increased (t=3.87,4.20,2.44,2.58; P＜0.05).At day 5 after modeling,compared with the pre-modeling,the protein expression of VEGF,total TGF-β1,activeTGF-β1,total TGF-β2 and active TGF-β2 were significantly increased (t=2.57,3.37,2.45,3.83,2.74; P＜0.05).IL-6,IL-10 and IL-13 protein were not detected at pre-modeling eyeballs,but were found at day 2 after modeling.Conclusion The expression of VEGF,fibrogenic mediators and inflammatory mediators in RPE-choroid complex in mice with experimental subretinal fibrosis are increase significantly. Key words: Wet macular degeneration/etiology; Cicatrix/physiopathology; Animal experimentation