Effect of Cathepsin B and vascular endothelial growth factor on the retinal neovascularization induced by hyperoxia
10.3760/cma.j.issn.2095-0160.2017.10.003
- VernacularTitle:组织蛋白酶B和血管内皮生长因子在高氧诱导的视网膜新生血管中的作用
- Author:
Wenjuan WANG
1
;
Guohong ZHOU
Author Information
1. 山西省眼科医院泪道病科
- Keywords:
Cathepsin B;
Vascular endothelial growth factor;
Retinal neovascularization
- From:
Chinese Journal of Experimental Ophthalmology
2017;35(10):873-877
- CountryChina
- Language:Chinese
-
Abstract:
Background Vascular endothelial growth factor (VEGF) plays a key role in the vascular development and neovascularization.Studies have proved that Cathepsin B is related to the formation of neovascularization,but its mechanism is unclear.Objective This study was to investigate the expression of Cathepsin B and VEGF in the retinal neovascularization induced by hyperoxia and the relationship between them.Methods Forty-four 7-day-old C57BL/6J mice were randomly assigned into 4 groups:normal control group,hyperoxia-induced group,normal control (NC)-green fluorescent protein (GFP)-lentivirus (Lv) group and Cathepsin B-RNA interference (RNAi)-Lv group,with 11 mice 22 eyes for each group.The mice in normal control group were survival in natural environment.The other three groups of 7-day-old mice were put in a sealed box of oxygen volume fraction (75±2)％ for 5 days and then sent back to normal environment.The mice of hyperoxia-induced group did not have any drug intervention,while the 12-day-old mice of NC-GFP-Lv group and Cathepsin B-RNAi-Lv group received intravitreal injection of NC-GFP-Lv 1 μl or Cathepsin B-RNAi-Lv 1 μl.All 17-day-old mice were sacrificed and the retinas were collected.The mRNA expression levels of Cathepsin B and VEGF were performed by real-time PCR;the protein expression levels of Cathepsin B and VEGF were detected by Western blot.Results Fluorescence microscope results showed that the layer and branch of retinal neovascularization were less in the Cathepsin B-RNAi-Lv group than those in the NC-GFP-Lv group and hyperoxia-induced group.The relative expression levels of Cathepsin B and VEGF mRNA in each group were significantly different (F =444.89,P =0.00;F =519.78,P =0.00).The relative expression levels of Cathepsin B and VEGF mRNA in hyperoxia-induced group,NC-GFP-Lv group and Cathepsin B-RNAi-Lv group were higher than those in normal control group,and the relative expression levels of Cathepsin B and VEGF mRNA in Cathepsin B-RNAi-Lv group were lower than those in hyperoxia-induced group and NC-GFP-Lv group,with significant differences between them (all at P ＜ 0.05).The relative expression levels of Cathepsin B and VEGF protein in each group were significantly different (F =54.37,P =0.00;F =79.65,P =0.00).The relative expression levels of Cathepsin B and VEGF protein in hyperoxia-induced group,NC-GFP-Lv group and Cathepsin B-RNAi-Lv group were higher than those in normal control group,and the relative expression levels of Cathepsin B and VEGF protein in Cathepsin B-RNAi-Lv group were lower than those in hyperoxia-induced group and NC-GFP-Lv group,with significant differences between them (all at P＜0.05).Conclusions The expression levels of Cathepsin B and VEGF in retinal neovascularization of hyperoxia-induced group were significantly higher than those in normal control group,Cathepsin B-RNAi-Lv can inhibit the mRNA and protein expression of Cathepsin B and VEGF,the expression of VEGF may be influenced by the expression of Cathepsin B.
