Effect of minocycline for expression of vascular endothelial growth factor receptor-1 and vascular endothelial growth factor receptor-2 in hypoxia chorioretinal endothelial cells of monkeys / 中华眼底病杂志

ABSTRACT

Objective To observe the expression of vascular endothelial growth factor receptor-1 (VEGFR-1) and VEGFR-2 in hypoxic chorioretinal endothelial cells of monkeys (RF/6A),and to evaluate the effect of minocycline.Methods RF/6A was cultured and divided into four groupscontrol group,hypoxia group,hypoxia and low dose of minocycline group (0.5 μmol/L),hypoxia and medium dose of minocycline group (5 μmol/L),and hypoxia and high dose of minocycline group (50 μmol/L).Real-time reverse transcription-polymerase chain reaction (RT-PCR) and immunohistopathological staining were used to measure the mRNA and protein expression of VEGFR-1 and VEGFR-2,respectively.Results RT-PCR showed that the expression of VEGFR-1 mRNA did not vary significantly between groups (F24 h =0.17,F48 h =1.53,F72 h =2.04; P＞0.05).Compared with hypoxia group,the expression of VEGFR-2 mRNA in all minocycline treated groups were significantly down-regulated (low minocycline,medium minocycline,high minocyclinet=4.69,20.16,17.12; P＜0.001).The immunohistopathological study showed the cells with positive staining of VEGFR-1 can be observed in all groups,and the staining was relatively weak and mainly located in cell membrane and cytoplasm.The optical density value analysis showed that the protein expression of VEGFR-1 did not vary significantly between groups at all time points(F24 h =0.251,F48 h=0.340,F72 h =0.589; P＞0.05).The VEGFR-2 positive staining cells were also observed in all groups,and the staining was relatively high.Brown staining particles of VEGFR-2 were observed in the cell membrane with minor staining particles in cytoplasm.The staining density of VEGFR-2 was significantly higher in hypoxia group than control group.Compared with the hypoxia group,the protein expression of VEGFR 2 in minocycline treated groups was significantly lower (F24 h =19.147,F48 h =14.893,F72 h =11.984; P＜0.05).Conclusion The expression of VEGFR-2 is up-regulated in RF/6A,and minocycline somewhat shows an inhibition effect.