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
10.3760/cma.j.issn.1005-1015.2013.01.015
- VernacularTitle:米诺环素对缺氧猴脉络膜视网膜血管内皮细胞表达血管内皮生长因子受体的影响
- Author:
Dan ZHU
;
Xianming BAI
;
Yong TAO
- Publication Type:Journal Article
- Keywords:
Minocycline;
Endothelial cells/pathophysiology;
Vascular endothelial growth factor receptor-l;
Vascular endothelial growth factor receptor-2;
Cell hypoxia
- From:
Chinese Journal of Ocular Fundus Diseases
2013;(1):62-66
- CountryChina
- Language:Chinese
-
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 groups:control 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 minocycline:t=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.