Objective To investigate the role of adenosine A2A receptor plays in retinal pathological neovascularization in mice. Methods A total of 202 mice were divided into room-air group (n=66) and oxygen induced retinopathy (OIR) group (n=136). Among room-air group, there were 18 A2A knock-out (KO) mice (KO subgroup) and 24 C57BL/6 mice as wide type (wide type subgroup). OIR group were divided into OIR control subgroup (n=48), A2A-OIR subgroup (n=24) and Caffeine-OIR subgroup (n=64). The retinal neovascularization of OIR group was induced by oxygen. The pathological neovascularization was determined by retinal sections. Fluorescent quantitative polymerase chain reaction (PCR) was used to measure the mRNA expression of A2A and vascular endothelial growth factor (VEGF). 0.1, 0.3, 1.0 g/L Caffeine was dissolve in drinking water of lactating females in Caffeine-OIR subgroup, non-perfusion areas of retina in mice at the age of 0 - 17, 0 - 7, 7- 17, 7-12, and 12- 17 days were analyzed in different dosage and when the dosage as 1.0 g/L. Results Compared with OIR control subgroup, the retinal non-perfusion areas and the numbers of endothelial cell nuclei breaking through the internal limiting membrane in A2A- OIR subgroup were reduced significantly (t=7.694, 7.747;P＜0.001). Compared with wide type subgroup, the level of A2A and VEGF mRNA in OIR control subgroup increased significantly (t=4.036, 2.230;P＜0.05). Compared with OIR control subgroup, the level of VEGF mRNA in A2A- OIR subgroup decreased significantly (t=3.122,P＜0.01). Compared with OIR control subgroup, the retinal non-perfusion areas in mice at the dosage of 0.1 and 1.0 g/L (t=2.397, 4.533) and at the age of 0 -17, 0 -7 days when the dosage as 1.0 g/L (t=4.070, 2.399) were reduced significantly (P＜0.05). Conclusions The expression of adenosine A2A receptor increases in oxygen-induced retinal pathological neovascularization. Adenosine A2A receptor may regulate the expression of VEGF. A2A receptor inactivation can inhibit oxygen-induced retinal pathological neovascularization.
Objective To investigate the role of ephrin A genes in the development of oxygen induced retinalneovascularization (OIR) in mice.Methods The OIR model was established by oxygen induction in new born C57BL/6J mice.Reversed transcript polymerase chain reaction (RT-PCR) was used to measure the expression levels of ephrin A1-A5 in retinas of mice in experimental and normal control group.Results All of the ephrin A family genes expressed in normal retinas. Ephrin A1 mRNA was significantly higher in OIR group(t=3.19,P=0.019); ephrin A2 mRNA was higher in the 15-day-old OIR retinas(t=3.71,P=0.033）; ephrin A3-A5 mRNA decreased or disappeared in 12 and 13-day-old RNV mice, and increased in 15-day-old OIR mice. Conclusion Ephrin A genes are involved in the development of retina and OIR.
Retinal neovascularization is a complicated pathophysiological process as a result of imbalance between angiogenic and antiangiogenic factors. Correct understanding of the signaling pathways, exploring the critical factors involved in retinal angiogenesis, looking for new strategies by reconstructing the new vessels are helpful for knowing the mechanism of the occurrence and development of reitnal neovascularization, which would be good for preventing and treating retinal neovascularization diseases.
ObjectiveTo observe the effects of angiostatin on the activity of extra-cellular signal-regulated protein kinase (ERK) of retinal microvascular endothelial cells of mice.MethodsAngiostatin was separated and purified by l-lysine sepharose 4B from human plasma. The primary retinal microvascular endothelial cells were divided into 4 groups: the control group, vascular endothelial growth factor (VEGF) 10 ng/ml group, angiostatin 130 μg/ml group, and VEGF (10 ng/ml) + angiostatin (130 μg/ml) group. The expression of ERK1 was assayed by Westernblotting method 1, 2, 5, 10, 15, and 30 minutes after the treatment of angiostatin.ResultsCompared with the control group, the expression of ERK-1 reduced 1 minute after treatment, reduced markedly after 10 minutes. After 30 minutes, no differences of the expression of ERK were seen between the control group and angiostatin group. The activation of ERK-1 of retinal microvascular endothelial cells occurred after stimulated by VEGF, and at the pitch at the peak after 5 minutes. The level of ERK in VEGF group increased 210% than that in the control (P＜0.05). After 30 minutes, no significant difference of the level of ERK between VEGF and the control group. And because of angiostatin, the expression of ERK-1 decreased 11.9%(1 minute)、17.9%(2 minutes)、38.7%(5 minutes)、49.3%(10 minutes) （P＜0.05）、27.9%(15 minutes)、1.12%(30 minutes) respectively.ConclusionsAngiostatin can effectively block the signal path through which VEGF transmits from outside of the cell to cellular nuclei. (Chin J Ocul Fundus Dis, 2005,21:170-173)