|
Objective: To establish the C57BL / 6 mouse oxygen-induced retinal neovascularization lesion model (OxygenInduced Retinopathy, OIR), observed the change of the content of the eye erythropoietin EPO (Erythropoietin, EPO), and to explore the EPO specific blocker of retinal neovascularization (Retinal Neovascularization, RNV) formed. Method: 1. Establish a mouse OIR model, observe and count groups retinal neovascularization. 2 newborn C57BL / 6 mice a total of 60, were randomly divided into experimental group (n = 50) and control group (n = 10). (1) experimental mice and female rats after the birth of the 7th (P7) were placed in a sealed box of oxygen, the oxygen partial pressure of 75% ± 5%, were cultured for 5 days (P12) to return to normal oxygen environment, oxygen partial pressure of 20% ± 1%, to create the OIR model. The control group of mice into the oxygen tank, together with female rats reared in normal oxygen environment. (2) All mice born after P12-P16 intravitreal injection every other day line right eye. Experimental group survived to 40, were randomly divided into four groups of 10 each containing 25ng (25ng group) were injected with soluble EPO receptor in PBS 0.5μl 50ng (50 ng group), 250ng (250ng group) as well as volume the EPO receptor PBS solution (PBS group), control group 10 mice injected with non-EPO receptor in PBS 0.5μl of. (3) mice in each group were sacrificed at P17, the removal of the right eye, the eye produced tissue sections, HE staining underwent histopathological examination, counting the internal limiting membrane the RNV endothelial cell nucleus number. Measured by radioimmunoassay PBS group and the control group of mice left eye retinal tissue EPO levels. Results: (1) retinal tissue sections: oxygen-induced P17 mice break through the internal limiting membrane RNV endothelial cell nuclei count was 80.0 ± 6.2, and mice under normoxic environment only 1.0 ± 0.9, and the difference was statistically significant (P <0.001). (2) EPO were measured by radioimmunoassay results: the PBS groups retinal EPO levels of 80.82 ± 20.70 IU / L, the control group was 14.42 ± 6.80 IU / L, and the difference was statistically significant (P <0.001). (3) the retinal tissue EPO expression RNV degree of proliferation was significantly positively correlated (rs = 0.58, P <0.01). (4) each model mice RNV number of endothelial cell nuclei were significantly more than the control group (1.3 ± 0.5 unit), the OIR model group, PBS group RNV endothelial cell nuclei count (82.0 ± 5.5) than various concentrations of EPO receptor group (P <0.001), RNV endothelial cell nuclei count 25ng group (62.2 ± 4.5 unit) and 50ng group RNV endothelial cell nuclei count (43.3 ± 3.2 unit) were more than 250ng group (22.2 ± 2.1) (P <0.001 ) and 25ng group RNV endothelial cell nuclei count more than 50ng group (P <0.001). Conclusion: 1. Using oxygen-induced retinopathy in mice can be successfully established model of retinal vascular lesions. 2 retinal neovascularization eye local EPO upregulation. Specific inhibitor of EPO can inhibit retinal neovascularization, might be able to become a new therapeutic targets.
|