Figure 2|Retinal angiographs in mice at the age of 17 days.
Figure 3|Morphology of retinal tissue in mice at the age of 17 days.
The whole retina flat-mounted angiography was used to determine whether MEG3 overexpression can reduce RNV morphologically (Figure 2). The retinal vessels of the OIR-C group were clearly dilated and the non-perfusion area is obvious. The control group and OIR-T group had fewer neovascular clusters and non-perfusion areas (P < 0.05; Figure 2). This result suggests that the overexpression of MEG3 inhibits the neovascularization. Hematoxylin-eosin staining (Figure 3) demonstrated that the number of preretinal neovascular cells in the OIR-T group decreased significantly compared with the OIR-C group (P < 0.01).
Figure 4|Immunohistochemical results of p-PI3K and p-Akt in the retina of mice at the age of 17 days.
Figure 5|Immunofluorescence results of the retina of mice at the age of 17 days.
Immunohistochemical of retinal sections revealed that p-PI3K and p-Akt were more highly expressed in the OIR-C group than in the control group (P < 0.01) (Figure 4). Immunofluorescence of retinal sections revealed that VEGF was highly expressed in the OIR-C group (Figure 5). However, VEGF expressions were significantly lower in the OIR-T group than in the OIR-C group (P < 0.01).
Figure 6|Protein expression levels of p-PI3K, p-AKT and VEGF in the retina of mice at the age of 17 days.
Western blotting showed that protein expression levels of p-PI3K, p-Akt, and VEGF in the OIR-T group were significantly reduced compared with the OIR-C group (P < 0.01) (Figure 6).
Figure 7|Protein expression level of IL-1β, IL-6 and TNF-α in the retina of mice at the age of 17 days.
We measured the expression of IL-1β, IL-6, and TNF-α to explore the effects of MEG3 on inflammation. The OIR-T group exhibited a significant decrease in inflammatory factors compared with the OIR-C group (P < 0.01) (Figure 7).