Objective: To investigate the effect of E2F1 on the survival and apoptosis of retinal ganglion cells (RGCs) induced by high glucose and its mechanism. Methods: A high-glycemic injury model was constructed with. RGC-5 cells used as the research object. After transfection, RGC-5 cells were divided into a blank control group, a high-sugar group, a high-sugar + si-E2F1 group, a high-sugar + si-con group, a high glucose + si-E2F1 + PBS group, and a high glucose + si-E2F1 + SD19 group. Real-time RT-PCR and Western blotting were used to detect the expression of E2F1 mRNA in cells; methyl thiazolyl tetrazolium (MTT) was used to detect cell viability; TUNEL and flow cytometry (FCM) were used to detect cell apoptosis. E2F1, p-signal transducer and activator of transcription 3 (p-STAT3), STAT3, p-Janus kinase 2 (p-JAK2), JAK2, cleaved caspase-3, and cleaved poly-ADP-ribose polymerase (cleaved PARP) protein expression in cells detected by Western blotting. Results: Compared with blank control group, E2F1 expression level increased in RGC-5 cells in the high glucose group, cell survival rate was significantly decreased, p-STAT3, p-JAK2, cleaved caspase-3, cleaved PARP, E2F1 expression, and apoptotic rate were increased. After silencing E2F1 expression, the cell survival rate was significantly increased, while cleaved caspase-3 protein expression, cleaved PARP protein expression, and apoptotic rate were decreased. SD19 partially reversed the protective effect of silencing E2F1 on RGC-5 cells. Conclusion: Inhibition of E2F1 expression can reduce high glucose-induced apoptosis in RGCs. The mechanism may be related to the inhibition of the JAK2/STAT3 signaling pathway.