AIM: To analyze the relationship between vitreous transthyretin (TTR) levels, high myopia and diabetic retinopathy (DR). METHODS: We selected 6722 individuals from the southern Jiangsu Province for diabetes and ophthalmic examinations. The TTR concentration in the vitreous of 50 patients with high myopia and diabetes, 50 patients with only DR, and 20 healthy controls were determined by ELISA. Key factors in Tie2 pathway in DR development including vascular endothecial growth factor (VEGF), Tie2, Angpt1, Angpt2, vascular endothelial growth factor receptor (VEGFR) 1 and VEGFR2 were also detected by ELISA. RESULTS: The prevalence of DR in patients with diabetes and myopia [<6.00 diopter (D)], diabetes and high myopia (>6.00 D), and diabetes without myopia were 11.1%, 2.5%, and 60.0%, respectively. The vitreous TTR concentration of patients with diabetes and high myopia was approximately 6.5- and 4.2-times higher than those of patients with DR and healthy controls, respectively (P<0.05). Following the vitreous TTR concentration, the levels of VEGF, Tie2, Angpt1, Angpt2, VEGFR1 and VEGFR2 in vitreous of diabetes and high myopia patients, DR patients and healthy controls were detected as dramatically fluctuated. CONCLUSION: The results suggest that TTR can affect the vitreous contents of key factors in Tie2 pathway for neovascularization, and there should be a protective association between abundant TTR levels in the vitreous of highly myopic patients and a decreased risk of DR.