Purpose:The purpose of this study is to investigate peripapillary retinal nerve fiber layer (RNFL) and macular retinal layer thickness according to serum homocysteine concentration. Methods: Serum homocysteine levels of 120 retinal vein occlusion patients were measured. Peripapillary RNFL thickness and macular retinal layer thickness of unaffected eyes were measured by spectral domain optical coherence tomography and analyzed. Results: Average peripapillary RNFL thickness thinning (86.18 ± 19.78 µm vs. 99.61 ± 17.21 µm) (p < 0.001) and all quadrant (superior, nasal inferior, temporal) of peripapillary RNFL thickness thinning were observed in hyperhomocysteinemia patients. Average central macular thickness (CMT) (266.01 ± 29.59 µm vs. 278.71 ± 25.17 µm) was thinner in hyperhomocysteinemia patients (p = 0.029). Thinning of the macular nerve fiber layer (11.71 ± 3.46 µm vs. 14.35 ± 9.03 µm) and inner nuclear layer (25.45 ± 9.84 µm vs. 30.94 ± 13.88 µm) were also observed (p = 0.022, p = 0.041, respectively). There was a negative correlation between serum homocysteine and peripapillary RNFL thickness (r = -0.223, p = 0.015), and CMT (r = -0.230, p = 0.012), especially of the inner retinal layer. Conclusions: This study found associations of serum homocysteine with peripapillary RNFL and macular retinal layer thickness.