Objective To evaluate the association between serum total bilirubin concentration (STBC) and diabetic retinopathy (DR) among the Chinese northeastern population. Methods A cross-sectional study was conducted in Liaoning between January 2015 and May 2017. Results A total of 742 subjects (419 men and 323 women) with type 2 diabetes mellitus (DM) who visited an ophthalmic clinic were included in this study. The mean age of the subjects was 59.55 ± 10.63 years, and 43.5% of the subjects were women. The mean of DM duration was 11.01 ± 7.35 years. STBC were negatively correlated with DM duration, urea nitrogen, serum creatinine, uric acid, and urine microalbumin. After adjusting for confounding factors, as a continuous variable, STBC was inversely associated with the risk of DR in total subjects (OR: 0.95, 95% CI: 0.93–0.99). When STBC was used as a tertiary variable, compared with the first tertile, the OR in the third tertile was 0.37 (95% CI: 0.22–0.64) in total subjects. Conclusion Our results demonstrate that a significant negative association was found between STBC and DR. STBC might be an early clinical marker for predicting the occurrence of DR.