Objective:To assess the relationship between serum total bilirubin and fundus arteriosclerosis in different genders.Methods:The physical examination data of Huadong Sanatorium in 2018 were analyzed, and a total of 26 275 people were included in this retrospective cross-sectional study. The age of this study was 18-86 (47.7±11.1) years old. Among them, there were 15 244 males (58.02%) and 11 031 females (41.98%). Participants were divided into 4 groups according to total bilirubin quartile values: Q1<11.50 μmol/L, Q2∶11.50-13.93 μmol/L, Q3∶13.94-17.14 μmol/L and Q4>17.14 μmol/L. The relationship between total serum bilirubin and fundus arteriosclerosis is determined using univariate and multivariate logistic regression analysis methods. The restricted cubic spline method was used to detect the dose-response relationship between total bilirubin and fundus arteriosclerosis. Results:In males, univariate analysis showed that high level of total bilirubin was a protective factor for fundus arteriosclerosis ( OR=0.87, 95% CI 0.78-0.97, P=0.012). After adjusting for other confounding factors, multivariate analysis showed that high level of total bilirubin remained as an independent protective factor for fundus arteriosclerosis ( OR=0.86, 95% CI 0.74-0.99, P=0.047). There was a linear dose-response relationship between total bilirubin level and fundus arteriosclerosis ( P=0.012). In females, univariate analysis showed that there were no statistically significant association between high level of total bilirubin and fundus arteriosclerosis ( OR=0.96, 95% CI 0.80-1.17, P=0.709). After adjusting for other confounding factors, multivariate analysis showed no statistically significant association between high level of total bilirubin and fundus arteriosclerosis ( OR=0.98, 95% CI 0.76-1.27, P=0.888). No linear dose-response relationship between total bilirubin level and fundus arteriosclerosis was found in females ( P=0.253). Conclusion:There are gender differences in the relationship between total bilirubin and fundus arteriosclerosis in this cohort. Elevated levels of total bilirubin are associated with fundus arteriosclerosis in males but not in females.