Objective: Different pathways may lead from night work to metabolic diseases, including type 2 diabetes. This study aimed to explore the direct and indirect pathways from night work to glycemic levels, considering the role of physical activity, waist circumference and snacking using data from ELSA-Brasil. Materials and methods: A structural equation model was used to confirm the pathways from night work to glycemic levels. The latent variable, “glycemic levels”, included fasting glucose, glycated hemoglobin and 2-hour plasma glucose. Results: A total of 10.396 participants were included in the analyses. The final model showed that among women, night work was associated with increased glycemic levels. A statistical significant association between night work and glycemic levels mediated by waist circumference was observed among women and men. Conclusions: The association between night shift and glycemic levels can be interpreted as an important step toward understanding the pathways that could explain night work as a risk factor for diabetes using epidemiological data.