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Background: Long working hours and effort-reward imbalance (ERI) among medical workers may contribute to poor mental health and reduced productivity. However, the potential mechanisms among them are not well understood. This study aimed to explore the role of depressive symptoms and ERI in the relationship between long working hours and presenteeism among village doctors. Methods: We conducted a cross-sectional study in Jiangsu Province, eastern China. 705 village doctors were assessed for working hours, ERI (the Effort-Reward Imbalance Questionnaire, ERI questionnaire), presenteeism (6-item Stanford Presenteeism Scale, SPS-6 Scale), and depressive symptoms (12-item General Health Questionnaire, GHQ-12). A moderated mediation model was used to test the role of depressive symptoms (M), and ERI (W) in the relationship between long working hours (X) and presenteeism (Y). Results: 45.11% of the village doctors worked more than 55 h per week, and 55.89% were exposed to ERI. The prevalence of depressive symptoms among Chinese village doctors was 40.85%. Long working hours (≥ 55 h per week) were significantly associated with presenteeism behaviors (β = 2.17, P < 0.001). Mediation analysis demonstrated that depressive symptoms (GHQ score > 3) could partly mediate the relationship between long working hours and presenteeism (indirect effect β = 0.64, P < 0.001). Moderated mediation further indicated that the interaction of long working hours and ERI was significantly and positively associated with depressive symptoms, which in turn predicted elevated presenteeism behaviors. Conclusions: Depressive symptoms had a mediating role in the association of long working hours with presenteeism behaviors among Chinese village doctors and ERI augment their negative effects. [ABSTRACT FROM AUTHOR]