This study aimed to explore the relationship of work stress with anxiety and depression and the mediation of job satisfaction. We conducted a survey among gas station workers in Wuhan, China. Multivariate and bootstrap mediating analyses were conducted. The rate of effort-reward imbalance, overcommitment (OC), anxiety, and depression were 22.9%, 29.2%, 20.1%, and 19.9%. Work stress was positively associated with anxiety ( β = 0.213 [95% confidence interval (CI), 0.156-0.270] for logarithmic effort-reward ratio [LgERR] and β = 0.563 [95% CI, 0.505-0.621] for OC) and depression ( β = 0.291 [95% CI, 0.227-0.356] for LgERR and β = 0.417 [95% CI, 0.351-0.483] for OC). The mediation of job satisfaction from LgERR and OC to anxiety and depression were 14.93%, 9.05%, 24.34%, and 19.85%. We should concern the anxiety and depression, and work stress status of workers and improve the situation by improving job satisfaction.