Work fatigue refers to physical, mental, and emotional exhaustion, resulting in the inability to work. Hitherto research indicate a close relationship between high-performance work systems and work fatigue, and there may be a double-edged sword effect of high-performance work systems on work fatigue. However, a comprehensive theoretical framework has not been developed to understand the relationship between them. Based on the challenge-hindrance stress model, this study employs role overload and job responsibility as mediating variables in a conceptual framework to understand the impact of high-performance work systems on work fatigue. Using the partial least square structural equation model and a sample of 360 employees in China, the mediating effects of role overload and job responsibility were confirmed. Further, the internal mechanisms of how high-performance work systems affect work fatigue are discussed, its adverse effects are confirmed, and its practical implications are proposed.