Primary health care (PHC) serves as the gatekeeper of health system and PHC physicians take on significant obligations to provide health care services in the pursuit of Universal Health Coverage (UHC). PHC physicians' health-related quality of life (HRQoL) can have a strong impact on patients, physicians and the health care system. Lifestyle interventions are found to be effective to improve HRQoL. The purpose of this study was to evaluate the association between lifestyle behaviors and HRQoL among PHC physicians, so that lifestyle intervention can be tailored by policy makers for health promotion. A survey covering 31 provinces and administrative regions in China was conducted in 2020 using a stratified sampling strategy. Data on sociodemographic characteristics lifestyle behaviors and HRQoL were collected by a self-administered questionnaire. HRQoL was measured through EuroQol-five dimension-five level (EQ-5D-5L) instrument. A Tobit regression model was performed to evaluate the association between sociodemographic characteristics, lifestyle behaviors and HRQoL. Among 894 PHC physicians who completed the survey, Anxiety/Depression (AD) was the dimension with the most problems reported (18.1%). Regular daily routine (β = 0.025, 95%CI 0.004 to 0.045) and good sleep quality (β = 0.049, 95% CI = 0.029 to 0.069) were protective factors for HRQoL, while smoking (β = -0.027, 95% CI = -0.079 to -0.003) and frequency of eating breakfast (β = -0.041, 95%CI = -0.079 to -0.003) were negatively associated with HRQoL. Physical activity and alcohol drinking were not significantly associated with HRQoL. These findings suggest that tailored interventions on daily routine, improving sleep quality, and tobacco control among PHC physicians may be effective strategies to improve their HRQoL.