This is the first study in China to investigate the prevalence of workplace violence (WPV) and its long-term association with post-traumatic stress disorder (PTSD) among healthcare workers. Conducted across various time points from March 27, 2020, to April 10, 2023, the research employed a four-wave repeated cross-sectional design involving 14,993 healthcare workers from 32 provincial administrative regions across the country. Nurses accounted for 34.3% of the sample, while doctors accounted for 65.7% of the sample, with a higher concentration in western China. Multivariate logistic regression analysis was employed to identify the risk factors for WPV, and hierarchical logistic regression models were used to examine the association between PTSD and WPV. The findings revealed that 71.4% of healthcare workers reported experiencing or witnessing WPV, with nearly half (47.5%) indicating exposure to violent incidents within the past year. Further analysis revealed significant associations between WPV incidence and various factors, including age, gender, work experience, work setting, and a history of contact with Coronavirus Disease 2019 (COVID-19) patients (the odds ratio [OR] of these influencing factors ranged from 1.10 to 2.03. All the p-values were less than .05). 28.5% of healthcare workers displayed symptoms of PTSD, with WPV identified as a major risk factor contributing to these symptoms (lifetime WPV: OR = 1.61, 95% CI [1.47-1.76]; 1-year WPV: OR = 1.75 [1.61-1.90]). Therefore, it is imperative to prioritize the mental health of healthcare workers and implement effective measures to prevent and mitigate WPV, ensuring their physical and psychological well-being as well as occupational safety.