Objective: To evaluate the diagnostic potential of serum DIA in detecting differential proteins in athletic patients with aortic dissection (AD) complicated by acute myocardial infarction (AMI). Methods: A cohort consisting of AD and AMI patients, including athletes, was assessed from July 2018 to August 2021. Serum DIA levels were compared among 30 participants divided into AD, MI, and control groups. High-throughput DIA quantification and Western blotting were used for protein marker analysis. Results: DIA analysis identified significant differences in protein expression among the groups. Proteomic profiling indicated distinct patterns in athletes, with specific markers showing potential for differential diagnosis. The diagnostic value of these markers was further validated using ROC curves. Conclusion: The study underscores the diagnostic relevance of serum DIA in athletic patients with AD and AMI, highlighting specific protein markers as potential diagnostic tools. This research contributes to the understanding of cardiovascular risks and diagnostic strategies in athletic populations.