Abstract: To address the challenge of evaluating the practical application effect of long-lasting phosphorescent road marking materials, this study determined the afterglow luminance decay law of the materials through indoor experiments. The test results showed that the luminance of the materials at 18:00, 20:00, and 22:00 was 16.02 cd/m², 6.51 cd/m², and 1.48 cd/m², respectively. Referring to the measured luminance, a typical road scene was constructed using Twin motion software to conduct a simulation analysis of the luminous effect of the long-lasting phosphorescent road markings. The results showed that the markings provided good visual guidance for drivers at 18:00 and 20:00 due to their high afterglow luminance, and at 22:00, the afterglow luminance remained visible to the naked eyes, assisting drivers in road recognition. Comparison with field construction effects in practical engineering applications showed that the simulation model realistically reproduced the actual application effect of the markings by simulating the afterglow luminance at different time points and the surrounding environment. The application of long-lasting phosphorescent road markings can display road alignment and direction over long distances, indicate road environments, significantly enhance drivers’ ability to recognize roads at night, and improve night-time driving safety, verifying the effectiveness of the simulation model. The method provides a quantitative basis for construction prediction and effect evaluation of long-lasting phosphorescent road marking projects, reducing the cost of trial and error for large-scale material application.