Night running is becoming an important physical activity during nighttime hours. However, the formal running spaces available in cities are often insufficient to meet the daily fitness needs of urban populations. Consequently, street spaces are frequently incorporated into the choices of night runners. Artificial lighting is an indispensable factor in the street running environment, however, the research on how artificial lighting in urban streets affects the comfort of night running spaces remains unclear. This study selected three classic night running routes in Suzhou City for evaluation using a fitness app for running route recommendations and conducted on-site photography using a panoramic camera. Subsequently, individuals experienced in night running were recruited to conduct a questionnaire survey on the artificial lighting factors depicted in the photographs. The results indicate that the brightness of environmental lighting in streets has the greatest impact on the comfort of night runners, followed by lighting coverage and color, while the type of environmental lighting has the least impact on the comfort of night runners. The final results can provide a theoretical basis for optimizing the lighting conditions of urban night running spaces, thereby contributing to the creation of a more comfortable urban night running environment.