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Interest in integrating 24-hour movement behaviors (sleep, physical activity, and screen time) has increased in recent years, supporting global guidelines that emphasize their relevance for adolescent health. However, there remains a need for studies that assess the effectiveness of interventions aimed at improving these behaviors. This study aimed to evaluate the effectiveness of a school-based intervention grounded in Self-Determination Theory on: (1) 24-hour movement behaviors; (2) adherence to 24-hour movement behaviors recommendations; (3) health-related quality of life; and (4) Self-Determination Theory constructs associated with 24-hour movement behaviors. A quasi-experimental design with experimental and control groups was conducted, including pre-test, post-test, and follow-up assessments. The study involved 82 adolescents aged 14–17 from two conveniently selected schools in the city of Bello, Colombia. The 12-week intervention included educational, family, and extracurricular components. Effectiveness was analyzed using mixed models and relative risk. The results revealed significant effects of the intervention on screen time (weekdays: F(4.68, 156)=2, p = .01; weekend: F(5.51, 156)=2, p = .005; total week: F(6.32, 156)=2, p = .002) and sleep (weekend: F(6.09, 156)= 2, p = .003; total week: F(3.88, 156)=, p = .02). A significant increase in meeting the weakday sleep recommendation was observed in the experimental group (RR: 1.85, IC95% 1.05–3.25), along with improvements in the Self-Determination Theory constructs of competence and relatedness in physical activity. The intervention demonstrated partial effectiveness, achieving improvements in screen time and sleep. This study contributes to the understanding of the design and implementation of educational interventions targeting 24-hour movement behaviors. Future research should refine the integration of motivational strategies to improve physical activity and overall adherence to recommendations, while also exploring the scalability and sustainability of these school-based interventions.近年来，学界对整合24小时运动行为（睡眠、身体活动与屏幕使用时间）的关注度持续提升，相关研究也为强调此类行为与青少年健康息息相关的全球指南提供了支撑。然而，当前仍需开展相关研究以评估旨在改善此类行为的干预措施的实际效果。本研究旨在评估一项基于自我决定理论（Self-Determination Theory）的校园干预措施对以下四方面的效果：（1）24小时运动行为；（2）24小时运动行为指南依从性；（3）健康相关生活质量；（4）与24小时运动行为相关的自我决定理论构念。本研究采用包含前测、后测与追踪测评的准实验设计，设置实验组与对照组。研究对象为哥伦比亚贝洛市两所便利抽样选取的学校中的82名14至17岁青少年。本次为期12周的干预涵盖教育、家庭与课外三大模块。研究采用混合效应模型与相对危险度对干预效果进行分析。研究结果显示，该干预措施对屏幕使用时间（工作日：F(4.68, 156)=2，p=0.01；休息日：F(5.51, 156)=2，p=0.005；全周总量：F(6.32, 156)=2，p=0.002）与睡眠（休息日：F(6.09, 156)=2，p=0.003；全周总量：F(3.88, 156)=，p=0.02）均存在显著影响。实验组中符合工作日睡眠指南的比例显著提升（相对危险度RR=1.85，95%置信区间IC95%=1.05~3.25），同时身体活动领域的自我决定理论构念——能力感与归属感也得到显著改善。该干预措施展现出部分有效性，仅在屏幕使用时间与睡眠层面实现了改善。本研究有助于深化对面向24小时运动行为的教育干预措施的设计与实施的认知。未来研究应优化动机策略的整合方式，以提升青少年的身体活动水平与整体指南依从性，同时探索此类校园干预措施的可推广性与可持续性。