肛提肌是盆底支持组织的关键结构，其损伤和再生修复与女性压力性尿失禁（SUI）的发病密切相关，但机制尚未明确。申请人的前期研究发现小鼠源成肌细胞（C2C12）损伤修复过程中ArfGAP3表达明显增高，细胞内囊泡分泌及GLUT4质膜聚集增多，提示ArfGAP3及GLUT4囊泡转运可能参与肌肉损伤修复过程。本研究拟构建C2C12细胞机械力损伤模型、小鼠肛提肌机械力和低雌激素损伤模型，应用CRISPR/Cas9技术进行ArfGAP3、GLUT4、CP/AP基因干预；通过透射电镜进行亚细胞结构分析，并结合肌肉电生理检测、PET/CT以及免疫标记、激光共聚焦观察等分子生物学方法在细胞水平和整体水平探讨ArfGAP3调控GLUT4质膜转运在肛提肌损伤修复中的作用，明确参与该过程的CP/AP类型和cAMP相关信号通路，从而阐明ArfGAP3调控盆底肛提肌损伤修复的机制，为SUI的防治提供新思路。

The levator ani muscle is a vital structure of pelvic supporting tissues. The injury and repair of levator ani muscle is strongly correlated with the pathogenesis of female stress urinary incontinence (SUI), but the exact mechanism is still not understood. Preliminary studies show that the expression levels of ArfGAP3, the secretion of intracellular vesicles and the content of plasma membrane GLUT4 are all increased significantly in regenerating mouse-derived myoblasts (C2C12) after mechanical injury, which suggests that ArfGAP3 and vesicle trafficking of GLUT4 may be involved in the repair process of muscle. In our study, a C2C12 cell model of mechanical damage, a mechanical traction-induced and an estrogen withdraw-induced mouse model of levator ani muscle injury will be established, CRISPR/Cas9 technology will be used for the gene intervention of ArfGAP3, GLUT4 and CP/AP. Besides, transmission electron microscopy will be utilized for subcellular structure analysis, and in combination with muscle electrophysiological measurement, PET/CT, immunolabelling, laser confocal observation and other molecular biological techniques to explore the role and related signaling pathways of ArfGAP3 in the process of levator ani muscle regeneration via regulating plasma membrane translocation of GLUT4 at cellular and integral levels, identify the subtypes of CP/AP and cAMP-related signaling pathways involved in this process, and further elucidating the mechanism that ArfGAP3 regulates levator ani muscle repair and providing innovative ideas for the prevention and therapy of SUI.