Wang Jingjing, Ji Lin, Gao Yingbo, Sun Jingyu, Zhou Xiaobin, Ding Yujia, Zhou Zihan, Guo Xiaofan, Liu Chao, Wang Yujie, Zhang Qingfu, Lv Zhenmu, Ma Dong
Department of Biochemistry and Molecular Biology, Key Laboratory of Neural and Vascular Biology, Ministry of Education, and Hebei Key Laboratory of Cardiovascular Homeostasis and Aging, Hebei Medical University, Shijiazhuang, 050017, Hebei, China.
The Fourth Department of Bone Injury, The First Affiliated Hospital of Hebei University of Traditional Chinese Medicine, Shijiazhuang, 050011, China.
Sci Rep. 2025 May 27;15(1):18509. doi: 10.1038/s41598-025-02488-9.
Cyclin-dependent kinase 5 (CDK5) plays a critical role in the inflammatory response. Macrophages are pivotal orchestrators of inflammation, fibrosis, and wound repair. However, the effectiveness of CDK5 in macrophages on cutaneous wound healing remains inadequately characterized. We determined the role of CDK5 signaling pathway in macrophages in mouse cutaneous wound healing through the established macrophage-specific deletion of CDK5 (myeCDK5) mice and the pharmacological CDK5 inhibitor Roscovitine. Phosphorylated proteomics, western blotting, Masson staining, and dualimmunofluorescence staining were performed to investigate the potential mechanisms underlying CDK5-mediated inflammatory regulation in macrophages in wound healing. CDK5 expression and phosphorylation were both elevated significantly in cutaneous wound healing process in mice. Moreover, an accelerated wound healing in myeCDK5 mice was exhibited with the reduced pro-inflammatory mediators (IL-1β and iNOS) and the elevated anti-inflammatory markers (IL-10 and CD163) expression significantly. CDK5 deficiency in macrophages enhanced tissue remodeling, evidenced by increased collagen deposition and capillary density (CD31 cells). Consistently, Roscovitine-treated mice also showed accelerated wound healing, accompanied by decreased pro-inflammatory factors and increased anti-inflammatory markers at the wound site. Mechanistically, the decreased phosphorylation of SIRT1 at the Ser14 and Ser47 sites, as a substrate of CDK5, was confirmed in myeCDK5 mice. These data are the first to indicate that CDK5 signaling-dependent regulation of SIRT1 phosphorylation in macrophage-mediated inflammation is required for the wound healing process, warranting consideration of the CDK5-SIRT1 pathway as a therapeutic target for cutaneous wound healing.
细胞周期蛋白依赖性激酶5（CDK5）在炎症反应中起关键作用。巨噬细胞是炎症、纤维化和伤口修复的关键协调者。然而，CDK5在巨噬细胞中对皮肤伤口愈合的作用仍未得到充分表征。我们通过建立巨噬细胞特异性缺失CDK5（myeCDK5）小鼠和使用药理学CDK5抑制剂Roscovitine，确定了CDK5信号通路在小鼠皮肤伤口愈合巨噬细胞中的作用。进行磷酸化蛋白质组学、蛋白质印迹、Masson染色和双免疫荧光染色，以研究伤口愈合中巨噬细胞中CDK5介导的炎症调节的潜在机制。在小鼠皮肤伤口愈合过程中，CDK5的表达和磷酸化均显著升高。此外，myeCDK5小鼠伤口愈合加速，促炎介质（IL-1β和iNOS）减少，抗炎标志物（IL-10和CD163）表达显著升高。巨噬细胞中CDK5的缺乏增强了组织重塑，表现为胶原蛋白沉积增加和毛细血管密度（CD31细胞）增加。同样，Roscovitine处理的小鼠也显示伤口愈合加速，同时伤口部位促炎因子减少，抗炎标志物增加。机制上，在myeCDK5小鼠中证实了作为CDK5底物的SIRT1在Ser14和Ser47位点的磷酸化减少。这些数据首次表明，巨噬细胞介导的炎症中CDK5信号依赖的SIRT1磷酸化调节是伤口愈合过程所必需的，这使得CDK5-SIRT1通路有望成为皮肤伤口愈合的治疗靶点。
