NETs activate the GAS6-AXL-NLRP3 axis in macrophages to drive morphine tolerance

作者：Qingyan Tian, Haiyue Guo, Mengyao Zhang, Kunmao Jiang, Fan Hu, Yan Xu, Li Wan, Xiaokai Zhou, Yinbing Pan, Wentao Liu, Chun‐Yi Jiang · 发表于：Cell Communication and Signaling · 年份：2025 · DOI：10.1186/s12964-025-02181-4 · 被引用次数：4 · 研究领域：Phagocytosis and Immune Regulation、Immune cells in cancer、Immune responses and vaccinations

BACKGROUND: The development of morphine tolerance presents a major clinical challenge in the effective management of severe pain. This study aims to explore the mechanisms underlying morphine tolerance from a novel perspective, with the ultimate goal of uncovering new insights and identifying promising therapeutic targets for its treatment. METHODS: C57BL/6J mice were used in the tail-flick test to evaluate morphine tolerance. Neutrophils derived from mouse bone marrow were employed to investigate the mechanisms underlying morphine-induced NETs formation. Bone marrow-derived macrophages (BMDMs) were harvested from the femur and tibia to study the role of NETs-induced inflammation in analgesic tolerance. Proinflammatory cytokines were measured using Western blotting and real-time PCR. The levels of NETs and the TLR7/9-NLRP3-related signaling pathway were assessed through Western blotting, real-time PCR, and ELISA. Confocal laser scanning microscopy was utilized to visualize NETs in the dorsal root ganglion (DRG) and in cells. RESULTS: Our experiments demonstrated that the levels of NETs in the plasma of patients using morphine for analgesia, as well as in morphine-tolerant animals, were significantly elevated. Genetic elimination of Pad4, neutrophil depletion, and treatment with DNase 1 and RNase A to disrupt NETs formation all effectively alleviated morphine tolerance. These findings indicate that NETs play a critical role in the development of morphine tolerance. Mechanistic...