The establishment of an early pro-regenerative niche is crucial for tissue regeneration^1,2. Gasdermin D (GSDMD)-dependent pyroptosis accounts for the release of inflammatory cytokines upon various insults^3-5. However, little is known about its role in tissue regeneration followed by homeostatic maintenance. Here, we show that macrophage GSDMD deficiency delayed tissue recovery, with little impact on the local inflammatory milieu or the lytic pyroptosis process. Metabolite secretome profiling of hyperactivated macrophages unveiled the non-canonical metabolite-secreting function of GSDMD. And we further identified 11,12-epoxyeicosatrienoic acid (11,12-EET) as a bioactive pro-healing oxylipin, secreted from hyperactive macrophages in a GSDMD-dependent manner. Indeed, accumulation of 11,12-EET by direct supplementation or deletion of its hydrolytic enzyme Ephx2 accelerated muscle regeneration. We further demonstrated that the Ephx2 level accumulated within aged muscle. And consecutive 11,12-EET treatment rejuvenated aged muscle. Mechanistically, 11,12-EET amplifies FGF-FGFR signaling by modulating FGF liquid-liquid phase separation, hence boosting the activation and proliferation of muscle stem cells (MuSCs). These data depict a GSDMD-guided metabolite crosstalk between macrophages and MuSCs that governs the repair process, which offers new therapeutic insights for the regeneration of injured or aged tissues.

早期促再生生态位的建立对于组织再生至关重要^1,2。Gasdermin D （GSDMD） 依赖性焦亡导致炎性细胞因子在各种损伤后释放^3-5。然而，人们对它在组织再生和稳态维持中的作用知之甚少。在这里，我们表明巨噬细胞 GSDMD 缺陷延迟了组织恢复，对局部炎症环境或裂解焦亡过程几乎没有影响。高活化巨噬细胞的代谢物分泌组分析揭示了 GSDMD 的非经典代谢物分泌功能。我们进一步鉴定出 11,12-环氧二十碳三烯酸 （11,12-EET） 是一种具有生物活性的促愈合氧磷脂，以 GSDMD 依赖性方式从过度活跃的巨噬细胞分泌。事实上，通过直接补充或缺失其水解酶 Ephx2 积累 11,12-EET 加速了肌肉再生。我们进一步证明 Ephx2 水平在衰老的肌肉中积累。和连续的 11,12-EET 治疗使衰老的肌肉恢复活力。从机制上讲，11,12-EET 通过调节 FGF 液-液相分离来扩增 FGF-FGFR 信号传导，从而促进肌肉干细胞 （MuSCs） 的活化和增殖。这些数据描述了巨噬细胞和 MuSC 之间 GSDMD 引导的代谢物串扰，它控制修复过程，这为受伤或老化组织的再生提供了新的治疗见解。