Xuebijing injection (XBJ) is widely used in China as an adjunctive therapy for sepsis and sepsis-associated acute kidney injury (SAKI). Here, we evaluated its immunometabolic and inflammatory effects in macrophages and identified candidate bioactive monomers to define the underlying mechanisms. An inflammatory model was established in RAW264.7 macrophages using lipopolysaccharide, and targeted energy metabolomics identified succinate (SA) as a key responsive metabolite. XBJ-derived monomer screening prioritized Albiflorin for mechanistic investigation. Exogenous SA replenishment, pharmacological inhibition, and siRNA-mediated knockdown were used to investigate key glycolytic nodes and examine causal relationships. Key findings were validated in bone marrow-derived macrophages (BMDMs) and in a cecal ligation and puncture (CLP)-induced SAKI rat model. In RAW264.7 macrophages, XBJ markedly reduced SA, and phenotype-guided screening prioritized Albiflorin as a representative monomer for mechanistic investigation. Albiflorin improved mitochondrial respiration, attenuated oxidative stress, and restored succinate dehydrogenase activity, consistent with reduced SA accumulation. Exogenous SA replenishment partially reversed Albiflorin's anti-glycolytic and anti-inflammatory effects and restored PFKFB3 expression, whereas HIF-1α inhibition or knockdown did not block SA-induced PFKFB3 upregulation. PFKFB3 inhibition or knockdown reduced inflammatory outputs and lactate production and constrained SA-driven glycolytic enhancement. These findings were further supported in BMDMs and in a CLP-induced SAKI rat model. This study identified SA as an XBJ-responsive metabolite linked to macrophage inflammation and prioritized Albiflorin as a representative effector monomer for mechanistic investigation. Albiflorin lowered SA burden and suppressed PFKFB3-associated glycolytic and inflammatory activation in macrophages, supporting its contribution to SAKI protection. Albiflorin alleviates SAKI by modulating the macrophage SA-PFKFB3 axis. As a representative effector monomer identified through formula deconvolution of XBJ, Albiflorin reduces mitochondrial ROS accumulation, restores SDH activity, and decreases SA accumulation. Reduced SA accumulation dampens PFKFB3-mediated glycolytic activation, thereby attenuating pro-inflammatory macrophage activation and inflammatory cytokine release, ultimately decreasing renal macrophage infiltration and ameliorating SAKI. Additional HIF-1α inhibition and knockdown experiments indicate that SA-induced PFKFB3 upregulation is not primarily dependent on HIF-1α signaling. • SA is identified as an XBJ-responsive effector metabolite linked to inflammatory outputs. • Albiflorin is prioritized as the core monomer mediating XBJ-driven SA reduction and anti-inflammatory effects. • Albiflorin restores SDH activity and mitochondrial redox-respiratory function, limiting intracellular SA accumulation. • SA upregulates PFKFB3 to drive glycolysis-linked inflammation, and Albiflorin disrupts the SA–PFKFB3 axis. • These findings were established in RAW264.7 cells and BMDMs and validated in CLP-induced SAKI in vivo.
Xu et al. (Thu,) studied this question.