Ginseng and Rhubarb combination ameliorates ferroptosis in persistent inflammation, immunosuppression, and catabolism syndrome via the CaMKII/HO-1 signaling pathway

Abstract

Objective: This study aimed to investigate the therapeutic effects and underlying mechanisms of the Ginseng and Rhubarb combination (GRC) on sepsis-induced persistent inflammation, immunosuppression, and catabolism syndrome (PICS). Methods: The mouse model of sepsis-induced PICS was established using cecal ligation and puncture (CLP). Mice were randomly assigned to five groups: sham, model, model + GRC, model + GRC + oleic acid (OA), and model + GRC + celastrol (CELA). GRC was administered by oral gavage once daily for 7 days, whereas the agonists were injected intraperitoneally for 3 days. Additionally, KN-93 (a calcium/calmodulin-dependent protein kinase II [CaMKII] inhibitor; 5 mg/kg) was used to verify the inhibitory regulation of CaMKII signaling. Seventy-two hours after surgery, serum and tissue samples were collected to evaluate the therapeutic effects of GRC. Outcome measures included survival, body weight, splenic immune cell distribution, histopathology, inflammatory cytokine levels, and ferroptosis-related indicators. Targeted metabolomics and data-independent acquisition (DIA)–based proteomics were performed to identify differential molecules and pathways associated with GRC treatment. Results: Compared to the model group, GRC markedly improved the survival rate and mitigated body weight loss in septic PICS mice. GRC reduced splenic neutrophil and macrophage infiltration and increased the proportions of CD4 + and CD8 + T cells. Histopathological examination revealed attenuation of organ damage. Levels of inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor-alpha (TNF-α), IL-4, IL-13) were significantly decreased, and metabolic disturbances were ameliorated. Biochemical assays showed that GRC reduced Fe 2+ and malondialdehyde (MDA) levels while increasing glutathione peroxidase (GSH-PX) and GPX4, indicating inhibition of ferroptosis. Western blot (WB) analysis demonstrated that GRC downregulated the CaMKII/heme oxygenase-1 (HO-1) pathway, and this regulatory effect was reversed by CaMKII and HO-1 agonists. Multi-omics analysis further revealed that GRC modulated metabolic and proteomic networks associated with immune regulation and ferroptosis, confirming that its therapeutic benefits were mediated through suppression of the CaMKII/HO-1/GPX4 axis. Conclusions: GRC exhibited significant anti-inflammatory, immunomodulatory, and organ-protective effects in the murine model of sepsis-induced PICS. These benefits were mediated through the suppression of ferroptosis via modulation of the CaMKII/HO-1 pathway, providing a novel experimental foundation and a potential therapeutic strategy for managing sepsis-associated PICS.

Keywords

Ferroptosis / Ginseng / PICS / Rhubarb / Sepsis / CaMKII/HO-1/GPX4 axis

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