Tongfu Pingchuan decoction enhances Chrm4 expression in intestinal macrophages to alleviate sepsis in rats

This study aims to explore the protective role of Tongfu Pingchuan decoction (TFP) on sepsis. Thirty rats were randomly assigned to Sham, sepsis (cecal ligation and puncture, CLP), and CLP + Tongfu Pingchuan decoction (TFP) groups (n = 10 per group). TFP (1 mL/kg) or PBS was administered by gavage twice daily for 7 days. Cecum tissues were subjected to transcriptome sequencing (n = 6 per group), followed by network pharmacology and WGCNA analyses. Key findings were validated in rat cecum tissues and LPS-stimulated RAW264.7 macrophages. Compared with the Sham group, the Model group showed activation of TNF signaling, complement and coagulation cascades, fatty acid metabolism, and PPAR signaling pathways, along with suppression of cholinergic synapse, ECM-receptor interaction, and vascular smooth muscle contraction pathways. These alterations were largely reversed by TFP treatment. Integrated bioinformatics analyses identified Chrm4 and Pygm as overlapping differentially expressed genes between predicted TFP targets and a key WGCNA module. Chrm4 expression was significantly downregulated in septic rats and restored by TFP administration. Sepsis induced M1 macrophage polarization and suppressed M2 polarization in cecum tissues, whereas TFP reduced M1 markers and enhanced M2 markers. In LPS-stimulated RAW264.7 cells, TFP similarly promoted M2 polarization, reduced IL-6 and TNF-α levels, increased IL-10 expression, and suppressed IL1RN, p-ERK2/ERK2, and p-AKT/AKT signaling. These effects were abolished by Chrm4 silencing or treatment with the PCTR1 inhibitor Boc-2. These findings demonstrate that TFP alleviates sepsis by restoring intestinal Chrm4 expression, promoting macrophage polarization toward a pro-resolving phenotype, and modulating ERK/AKT-associated inflammatory signaling pathways.

The online version contains supplementary material available at 10.1007/s13205-026-04871-3.