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| Effect of Electroacupuncture Combined with Spleen-Invigorating and Qi-Regulating Method on Intestinal Flora Disorder in Sepsis Rats via TLR4/NF-κB Signaling Pathway |
| ZANG Hongmin, CUI Ying, et al |
| Shijiazhuang Hospital of Traditional Chinese Medicine, Hebei Shijiazhuang 050000, China |
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Abstract Objective: To explore the improvement effect of electroacupuncture combined with spleen-invigorating and qi-regulating formula on intestinal flora disorder in sepsis rats and its possible molecular mechanism based on the TLR4/NF-κB signaling pathway. Methods: SD rats were divided into sham operation group, model group, electroacupuncture group, spleen and qi formula group, and electroacupuncture + spleen and qi formula group, and the sepsis rat model was constructed by cecal ligation and perforation. HE staining was used to detect pathological damage of colon tissues. ELISA was applied to detect serum levels of TNF-α, IL-6, and IL-1β. 16S rRNA sequencing was performed to analyze intestinal flora. Western blot was used to detect the expression of TLR4, p-p65 NF-κB/p65 NF-κB, p-IκB and other proteins related to the TLR4/NF-κB signaling pathway in colon tissues. Results: Compared with the sham operation group, the model group showed obvious pathological damage in colon tissues, significantly increased serum levels of TNF-α, IL-6, and IL-1β (P<0.05), and decreased alpha diversity indices (observed_species, Chao1, Shannon, Simpson) of intestinal flora (P<0.05). At the phylum level, the relative abundance of Proteobacteria increased, while that of Firmicutes and Bacteroidetes decreased. At the genus level, the relative abundances of Alloprevotella and Bifidobacterium (related to short-chain fatty acid production and anti-inflammation) decreased, while those of Escherichia-Shigella and Klebsiella (related to lipopolysaccharide production) increased. The expression of TLR4, p-p65 NF-κB/p65 NF-κB, and p-IκB proteins in colon tissues significantly increased (P<0.05). Compared with the model group, the electroacupuncture group and spleen-invigorating and qi-regulating formula group showed alleviated pathological damage in colon tissues, significantly decreased serum levels of TNF-α, IL-6, and IL-1β (P<0.05), increased alpha diversity indices of intestinal flora (P<0.05), increased relative abundances of Firmicutes, Bacteroidetes, Alloprevotella, and Bifidobacterium, decreased relative abundances of Proteobacteria, Escherichia_Shigella, and Klebsiella, and significantly decreased expression of TLR4, p-p65 NF-κB/p65 NF-κB, and p-IκB proteins in colon tissues (P<0.05). The effect of electroacupuncture combined with spleen-invigorating and qi-regulating formula was significantly better than that of single electroacupuncture or formula (P<0.05). Conclusion: Electroacupuncture combined with spleen-invigorating and qi-regulating formula may improve intestinal flora disorder and inhibit inflammation by inhibiting the TLR4/NF-κB signaling pathway, thereby reducing intestinal damage in sepsis rats, and the combination of the two therapeutic methods has a synergistic effect.
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2025, Vol. 31 
