Component identification and integrated ileal single-cell sequencing analysis reveal the protective role of Sijunzi Decoction against stress-induced intestinal injury via targeting the TLR4/NF-κB pathway
XU Hao
XIAO Guo-zhong
CHEN Hua-xian
LIN Hong-cheng
HE Qiu-lan
Abstract：Objective To investigate the molecular mechanisms by which Sijunzi Decoction(SJZ)ameliorates stress-induced intestinal mucosal injury(SII)in a severe burn mouse model through a combined strategy of mass spec-trometry-based component identification and ileal single-cell RNA sequencing analysis.Methods A mouse model of stress-induced intestinal injury following severe burn was established.Active compounds in SJZ were identified via mass spectrometry,and ileal single-cell transcriptomic analysis was performed.Potential molecular targets of SJZ in SII were screened through integrated bioinformatics analysis,and validated by histopathological examination,immunofluorescence,and qPCR.Results A total of 2 415 chemical constituents were detected in SJZ,including peptides,tri terpenoids,and flavonoids.Specifically,28 components were identified from Panax ginseng,7 from Atractylodes macrocephala,12 from Poria cocos,and 13 from Glycyrrhiza uralensis.Severe burn led to significant reduction in intestinal epithelial cell popula-tions and upregulation of the NF-κB signaling pathway.Integrated analysis identified TLR4 and NF-κB as key targets regulated by SJZ.SJZ treatment reduced epithelial cell loss,increased mucin secretion,and downregulated TLR4 and NF-κB at both mRNA and protein levels compared with the sterile water group.Conclusion SJZ may exert protective effects against stress-induced intestinal injury by suppressing the TLR4/NF-κB signaling pathway and preserving intes-tinal epithelial integrity.
Keywords：Sijunzi Decoctionstress-induced intestinal mucosal injurysingle-cell RNA sequencingTLR4/NF-κB signaling pathway
Publication Date：2025-06-15
Online Publishing Date：2025-08-18(First online date of this platform, not the publication date of the document)
Pages：8( 818-825 )