关键词:
Tiaochang Xiaoliu Decoction
colorectal tumor
CXCR2^(+)neutrophils
gut microbiota
bile acid metabolism
lipid metabolism
摘要:
OBJECTIVE To investigate the therapeutic efficacy of Tiaochang Xiaoliu Decoction(TXD)in colorectal adenomas and to elucidate its underlying mechanisms.METHODS The chemical constituents of TXD were characterized using liquid chromatography-mass spectrometry(LC-MS),and potential therapeutic targets were identified through network pharmacology analysis.The Apc^(Min/+)mice were treated with low-or high-dose TXD,with regorafenib serving as a positive control.Tumor burden was quantified,and epithelial proliferation was assessed by histopathological staining.Neutrophil infiltration,including the CXCR2^(+)neutrophil subset,was evaluated by immunofluorescence and flow cytometry.Transcriptomic profiling(RNA-seq)was performed to determine gene expression changes induced by TXD.Intestinal barrier integrity,gut microbiota composition(16S rRNA sequencing),and fecal metabolomic profiles(untargeted metabolomics)were comprehensively analyzed.RESULTS LC-MS analysis revealed the complex chemical composition of TXD under both positive and negative ion modes.Network pharmacology identified multiple active components targeting key molecules implicated in colorectal adenoma progression,including CXCR2,IL-6,TNF,and MMP9,as well as inflammation-and cancer-related signaling pathways.TXD treatment significantly reduced tumor burden and suppressed epithelial proliferation in Apc^(Min/+)mice.Immunofluorescence analysis demonstrated a marked decrease in Ly6G^(+)MPO^(+)neutrophil infiltration in colonic and tumor tissues(P<0.001).Flow cytometry further showed that TXD significantly reduced the proportions of CD45^(+)Ly6G^(+)CD11b^(+)neutrophils and CXCR2^(+)neutrophil subsets(P<0.001,P<0.01),accompanied by decreased levels of CXCL1/2/5 and IL-6(all P<0.01).Transcriptomic analysis revealed that TXD downregulated genes associated with neutrophil chemotaxis and activation,including CXCR2,Ly6G,and MPO.In addition,TXD significantly restored intestinal epithelial barrier integrity and reshaped gut micr