Dachengqitang mitigates endoscopic retrograde cholangiopancreatography-induced pancreatitis

Figure 1 Illustrates the data processing and identification of differentially expressed genes. A: A heatmap displaying the sample-to-sample correlation. The diabetes group displayed a stronger intra-group correlation compared to the control group; B: A volcano plot generated using the limma R tool, showcasing all the differentially expressed genes in both the control and diabetes groups.

Figure 2 Functional and pathway enrichment. A: Gene Ontology enrichment analysis showing significant enrichment in blood microparticle, primary lysosome, azurophil granule, and vacuolar lumen; B: Molecular function analysis displaying enrichment in endopeptidase inhibitor activity, peptidase inhibitor activity, endopeptidase regulator activity, and peptidase regulator activity.

Figure 3 Functional and pathway enrichment analyses were performed on the gene set enrichment analysis results. A: Enrichment in lymphoid cell-related pathways; B: Enrichment in non-lymphoid cell-related pathways; C: Wiki pathways analysis showing extrafollicular B cell activation by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2); D: Wiki pathways analysis displaying network map of SARS-CoV-2 signaling pathway; E: Kyoto Encyclopedia of Genes and Genomes pathway analysis revealing enrichment in cytokine-cytokine receptor interaction and chemokine signaling pathway.

Figure 4 The extent of AP-induced intestinal injury was found to be positively correlated with Dachengqitang in mice. A-C: Hematoxylin and eosin staining showed milder injury in mice treated with Dachengqitang (DCQT); D-I: Immunofluorescence analysis revealed a significant decrease in the intestinal injury marker following 24 hours of AP treatment; J and K: Furthermore, the protein expression of occludin and ZO-1 notably increased post-DCQT treatment, as illustrated in the immunofluorescence images. ^aP < 0.05. A-I features a scale bar of 50 μm. DCQT: Dachengqitang.

Figure 5 In vivo experiments have demonstrated that clearance of macrophages can improve intestinal function and the integrity of the epithelium. A: Histological examination staining showing intestinal morphology in Control group; B: In Control group, immunofluorescence staining displaying ZO-1 expression in intestinal tissues; C: In Control group, immunofluorescence staining showing CD86 expression; D: Histological examination staining showing intestinal morphology in SAP group; E: In SAP group, immunofluorescence staining displaying ZO-1 expression in intestinal tissues; F: In SAP group, immunofluorescence staining showing CD86 expression; G: Histological examination staining showing intestinal morphology in SAP model treated with DCQT (SAP + DCQT) group; H: In SAP + DCQT group, immunofluorescence staining displaying ZO-1 expression in intestinal tissues; I: In SAP + DCQT group, immunofluorescence staining showing CD86 expression; J: Histological examination staining showing intestinal morphology in SAP + DCQT + CI group; K: In SAP + DCQT + CI group, immunofluorescence staining displaying ZO-1 expression in intestinal tissues; L: In SAP + DCQT + CI group, immunofluorescence staining showing CD86 expression; M: Quantitative analysis of ZO-1 fluorescence intensity; N: Quantitative analysis of CD86 fluorescence intensity. SAP significantly decreased ZO-1 expression and increased CD86 expression, while DCQT treatment restored ZO-1 levels and reduced CD86 expression. ^aP < 0.05. DCQT: Dachengqitang.

Figure 6 In vivo studies have shown that Dachengqitang leads to the upregulation of Arg-1 and IL-10. A-J: Immunofluorescence staining showing Arg-1 expression in intestinal tissues across control, diabetes, and Dachengqitang (DCQT) treatment groups; K-T: Immunofluorescence staining displaying IL-10 expression levels across different experimental groups. Diabetes induction significantly decreased Arg-1 and IL-10 fluorescence intensity, while DCQT treatment restored their expression levels DCQT. The images are accompanied by a scale bar of 50 μm. ^aP < 0.05. DCQT: Dachengqitang.

Figure 7 In vitro models demonstrated that oxygen-glucose deprivation inhibits injury to the intestinal epithelium. A-C: Immunofluorescence staining showing decreased Occludin expression following OGD treatment; D-F: Immunofluorescence staining displaying reduced ZO-1 expression after OGD treatment; G: DCQT administration restored Occludin fluorescence intensity in OGD group; H: DCQT treatment improved ZO-1 fluorescence intensity in OGD group. ^aP < 0.05. Scale bar: 50 μm. OGD: Oxygen-glucose deprivation; DCQT: Dachengqitang.

Figure 8 Dachengqitang induces mitochondrial impairment in Caco-2 cells in vitro. A-C: Mitochondrial membrane potential imaging in Control group after JC-1 staining; D-F: Mitochondrial membrane potential imaging in oxygen-glucose deprivation (OGD) group after JC-1 staining; G-I: Mitochondrial membrane potential imaging in Dachengqitang (DCQT) treatment group after JC-1 staining. Figure showing mitochondrial membrane potential across three groups; J: DCQT protects against OGD-induced mitochondrial membrane potential loss. DCQT treatment significantly reduced JC-1 monomer formation and improved mitochondrial membrane potential compared to OGD treatment alone. ^aP < 0.05. OGD: Oxygen-glucose deprivation; DCQT: Dachengqitang.