图1 1,3-DA对DSS诱导的小鼠肠炎症状的影响

Fig.1 Effect of 1,3-DA intervention on DSS-induced colitis in mice (n=10). A: Schematic of colitis modeling and treatments of the mice. B: Body weight changes of the mice in each group. C, D: Comparison of colon length of the mice among the groups. E: Disease activity index (DAI) score of the mice after different treatments. *P<0.05 vs CON group, #P<0.05 vs DSS group.

图2 1,3-DA对DSS小鼠肠组织病理学的影响

Fig.2 Effect of 1,3-DA on intestinal histopathology in DSS-treated mice (n=10). A: HE staining and inflammatory scores of intestinal tissues of the mice in each group (Original magnification: ×200). B: Histopathological scores of the mice in each group. C: AB-PAS staining of the intestinal tissues in each group (×200). D: Goblet cell counts in the visual field of each group. *P<0.05 vs CON group, #P<0.05 vs DSS group.

图 3 1,3-DA 对 DSS 小鼠结肠组织炎症因子的影响

Fig.3 Effect of 1,3-DA on inflammatory factors in the intestinal tissue of DSS-treated mice (n=10). A: Immunohistochemical staining for IFN-γ and relative OD values in different groups. B: Immunohistochemical staining for MPO and relative OD values in different groups (Scale bar=100 μm). *P<0.05 vs CON group, #P<0.05 vs DSS group.

图4 1,3-DA对DSS小鼠结肠组织中紧密连接蛋白的影响

Fig.4 Effect of 1,3-DA on tight junction proteins in the colonic tissues of DSS-treated mice (n=10). A: Immunofluorescence staining in each group. B: Western blotting results in each group. *P<0.05 vs CON group, #P<0.05 vs DSS group.

图5 1,3-DA对DSS小鼠结肠组织中氧化应激的调控作用

Fig.5 Regulatory effect of 1,3-DA on oxidative stress in the colonic tissues of DSS-treated mice (n=10). A: GSH-PX levels in each group. B: SOD levels in each group. C: MDA levels in each group. D: CAT levels in each group. *P<0.05 vs CON group, #P<0.05 vs DSS group.

图6 1,3-DA对H₂O₂诱导的NCM460细胞肠屏障的影响

Fig.6 Effect of 1,3-DA on intestinal barrier of H2O2-induced NCM460 cells (n=6). A: Immunofluorescence staining of tight junction proteins claudin-1 and ZO-1 in the cells. B: Western blotting of claudin-1 and ZO-1 in the cells in each group. *P<0.05 vs CON-C group, #P<0.05 vs H2O2 group.

图7 1,3-DA对H2O2诱导的NCM460细胞活性氧的影响

Fig.7 Effect of 1,3-DA on ROS induced by H2O2 in NCM460 cells (n=5). A, B: Flow cytometry for analyzing ROS in the cells in each group. C: ROS staining with fluorescent probe DCFH-DA in the cells in each group. *P<0.05 vs CON-C group, #P<0.05 vs H2O2 group.

图8 1,3-DA网络药理学分析

Fig.8 1,3-DA network pharmacology. A: Venn diagram. B: PPI network diagram. C: KEGG pathway enrichment analysis. D, E: Molecular docking results of the interaction between PI3Kδ and 1,3-DA. F, G: Results of molecular docking between AKT1 and 1,3-DA.

图9 1,3-DA调控PI3K/Akt通路改善结肠炎小鼠肠道氧化应激损伤

Fig.9 1,3-DA regulates the PI3K/Akt pathway to improve intestinal oxidative stress damage in mice with colitis (n=10 for in vivo experiments and n=6 for in vitro experiment). A, B: Western blotting for detecting intestinal PI3K and Akt expressions of the mice. C, D: Western blotting for detecting PI3K and Akt expressions in H2O2-induced NCM460 cells with different treatments. E-H: Intestinal oxidative stress indexes (GSH-PX, SOD, MDA, and CAT). I, J: Intestinal tight junction proteins detected by immunofluorescence staining and Western blotting. K, L: Intracellular ROS levels detected by flow cytometry (drug-treated groups vs 740Y-P activator group). M: Intracellular ROS detected using DCFH-DA staining (drug-treated vs 740Y-P activator group). *P<0.05 vs CON or CON-C group, #P<0.05 vs H2O2 or DSS group.