图1 AVI干预对TNBS模型小鼠疾病状态的影响

Fig.1 Effect of AVI intervention on body weight and disease activity in mice with TNBS-induced CD-like colitis. A: Changes of body weight. B: Changes of DAI scores. WT: Wild type group; TNBS: TNBS-induced model group; AVI: AVI treatment group. *P<0.05 vs WT group. #P<0.05 vs TNBS group.

图2 AVI干预对TNBS模型小鼠肠道炎症的影响

Fig.2 Effect of AVI on intestinal inflammation in the mouse models. A, B: Comparison of colon lengths among the groups. C: Histopathological score of the colon. D: HE staining of colon tissue in different groups. E: AB-PAS staining of colon tissue in different groups. *P<0.05 vs WT group. #P<0.05 vs TNBS group. Scale bar=50 μm.

图3 AVI干预对TNBS模型小鼠肠道炎症因子的影响

Fig.3 Effect of AVI on intestinal inflammatory factors in the mouse models. A, B: ELISA results of TNF‑α and IL-1β in the intestinal mucosa of the mice. C, D: TNF-α and IL-1β mRNA expression in the intestinal mucosa of the mice detected by RT-qPCR. *P<0.05 vs WT group. #P<0.05 vs TNBS group.

图4 不同浓度的AVI作用24 h对Caco-2细胞活力的影响

Fig.4 Changes in viability of Caco-2 cells after treatment with different concentrations of AVI for 24 h. *P<0.05 vs 0 μmol/L.

图5 AVI干预对TNBS模型小鼠结肠炎肠屏障的影响

Fig.5 Effect of AVI on intestinal barrier function in mice with TNBS-induced colitis. A: Immunofluorescence staining for detecting ZO-1, claudin-1 and MUC2 in the mouse colon (Scale bar=50 μm). B, C: Relative expression levels of ZO-1, Claudin-1 and MUC2 proteins in intestinal mucosa detected by Western blotting. *P<0.05 vs WT group. #P<0.05 vs TNBS group.

图6 AVI干预对TNF-α诱导的Caco-2细胞屏障损伤的影响

Fig.6 Effect of AVI on the barrier damage of Caco-2 cells induced by TNF-α. A: Immunofluorescence staining for detecting expressions of ZO-1 and claudin-1 in Caco-2 cells (Scale bar=50 μm). B, C: Relative expression levels of ZO-1 and claudin-1 proteins in Caco-2 cells detected by Western blotting. *P<0.05 vs Control group. #P<0.05 vs TNF-α group.

图7 AVI干预对TNBS模型小鼠肠上皮细胞凋亡的影响

Fig.7 Effect of AVI on intestinal epithelial cell apoptosis in mice with TNBS-induced colitis. A: TUNEL staining of the colon tissue. B: Apoptosis rate of the intestinal epithelial cells (Scale bar=50 μm). C, D: Western blotting for detecting relative expression levels of Bcl-2, Bax and C-caspase3 in colonic mucosa. *P<0.05 vs WT group. #P<0.05 vs TNBS group.

图8 AVI干预对TNF-α诱导的凋亡模型中Caco-2细胞凋亡的影响

Fig.8 Effect of AVI on apoptosis in TNF-α-induced Caco-2 cells. A: TUNEL staining of Caco-2 cells (Scale bar=50 μm). B: Apoptosis rate of Caco-2 cells. C, D: Relative expression levels of Bcl-2, Bax and C-caspase-3 in Caco-2 cells detected by Western blotting.*P<0.05 vs Control group. #P<0.05 vs TNF-α group.

图9 网络药理学分析结果

Fig.9 Network pharmacology analysis results. A: Venn diagram of the intersection between CD genes and AVI genes. B, C: PPI network diagram. D: Results of KEGG pathway enrichment analysis for the intersection genes between AVI and CD.

图10 AVI干预对体内和体外PI3K/AKT/NF-κB通路的影响

Fig.10 Effect of AVI on the PI3K/AKT/NF-κB pathway in the mouse models and in Caco-2 cells. A, B: Relative expression levels of PI3K, p-PI3K, AKT, p-AKT, p65 and p-p65 proteins in mouse colon tissue detected by Western blotting (*P<0.05 vs WT group; #P<0.05 vs TNBS group). C, D: Relative expression levels of PI3K, p-PI3K, AKT, p-AKT, p65 and p-p65 proteins in Caco-2 cells detected by Western blotting (*P<0.05 vs Control group. #P<0.05 vs TNF-α group).

图11 Recilisib对AVI治疗肠上皮细胞凋亡及AKT1 siRNA对体外凋亡模型的影响

Fig.11 Effect of Recilisib and AKT1 siRNA on apoptosis of AVI-treated Caco-2 cells. A, B: Relative expression levels of PI3K, p-PI3K, AKT, p-AKT, p65 and p-p65 proteins detected by Western blotting. C: TUNEL staining of Caco-2 cells (Scale bar=50 μm). D: Apoptosis rate of Caco-2 cells. E, F: Relative expression levels of Bcl-2, Bax, and C-caspase3 proteins detected by Western blotting. *P<0.05 vs AVI group, #P<0.05 vs Recilisib group.