Wumei Pills enhance intestinal stem cell - mediated repair in chemotherapy-induced mucositis via Lactobacillus reuteri - dependent modulation of the gut microbiota

Figure 1 The flow chart of mouse experiment. 5-FU: 5-fluorouracil; ABX: Antibiotic mixture; WMP: Wumei Pills; L. reuteri: Lactobacillus reuteri; CFU: Colony-forming unit.

Figure 2 Effects of Wumei Pills and Lactobacillus reuteri on general condition, intestinal inflammation, villus height and crypt depth in 5-fluorouracil and pseudo-germ-free mice. A: Body weight; B: Food intake; C and D: Diarrhea scores; E: Crypt depth; F: Villus height; G: Representative hematoxylin and eosin-stained images in the colon and jejunum. Values represent the mean ± SEM. ^aP < 0.05 vs control group; ^bP < 0.01 vs control group; ^cP < 0.001 vs control group; ^dP < 0.0001 vs 5-fluorouracil + antibiotic mixture group; ^eP < 0.05 vs 5-fluorouracil group. 5-FU: 5-fluorouracil; ABX: Antibiotic mixture; WMP: Wumei Pills; L. reuteri: Lactobacillus reuteri.

Figure 3 Effect of Wumei Pills and Lactobacillus reuteri on inflammatory cytokine and lipopolysaccharides. A: Inflammatory cytokine level; B: The expression of lipopolysaccharides; C: Colonization of Lactobacillus reuteri (L. reuteri) [M: 5-fluorouracil (5-FU) group; M_Ler: 5-FU + L. reuteri; M_WMP: 5-FU + Wumei Pills (WMP); M_ABX; 5-FU + antibiotic mixture (ABX) group; M_ABX_Ler: 5-FU + ABX + L. reuteri; M_ABX_WMP: 5-FU + ABX + WMP]. ^aP < 0.05; ^cP < 0.001. 5-FU: 5-fluorouracil; ABX: Antibiotic mixture; WMP: Wumei Pills; L. reuteri: Lactobacillus reuteri; IL: Interleukin; TNF: Tumor necrosis factor; LPS: Lipopolysaccharides.

Figure 4 Concentration of dextran measured by mean fluorescence intensity of fluorescein isothiocyanate in serum. Values represent the mean ± SEM. ^cP < 0.001. FITC: Fluorescein isothiocyanate; 5-FU: 5-fluorouracil; ABX: Antibiotic mixture; L. reuteri: Lactobacillus reuteri; WMP: Wumei Pills.

Figure 5 Wumei Pills and Lactobacillus reuteri enhance the proliferation of intestinal cells in chemotherapy-induced intestinal mucositis mice. Images showing immunofluorescence staining with proliferating cell nuclear antigen antibodies in the jejunum. 5-FU: 5-fluorouracil; ABX: Antibiotic mixture; WMP: Wumei Pills; L. reuteri: Lactobacillus reuteri; PCNA: Proliferating cell nuclear antigen.

Figure 6 Wumei Pills and Lactobacillus reuteri increase intestinal stem cell activity and activates the Wingless-type/β-catenin signaling pathway in chemotherapy-induced intestinal mucositis mice. A: The gene expression of Wingless-type 3, β-catenin analyzed by immunohistochemistry staining; B: Images showing immunohistochemistry staining with leucine-rich-repeat-containing G-protein-coupled receptor 5 antibodies in the jejunum. 5-FU: 5-fluorouracil; ABX: Antibiotic mixture; L. reuteri: Lactobacillus reuteri; WMP: Wumei Pills; Wnt3: Wingless-type 3; Lgr5: Leucine-rich-repeat-containing G-protein-coupled receptor 5.

Figure 7 Wumei Pills regulates Lactobacillus reuteri promoting intestinal stem cells activity by reactivating the Wingless-type/β-catenin signaling pathway to attenuate 5-fluorouracil induced intestinal epithelial injury. 5-FU: 5-fluorouracil; CIM: Chemotherapy-induced intestinal mucositis; CSM: Crypt-specific microbiota; ISC: Intestinal stem cell; Wnt: Wingless-type.