Figure 4. The novel P2X2 antagonist ambroxol prevents IL‐6 release from EGCs.

• A
  Ambroxol treatment scheme. Mice were treated with ambroxol or vehicle and underwent a sham operation (laparotomy) or intestinal manipulation (IM). Small bowel muscularis externa (ME) was isolated and analyzed 3 or 24 h after surgery.
• B–D
  Postoperative gene expression analyses of indicated gliosis marker in the ME.
• E
  Histological analysis of EGC proliferation by quantification of Ki67 (violet)‐ and Sox10 ‐positive EGCs at IM24h. Mice were treated as seen in (A).
• F
  Histological counting of infiltrating myeloperoxidase (MPO)‐positive leukocytes in the postoperative ME (n = 6–8).
• G
  FACS analysis of infiltrating cells in the ME of mice treated with ambroxol or vehicle. CD45, Ly6C, and CX3CR1 were used to distinguish resident macrophages (CD45⁺/Ly6C⁻/CX3CR1⁺), infiltrating monocytes (CD45⁺/Ly6C⁺/CX3CR1⁻) and infiltrated monocyte‐derived macrophages (CD45⁺/Ly6C⁺/CX3CR1⁺).
• H
  Postoperative in vivo GI transit measurement in mice treated with ambroxol or vehicle.

Data information: In (B–D), data are represented as fold change + SEM vs the sham groups (n = 6–8, POI mice per group). In (E), data are represented as mean of double‐positive cells per total Sox10‐positive cells + SEM; six whole mount specimens per conditions; n = 11 mice per IM group and n = 3 per sham group. In (F), data are presented as mean + SEM MPO⁺ cells/mm² small intestine ME tissue. In (G), data are presented as cells per 100 mg ME tissue + SEM n = 3–5 POI mice per group. In (H), data are presented as mean + SEM; n = 12 mice per group. Statistics were performed by applying unpaired Student's t‐test and one‐way ANOVA with a subsequent Bonferroni test (B‐H). * indicates significance compared to sham animals, and ^# indicates significance between vehicle and ambroxol treatment with */^# P < 0.05, **/^## P < 0.01, and ***/^### P < 0.001.