Figure 9. Andrographolide-driven mitophagy-mediated NLRP3 inflammasome inactivation is responsible for amelioration of murine models for colitis and CAC. (A–E) Mice were treated with 2.5% DSS in their drinking water for 7 d to induce colitis. Andro (5 mg/kg) was administered i.p. daily. CQ (50 mg/kg) was administered i.p. every 2 d. Mice were sacrificed on d 10 after colitis induction. Values are mean ± SEM of 6 mice/group. (A) The body weight of the mice was measured and presented as a percentage of the original body weight. (B) The colon was photographed. (C) The length of the colon was measured when the mice were sacrificed. (D) Protein levels of various cytokines in colon homogenates from DSS-induced mice at d 10 were examined by ELISA. *P < 0.05, **P < 0.01. (E) Expression of PTGS2/COX2 and p-RELA/p-p65 were examined by immunohistochemical staining of paraffin-embedded colon sections from DSS-induced mice at d 10. (F–I) Mice were injected i.p. with a single dose (7.5 mg/kg) of AOM followed by 3 cycles of 2.5% DSS given in the drinking water for 5 d. Andro (15 mg/kg) was given i.g. daily and CQ (50 mg/kg) was i.p. administered every 2 d during the interval between DSS cycles. Mice were sacrificed on d 95 after CAC induction. Values are mean ± SEM of 6 mice/group. (F) The inside of the colon was photographed. (G) Tumor numbers, size, and load were measured. *P < 0.05, **P < 0.01. (H) Macrophages were isolated from the spleen of AOM-DSS mice on d 95 using commercial magnetic beads as described in Materials and Methods. After stimulation with 5 mM ATP for 30 min, proteins were collected for western blotting. (I) Statistical data of the expressions of CASP1 and LC3 from 6 mice were shown. *P < 0.05. Andro, andrographolide.