Figure 5.

Sustained autophagy stimulated by GCM contributes to its protective effect against acetaminophen (APAP) hepatotoxicity. (A) C57BL/6N mice received 50 mg·kg⁻¹ GCM at 2 h post‐acetaminophen challenge, and liver tissue were collected at 3, 6 and 12 h. LC3II proteins were measured by Western blot at various time points as indicated. Bar graphs show densitometry quantification of LC3II/LC3I. (B–E) Wild‐type C57BL/6N mice were injected (i.p.) with chloroquine (CQ; 60 mg·kg⁻¹) and ML385 (40 mg·kg⁻¹) 2 h before acetaminophen (300 mg·kg⁻¹). Mice were then further treated with or without GCM at 2 h post‐acetaminophen administration. Blood and liver tissue were collected at 6, 12 and 24 h. (B) Western blot of whole liver homogenate for LC3 in mice with or without chloroquine (60 mg·kg⁻¹) at 12 h. Bar graphs show densitometry of LC3II/LC3I. (C) Serum ALT levels were quantified at 24 h. (D) Representative images of haematoxylin and eosin staining are presented. (E) Western blot of whole liver homogenate for p‐JNK and 3‐NT in mice with or without chloroquine (60 mg·kg⁻¹) at 6 h. Bar graphs show densitometry of p‐JNK/JNK or 3‐NT/β‐actin. Data are presented as mean ± SD for n = 6 mice per group. *P < 0.05 compared with the corresponding acetaminophen group.