FIGURE 4.

Xanthone treatment ameliorates LPS‐induced sepsis and mice keratitis. (A and B) ELISA of IL‐1β (A) and TNFα (B) in sera from mice intraperitoneally injected with saline or xanthone (20 mg/kg body weight) with LPS (25 mg/kg body weight) for 4 h. Data are the mean ± SEM (n = 6 mice/group). (C) Survival of mice intraperitoneally injected with saline or xanthone (20 mg/kg body weight) with LPS (20 mg/kg body weight, n = 11 mice/group). (D–G) Mice were sham operated or intrastromal injected with LPS and treated with xanthone or DMSO control three times a day for 24 or 48 h. Representive clinical images were taken with stereomicroscope (D, upper panel), and H&E staining of mice corneal (D, lower panel). Mice corneal clinical scoring was based on a 4‐point system (E). Average infiltrated inflammatory cells number was counted at least three slides manually (F), while stroma thickness was determined by ImageJ (G). (H) Intrastromal IL‐1β level was determined by ELISA 24 and 48 h after LPS injection. (I and J) The level of NLRP3 inflammasome activation level of mice corneal infiltrated macrophage was determined with FLICA staining (I), and representative histogram was shown (J). (K and L) Lysates of mice cornea was determined by Western blot (K) and quantifications were normalized determined by ImageJ (L). (M) Schematic representation of the inhibition effect of xanthone upon NLRP3 inflammasome activation. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, two‐tailed unpaired Student's t‐test and one‐way ANOVA, Log‐rank (Mantel–Cox) test for survival