Figure 6.

Disulfiram protects against LPS-induced sepsis. (a-f) Mice were pretreated with disulfiram (50 mg/kg) or vehicle (Ctrl) by intraperitoneal injection 24 and 4 hrs before intraperitoneal challenge with 15 (a-d), 25 (e), or 50 (f) mg/kg LPS and followed for survival (a,e,f). Statistical analysis was performed using the log-rank (Mantel-Cox) test (a, n = 12 mice/group, e,f, n = 8 mice/group). Serum IL-1β, TNF and IL-6 were measured by ELISA (n= 5 mice/group) 12 hrs post LPS challenge (b-d). Data were analyzed using two-tailed Student’s t-test. Shown are mean ± s.d. (g) Mice were pretreated with disulfiram (50 mg/kg) or vehicle (Ctrl) by intraperitoneal injection 4 hrs before and daily after intraperitoneal LPS challenge (25 mg/kg) and followed for survival. Statistical analysis was performed using the log-rank (Mantel-Cox) test (n = 8 mice/group). (h) Peritoneal macrophages from four indicated groups of mice were analyzed for NLRP3, GSDMD and HMGB1 by immunoblot. (i) Mice were challenged with 25 mg/kg LPS intraperitoneally and then treated with vehicle (Ctrl) or 50 mg/kg disulfiram given 0 and 12 hrs later. Indicated mice also received copper gluconate (0.15 mg/kg). Statistical analysis was performed using the log-rank (Mantel-Cox) test (n = 8 mice/group). Experiments were repeated three independent times.