Figure 4

Choline treatment suppresses HMGB1 release and improves survival in severe sepsis. (A) Choline suppresses HMGB1 release from endotoxin-stimulated RAW cells. RAW cells were exposed to the indicated concentration of choline or vehicle 10 min prior to LPS (100 ng/mL) addition for 24 h. Culture supernatants were harvested, and secreted HMGB1 was detected by Western blot analysis. HMGB1 was not detected in the supernatant from cells that were not treated with LPS. Data represent the mean ± SEM of four experiments conducted in duplicate (*P < 0.05, **P < 0.02, ***P < 0.001 as compared with lowest choline concentration tested). (B) Mice (n = 12) were administered i.p. with either saline or choline (25 mg/kg) 24 h after CLP. Mice received additional treatments at 30 h and 44 h after CLP. Serum HMGB1 levels were determined in surviving mice (n = 11 for choline treatment, n = 7 for control treatment) in blood obtained at 45 h after CLP (*P < 0.0008). (C) Mice (n = 26 per group) were subjected to CLP surgery. 24 h after CLP, mice were randomized and injected i.p. with either saline or choline (25 mg/kg). This treatment was repeated 6 h later (30 h after CLP) and twice daily for 2 d more for a total of six treatments, and survival was monitored for 2 wks (*P < 0.002).