The disaccharide trehalose inhibits proinflammatory phenotype activation in macrophages and prevents mortality in experimental septic shock

Proinflammatory phenotype activation in macrophages (M6s) after sepsis orchestrates an inflammatory response leading to multiple organ dysfunction. Trehalose preserves cell viability during exposure to a range of environmental stresses. We investigated whether trehalose may inhibit endotoxin-induced activation of the inflammatory phenotype in M6s. Rat peritoneal M6s were stimulated with 50 2g/mL of Salmonella enteritidis lipopolysaccharide (LPS). Stimulated M6s were coincubated with trehalose (25, 50, and 100 mmol), sucrose (100 mmol), or RPMI alone. Macrophages cultures were used for Western blot analysis of extracellular-regulated kinase, c-jun-N terminal kinase, and inducible nitric oxide synthase; interleukin (IL) 1", IL-6, and tumor necrosis factor ! (TNF-!) gene expression by real-time reverse transcriptaseYpolymerase chain reaction, and supernatants for measuring the release of inflammatory cytokines and nitrite content. In vitro trehalose significantly blunted LPS-induced extracellular-regulated kinase (LPS = 21 T 6 integrated intensity; LPS + trehalose 100 mmol = 2 T 0.3 integrated intensity), c-jun-N terminal kinase (LPS = 15 T 5 integrated intensity; LPS + trehalose 100 mmol = 3.5 T 0.9 integrated intensity), and inducible nitric oxide synthase activation (LPS = 12 T 3 integrated intensity; LPS + trehalose 100 mmol = 1 T 0.09 integrated intensity), blunted IL-1" (LPS = 5 T 1.9 n-folds/"-actin; LPS + trehalose 100 mmol = 1.5 T 0.8 n-folds/"-actin), IL-6 (LPS = 4 T 1.5 n-folds/"-actin; LPS + trehalose 100 mmol = 1.4 T 0.5 n-folds/"-actin), and TNF-! (LPS = 4.2 T 1.6 n-folds/"-actin; LPS + trehalose 100 mmol = 1.1 T 0.7 n-folds/"-actin) gene expression, and markedly reduced the release of inflammatory cytokines and nitrite content. Furthermore, in vivo trehalose prevented mortality in rats challenged with a lethal dose (20 mg/kg; LD90) of LPS (80% survival rate and 70% survival rate 24 and 72 h after LPS injection, respectively) and reduced serum TNF-!. Sucrose did not modified inflammatory phenotype in vitro nor in vivo protected against endotoxin-induced mortality. Our study suggests that trehalose inhibits proinflammatory phenotype activation in M6s and prevents endotoxin-induced mortality.