title: The disaccharide trehalose inhibits proinflammatory phenotype activation in macrophages and prevents mortality in experimental septic shock.
abstract: Proinflammatory phenotype activation in macrophages (MPhis) 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 MPhis. Rat peritoneal MPhis were stimulated with 50 microg/mL of Salmonella enteritidis lipopolysaccharide (LPS). Stimulated MPhis 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) 1beta, IL-6, and tumor necrosis factor alpha (TNF-alpha) gene expression by real-time reverse transcriptase-polymerase 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 +/- 6 integrated intensity; LPS + trehalose 100 mmol = 2 +/- 0.3 integrated intensity), c-jun-N terminal kinase (LPS = 15 +/- 5 integrated intensity; LPS + trehalose 100 mmol = 3.5 +/- 0.9 integrated intensity), and inducible nitric oxide synthase activation (LPS = 12 +/- 3 integrated intensity; LPS + trehalose 100 mmol = 1 +/- 0.09 integrated intensity), blunted IL-1beta (LPS = 5 +/- 1.9 n-folds/beta-actin; LPS + trehalose 100 mmol = 1.5 +/- 0.8 n-folds/beta-actin), IL-6 (LPS = 4 +/- 1.5 n-folds/beta-actin; LPS + trehalose 100 mmol = 1.4 +/- 0.5 n-folds/beta-actin), and TNF-alpha (LPS = 4.2 +/- 1.6 n-folds/beta-actin; LPS + trehalose 100 mmol = 1.1 +/- 0.7 n-folds/beta-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-alpha. 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 MPhis and prevents endotoxin-induced mortality.
authors.affiliation: Departments of Clinical and Experimental Medicine and Pharmacology, University of Messina, Messina, Italy.
authors.forename: Letteria
authors.initials: L
authors.lastname: Minutoli
authors.forename: Domenica
authors.initials: D
authors.lastname: Altavilla
authors.forename: Alessandra
authors.initials: A
authors.lastname: Bitto
authors.forename: Francesca
authors.initials: F
authors.lastname: Polito
authors.forename: Ersilia
authors.initials: E
authors.lastname: Bellocco
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Laganà
authors.forename: Daniela
authors.initials: D
authors.lastname: Giuliani
authors.forename: Tiziana
authors.initials: T
authors.lastname: Fiumara
authors.forename: Salvatore
authors.initials: S
authors.lastname: Magazù
authors.forename: Pietro
authors.initials: P
authors.lastname: Ruggeri
authors.forename: Salvatore
authors.initials: S
authors.lastname: Guarini
authors.forename: Francesco
authors.initials: F
authors.lastname: Squadrito
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D016130:Immunophenotyping; D018836:Inflammation Mediators / Q000502:physiology*; D008264:Macrophages / Q000145:classification / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012772:Shock, Septic / Q000378:metabolism* / Q000401:mortality; D014199:Trehalose / Q000502:physiology*
chemical_list: D018836:Inflammation Mediators; D014199:Trehalose