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Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues
Published on: February 17, 2017
The disaccharide trehalose inhibits proinflammatory phenotype activation in macrophages and prevents mortality in
Letteria Minutoli1, Domenica Altavilla, Alessandra Bitto
1Departments of Clinical and Experimental Medicine and Pharmacology, University of Messina, Messina, Italy.
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.
Insights
Trehalose effectively inhibits the activation of macrophages
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Sepsis-induced macrophage activation drives multiple organ dysfunction.
- Trehalose is known to preserve cell viability under stress.
Purpose of the Study:
- To investigate trehalose's potential to inhibit endotoxin-induced inflammatory activation in macrophages.
- To evaluate trehalose's efficacy in preventing sepsis-related mortality in vivo.
Main Methods:
- Primary rat macrophages were stimulated with lipopolysaccharide (LPS).
- Cells were treated with varying concentrations of trehalose or sucrose.
- Assessed inflammatory markers via Western blot, RT-PCR, and cytokine/nitrite measurements.
- In vivo studies involved challenging rats with a lethal LPS dose and administering trehalose.
Main Results:
- Trehalose significantly reduced LPS-induced activation of key inflammatory pathways (ERK, JNK, iNOS) in macrophages.
- Trehalose suppressed the gene expression of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α).
- In vivo, trehalose administration increased survival rates in rats challenged with lethal LPS and reduced serum TNF-α levels.
Conclusions:
- Trehalose demonstrates potent anti-inflammatory effects by inhibiting macrophage activation.
- Trehalose offers significant protection against endotoxin-induced mortality in a sepsis model.
- Sucrose did not exhibit similar protective effects, highlighting trehalose's specific mechanism.
