Figure 1

Effects of trehalose on inflammatory responses in cultured cells. As indicated in the graphs, culture media, hemolysate, or both contained saline (Sal), 5% trehalose (Tre), or 5% maltose (Mal). (A, B) Effect of trehalose on the induction of eicosanoids in hemolysate-treated cells. RAW 264.7 cells were treated with or without 10% hemolysates for 8 h. Prostaglandin E₂ (PGE₂) (A) and cysteinyl leukotriene (B) levels were measured by enzyme immunoassay (EIA). n = 4 (A) and n = 5 (B) in each group, respectively. (C) Effect of trehalose on production of cyclooxygenase-2 (COX-2) in hemolysate-treated cells. RAW 264.7 cells were treated with 10% hemolysates, vehicle, or 20 ng/ml LPS (used as a positive control) for 8 h. The amounts of COX-2 and β-actin (used as an internal control) were analyzed by western blotting. The graph shows the band intensities of COX-2, adjusted to the corresponding β-actin bands and calculated as intensity ratio relative to that of the saline control. n = 5 in each group. (D) Effect of trehalose on the release of arachidonic acid in hemolysate-treated cells. RAW 264.7 cells were prelabeled with [³H]-arachidonic acid and then treated with or without 10% hemolysate for 3 h. The radioactivity was measured to quantify arachidonic acid release. n = 7 in each group. (E) Effect of trehalose on the induction of endothelin-1 in hemolysate-treated cells. Human umbilical vein endothelial cells (HUVECs) were treated with or without 10% hemolysates for 18 h. Endothelin-1 levels were measured by EIA. n = 5 in each group. (F, G) Effect of trehalose on the induction of pro-inflammatory cytokines in hemolysate-treated cells. RAW 264.7 cells were treated with or without 10% hemolysates for 8 h. Tumor necrosis factor-α (TNF-α) (F) and interleukin-1β (IL-1β) (G) levels were measured by EIA. n = 3 (F) and n = 4 (G) in each group, respectively. (H) Effect of trehalose on the production of inducible nitric oxygen species (iNOS) in hemolysate-treated cells. RAW 264.7 cells were treated with vehicle, 10% hemolysates, or 20 ng/ml LPS for 8 h. Amounts of iNOS and β-actin were analyzed by western blotting. The graph shows the band intensities of iNOS, adjusted to the corresponding β-actin bands and calculated as intensity ratio relative to that of the saline control. n = 7 in each group. Whole experiments were repeated more than three times. All data are shown as means ± SD. *P < 0.05 (Kruskal-Wallis test).