Objective: To evaluate the effects of O-linked β-N-acetylglucosamine (O-GlcNAc) levels on survival, inflammation, and organ damage 24 hrs after trauma-hemorrhage. We have previously shown that increasing protein O-GlcNAc levels by different mechanisms reduced inflammatory responses and improved organ function 2 hrs after trauma-hemorrhage. Design: Prospective, randomized, controlled study. Setting: Animal research laboratory. Subjects: Male, adult Sprague-Dawley rats. Interventions: Overnight fasted animals were subjected to either sham surgery or trauma-hemorrhage and during the resuscitation phase received glucosamine (270 mg/kg) to increase O-GlcNAc synthesis or O-(2-acetamido-2-deoxy-D-glucopyranosylidene) amino N-phenyl carbamate (PUGNAc, 7 mg/kg) to inhibit O-GlcNAc removal, or mannitol as control. Measurements and Main Results: Survival was followed up for 24 hrs. Surviving rats were euthanized and inflammatory responses, and end organ injuries were assessed. Both glucosamine and PUGNAc increased 24-hr survival compared with controls (control: 53%, GN: 85%, PUGNAc: 86%, log-rank test, p <.05). PUGNAc attenuated the trauma-hemorrhage-induced increase in serum interleukin-6 (sham surgery: 8 ± 6, control: 181 ± 36, PUGNAc: 42 ± 22 pg/mL, p <.05), alanine transaminase (sham surgery: 95 ± 14, control: 297 ± 56, PUGNAc: 126 ± 21 IU, p <.05), aspartate transaminase (sham surgery: 536 ± 110, control: 1661 ± 215, PUGNAc: 897 ± 155 IU, p <.05), and lactate dehydrogenase (sham surgery: 160 ± 18, control: 1499 ± 311, PUGNAc: 357 ± 99 IU, p <.05); however, glucosamine had no effect on these serum parameters. Furthermore, PUGNAc but not glucosamine maintained O-GlcNAc levels in liver and lung and significantly attenuated the NF-κB DNA activation in the liver. In the liver and heart, increased inducible nitric oxide synthase expression was also attenuated in the PUGNAc-treated group. Conclusions: These results demonstrate that increasing O-GlcNAc with either glucosamine or PUGNAc improved 24-hr survival after trauma-hemorrhage. However, only PUGNAc treatment attenuated significantly the subsequent tissue injury and inflammatory responses, suggesting that inhibition of O-GlcNAc removal may represent a new therapeutic approach for the treatment of hypovolemic shock. Copyright © 2010 by the Society of Critical Care Medicine and Lippincott Williams & Wilkins.