Although the barrier function of the intestinal mucosa is impaired after hemorrhage, it remains unclear whether this is associated with a deficit in mucosal function. The aim of this study, therefore, was to determine whether trauma-hemorrhage affects the in vivo lipid absorptive capacity of the gut and, if so, to characterize the uptake process of free fatty acids in isolated enterocytes. To study this, rats were anesthetized, a laparotomy was performed (i.e., trauma was induced), and various blood vessels were cannulated. For in vivo lipid absorption, the main intestinal lymph vessel was cannulated and a jejunostomy feeding tube was inserted. The animals were bled to and maintained at a mean arterial pressure of 40 mmHg until 40% of shed blood volume was returned in the form of Ringer lactate. They were then resuscitated with four times the volume of maximal bleed out with Ringer lactate. The in vivo and in vitro lipid absorptive capacities were assessed by measuring lymph triglyceride output after a fat load and by determining the linoleic acid uptake rates on isolated enterocytes, respectively. The results show that the in vivo lipid absorption capacity of the gut is severely depressed after trauma-hemorrhage and resuscitation. Similarly, in enterocytes isolated from hemorrhaged rats, fatty acid uptake capacity, as reflected by the decreased maximal uptake rates, was significantly reduced: 1.2 ± 0.2 and 2.6 ± 0.6 nmol · min-1 · 106 cells-1 for hemorrhaged and sham, respectively. Thus gut lipid absorptive function is depressed after trauma-hemorrhage and resuscitation, which is at least partially due to the depressed uptake mechanism of the enterocyte.