Nucleotide oligomerization domain (NOD) proteins recognize peptidoglycan fragments, resulting in up-regulation of transcription factors, and may enhance the inflammatory response to infection. Specifically, NOD2 has been shown to sense muramyl dipeptide (MDP), which is released during bacterial cell growth and replication. Activation of NOD2 by MDP enhances the inflammatory response caused by LPS (endotoxin). Here, we investigated the effects of MDP on the organ injury/dysfunction caused by systemic administration of a low dose of LPS. Male Wistar rats were coadministered with either MDP (1 - 10 mg kg(-1), i.v.) or vehicle (0.5 mL kg(-1) saline, i.v.), and a low dose of LPS (1 mg kg(-1), i.v.) or vehicle (1 mL kg(-1), saline, i.v.). MAP and heart rate were continuously monitored for 6 h. Markers of organ dysfunction/injury, plasma cytokine levels, and lung myeloperoxidase activity were measured 6 h after MDP and LPS coadministration. In a separate study, MDP (3 or 10 mg kg(-1), i.v.) or vehicle (0.5 mL kg(-1) saline, i.v.) was administered 24 h before LPS infusion. When compared with animals receiving low-dose LPS alone, coadministration of MDP (10 mg kg(-1), i.v.) and LPS, or administration of MDP (10 mg kg(-1), i.v.) 24 h before LPS resulted in a significant increase in the degree of organ injury, cytokine release, and lung injury caused by LPS alone. Thus, our results demonstrate that the two bacterial wall components MDP and LPS work in concert to cause multiple organ injury and systemic inflammation. We hope that our results stimulate other studies designed to evaluate the effects of NOD ligands in animal models of inflammation.