Mesenteric lymph contains unidentified proinflammatory mediators that increase in concentration after hemorrhage. In the search for candidate mediators, we examined mesenteric lymph for the presence of proinflammatory substances that are known to be produced in the gut: (a) antimicrobial peptides and antimicrobial proteins produced in the Paneth cells of the intestine (alpha-defensin 4, secretory phospholipase A2 [sPLA2], and Reg 2 protein) and (b) asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NOS. Anesthetized male rats were hemorrhaged to 40 mmHg and maintained at that pressure by intermittent blood withdrawal until the pressure fell to less than 40 mmHg (decompensation) at which point they were resuscitated with three times the shed blood volume of Ringer's lactate solution administered over 1 h. Mesenteric lymph samples were analyzed for ADMA by enzyme-linked immunosorbent assay and for alpha-defensin 4, sPLA2, and Reg2 by Western blotting. Protein concentration in lymph was unchanged by hemorrhage, but alpha-defensin 4 increased significantly (12-fold greater than control) as did ADMA (2-fold greater than control). The sPLA2 could not be detected in lymph, and Reg 2 was unchanged during hemorrhage. During resuscitation, lymph flow tended to increase, but the concentration of ADMA and alpha-defensin 4 by volume did not increase. Reg 2 decreased during resuscitation. The results indicate that ADMA and immunoreactive product to alpha-defensin 4 may contribute to the increase in inflammatory activity of mesenteric lymph during hemorrhage, but they are unlikely to be the mediators responsible for the increase in the concentration of inflammatory mediators in postresuscitation lymph.