These experiments confirmed CXCR2 expression on wild-type hepatocytes and neutrophils and no CXCR2 expression on CXCR2 knockout

hepatocytes and neutrophils (data not shown). To exclude the possibility of contamination by genomic DNA, we designed a second pair of primers that crossed exon 1 and exon 2. These experiments confirmed that wild-type hepatocytes and neutrophils expressed CXCR2 (Fig. 7A); CXCR2 knockout hepatocytes selleck chemicals llc did not express any detectable CXCR2. Flow cytometry confirmed these results (Fig. 7B,C). Wild-type hepatocytes expressed low CXCR2 levels. There was no CXCR2 expression on knockout mouse hepatocytes. The mean fluorescence intensity (MFI) of CXCR2 on wild-type hepatocytes (5.26% ± 0.33%) was significantly increased (Fig. 7D) versus knockout mouse hepatocytes (3.42% ± 0.37%, P < 0.05). MFI of hepatocyte CXCR2 expression in wild-type mice 3 hours after APAP dosing was the same as that of untreated hepatocytes, and this suggested that APAP treatment does not change hepatocyte CXCR2 expression in wild-type mice (Fig. 7D). Our results suggest that CXCR2 signaling facilitates apoptosis after APAP dosing. CXCR2

activation requires that CXCR2 ligands, which include KC and MIP2, bind to this receptor. We hypothesized that APAP increases KC and MIP2 production, so hepatic KC and MIP2 protein expression was measured in wild-type and knockout mice after APAP administration by ELISA selleck kinase inhibitor (Fig. 8A,B). KC and MIP2 protein levels increased after APAP and peaked at 4 hours in both wild-type and knockout mice. KC and MIP2 levels in the CXCR2 knockout mice were significantly higher than those in the wild-type mice (P < 0.01) at every time point. These experiments show that CXCR2 knockout mice have a survival advantage over wild-type mice after a median

lethal dose of APAP. The liver injury following APAP in CXCR2 knockout mice is less than that seen in wild-type mice, and this results in significantly selleck chemical lower levels of serum liver enzymes and less liver injury. Further experiments have suggested that this is at least partially related to less apoptosis in knockout mice versus control animals, with no differences in hepatocyte proliferation. However, the role of apoptosis in APAP-induced liver injury is controversial, and it is possible that the less profound GSH depletion seen in this strain of mouse may allow apoptosis to proceed in a more significant fashion than that seen in other models. Although the CXCR2 receptor and its ligands, the CXC chemokines, are known to mediate the inflammatory response, these ligand/receptor interactions also modulate proliferation. For example, Bone-Larson and colleagues demonstrated increased hepatocyte proliferation after APAP injury, which was a CXCR2-dependent response.5 This beneficial proliferative response is dependent on increased CXCR2 expression.5 The CXC chemokines also have a therapeutic role in APAP-induced liver injury.