Role of EP4 prostaglandin E2 receptor in the ischemic liver.

Prostaglandin E(2) (PGE(2)) mediates a variety of both innate and adaptive immunity responses through 4 distinct receptors, EP1-4. Recent studies have suggested the physiological and pathological role of EP4 in various inflammatory diseases. In this study, we investigated the importance of the EP4 receptor, and the efficacy of a selective EP4 agonist to alter hepatic ischemia/reperfusion (I/R) injury, an important cause of damage in liver resection and transplantation. We used an established murine I/R injury model, 70% partial hepatic ischemia for 90 minutes in male C57BL/6 mice. The local expression of EP4 messenger RNA (mRNA) in the naive and the ischemic liver at 2 hours after reperfusion was examined using RT-PCR analysis. Some mice received the EP4 selective agonist during I/R. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured as markers of hepatic injury. EP4 expression in the liver was significantly up-regulated at 2 hours after reperfusion. Furthermore, treatment with EP4 agonist significantly inhibited hepatic injury at 6 hours after reperfusion. Our data suggest an inhibitory role of EP4 PGE(2) receptor in hepatic I/R injury and the therapeutic efficacy of a selective EP4 agonist for liver protection.

Modification of the single unpaired sulfhydryl group of beta-lactoglobulin under high pressure and the role of intermolecular S-S exchange in the pressure denaturation [single SH of beta-lactoglobulin and pressure denaturation].

Chemical modification reactions of the unpaired sulfhydryl group of beta-lactoglobulin (LG) under high pressure and the role of this group in the pressure-induced denaturation were investigated. When LG was incubated at 400 MPa (pH 6.8) for 1 h, dimerization through intermolecular reaction of SH was observed. The generation of the covalently linked dimers were prevented by the presence of N-ethylmaleimide (NEM), an agent for SH-specific modification. The reactivity of the SH group of LG, which is buried inside in its native state, was increased by high pressure, as a result of its exposure to the protein surface accompanied by the pressure denaturation. The effect of NEM was also observed in the fluorescence change caused by high pressure, in both the intrinsic fluorescence of LG and the retinol fluorescence of the LG-retinol complex. The control showed an irreversible change at neutral pH, but it became mostly reversible in the presence of NEM. Compatible results were obtained by CD spectroscopy. Inter- and intramolecular reactions of the SH group are suggested to be main causes for the pressure-induced irreversible denaturation of LG.