HEPATOCYTE INJURY BY ACTIVATED NEUTROPHILS IN-VITRO IS MEDIATED BY PROTEASES

Objective This study determined the mechanism used by neutrophils (PMN s) to induce hepatocellular injury Summary Background Data Neutrophils have been shown to be potent mediators of cell and tissue injury and have been hypothesized to contribute to the hepatic injury that occurs after trauma and infection. Oxygen radical scavengers protect the liv er in vivo from inflammatory injury and it has been suggested that PMN s are the source of these toxic oxygen radicals. The specific mechanis m used by PMNs to produce hepatocellular damage, however, has not been determined Methods Neutrophils were cultured in vitro with hepatocyte s (HCs) and stimulated with phorbol 12-myristate 13-acetate (PMA) to i nduce HC injury in the presence of oxygen radical scavengers and prote ase inhibitors. Results PMA induced a PMN-mediated HC injury that was dependent on the number of PMNs present and the concentration of PMA. Protease inhibitors reduced the extent of HC injury, while oxygen radi cal scavengers had no effect. Hydrogen peroxide, directly applied, was able to injure HCs, but only at concentrations greater than those tha t could be produced by PMA-stimulated PMNs Conclusions PMNs are cytoto xic to cultured HCs, predominantly due to the release of proteolytic e nzymes, while HCs appear relatively resistant to oxidative injury, Inv olvement of neutrophil toxic oxygen radicals in hepatic damage in vivo may require impairment of HC antioxidant defenses or may involve inju ry to nonparenchymal liver cells with secondary effects on HCs.