Glutathione, lactobionate, and histidine: cryptic inhibitors of matrix metalloproteinases contained in university of wisconsin and histidine/tryptophan/ketoglutarate liver preservation solutions

Abstract

UW solution and HTK solution are both used for cold preservation of liver allografts. Although they are about equally effective, their compositions are very different, and they were formulated using different rationales. The authors recently showed an important role for MMPs in liver preservation injury and consequently postulated that these preservation solutions contain cryptic inhibitors of MMP activity. To determine this possibility, the ability of these solutions to inhibit MMP activity was studied. The source of MMP2 and MMP9 was human liver effluents obtained at the time of liver transplantation or commercially available human recombinant MMP2 and MMP9. MMP2 and MMP9 showed gelatinolytic activity at 37ЊC and also at 4ЊC, although activity at 4ЊC was reduced. Activity was inhibited by University of Wisconsin (UW) and Histidine/Tryptophan/ Ketoglutarate (HTK) solutions. Examination of individual ingredients disclosed that reduced glutathione (GSH) and lactobionate in UW solution and histidine in HTK solution were the cryptic inhibitors. HTK solution was a more effective inhibitor than UW solution. GSH inhibited the activity of both enzymes, but was a much more effective inhibitor of MMP9 than MMP2. Oxidized glutathione(GSSG) was a much less effective inhibitor of the enzymes. The inhibitor constants (K i ) of GSH for MMP2 and MMP9 were 34 mol/L and 3 mol/L, respectively. The authors conclude that MMP inhibition is a cryptic property of both commonly used liver preservation solutions and contributes importantly to their action. Furthermore, GSH appears to be an effective inhibitor of gelatinases at concentrations at which it is normally present in extracellular fluid. (HEPATOLOGY 2000;31:1115-1122.)University of Wisconsin (UW) solution became the standard liver preservation solution more than 10 years ago. 1,2 It greatly improved the results of hepatic preservation compared with Eurocollins solution, which was in common use before that time. 1,2 UW solution was formulated with specific ingredients to retard the 4 classical effects of hypothermiacell swelling, impaired energy metabolism, acidosis, and accumulation of precursors of reactive oxygen intermediates. 2 Metalloproteinase inhibition was not a goal of its formulation because the role of metalloproteinases in preservation injury 3 was unknown at the time that UW solution was devised.Bretschneider' s solution, also known as histidine/tryptophan/ketoglutarate solution, or HTK solution 4 is also used for liver preservation, mainly in Germany. HTK solution was originally introduced as a cardioplegic solution and was later adapted to cold preservation of organs. It was formulated to retard acidosis (histidine), to prevent membrane injury (tryptophan), and to provide a substrate for energy metabolism (ketoglutarate). Like UW solution, inhibition of metalloproteinases was not a goal of its formulation.Two recent human studies, 5,6 one of which was a randomized controlled trial, 6 compared effectiveness of UW and HTK solutions. These studies found that the sol...

Histidine-tryptophan-ketoglutarate versus University of Wisconsin solution in living donor liver transplantation: Results of a prospective study

“…9,15,16 Experimental and clinical studies have shown that both UW and HTK solutions are effective despite the fact that their compositions are very different and their mechanisms of action are not completely known. 3,17,18,19 The controlled settings of LDLT offer a unique opportunity to study the efficacy of these preservation solutions. In fact, most of the variables found in cadaveric liver donors are not present.…”

“…1,2 Despite their very different compositions, both solutions seem to be equally effective and safe in the long-term preservation of the cadaveric graft. 3 Previous studies comparing the two solutions in cadaveric liver grafts were not randomized, and the results may have been influenced by different factors. Specifically donor age, past medical history, intensive care stay, clinical conditions at the time of the harvest, and ischemic time [4][5][6] influence the quality of the graft and play a role in the functional recovery of the liver and outcome of the patient.…”

The utility of marginal donors in liver transplantation

“…Its formulation was compounded to retard acidosis (histidine), prevent membrane injury (tryptophan), and provide a substrate for energy metabolism (ketoglutarate). 78 Recent studies comparing the effectiveness of the two solutions have shown superior outcomes with the HTK solution when the cold ischemic times were below average. 78,80 On the other hand, with long cold ischemic times, UW seems to represent a better solution for preservation purposes.…”

“…78 Recent studies comparing the effectiveness of the two solutions have shown superior outcomes with the HTK solution when the cold ischemic times were below average. 78,80 On the other hand, with long cold ischemic times, UW seems to represent a better solution for preservation purposes. 77 …”

Effect of cold preservation on intracellular calcium concentration and calpain activity in rat sinusoidal endothelial cells

“…[1][2][3][4] SEC are the main targets of cold preservation injury, 5,6 and understanding the mechanisms by which they are injured might provide an avenue to development of improved preservation solutions. Our studies show that cold causes actin disassembly 2 and that this is associated with rounding of the cells 2 and release of matrix metalloproteases (MMPs) into the supernatants of SEC cell cultures. 1,3 Actin disassembly and MMP secretion lead to activation of the cell surface, as shown by increased expression of von Willebrand factor and increased adhesiveness for platelets.…”