Glutathione protects the rat liver against reperfusion injury after hypothermic preservation  Manfred Bilzer, Gustav Paumgartner, Alexander L. Gerbes  Gastroenterology  Volume 117, Issue 1, Pages 200-210 (July 1999) DOI: 10.1016/S0016-5085(99)70568-8 Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 1 Experimental protocol. For detailed explanation see Materials and Methods. Gastroenterology 1999 117, 200-210DOI: (10.1016/S0016-5085(99)70568-8) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 2 Sinusoidal washout of (A) LDH and (B) PNP after cold liver preservation. After 24 hours of preservation in UW solution, livers were reperfused in the absence (•, n = 6; group 3) or presence of 2 mmol/L (▴, n = 6; group 6) or 4 mmol/L GSH (▵, n = 4; group 7). Results are means ± SD. *P < 0.05 compared with untreated livers. Gastroenterology 1999 117, 200-210DOI: (10.1016/S0016-5085(99)70568-8) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 3 Influence of GSH on sinusoidal efflux of (A) LDH and (B) PNP during 2 hours of reperfusion after cold ischemia. Livers were reperfused with 2 mmol/L (▴, n = 6; group 6) or 4 mmol/L GSH (▵, n = 4; group 7). Results are means ± SD. *P < 0.05 compared with untreated livers (•, n = 6; group 3). Gastroenterology 1999 117, 200-210DOI: (10.1016/S0016-5085(99)70568-8) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 4 Trypan blue uptake by cold-preserved livers. (A) Compared with rat livers after 150 minutes of continuous perfusion, (B) a marked trypan blue staining occurred in cold-preserved livers after 2 hours of reperfusion (total perfusion time, 150 minutes). Trypan blue was predominantly located in periportal and midzonal cells. (C) Staining was less eminent in livers reperfused with 4 mmol/L GSH (original magnification 160×). (D) Higher magnification (600×) showed trypan blue staining of hepatocytes and nonparenchymal liver cells. In GSH-treated livers, staining was mainly seen in nonparenchymal cells, but only to a less extent in hepatocytes (E ). PP, periportal; PC, pericentral; H, hepatocytes; NC, nonparenchymal liver cells. Gastroenterology 1999 117, 200-210DOI: (10.1016/S0016-5085(99)70568-8) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 5 Effect of GSH treatment on bile flow after cold-liver preservation. Livers were reperfused with 2 mmol/L GSH (▴, n = 6; group 6) or 4 mmol/L GSH (▵, n = 4; group 7). Results are ± SD. *P < 0.05 compared with untreated livers (•, n = 6; group 3). Gastroenterology 1999 117, 200-210DOI: (10.1016/S0016-5085(99)70568-8) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 6 Effect of GSH on portal pressure during reperfusion. After starting reperfusion, portal pressure of untreated livers (•, n = 6; group 3) markedly increased and remained elevated until the end of reperfusion. The increase of portal pressure was significantly attenuated by 2 mmol/L GSH (▴, n = 6; group 6) or 4 mmol/L GSH (▵, n = 4; group 7). GSH (1 mmol/L) (○, n = 6; group 5) did not show hemodynamic effects. *P < 0.05 compared with untreated livers. Gastroenterology 1999 117, 200-210DOI: (10.1016/S0016-5085(99)70568-8) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 7 Intracellular GSH content of untreated and GSH-treated livers before and after 2 hours of reperfusion. The intracellular GSH content of untreated preserved livers (■, n = 6) was significantly reduced after 2 hours of reperfusion (*P < 0.05). In livers that were reperfused with 4 mmol/L GSH (▩, n = 5), this loss of intracellular GSH was significantly attenuated. **P < 0.05 compared with untreated livers. Gastroenterology 1999 117, 200-210DOI: (10.1016/S0016-5085(99)70568-8) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 8 Sinusoidal GSH efflux during reperfusion after hypothermic ischemia. During the first 80 minutes of reperfusion, sinusoidal GSH efflux of preserved livers (▴, n = 6; group 3) was similar to efflux rates of continuously perfused livers (▵, n = 6; group 1). During further reperfusion, GSH efflux significantly increased. *P < 0.05 compared with continuously perfused livers. Gastroenterology 1999 117, 200-210DOI: (10.1016/S0016-5085(99)70568-8) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 9 Relation between sinusoidal GSH efflux and postischemic cell damage. Sinusoidal (A) LDH and (B) PNP efflux rates between 20 and 120 minutes of reperfusion were plotted against the corresponding efflux rates of GSH. A linear correlation was found, described by the following equations: A, y = 0.073 × +4.55, r2 = 0.81; B, y = 0.808 × +2.05, r2 = 0.65. Gastroenterology 1999 117, 200-210DOI: (10.1016/S0016-5085(99)70568-8) Copyright © 1999 American Gastroenterological Association Terms and Conditions