Volume 127, Issue 5, Pages (November 2004)

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Volume 127, Issue 5, Pages 1488-1496 (November 2004) Ischemic preconditioning protects hepatocytes via reactive oxygen species derived from Kupffer cells in rats  Kazuaki Tejima, Masahiro Arai, Hitoshi Ikeda, Tomoaki Tomiya, Mikio Yanase, Yukiko Inoue, Kayo Nagashima, Takako Nishikawa, Naoko Watanabe, Masao Omata, Kenji Fujiwara  Gastroenterology  Volume 127, Issue 5, Pages 1488-1496 (November 2004) DOI: 10.1053/j.gastro.2004.07.023 Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 1 Prevention by GdCl3 pretreatment with ischemic preconditioning of warm ischemia/reperfusion injury in vivo. Rats were injected with GdCl3 or vehicle as described in Materials and Methods. Their livers were then preconditioned by 10 minutes of ischemia and 10 minutes of reperfusion, or a sham operation was performed.Subsequently, livers were subjected to 40 minutes of ischemia followed by 60 minutes of reperfusion, and serum ALT activities were determined. Bars represent means ± SE from 6 rats in each group. #P < .01 vs. no ischemia/reperfusion (I/R); *P < .05 vs. sham operation; $P < .05 vs. vehicle treatment and ischemic preconditioning. KU, karmen unit. Gastroenterology 2004 127, 1488-1496DOI: (10.1053/j.gastro.2004.07.023) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 2 Serum HA concentrations after in vivo warm ischemia/reperfusion. Rat livers were preconditioned by 10 minutes of ischemia and 10 minutes of reperfusion, or a sham operation was performed. Subsequently, livers were subjected to 40 minutes of ischemia followed by 60 minutes of reperfusion, and serum HA concentrations were determined. Bars represent means ± SE from 6 rats in each group. IP, ischemic preconditioning. #P < .05 vs. no ischemia/reperfusion (I/R). Gastroenterology 2004 127, 1488-1496DOI: (10.1053/j.gastro.2004.07.023) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 3 Prevention by GdCl3 pretreatment with ischemic preconditioning of warm ischemia/reperfusion injury in the isolated perfused liver system. Rats were injected with GdCl3 or vehicle as described in Materials and Methods. Their livers were then preconditioned by 10 minutes of ischemia and 10 minutes of reperfusion, or a sham operation was performed. Subsequently, livers were perfused with KHB and subjected to 40 minutes of ischemic storage followed by 60 minutes of reperfusion. At the end of reperfusion, ALT activities in the perfusate were determined. Bars represent means ± SE from 6 rats in each group. #P < .01 vs. no ischemia/reperfusion (I/R); *P < .01 vs. sham operation; $P < .01 vs. vehicle treatment and ischemic preconditioning. KU, karmen unit. Gastroenterology 2004 127, 1488-1496DOI: (10.1053/j.gastro.2004.07.023) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 4 Hepatic HA uptake during 60 minutes of reperfusion after 40 minutes of ischemic storage. Rat livers were preconditioned by 10 minutes of ischemia and 10 minutes of reperfusion, or a sham operation was performed. Subsequently, livers were perfused with KHB and subjected to 40 minutes of ischemic storage followed by 60 minutes of reperfusion. HA concentrations in the perfusate were determined before and after reperfusion, and hepatic HA uptake was calculated. Bars represent means ± SE from 6 rats in each group. IP, ischemic preconditioning; I/R, ischemia/reperfusion. Gastroenterology 2004 127, 1488-1496DOI: (10.1053/j.gastro.2004.07.023) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 5 ROS formation in Kupffer cells after ischemic preconditioning. Rat livers were preconditioned by 10 minutes of ischemia, or a sham operation was performed. Livers were then perfused with KHB containing NBT as described in Materials and Methods. (A) In sham control livers, formazan deposition was not developed. (B) In livers after 10 minutes of ischemia, formazan deposition appeared in nonparenchymal cells (arrows). Gastroenterology 2004 127, 1488-1496DOI: (10.1053/j.gastro.2004.07.023) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 6 Prevention by ROS scavenger pretreatment with ischemic preconditioning of warm ischemia/reperfusion injury in the isolated perfused liver system. Rats were injected with NAC, SOD, or catalase as described in Materials and Methods. Their livers were then preconditioned by 10 minutes of ischemia and 10 minutes of reperfusion, or a sham operation was performed. Subsequently, livers were perfused with KHB and subjected to 40 minutes of ischemic storage followed by 60 minutes of reperfusion. At the end of reperfusion, ALT activities in the perfusate were determined. Bars represent means ± SE from 6 rats in each group. KU, karmen unit. Gastroenterology 2004 127, 1488-1496DOI: (10.1053/j.gastro.2004.07.023) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 7 Hepatocyte protection by preconditioning with H2O2 perfusion. Rat livers were perfused with KHB containing H2O2 instead of ischemic preconditioning. Livers were then subjected to 40 minutes of ischemic storage followed by 60 minutes of reperfusion. At the end of reperfusion, ALT activities in the perfusate were determined. Bars represent means ± SE from 6 rats in each group. *P < .01 vs. 0 mmol/L. KU, karmen unit. Gastroenterology 2004 127, 1488-1496DOI: (10.1053/j.gastro.2004.07.023) Copyright © 2004 American Gastroenterological Association Terms and Conditions