Volume 42, Issue 6, Pages 870-879 (June 2005) Insulin prevents liver damage and preserves liver function in lipopolysaccharide-induced endotoxemic rats  Marc G. Jeschke, Hauke Rensing, Dagmar Klein, Thomas Schubert, Anke EM Mautes, Ulrich Bolder, Roland S Croner  Journal of Hepatology  Volume 42, Issue 6, Pages 870-879 (June 2005) DOI: 10.1016/j.jhep.2004.12.036 Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

Fig. 1 Serum glucose decreased in both groups LPS and LPS+insulin compared to normal levels. There were no significant differences between LPS vs. LPS+insulin. Data presented as means±SEM with n=7 each group and each time point. Journal of Hepatology 2005 42, 870-879DOI: (10.1016/j.jhep.2004.12.036) Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

Fig. 2 A–D: (A) Serum albumin decreased immediately after LPS administration and remained low throughout the study. Insulin increased serum albumin at days 1, 2, 5 and 7 post LPS when compared to LPS. *Significant difference between LPS+insulin vs. LPS, P<0.05. Data presented as means±SEM with n=7 each group and each time point. (B) Serum CRP increased after LPS injection and remained elevated. Insulin administration significantly decreased serum CRP levels at all days when compared to controls. *Significant difference between LPS+insulin vs. LPS, P<0.05. Data presented as means±SEM with n=7 each group and each time point. (C) Endotoxemia caused a significant increase in serum AST immediately after LPS administration. Insulin significantly decreased serum AST at days 1 and 2 compared to LPS. *Significant difference between LPS+insulin vs. LPS, P<0.05. Data presented as means±SEM with n=7 each group and each time point. (D) Insulin decreased serum ALT at the first two days after LPS injection. *Significant difference between LPS+insulin vs. LPS, P<0.05. Data presented as means±SEM with n=7 each group and each time point. Journal of Hepatology 2005 42, 870-879DOI: (10.1016/j.jhep.2004.12.036) Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

Fig. 3 ATP was found to be present in normal animals. Endotoxemia caused decreased levels of intracellular hepatic ATP. There were no significant differences between LPS and LPS+insulin. Intrahepatic glucose levels are low. LPS caused an increase of intrahepatic glucose levels, which were decreased to normal concentration with insulin administration. Rats receiving LPS+insulin had normal hepatic glucose levels, P<0.05. Endotoxemia caused a significant increase in lactate, whereas animals receiving LPS+insulin had normal lactate levels, P<0.05. Absorbance was measured and values are shown in Table 1. n=3 for normal and n=4 for LPS and LPS+insulin in each group. Color interpretation from highest to lowest concentration: red>orange>yellow>green>light blue>dark blue. Journal of Hepatology 2005 42, 870-879DOI: (10.1016/j.jhep.2004.12.036) Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

Fig. 4 Hepatic genes in endotoxemic and endotoxemic plus insulin treated animals 24h after endotoxemia with n=3 for each group. Out of 15,000 genes, 88 genes were significantly up-regulated (red color), and 51 were significantly down-regulated (green color). Details are given in Table 2. Journal of Hepatology 2005 42, 870-879DOI: (10.1016/j.jhep.2004.12.036) Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

Fig. 5 A--C: (A) Endotoxemia caused a significant increase in hepatocyte apoptosis compared to normal. Insulin administration decreased hepatocyte apoptosis at all time points. *Significant difference between insulin and control, P<0.05. Data presented as means±SEM with n=7 for each group and each time point. (B) Insulin significantly increased hepatocyte proliferation on days 1, 5 and 7 compared to controls. *Significant difference between insulin and control, P<0.05. Data presented as means±SEM with n=7 for each group and each time point. (C) In order to determine net cell balance proliferation was divided by apoptosis. Insulin significantly improved cellular net balance by stimulating cell survival compared to controls on all study days. *Significant difference between insulin and control, P<0.05. Data presented as means±SEM with n=7 for each group and each time point. Journal of Hepatology 2005 42, 870-879DOI: (10.1016/j.jhep.2004.12.036) Copyright © 2005 European Association for the Study of the Liver Terms and Conditions

Fig. 6 A–B: (A) Hepatic caspase-3 increased compared to normal. Insulin prevented the increase on the first and second day after LPS compared to controls. *Significant difference between insulin and control, P<0.05. Data presented as means±SEM with n=7 for each group and each time point. (B) Bcl-2 was decreased during endotoxemia. Insulin significantly increased Bcl-2 protein levels in the liver 2 and 5 days after LPS. *Significant difference between insulin and control, P<0.05. Data presented as means±SEM with n=7 for each group and each time point. Journal of Hepatology 2005 42, 870-879DOI: (10.1016/j.jhep.2004.12.036) Copyright © 2005 European Association for the Study of the Liver Terms and Conditions