Pretreatment with intrathecal or intravenous morphine attenuates hepatic ischaemia– reperfusion injury in normal and cirrhotic rat liver Y. Wang, G.T.C.

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Pretreatment with intrathecal or intravenous morphine attenuates hepatic ischaemia– reperfusion injury in normal and cirrhotic rat liver Y. Wang, G.T.C. Wong, K. Man, M.G. Irwin British Journal of Anaesthesia Volume 109, Issue 4, Pages 529-539 (October 2012) DOI: 10.1093/bja/aes209 Copyright © 2012 The Author(s) Terms and Conditions

Fig 1 Dose–response of morphine preconditioning on hepatic IRI in normal liver. (a) Representative haematoxylin and eosin-stained sections of ischaemic livers at different morphine doses and their corresponding Suzuki score. IVCON, i.v. control; IVMPC, i.v. morphine preconditioning; ITCON, i.t. control; ITMPC, i.t. morphine preconditioning; n=6 per group. Data expressed as mean (sd). *P<0.05 vs IVCON; **P<0.05 vs IVCON; #P<0.05 vs ITCON; ##P<0.05 vs ITCON. British Journal of Anaesthesia 2012 109, 529-539DOI: (10.1093/bja/aes209) Copyright © 2012 The Author(s) Terms and Conditions

Fig 2 Effect of opioid receptor and Akt inhibition on the effects of morphine pretreatment in normal livers evaluated by Suzuki scores (a), apoptotic index (b), and expression of cleaved caspase-3 (c). The mean band density was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). IVCON, i.v. control; IVMPC, i.v. morphine preconditioning at 100 µg kg−1; ITCON, i.t. control; ITMPC, i.t. morphine preconditioning at 10 µg kg−1; NM, naloxone methiodide (50 mg kg−1); W, wortmannin (15 µg kg−1); RD, relative density. Data expressed as mean (sd). n=6 per group, *P<0.05 vs IVCON; #P<0.05 vs ITCON. British Journal of Anaesthesia 2012 109, 529-539DOI: (10.1093/bja/aes209) Copyright © 2012 The Author(s) Terms and Conditions

Fig 3 Effect of morphine preconditioning on expression of total and phosphorylated Akt, JAK2, and STAT3, in the presence or absence of wortmanin. (a) I.V. morphine preconditioning and (b) i.t. morphine preconditioning groups. The mean band density was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). IVCON, i.v. control; IVMPC, i.v. morphine preconditioning at 100 µg kg−1; ITCON, i.t. control; ITMPC, i.t. morphine preconditioning at 10 µg kg−1; W, wortmannin (15 µg kg−1). Data expressed as mean (sd). *P<0.05 vs IVCON; **P<0.01 vs IVCON; #P<0.05 vs ITCON; ##P<0.01 vs ITCON. British Journal of Anaesthesia 2012 109, 529-539DOI: (10.1093/bja/aes209) Copyright © 2012 The Author(s) Terms and Conditions

Fig 4 Effect of morphine preconditioning on hepatic IRI in cirrhotic liver. (a) Representative haematoxylin and eosin and TUNEL-stained sections of ischaemic cirrhotic liver and their corresponding Suzuki score and apoptotic index. (b) ALT and AST levels at 6 h reperfusion. (c) Cleaved caspase-3 levels, with the mean band density normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Data expressed as mean (sd). IVMPC, i.v. morphine preconditioning at 100 µg kg−1; ITCON, i.t. control; ITMPC, i.t. morphine preconditioning at 10 µg kg−1; *P<0.05 vs IVCON; #P<0.05 vs ITCON. British Journal of Anaesthesia 2012 109, 529-539DOI: (10.1093/bja/aes209) Copyright © 2012 The Author(s) Terms and Conditions

Fig 5 Effect of morphine preconditioning on the expression of total and phosphorylated Akt, JAK2, STAT3, and PKCα/βII, and also HO-1 and iNOS. (a) Immunoblot analysis of the expression of total and phosphorylated Akt, JAK2, and STAT3. (b) Expression of phosphorylated PKC (α/βII subunit), HO-1, and iNOS. The mean band density was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Data expressed as mean (sd). *P<0.05 vs IVCON; **P<0.01 vs IVCON; #P<0.05 vs ITCON. British Journal of Anaesthesia 2012 109, 529-539DOI: (10.1093/bja/aes209) Copyright © 2012 The Author(s) Terms and Conditions