CRITICAL CARE MEDICINE Molecular Changes Induced in Rat Liver by Hemorrhage and Effects of Melanocortin Treatment Caterina Lonati, Ph.D.,* Andrea Sordi, Ph.D.,* Daniela Giuliani, Ph.D.,† Luca Spaccapelo, M.D.,‡ Patrizia Leonardi, B.Sc.,§ Andrea Carlin, B.Sc.,§ Alessandra Ottani, Ph.D.,† Maria Galantucci, Ph.D.,# Paolo Grieco, Ph.D., Anna Catania, M.D.,** Salvatore Guarini, Ph.D.†† ABSTRACT Background: Melanocortin peptides improve hemody- namic parameters and prevent death during severe hem- orrhagic shock. In the present research we determined influences of a synthetic melanocortin 1/4 receptor ago- nist on the molecular changes that occur in rat liver during hemorrhage. Methods: Controlled-volume hemorrhage was performed in adult rats under general anesthesia by a stepwise blood withdrawal until mean arterial pressure fell to 40 mmHg. Then rats received either saline or the synthetic melano- cortin 1/4 receptor agonist Butir-His-D-Phe-Arg-Trp-Sar- NH 2 (Ro27-3225; n = 6 – 8 per group). Hemogasanalysis was performed throughout a 60-min period. Gene expres- sion in liver samples was determined at 1 or 3 h using quan- titative real-time polymerase chain reaction. Results: At 1 h, in saline-treated shocked rats, there were significant increases in activating transcription factor 3 (Atf3), early growth response 1 (Egr1), heme oxygenase (de- cycling) 1 (Hmox1), FBJ murine osteosarcoma viral onco- gene homolog (Fos), and jun oncogene (Jun). These changes were prevented by Ro27-3225 (mean  SEM: Atf3 152.83  58.62 vs. 579.00  124.13, P = 0.002; Egr1 13.21  1.28 vs. 26.63  1.02, P = 0.001; Hmox1 3.28  0.31 vs. 166.54  35.03, P = 0.002; Fos 4.36  1.03 vs. 14.90  3.44, P  0.001; Jun 6.62  1.93 vs. 15.07  2.09, P = 0.005; respectively). Increases in alpha-2-macroglobulin (A2m), heat shock 70kD protein 1A (Hspa1a), erythropoie- tin (Epo), and interleukin-6 (Il6) occurred at 3 h in shocked rats and were prevented by Ro27-3225 treatment (A2m 6.90  0.82 vs. 36.73  4.00, P  0.001; Hspa1a 10.34  3.28 vs. 25.72  3.64, P = 0.001; Epo 0.49  0.13 vs. 2.37  0.73, P = 0.002; Il6 1.05  0.15 vs. 1.88  0.23, P  0.001; respectively). Further, at 3 h in shocked rats treated with Ro27-3225 there were significant increases in * Postdoctoral Fellow, Center for Surgical Research, ** Director, Center for Preclinical Investigation, Fondazione IRCCS Ca’ Granda - Ospedale Maggiore Policlinico, Milano, Italy. † Assistant Professor of Pharmacology, ‡ Postgraduate Student, # Postdoctoral Fellow, †† Full Professor of Pharmacology, Department of Biomedical Sciences, Sec- tion of Pharmacology, University of Modena and Reggio Emilia, Mod- ena, Italy. § Research Assistant, Department of Internal Medicine, Uni- versity of Milano, Milano, Italy.  Associate Professor of Pharmaceutical Chemistry, Department of Pharmaceutical Chemistry and Toxicology, University of Napoli “Federico II,” Napoli, Italy. Received from the Department of Biomedical Sciences, Section of Pharmacology, University of Modena and Reggio Emilia, Mod- ena, Italy. Submitted for publication August 5, 2011. Accepted for publication December 6, 2011. Supported by grant No. 20074Z8W3S from Ministero dell’Istruzione, dell’Universita ` e della Ricerca Scien- tifica, Roma, Italy; grant No. 060-08 from Fondazione Ca’ Granda Ospedale Maggiore Policlinico, Milano, Italy; and grant “Synthetic Melanocortins as a novel class of antiinflammatory drugs” from Fondazione Fiera, Milano, Italy. Address correspondence to Dr. Guarini: Department of Biomed- ical Sciences, Section of Pharmacology, University of Modena and Reggio Emilia, Modena, Italy. salvatore.guarini@unimore.it. Infor- mation on purchasing reprints may be found at www.anesthesiology. org or on the masthead page at the beginning of this issue. ANES- THESIOLOGY’S articles are made freely accessible to all readers, for personal use only, 6 months from the cover date of the issue. Copyright © 2012, the American Society of Anesthesiologists, Inc. Lippincott Williams & Wilkins. Anesthesiology 2012; 116:692–700 What We Already Know about This Topic • Melanocortin molecules, including adrenocorticotropic hor- mone, may protect organs against hemorrhagic shock-in- duced injury • This study in rats investigates if a synthetic melanocortin may alleviate hemorrhagic-induced systemic and hepatic injury What This Article Tells Us That Is New • The melanocortin receptor agonist Ro27-3225 restores mean arterial pressure; attenuates metabolic acidosis; and inhibits upregulation of interleukin-6, acute phase protein A2m, and several other injury mediators in the liver after a hemorrhagic challenge  Supplemental digital content is available for this article. Direct URL citations appear in the printed text and are available in both the HTML and PDF versions of this article. Links to the digital files are provided in the HTML text of this article on the Journal’s Web site (www.anesthesiology.org). 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