XWH - 06 - 1 - 0530 TITLE : Early Treatment in Shock PRINCIPAL

Background: Inflammatory factors play an important role in cellular damage after shock and resuscitation. Crocetin, a saffron-derived carotenoid, has been shown to improve postshock recovery of cellular adenosine triphosphate (ATP) and to increase overall survival in an experimental model of hemorrhagic shock. The hypothesis of the present study is that treatment with crocetin at the beginning of resuscitation suppresses subsequent expression of messenger ribonucleic acid (mRNA) for tumor necrosis factor (TNF), interleukin-1 (IL-1 ) and inducible nitric oxide synthase (iNOS). Methods: Male Sprague-Dawley rats (n 45, 350 30 g) were randomly assigned to 5 groups of 9 animals each. After anesthesia with isoflurane, the femoral artery and vein were surgically cannulated. Hemorrhagic shock was induced by withdrawing blood through the arterial cannula until the mean arterial pressure (MAP) was 25–30 mm Hg and maintained at the level for 30 minutes with further withdrawals. Resuscitation was carried out by giving 21 mL/kg Ringer’s lactate (LR) and returning the shed blood, with or without the initial administration of crocetin (2 mg/kg). Controls were normal (anesthesia only), sham (surgical preparation), and shock (preparation and shock). Rats were killed 30 minutes after completion of resuscitation. Liver samples were collected for reverse transcription-polymerase chain reaction (RT-PCR) analysis of mRNA (TNF, IL-1 , iNOS, and -actin). Results: Liver mRNA expression for TNF, IL-1 , and iNOS was found in more animals in the shock and shock-plus-resuscitation groups than in the sham control group. The group resuscitated from shock with crocetin had mRNA expression for TNF, IL-1 , and iNOS in fewer animals than either of the other shock groups and was no different from the sham control group. Conclusions: Crocetin modified the hepatic mRNA expression of cytokines and iNOS in a shock model. This agent continues to show promise as a potential treatment for hemorrhagic shock. ( Journal of Parenteral and Enteral Nutrition 30:297–301, 2006) Advances in shock therapy have made it possible to resuscitate all but the most severely injured patients with blood, blood components, and intravenous (IV) fluids. However, the sequelae of shock and resuscitation include widespread tissue and organ injury, which can lead to multiple-organ failure and death. The current focus of resuscitative efforts is on restoration of blood volume and control of hemorrhage, not on prevention of subsequent injury. Inflammatory factors play an important role in the response to reperfusion and are instrumental in the induction of organ failure after shock. Uncontrolled systemic activation of the inflammatory response may lead to cellular damage and organ dysfunction. Studies have shown that hemorrhagic shock and resuscitation are associated with the activation of several proinflammatory genes in the liver and the development of hepatocellular injury. In addition, hepatic inducible nitric oxide synthase (iNOS) expression is an early molecular response to hemorrhagic shock in experimental animals and in injured patients. Crocetin, a constituent of saffron, has long been used in traditional Chinese medicine as a treatment for many diseases. It is a 16-carbon chain with 8 double bonds and 4 attached methyl groups and a carboxylic acid ( COOH) group at each end of the molecule (Figure 1). The molecular weight is 328. It was reported by Gainer and coworkers to increase oxygen diffusion and to improve tissue oxygenation postresuscitation. It has recently been shown by us that crocetin improves the recovery of cellular ATP and improves survival in the rat model of hemorrhagic shock. Received for publication December 9, 2005. Accepted for publication March 8, 2006. Correspondence: Charles W. Van Way III, MD, Department of Surgery, Shock/Trauma Research Center, UMKC, 2301 Holmes Street, Kansas City, Missouri. Electronic mail may be sent to charles. [email protected]. FIGURE 1. Structure diagram of the crocetin molecule. Reprinted from Tarantilis PA, Morjani H, Polissiou M, Manfait M. Inhibition of growth and induction of differentiation of promyelocytic leukemia (HL-60) by carotenoids from C. Sativus L. Anticancer Res. 1994;14: 1913–1918, with permission from the International Institute of Anticancer Research (IIAR). 0148-6071/06/3004-0297$03.00/0 Vol. 30, No. 4 JOURNAL OF PARENTERAL AND ENTERAL NUTRITION Printed in U.S.A. Copyright © 2006 by the American Society for Parenteral and Enteral Nutrition