Alanine-Glutamine Dipeptide (AGD) Inhibits Expression of Inflammation-Related Genes in Hemorrhagic Shock

Abstract

Background: Inflammatory factors play an important role in the production of cellular damage after shock and reperfusion. Glutamine has been used to modulate the inflammatory response. Alanine-glutamine dipeptide (AGD) is a glutamine source. The hypothesis of the present study is that AGD given during resuscitation will suppress postshock expression of messenger ribonucleic acid (mRNA) for tumor necrosis factor (TNF-α), interleukin-1 (IL-1β) and inducible nitric oxide synthase (iNOS). Methods: Male Sprague-Dawley rats (n = 74, 350 g ± 30 g) were randomly assigned to 5 groups. Under isoflurane anesthesia, the femoral artery and vein were 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) with or without the administration of AGD (936 mg/kg) and returning the shed blood. Controls were normal (anesthesia only), sham (surgical preparation), and shock (preparation and shock). Rats (n = 45, 9 per group) were killed 30 minutes after completion of resuscitation. Liver samples were collected, and total RNA was isolated for reverse transcription–polymerase chain reaction analysis of mRNA (TNF-α, IL-1β, iNOS, and β-actin). Results: MAP recovered more quickly in the AGD group than in the LR group. Increased expression of liver mRNA for TNF-α, IL-1β, and iNOS was seen after hemorrhagic shock and resuscitation. AGD treatment significantly reduced mRNA expression for all 3. Conclusions: AGD modified the expression of genes controlling cytokines and iNOS in the liver. This agent is a potential treatment for hemorrhagic shock.

The use of alanine-glutamine dipeptide (AGD), a glutamine source administered in resuscitation for hemorrhagic shock, helped restore blood pressure quickly and inhibited mRNA expression for proinflammatory cytokines and inducible nitric oxide synthase.

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References

Yang R, Gallo DJ, Baust JJ, Watkins SK, Delude RL, Fink MP. Effect of hemorrhagic shock on gut barrier function and expression of stress-related genes in normal and gnotobiotic mice. Am J Physiol Regul Integr Comp Physiol. 2002;283:R1263– R1274.

Collins JL, Vodovotz Y, Hierholzer C, et al. Characterization of the expression of inducible nitric oxide synthase in rat and human liver during hemorrhagic shock. Shock. 2003 ;19:117– 122.

Goeters C, Wenn A, Mertes N, et al. Parenteral L-alanyl-L-glutamine improves 6-month outcome in critically ill patients. Crit Care Med. 2002;30:2153– 2154.

Griffiths RD, Allen KD, Andrews FJ, Jones C. Infection, multiple organ failure, and survival in the intensive care unit: influence of glutamine-supplemented parenteral nutrition on acquired infection.Nutrition. 2002;18:546– 552.

Van Way CW 3rd, Dhar A, Morrison D. Hemorrahagic shock: a new look at an old problem. Mo Med. 2003 ;100:518– 523.

Jerry LH. NCSS 97 User's Guide-I. Kaysville, UT: Number Cruncher Statistical Systems; 1997.

Scharte M, Baba HA, Van Aken H, et al. Alanyl-glutamine dipeptide does not affect hemodynamics despite a greater increase in myocardial heat shock protein 72 immunoreactivity in endotoxemic sheep. J Nutr. 2001 ;131:1433– 1437.

Flynn WJ Jr, Gosche JR, Garrison RN. Intestinal blood flow is restored with glutamine or glucose suffusion after hemorrhage. J Surg Res. 1992;52:499– 504.

Rhee P, Waxman K, Clark L, et al. Tumor necrosis factor and monocytes are released during hemorrhagic shock. Resuscitation. 1993 ;25:249– 255.

10.

Gaddipati JP, Sundar SV, Calemine J, Seth P, Sidhu GS, Maheshwari RK. Differential regulation of cytokines and transcription factors in liver by curcumin following hemorrhage/resuscitation. Shock. 2003 ;19:150– 156.

11.

Anaya-Prado R, Toledo-Pereyra LH, Walsh J, Guo RF, Reuben J, Ward PA. Exogenous nitric oxide donor and related compounds protect against lung inflammatory response after hemorrhagic shock and resuscitation. J Trauma. 2004;57:980– 988.

12.

Bhatia M, Moochhala S. Role of inflammatory mediators in the pathophysiology of acute respiratory distress syndrome. J Pathol. 2004;202:145– 156.

13.

Cohen J. The immunopathogenesis of sepsis. Nature. 2002 ;420:885– 891.

14.

Schroder J, Kahlke V, Fandrich F, et al. Glutamine dipeptides-supplemented parenteral nutrition reverses gut mucosal structure and interleukin-6 release of rat intestinal mononuclear cells after hemorrhagic shock. Shock. 1998 ;10:26– 31.

15.

Angele MK, Faist E. Clinical review: immunodepression in the surgical patient and increased susceptibility to infection. Crit Care. 2002;6:298– 305.

16.

Menezes JM, Hierholzer C, Watkins SC, Billiar TR, Peitzman AB, Harbrecht BG. The modulation of hepatic injury and heat shock expression by inhibition of inducible nitric oxide synthase after hemorrhagic shock.Shock. 2002;17:13– 18.

17.

Hierholzer C, Harbrecht B, Menezes JM, et al. Essential role of induced nitric oxide in the initiation of the inflammatory response after hemorrhagic shock. J Exp Med. 1998 ;187:917– 928.

18.

McDonald MC, Izumi M, Cuzzocrea S, Thiemermann C. A novel, potent and selective inhibitor of the activity of inducible nitric oxide synthase (GW274150) reduces the organ injury in hemorrhagic shock. J Physiol Pharmacol. 2002;53:555– 569.

19.

Hierholzer C, Menezes JM, Ungeheuer A, Billiar TR, Tweardy DJ, Harbrecht BG. A nitric oxide scavenger protects against pulmonary inflammation following hemorrhagic shock. Shock. 2002 ;17:98– 103.

20.

Taylor BS, Alarcon LH, Billiar TR. Inducible nitric oxide synthase in the liver: regulation and function. Biochemistry (Mosc). 1998 ;63:766– 781.

21.

Hierholzer C, Kalff JC, Billiar TR, Bauer AJ, Tweardy DJ, Harbrecht BG. Induced nitric oxide promotes intestinal inflammation following hemorrhagic shock. Am J Physiol Gastrointest Liver Physiol. 2004 ;286:G225– G233.

22.

Morlion BJ, Stehle P, Wachtler P, et al. Total parenteral nutrition with glutamine dipeptide after major abdominal surgery: a randomized, double-blind, controlled study. Ann Surg. 1998 ;227:302– 308.

23.

Musch MW, Hayden D, Sugi K, Straus D, Chang EB. Cell-specific induction of hsp72-mediated protection by glutamine against oxidant injury in IEC18 cells. Proc Assoc Am Physicians. 1998 ;110:136– 139.

24.

Ogle CK, Ogle JD, Mao JX, et al. Effect of glutamine on phagocytosis and bacterial killing by normal and pediatric burn patient neutrophils. JPEN J Parenter Enteral Nutr. 1994 ;18:128– 133.

25.

Kircher B, Eibl G, Enrich B, Latzer K, Herold M, Niederwieser D. The role of L-alanyl-L-glutamine in the immune response in vitro. Wien Klin Wochenschr. 2002 ;114:702– 708.

26.

Lin MT, Saito H, Furukawa S, et al. Alanyl-glutamine enriched total parenteral nutrition improves local, systemic, and remote organ responses to intraperitoneal bacterial challenge. JPEN J Parenter Enteral Nutr. 2001;25:346– 351.

27.

Spittler A, Winkler S, Gotzinger P, et al. Influence of glutamine on the phenotype and function of human monocytes. Blood. 1995 ;86:1564– 1569.