Crocetin Inhibits mRNA Expression for Tumor Necrosis Factor-α,Interleukin-1β,and Inducible Nitric Oxide Synthase in Hemorrhagic Shock

Abstract

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-α), interleukin-1 (IL-1β) 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-α, IL-1β, iNOS, and β-actin). Results: Liver mRNA expression for TNF-α, IL-1β, 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-α, IL-1β, 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.

Crocetin, a constituent of saffron, was administered in resuscitation after hemorrhagic shock in the rat model. It inhibited expression of mA for proinflammatory cytokines and inducible nitric oxide synthase in the rat model.

Get full access to this article

View all access options for this article.

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.

Gainer JL, Rudolph DB, Caraway DL. The effect of crocetin on hemorrhagic shock in rats. Circ Shock. 1993 ;41:1– 7.

Singer M, Stidwill RP, Nathan A, Gainer JL. Intravenous crocetinate prolongs survival in a rat model of lethal hypoxemia. Crit Care Med. 2000;28:1968– 1972.

Van Way CW 3rd, Dhar A, Morrison DC, Longorio MA, Maxfield DM. Cellular energetics in hemorrhagic shock: restoring adenosine triphosphate to the cells. J Trauma. 2003 ;54(5 Suppl):S169– S176.

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

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

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.

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.

10.

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

11.

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

12.

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

13.

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.

14.

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.

15.

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.

16.

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.

17.

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

18.

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.

19.

Hosseinzadeh H, Younesi HM. Antinociceptive and anti-inflammatory effects of Crocus sativus L. stigma and petal extracts in mice [BMC Pharmacology Web site]. Available at: http://www.biomedcentral.com/1471-2210/2/7. Accessed April 15, 2005.

20.

Wang CJ, Lee MJ, Chang MC, Lin JK. Inhibition of tumor promotion in benzo[a]pyrene-initiated CD-1 mouse skin by crocetin.Carcinogenesis. 1995;16:187– 191.

21.

Wang CJ, Hsu JD, Lin JK. Suppression of aflatoxin B1-induced hepatotoxic lesions by crocetin (a natural carotenoid).Carcinogenesis. 1991;12:1807– 1810.

22.

Hsu JD, Chou FP, Lee MJ, et al. Suppression of the TPA-induced expression of nuclear-protooncogenes in mouse epidermis by crocetin via antioxidant activity. Anticancer Res. 1999 ;19:4221– 4227.

23.

Tseng TH, Chu CY, Huang JM, Shiow SJ, Wang CJ. Crocetin protects against oxidative damage in rat primary hepatocytes. Cancer Lett. 1995;97:61– 67.

24.

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.