Antithrombin reduces reperfusion-induced liver injury in mice by enhancing sensory neuron activation

Summary

We recently demonstrated that antithrombin (AT) reduces ischemia/reperfusion (I/R)-induced liver injury in rats by increasing hepatic tissue levels of calcitonin gene-related peptide (CGRP),a neuropeptide released from the sensory nerve endings. In the present study, we examined the effect of AT on I/Rinduced liver injury in wild type mice (CGRP^+/+) and congenitally αCGRP-deficient mice (CGRP^−/−). We also investigated any effects of AT on CGRP release from dorsal root ganglion neurons (DRG) isolated from CGRP^+/+. Based on results obtained in the present study, we attempted to determine if the anti-inflammatory activity of AT in vivo is dependent mainly on sensory neuron activation. AT enhanced ischemia/reperfusion-induced increases in hepatic tissue levels of CGRP and 6-keto-PGF_{1α ,} a stable metabolite of PGI₂, in CGRP^+/+, but it did not enhance these increases in CGRP^−/−. AT inhibited reperfusion-induced increases in serum alanine aminotransferase levels by increasing hepatic tissue blood flow and by attenuating increases in hepatic levels of tumor necrosis factor and myeloperoxidase in CGRP^+/+,although it showed neither of these therapeutic effects in CGRP^−/−. AT increased CGRP release from cultured DRGs only in the presence of anandamide, and AT-induced increase in CGRP release was not observed in the presence KT5720, an inhibitor of protein kinase A (PKA).AT markedly increased intracellular levels of cAMP in the presence of anandamide. These results strongly suggest that AT might reduce I/R-induced liver injury by enhancing activation of the sensory neurons through activation of PKA in sensory neurons.

Keywords

Antithrombin - capsaicin-sensitive sensory neurons - calcitonin gene-related peptide - liver injury - protein kinase A

• References

• 1 Rosenberg RD. Biochemistry of heparin antithrombin interactions, and the physiological role of this natural anticoagulant mechanism. Am J Med 1989; 87 Suppl 3B 2S-9S.
• 2 Okajima K, Ueyama H, Hashimoto Y. et al. Homozygous variant of antithrombin III that lacks affinity for heparin: AT III Kumamoto. Thromb Haemost 1989; 61: 20-4.
• 3 Okajima K, Abe H, Maeda S. et al. Antithrombin III Nagasaki (Ser116-Pro): a heterozygous variant with defective heparin binding associated with thrombosis. Blood 1993; 81: 1300-5.
• 4 Coccheri S, Palareti G. Antithrombin III replacement in orthotopic liver transplantation. Semin Thromb Haemost 1993; 19: 268-72.
• 5 Okajima K. Regulation of inflammatory responses by natural anticoagulants. Immunol Rev 2001; 184: 258-74.
• 6 Keller GA, West MA, Cerra FB. et al. Macrophagemediated modulation of hepatic function in multi-system failure. J Surg Res 1985; 39: 555-63.
• 7 Jaeschke H, Farhood A, Smith CW. Neutrophils contribute to ischemia/reperfusion injury in rat liver in vivo . FASEB J 1990; 04: 3355-9.
• 8 Eisenhut T, Sinha B, Grottrup-Wolfers E. et al. Prostacyclin analogs suppress the synthesis of tumor necrosis factor-α in LPS-stimulated human peripheral blood mononuclear cells. Immunopharmacology 1993; 26: 259-64.
• 9 Kainoh M, Imai R, Umetsu T. et al. Prostacyclin and beraprost sodium as suppressors of activated rat polymorphonuclear leukocytes. Biochem Pharmacol 1990; 39: 477-84.
• 10 Dray A. Inflammatory mediators of pain. Br J Anaesth 1995; 75: 125-31.
• 11 Szallasi A, Blumberg PM. Vanilloid receptors: new insights enhance potential asa therapeutic target. Pain 1996; 68: 195-208.
• 12 Crossman D, McEwan J, MacDermot J. et al. Human calcitonin gene-related peptide activates adenylate cyclase and releases prostacyclin from human umbilical vein endothelial cells. Br J Pharmacol 1987; 92: 695-701.
• 13 Fields HL. Pain. New York: McGraw-Hill; 1987
• 14 Harada N, Okajima K, Uchiba M. et al. Ischemia/ reperfusion-induced increase in the hepatic level of prostacyclin is mainly mediated by activation of capsaicin-sensitive sensory neurons in rats. J Lab Clin Med 2002; 139: 218-26.
• 15 Harada N, Okajima K, Yuksel M. et al. Contribution of capsaicin-sensitive sensory neurons to antithrombin-induced reduction of ischemia/reperfusion-induced liver injury in rats. Thromb Haemost 2005; 93: 48-56.
• 16 Uchiba M, Okajima K, Kaun C. et al. Inhibition of the endothelial cell activation by antithrombin in vitro . Thromb Haemost 2004; 92: 1420-7.
• 17 De Petrocellis L, Harrison S, Bisogno T. et al. The vanilloid receptor (VR-1)-mediated effects of anandamide are potently enhanced by the cAMP-dependent protein kinase. J Neurochem 2001; 77: 1660-3.
• 18 Oh-hashi Y, Shindo T, Kurihara Y. et al. Elevated sympathetic nervous activity in mice deficient in aCGRP. Circ Res 2001; 89: 983-90.
• 19 Miller-Anderson M, Borg H, Anderson LO. Purification of antithrombin III by affinity chromatography. Thromb Res 1974; 05: 439-52.
• 20 Uchiba M, Okajima K, Murakami K. et al. Effect of antithrombin III (AT III) and Trp⁴⁹-modified AT III on plasma level of 6-keto-PGF1α in rats. Thromb Res 1995; 80: 201-8.
• 21 Khandoga A, Biberthaler P, Messmer K. et al. Platelet-endothelial cell interactions during hepatic ischemia-reperfusion in vivo:a systematic analysis. Microvasc Res 2003; 65: 71-7.
• 22 Hayashi H, Chaudry IH, Clemens MG. et al. Hepatic ischemia models for determining the effects of ATP-MgCl2 treatment. J Surg Res 1986; 40: 167-75.
• 23 Liu L, Simon SA. Capsazepine, a vanilloid receptor antagonist, inhibits nicotinic acetylcholine receptors in rat trigeminal ganglia. Neurosci Lett 1997; 228: 29-32.
• 24 Akiyoshi H, Gonda T, Terada T. A comparative histochemical and immunohistochemical study of aminergic, cholinergic and peptidergic innervation in rat, hamster, guinea pig, dog and human livers. Liver 1998; 18: 352-9.
• 25 Harada N, Okajima K, Kushimoto S. et al. Antithrombin reduces ischemia/reperfusion injury of rat liver by increasing the hepatic level of prostacyclin. Blood 1999; 93: 157-64.
• 26 Shito M, Wakabayashi G, Ueda M. et al. Interleukin 1 receptor blockade reduces tumor necrosis factor production, tissue injury, and mortality after hepatic ischemia-reperfusion in the rat. Transplantation 1997; 63: 143-8.
• 27 Warren JS, Yabroff KR, Remick DG. et al. Tumor necrosis factor participates in the pathogenesis of acute immune complex alveolitis in the rat. J Clin Invest 1989; 84: 1873-82.
• 28 Kushimoto S, Okajima K, Uchiba M. et al. Role of granulocyte elastase in ischemia / reperfusion injury of rat liver. Crit Care Med 1996; 24: 1908-12.
• 29 Tognetto M, Amadesi S, Harrison S. et al. Anandamide excites central terminals of dorsal root ganglion neurons via vanilloid receptor-1 activation. J Neurosci 2001; 21: 1104-9.
• 30 Price TJ, Patwardhan A, Akopian AN. et al. Modulation of trigeminal sensory neuron activity by the dual cannabinoid-vanilloid agonists anandamide, N-arachi-donoyldopamine and arachidonyl-2-chloroethylamide. Br J Pharmacol 2004; 141: 1118-30.
• 31 Hou L, Wang X. PKC and PKA, but not PKG mediate LPS-induced CGRP release and [Ca2+]i elevation in DRG neurons of neonatal rats. J Neurosci Res 2001; 66: 592-600.
• 32 Bull HA, Hothersall J, Chowdhury N. et al. Neuropeptides induce release of nitric oxide from human dermal microvascular endothelial cells. J Invest Dermatol 1996; 106: 655-60.
• 33 Armstrong JM, Chapple D, Dusting GJ. et al. Cardiovascular actions of prostacyclin (PGI2) in chloralose anesthetized dogs. Br J Pharmacol 1977; 61: 136-41.
• 34 Harada N, Okajima K, Kushimoto S. Gabexate mesilate, a synthetic protease inhibitor, reduces ischemia/reperfusion injury of rat liver by inhibiting leukocyte activation. Crit Care Med 1999; 27: 1958-64.
• 35 Stoyanova II, Gulubova MV. Peptidergic nerve fibers in the human liver. Acta Histochem 1998; 100: 245-56.
• 36 Koj A, Regoeczi E. Effect of experimental inflammation on the synthesis and distribution of antithrombin III and alpha-1-antitrypsin in rabbits. Br J Exp Pathol 1978; 59: 473-81.
• 37 van Nieuw GPAmerongen, van Hinsbergh VW. Targets for pharmacological intervention of endothelial hyperpermeability and barrier function. Vascular Phamacol 2003; 39: 257-72.
• 38 Okajima K, Harada N, Uchiba M. et al. Neutrophil elasatase contributes to the development of ischemia/ reperfusion-induced liver injury by decreasing endothelial production of prostacyclin in rats. Am J Physiol Gastrointest Liver Physiol 2004; 287: G1116-G1123.
• 39 Harada N, Okajima K, Uchiba M. et al. Antithrombin reduces ischemia/reperfusion-induced liver injury in rats by activation of cyclooxygenase-1. Thromb Haemost 2004; 92: 550-8.