Arginase blockade lessens endothelial dysfunction after thrombosis

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Arginase blockade lessens endothelial dysfunction after thrombosis Chandani Lewis, MD, Weifei Zhu, PhD, Mircea L. Pavkov, MD, Corttrell M. Kinney, BS, Paul E. DiCorleto, PhD, Vikram S. Kashyap, MD Journal of Vascular Surgery Volume 48, Issue 2, Pages 441-446 (August 2008) DOI: 10.1016/j.jvs.2008.02.030 Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 1 A, Thrombin-induced arginase I messenger RNA (mRNA) expression in five separate experiments. B, Thrombin-induced arginine I protein expression in three separate experiments. *P = .001. Journal of Vascular Surgery 2008 48, 441-446DOI: (10.1016/j.jvs.2008.02.030) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 2 Effect of thrombosis on endothelial-dependent relaxation (EDR) of rat infrarenal aortic (IRA) ring segments. EDR is plotted as percentage relaxation of baseline on the y axis, with log increases in the dose of acetylcholine (Ach, 10−9M to 10−4M) on the x axis. EDR of control (nonthrombosed) rat IRA (diamonds) is compared with IRAs after 1 hour of thrombosis (squares). EDR is depressed in the thrombosis group for all acetylcholine doses (n = 5 in control and n = 4 thrombosed group). *P < .05, **P < .001 by two-way analysis of variance. Data are presented with the standard error. Journal of Vascular Surgery 2008 48, 441-446DOI: (10.1016/j.jvs.2008.02.030) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 3 A, Western blot analysis of arginase I (ARGI) in rat infrarenal arterial segments. Arginase I protein levels were upregulated in endothelium intact thrombosed infrarenal aortic ring segments compared with control (n = 3). B, Western blot analysis of denuded aortic segments. Arginase I protein is not detected (arrow indicates location of arginase I protein by size). GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Journal of Vascular Surgery 2008 48, 441-446DOI: (10.1016/j.jvs.2008.02.030) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 4 Effect of nonspecific arginase inhibition on endothelial-dependent relaxation (EDR) of thrombosed rat infrarenal aortic (IRA) ring segments is shown. EDR is plotted as percentage relaxation of baseline, with log increases in the dose of acetylcholine (Ach, 10−9M to 10−4M). EDR of nonthrombosed (control IRA, diamonds; n = 3) is compared with IRA segments after 1 hour of thrombosis untreated and treated with arginase inhibitors (thrombosed IRA, clear squares; n = 3) and treated with nonspecific arginase inhibitor difluoromethylornithine (DFMO IRA, filled squares; n = 9). Treatment with arginase inhibitor restores the EDR in thrombosed IRA. *P < .05, **P < .001 DFMO vs thrombosed IRA; #P < .01, control vs thrombosed IRA by two-way analysis of variance. Data are presented with the standard error. Journal of Vascular Surgery 2008 48, 441-446DOI: (10.1016/j.jvs.2008.02.030) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 5 Effect of specific arginase inhibition on endothelial-dependent relaxation (EDR) of thrombosed infrarenal aorta (IRA) is shown. EDR plotted as percentage relaxation of baseline, with log increases from 10−9M to 10−4M in the dose of acetylcholine (Ach). EDR of nonthrombosed (control IRA, triangle; n = 3) is compared with IRA after 1 hour of thrombosis untreated with arginase inhibitors (Thrombosed IRA, clear square; n = 3) and treated with the specific arginase inhibitor [S]-[2-boronoethyl]-L-Cysteine-HCl (BEC IRA, black square; n = 6). Treatment with arginase inhibitor restores the EDR in thrombosed IRA. **P < .001 BEC vs thrombosed IRA analysis of variance. #P < .01, ##P < .001 control vs thrombosed IRA by two-way analysis of variance. Data are presented with the standard error. Journal of Vascular Surgery 2008 48, 441-446DOI: (10.1016/j.jvs.2008.02.030) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions