Age-associated pro-inflammatory adaptations of the mouse thoracic aorta

Summary

Arterial ageing may be associated with a reduction in vasodilation due to increased reactive oxygen species (ROS) production, whereas endothelial cell activation induces procoagulant changes. However, little is known on the effect of ageing on expression of anticoagulant endothelial markers such as endothelial protein C receptor (EPCR). To study age-associated alterations in smooth muscle cell (SMC) and endothelial cell (EC) structure and function, the aorta was isolated from 10-week-and 12– and 24-month-old C57BL/6J mice and analysed for its expression of genes involved in senescence, oxidative stress production, coagulation and matrix remodelling. In addition, vasorelaxation experiments were performed using 10-week-and 24-month-old thoracic aortic ring segments in organ chamber baths. The media thickness of the thoracic aorta progressively increased with age, associated with hypertrophy of vascular SMCs. Basal nitric oxide production and sensitivity to acetylcholine-mediated vasodilation in thoracic aorta rings was reduced with age, whereas no significant differences in ROS production could be demonstrated. Gene expression of tissue factor, EPCR and von Willebrand factor was not affected by ageing of the aorta, whereas that of thrombomodulin was mildly reduced and that of xanthine dehydrogenase, NADPH oxidase 4, tumour necrosis factor-α and vascular cell adhesion molecule-1 significantly enhanced. In conclusion, a reduction in endothelial cell-mediated vasodilation in aged thoracic aortas of C57BL/6J mice was accompanied by a shift towards a pro-inflammatory state of the endothelium.

Keywords

Vascular ageing - C57BL/6J mice - endothelial dysfunction - EPCR

• References

• 1 Wilkerson WR, Sane DC. Aging and thrombosis. Semin Thromb Hemost 2002; 28: 555-568.
• 2 Makin A, Silverman SH, Lip GYH. Peripheral vascular disease and Virchow’s triad for thrombogenesis. Q J Med 2002; 95: 199-210.
• 3 van der Loo B, Labugger R, Skepper JN. et al. Enhanced peroxynitrite formation is associated with vascular aging. J Exp Med 2000; 192: 1731-1744.
• 4 Brandes RP, Fleming I, Russe R. Endothelial aging. Cardiovasc Res 2005; 66: 286-294.
• 5 Donato AJ, Eskurza I, Silver AE. et al. Direct evidence of endothelial oxidative stress with aging in humans in relation to impaired endothelium-dependent dilation and upregulation of nuclear factor-kappa B. Circulation Res 2007; 100: 1659-1666.
• 6 Kollander R, Solovey A, Milbauer LC. et al. Nuclear factor-kappa B (NFkappaB) component p50 in blood mononuclear cells regulates endothelial tissue factor expression in sickle transgenic mice: implications for the coagulopathy of sickle cell disease. Transl Res 2010; 155: 170-177.
• 7 Vaughan DE. PAI-1 and atherothrombosis. J Thromb Haemost 2005; 3: 1879-1883.
• 8 Reininger AJ. Function of von Willebrand factor in haemostasis and thrombosis. Haemophilia 2008; 14: 11-26.
• 9 Stämpfli SF, Akhmedov A, Gebhard C. et al. Aging induces endothelial dysfunction while sparing arterial thrombosis. Arterioscler Thromb Vasc Biol 2010; 30: 1960-1967.
• 10 Starr ME, Ueda J, Takahashi H. et al. Age-dependent vulnerability to endotoxemia is associated with reduction of anticoagulant factors activated protein C and thrombomodulin. Blood 2010; 115: 4886-4893.
• 11 Dahlback B, Villoutreix BO. The anticoagulant protein C pathway. FEBS Lett 2005; 579: 3310-3316.
• 12 Esmon CT. Protein C anticoagulant-anti-inflammatory effects. Semin Immunopathol 2012; 34: 127-132.
• 13 Chen XD, Tian L, Li M. et al. Relationship between endothelial cell protein C receptor gene 693A/G polymorphisms and deep venous thrombosis. Chin Med J (Engl) 2011; 124: 72-75.
• 14 Van de Water NS, French JK, McDowell J. et al. The endothelial protein C receptor (EPCR) 23 bp insert in patients with myocardial infarction. Thromb Haemost 2001; 85: 749-751.
• 15 Wang J, Yang L, Rezaie AR. et al. Activated protein C protects against myocardial ischaemic/reperfusion injury through AMP-activated protein kinase signalling. J Thromb Haemost 2011; 9: 1308-1317.
• 16 Hemmeryckx B, Emmerechts J, Bovill EG. et al. Effect of ageing on the murine venous circulation. Histochem Cell Biol 2012; 137: 537-546.
• 17 Zhang J, Burridge KA, Friedman MH. In vivo differences between endothelial transcriptional profiles of coronary and iliac arteries revealed by microarray analysis. Am J Physiol Heart Circ Physiol 2008; 295: H1556-H1661.
• 18 Hemmeryckx B, Van Hove CE, Fransen P. et al. Progression of the prothrombotic state in ageing Bmal1-deficient mice. Arterioscler Thromb Vasc Biol 2011; 31: 2552-2559.
• 19 Fransen P, Van Assche T, Guns PJ. et al. Endothelial function in aorta segments of apolipoprotein E-deficient mice before development of atherosclerotic lesions. Pflugers Arch 2008; 455: 811-818.
• 20 Luttun A, Lutgens E, Manderveld A. et al. Loss of matrix metalloproteinase-9 or matrix metalloproteinase-12 protects apolipoprotein E-deficient mice against atherosclerotic media destruction but differentially affects plaque growth. Circulation 2004; 109: 1408-1414.
• 21 Konsti J, Lundin M, Joensuu H. et al. Development and evaluation of a virtual microscopy application for automated assessment of Ki-67 expression in breast cancer. BMC Clin Pathol 2011; 11: 3.
• 22 Fleenor BS, Seals DR, Zigler ML. et al. Superoxide-lowering therapy with TEMPOL reverses arterial dysfunction with aging in mice. Aging Cell 2012; 11: 269-276.
• 23 Van Hove CE, Van der Donkt C, Herman AG. et al. Vasodilator efficacy of nitric oxide depends on mechanisms of intracelllar calcium mobilisation in mouse aortic smooth muscle cells. Br J Pharmacol 2009; 158: 920-930.
• 24 Kang LS, Reyes RA, Muller-Delp JM. Aging impairs flow-induced dilation in coronary arterioles: role of NO and H(2)O(2). Am J Physiol Heart Circ Physiol 2009; 297: H1087-H1095.
• 25 Lentz SR, Tsiang M, Sadler JE. Regulation of thrombomodulin by tumor necrosis factor - comparison of transcriptional and posttranscriptional mechanisms. Blood 1991; 77: 542-550.
• 26 Kume M, Hayashi T, Yuasa H. et al. Bacterial lipopolysaccharide decreases thrombomodulin expression in the sinusoidal endothelial cells of rats: a possible mechanism of intrasinusoidal microthrombus formation and liver dysfunction. J Hepatol 2003; 38: 9-17.
• 27 Conway EM, Rosenberg RD. Tumor necrosis factor suppresses transcription of the thrombomodulin gene in endothelial cells. Mol Cell Biol 1988; 8: 5588-5592.
• 28 Rajan S, Ye J, Bai S. et al. NF-kappaB, but not p38 MAP kinase, is required for TNF-alpha-induced expression of cell adhesion molecules in endothelial cells. J Cell Biochem 2008; 105: 447-486.
• 29 Zhang H, Park Y, Wu J. et al. Role of TNF-alpha in vascular dysfunction. Clin Sci 2009; 116: 219-230.
• 30 Lesniewski LA, Durrant JR, Connell ML. et al. Salicylate treatment improves age-associated vascular endothelial dysfunction: potential role of nuclear factor kappaB and forkhead Box O phosphorylation. J Gerontol A Biol Sci Med Sci 2011; 66: 409-418.
• 31 Lesniewski LA, Durrant JR, Connell ML. et al. Aerobic exercise reverses arterial inflammation with aging in mice. Am J Physiol Heart Circ Physiol 2011; 301: H1025-H1032.
• 32 Stein S, Schäfer N, Breitenstein A. et al. SIRT1 reduces endothelial activation without affecting vascular function in ApoE-/- mice. Aging 2010; 2: 353-360.
• 33 Eiserich JP, Baldus S, Brennan ML. et al. Myeloperoxidase, a leukocyte-derived vascular NO oxidase. Science 2002; 296: 2391-2394.
• 34 Walker AE, Seibert SM, Donato AJ. et al. Vascular endothelial function is related to white blood cell count and myeloperoxidase among healthy middle-aged and older adults. Hypertension 2010; 55: 363-369.