The prognostic values of leukocyte Rho kinase activity in acute ischemic stroke

Cheng-I Cheng, Yu-Chun Lin, Tzu-Hsien Tsai, Hung-Sheng Lin, Chia-Wei Liou, Wen-Neng Chang, Cheng-Hsien Lu, Chun-Man Yuen, Hon-Kan Yip, Cheng-I Cheng, Yu-Chun Lin, Tzu-Hsien Tsai, Hung-Sheng Lin, Chia-Wei Liou, Wen-Neng Chang, Cheng-Hsien Lu, Chun-Man Yuen, Hon-Kan Yip

Abstract

Objective: It has been reported that leukocyte ROCK activity is elevated in patients after ischemic stroke, but it is unclear whether leukocyte ROCK activity is associated with clinical outcomes following acute stroke events. The objective of this study is to investigate if leukocyte ROCK activity can predict the outcomes in patients with acute ischemic stroke.

Materials and methods: We enrolled 110 patients of acute ischemic stroke and measured the leukocyte ROCK activity and plasma level of inflammatory cytokines to correlate the clinical outcomes of these patients.

Results: The leukocyte ROCK activity at 48 hours after admission in acute ischemic stroke patients was higher as compared to a risk-matched population. The leukocyte ROCK activity significantly correlated with National Institute of Health Stroke Scale (NIHSS) difference between admission and 90 days after stroke event. Kaplan-Meier survival estimates showed lower stroke-free survival during follow-up period in patients with high leukocyte ROCK activity or plasma hsCRP level. Leukocyte ROCK activity independently predicted the recurrent stroke in patients with atherosclerotic stroke.

Conclusions: This study shows elevated leukocyte ROCK activity in patients with ischemic stroke as compared to risk-matched subjects and is an independent predictor for recurrent stroke.

Figures

Figure 1
Figure 1
Leukocyte ROCK activity in acute ischemic stroke patients and risk-matched subjects. The bar chart displays the relative leukocyte ROCK activity in acute ischemic stroke patients (n = 33) and risk-matched subjects (n = 33) as indicated. The representative blot in the panel demonstrates the Western blot of p-MBS and t-MBS of three patients in each group.
Figure 2
Figure 2
Correlation of hsCRP level and ROCK activity with neurological function scales in patients with stroke. (a) Correlation with NIHSS upon admission. (b) Correlation with NIHSS on 90th day. (c) Correlation with NIHSS changes between admission and 90th day. (d) Plasma hsCRP level and leukocyte ROCK activity grouped by modified Rankin score. (e) Barthel index in patients with high and low plasma hsCRP level. (f) Barthel index in patients with high and low leukocyte ROCK activity. ▲: leukocyte ROCK activity; ■: plasma hsCRP level; black solid line: regression curve of leukocyte ROCK activity; gray solid line: regression curve of plasma hsCRP level.
Figure 3
Figure 3
Kaplan-Meier survival estimates of stroke-free survival during follow-up. (a) Survival estimate of stroke-free survival in patients of high and low hsCRP groups. Solid line: low hsCRP group; dashed line: high hsCRP group. (b) Survival estimate of stroke-free survival in patients of high and low leukocyte ROCK activity groups. Solid line: low leukocyte ROCK activity group; dashed line: high leukocyte ROCK activity group. (c) Survival estimate of stroke-free survival by grouped by plasma hsCRP and leukocyte ROCK activity. Black solid line: low hsCRP and low ROCK activity; black dashed line: low hsCRP and high ROCK activity; gray solid line: high hsCRP and low ROCK activity; gray dashed line: high hsCRP and high ROCK activity.
Figure 4
Figure 4
Effect of cardiovascular medications on in vitro ROCK activity in human leukocytes. Representative immunoblotting and density plot of Rho kinase activity in human leukocytes (expressed as p-MBS/t-MBS ratio) under the treatment of pharmaceutical agents as indicated. LPA was used as positive control for Rho kinase activation. *: P < 0.05 versus vehicle. #: P < 0.05 versus LPA. LPA: lysophosphatidic acid.

References

    1. Rubattu S, Giliberti R, Volpe M. Etiology and pathophysiology of stroke as a complex trait. American Journal of Hypertension. 2000;13(10):1139–1148.
    1. Tuttolomondo A, Di Raimondo D, di Sciacca R, Pinto A, Licata G. Inflammatory cytokines in acute ischemic stroke. Current Pharmaceutical Design. 2008;14(33):3574–3589.
    1. Weber C, Zernecke A, Libby P. The multifaceted contributions of leukocyte subsets to atherosclerosis: lessons from mouse models. Nature Reviews Immunology. 2008;8(10):802–815.
    1. Johnsen SH, Fosse E, Joakimsen O, et al. Monocyte count is a predictor of novel plaque formation: a 7-year follow-up study of 2610 persons without carotid plaque at baseline the Tromsø study. Stroke. 2005;36(4):715–719.
    1. Gidday JM, Gasche YG, Copin J, et al. Leukocyte-derived matrix metalloproteinase-9 mediates blood-brain barrier breakdown and is proinflammatory after transient focal cerebral ischemia. American Journal of Physiology—Heart and Circulatory Physiology. 2005;289(2):H558–H568.
    1. Hiroki J, Shimokawa H, Higashi M, et al. Inflammatory stimuli upregulate Rho-kinase in human coronary vascular smooth muscle cells. Journal of Molecular and Cellular Cardiology. 2004;37(2):537–546.
    1. Luykenaar KD, El-Rahman RA, Walsh MP. Rho-kinase-mediated suppression of KDR current in cerebral arteries requires an intact actin cytoskeleton. American Journal of Physiology—Heart and Circulatory Physiology. 2009;296(4):H917–H926.
    1. Leong SY, Faux CH, Turbic A, Dixon KJ, Turnley AM. The Rho kinase pathway regulates mouse adult neural precursor cell migration. Stem Cells. 2011;29(2):332–343.
    1. Nishimura J, Bi D, Kanaide H. Dependence of proliferating dedifferentiated vascular smooth muscle contraction on Rho-Rho kinase system. Trends in Cardiovascular Medicine. 2006;16(4):124–128.
    1. Slotta JE, Braun OÖ, Menger MD, Thorlacius H. Fasudil, a Rho-kinase inhibitor, inhibits leukocyte adhesion in inflamed large blood vessels in vivo. Inflammation Research. 2006;55(9):364–367.
    1. Wang H, Liu P, Oyama N, et al. Deficiency of ROCK1 in bone marrow-derived cells protects against atherosclerosis in LDLR-/- mice. The FASEB Journal. 2008;22(10):3561–3570.
    1. Nakakuki T, Ito M, Iwasaki H, et al. Rho/Rho-kinase pathway contributes to C-reactive protein-induced plasminogen activator inhibitor-1 expression in endothelial cells. Arteriosclerosis, Thrombosis, and Vascular Biology. 2005;25(10):2088–2093.
    1. Hage FG, Szalai AJ. C-reactive protein gene polymorphisms, C-reactive protein blood levels, and cardiovascular disease risk. Journal of the American College of Cardiology. 2007;50(12):1115–1122.
    1. Liu P, Chen J, Lin L, Liao JK. Increased Rho kinase activity in a Taiwanese population with metabolic syndrome. Journal of the American College of Cardiology. 2007;49(15):1619–1624.
    1. Hidaka T, Hata T, Soga J, et al. Increased leukocyte rho kinase (ROCK) activity and endothelial dysfunction in cigarette smokers. Hypertension Research. 2010;33(4):354–359.
    1. Nohria A, Prsic A, Liu P, et al. Statins inhibit Rho kinase activity in patients with atherosclerosis. Atherosclerosis. 2009;205(2):517–521.
    1. Feske SK, Sorond FA, Henderson GV, et al. Increased leukocyte ROCK activity in patients after acute ischemic stroke. Brain Research. 2009;1257:89–93.
    1. Liu P, Liao JK. A method for measuring Rho kinase activity in tissues and cells. Methods in Enzymology. 2008;439:181–189.
    1. Atkins KB, Irey B, Xiang N, Brosius FC., III A rapid, PPAR-γ-dependent effect of pioglitazone on the phosphorylation of MYPT. American Journal of Physiology—Cell Physiology. 2009;296(5):C1151–C1161.
    1. Shima E, Katsube M, Kato T, et al. Calcium channel blockers suppress cytokine-induced activation of human neutrophils. American Journal of Hypertension. 2008;21(1):78–84.
    1. Alvarez Y, Municio C, Alonso S, San Román JA, Sánchez Crespo M, Fernández N. Cyclooxygenase-2 induced by zymosan in human monocyte-derived dendritic cells shows high stability, and its expression is enhanced by atorvastatin. Journal of Pharmacology and Experimental Therapeutics. 2009;329(3):987–994.
    1. Chen H, Xu Z, Chen L, Wang W, Fang Q, Yan X. Valsartan and telmisartan abrogate angiotensin II-induced down-regulation of ABCA1 expression via AT1 receptor, rather than AT2 receptor or PPARγ activation. Journal of Cardiovascular Pharmacology. 2012;59:570–575.
    1. Chen J, Baydoun AR, Xu R, et al. Lysophosphatidic acid protects mesenchymal stem cells against hypoxia and serum deprivation-induced apoptosis. Stem Cells. 2008;26(1):135–145.
    1. Johnston SC, Leira EC, Hansen MD, Adams HP., Jr. Early recovery after cerebral ischemia risk of subsequent neurological deterioration. Annals of Neurology. 2003;54(4):439–444.
    1. Kakuda W, Hamilton S, Thijs VN, et al. Optimal outcome measures for detecting clinical benefits of early reperfusion: insights from the DEFUSE Study. Journal of Stroke and Cerebrovascular Diseases. 2008;17(4):235–240.
    1. Tuttolomondo A, Pedone C, Pinto A, et al. Predictors of outcome in acute ischemic cerebrovascular syndromes: the GIFA study. International Journal of Cardiology. 2008;125(3):391–396.
    1. Maas MB, Furie KL. Molecular biomarkers in stroke diagnosis and prognosis. Biomarkers in Medicine. 2009;3(4):363–383.
    1. Whiteley W, Jackson C, Lewis S, et al. Inflammatory markers and poor outcome after stroke: a prospective cohort study and systematic review of interleukin-6. PLoS Medicine. 2009;6(9)e1000145
    1. Li ZG, Yu ZC, Yu YP, et al. Lysophosphatidic Acid level and the incidence of silent brain infarction in patients with nonvalvular atrial fibrillation. International Journal of Molecular Sciences. 2010;11:3988–3998.
    1. Li Z, Yu Z, Wang D, et al. Influence of acetylsalicylate on plasma lysophosphatidic acid level in patients with ischemic cerebral vascular diseases. Neurological Research. 2008;30(4):366–369.
    1. Chen X, Yang XY, Wang ND, et al. Serum lysophosphatidic acid concentrations measured by dot immunogold filtration assay in patients with acute myocardial infarction. Scandinavian Journal of Clinical and Laboratory Investigation. 2003;63(7-8):497–503.
    1. Siess W, Zangl KJ, Essler M, et al. Lysophosphatidic acid mediates the rapid activation of platelets and endothelial cells by mildly oxidized low density lipoprotein and accumulates in human atherosclerotic lesions. Proceedings of the National Academy of Sciences of the United States of America. 1999;96(12):6931–6936.
    1. Kim SJ, Moon GJ, Bang OY. Biomarkers for stroke. Journal of Stroke. 2013;15:27–37.
    1. Liu P, Liu Y, Lin L, Chen J, Liao JK. Evidence for statin pleiotropy in humans: differential effects of statins and ezetimibe on Rho-associated coiled-coil containing protein kinase activity, endothelial function, and inflammation. Circulation. 2009;119(1):131–138.
    1. Hata T, Soga J, Hidaka T, et al. Calcium channel blocker and Rho-associated kinase activity in patients with hypertension. Journal of Hypertension. 2011;29(2):373–379.
    1. Ocaranza MP, Gabrielli L, Mora I, et al. Markedly increased Rho-kinase activity in circulating leukocytes in patients with chronic heart failure. American Heart Journal. 2011;161(5):931–937.
    1. Yano K, Kawasaki K, Hattori T, et al. Demonstration of elevation and localization of Rho-kinase activity in the brain of a rat model of cerebral infarction. European Journal of Pharmacology. 2008;594(1–3):77–83.
    1. Rikitake Y, Kim H, Huang Z, et al. Inhibition of Rho kinase (ROCK) leads to increased cerebral blood flow and stroke protection. Stroke. 2005;36(10):2251–2257.
    1. Kajimoto H, Hashimoto K, Bonnet SN, et al. Oxygen activates the rho/rho-kinase pathway and induces RhoB and ROCK-1 expression in human and rabbit ductus arteriosus by increasing mitochondria-derived reactive oxygen species: a newly recognized mechanism for sustaining ductal constriction. Circulation. 2007;115(13):1777–1788.
    1. Takemoto M, Sun J, Hiroki J, Shimokawa H, Liao JK. Rho-kinase mediates hypoxia-induced downregulation of endothelial nitric oxide synthase. Circulation. 2002;106(1):57–62.
    1. Laschke MW, Dold S, Jeppsson B, Schilling MK, Menger MD, Thorlacius H. Rho-kinase inhibitor attenuates cholestasis-induced CXC chemokine formation, leukocyte recruitment, and hepatocellular damage in the liver. Journal of Surgical Research. 2010;159(2):666–673.
    1. Bardi G, Niggli V, Loetscher P. Rho kinase is required for CCR7-mediated polarization and chemotaxis of T lymphocytes. FEBS Letters. 2003;542(1–3):79–83.
    1. Lampl Y, Boaz M, Gilad R, et al. Minocycline treatment in acute stroke: an open-label, evaluator-blinded study. Neurology. 2007;69(14):1404–1410.
    1. Diener H, Schneider D, Lampl Y, Bornstein NM, Kozak A, Rosenberg G. DP-b99, a membrane-activated metal ion chelator, as neuroprotective therapy in ischemic stroke. Stroke. 2008;39(6):1774–1778.
    1. Kidwell CS, Lees KR, Muir KW, et al. Results of the MRI substudy of the intravenous magnesium efficacy in stroke trial. Stroke. 2009;40(5):1704–1709.
    1. Martin C, Göggel R, Ressmeyer A-R, Uhlig S. Pressor responses to platelet-activating factor and thromboxane are mediated by Rho-kinase. American Journal of Physiology—Lung Cellular and Molecular Physiology. 2004;287(1):L250–L257.
    1. Bivard A, Lin L, Parsonsb MW. Review of stroke thrombolytics. Journal of Stroke. 2013;15:90–98.
    1. Geeganage CM, Bath PMW. Relationship between therapeutic changes in blood pressure and outcomes in acute stroke: a metaregression. Hypertension. 2009;54(4):775–781.
    1. Wang CC, Reusch JE. Diabetes and cardiovascular disease: changing the focus from glycemic control to improving long-term survival. The American Journal of Cardiology. 2012;110:58B–68B.
    1. Lindholm LH, Carlberg B, Samuelsson O. Should β blockers remain first choice in the treatment of primary hypertension? A meta-analysis. The Lancet. 2005;366(9496):1545–1553.
    1. Verdecchia P, Reboldi G, Angeli F, et al. Angiotensin-converting enzyme inhibitors and calcium channel blockers for coronary heart disease and stroke prevention. Hypertension. 2005;46(2):386–392.
    1. Julius S, Kjeldsen SE, Weber M, et al. Outcomes in hypertensive patients at high cardiovascular risk treated with regimens based on valsartan or amlodipine: the VALUE randomised trial. The Lancet. 2004;363(9426):2022–2031.
    1. Fujimura N, Noma K, Hata T, et al. Mineralocorticoid receptor blocker eplerenone improves endothelial function and inhibits Rho-associated kinase activity in patients with hypertension. Clinical Pharmacology and Therapeutics. 2012;91(2):289–297.
    1. Kitayama J, Kitazono T, Ooboshi H, Takada J, Fujishima M, Ibayashi S. Long-term effects of benidipine on cerebral vasoreactivity in hypertensive rats. European Journal of Pharmacology. 2002;438(3):153–158.
    1. Failli P, Alfarano C, Franchi-Micheli S, et al. Losartan counteracts the hyper-reactivity to angiotensin II and ROCK1 over-activation in aortas isolated from streptozotocin-injected diabetic rats. Cardiovascular Diabetology. 2009;8, article 32
    1. Kaynar K, Ulusoy S, Ovali E, Vanizor B, Dikmen T, Gul S. TGF-β and TNF-α producing effects of losartan and amlodipine on human mononuclear cell culture. Nephrology. 2005;10(5):478–482.

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