Hypertension and diabetes are associated with a risk of cardiovascular disease. Clonidine is currently used as a fourth-line drug therapy for hypertension because of its rebound hypertensive effect and short half-life. The purpose of this study was to investigate the combined effect of an antihypertensive drug (clonidine) and a G-protein-coupled receptor kinase 2 (GRK2) inhibitor on rebound hypertension and endothelial dysfunction. The clonidine and/or GRK2 inhibitor were administered by continuous infusion for 14 days by using an osmotic pump that was implanted subcutaneously. To test the effects of GRK2 inhibitotr, we measured blood pressure by using a tail-cuff system in diabetic mice in which rebound hypertension was induced by withdrawal after clonidine treatment and measured vascular responses in isolated aortas from these mice. The mice were then euthanized 7 days later. We observed that, in diabetes mellitus (DM) mice, blood pressure began to decline after 3 days of clonidine or clonidine + GRK2-inhibitor infusion. However, 15 days after initiation of treatment, the blood pressure of the clonidine only-treated DM mice began to increase and resulted in a high final blood pressure. At 21 days, clonidine withdrawal triggered rebound hypertension together with impaired endothelium-dependent relaxation, increased GRK2 activity, and reduced Akt/endothelial NO synthase (eNOS)/NO production in aortas. Conversely, withdrawal of the combination clonidine/GRK2-inhibitor treatment did not cause rebound hypertension, and normal induction of endothelium-dependent relaxation, decreased GRK2 activity, and increased Akt/eNOS were observed in aortas from DM mice. These results suggest that suppression of GRK2 activity affects rebound hypertension-associated vascular endothelial dysfunction by targeting the Akt/eNOS signaling pathway.
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Campbell KL, Kushner H, Falkner B. Obesity and high blood pressure: a clinical phenotype for the insulin resistance syndrome in African Americans. J Clin Hypertens (Greenwich). 2004;6:364–70. quiz371–372
Chen G, McAlister FA, Walker RL, Hemmelgarn BR, Campbell NR. Cardiovascular outcomes in framingham participants with diabetes: the importance of blood pressure. Hypertension. 2011;57:891–7.
Endemann DH, Pu Q, De Ciuceis C, Savoia C, Virdis A, Neves MF, Touyz RM, Schiffrin EL. Persistent remodeling of resistance arteries in type 2 diabetic patients on antihypertensive treatment. Hypertension. 2004;43:399–404.
Colivicchi F, Mettimano M, Genovesi-Ebert A, Schinzari F, Iantorno M, Melina G, Santini M, Cardillo C, Melina D. Differences between diabetic and non-diabetic hypertensive patients with first acute non-ST elevation myocardial infarction and predictors of in-hospital complications. J Cardiovasc Med (Hagerstown). 2008;9:267–72.
Ballo P, Cameli M, Mondillo S, Giacomin E, Lisi M, Padeletti M, Bocelli A, Galderisi M. Impact of diabetes and hypertension on left ventricular longitudinal systolic function. Diabetes Res Clin Pract. 2010;90:209–15.
Inaba Y, Chen JA, Bergmann SR. Prediction of future cardiovascular outcomes by flow-mediated vasodilatation of brachial artery: a meta-analysis. Int J Cardiovasc Imaging. 2010;26:631–40.
Vlachopoulos C, Aznaouridis K, O’Rourke MF, Safar ME, Baou K, Stefanadis C. Prediction of cardiovascular events and all-cause mortality with central haemodynamics: a systematic review and meta-analysis. Eur Heart J. 2010;31:1865–71.
Henry RM, Ferreira I, Kostense PJ, Dekker JM, Nijpels G, Heine RJ, Kamp O, Bouter LM, Stehouwer CD. Type 2 diabetes is associated with impaired endothelium-dependent, flow-mediated dilation, but impaired glucose metabolism is not; the Hoorn study. Atherosclerosis. 2004;174:49–56.
Feldman RD, Gros R. Impaired vasodilator function in hypertension: the role of alterations in receptor-G protein coupling. Trends Cardiovasc Med. 1998;8:297–305.
Brinks HL, Eckhart AD. Regulation of GPCR signaling in hypertension. Biochim Biophys Acta. 2010;1802:1268–75.
Penela P, Murga C, Ribas C, Tutor AS, Peregrín S, Mayor F Jr. Mechanisms of regulation of G protein-coupled receptor kinases (GRKs) and cardiovascular disease. Cardiovasc Res. 2006;69:46–56.
Gros R, Chorazyczewski J, Meek MD, Benovic JL, Ferguson SS, Feldman RD. G-protein-coupled receptor kinase activity in hypertension: increased vascular and lymphocyte G-protein receptor kinase-2 protein expression. Hypertension. 2000;35:38–42.
Taguchi K, Kobayashi T, Takenouchi Y, Matsumoto T, Kamata K. Angiotensin II causes endothelial dysfunction via the GRK2/Akt/eNOS pathway in aortas from a murine type 2 diabetic model. Pharmacol Res. 2011;64:535–46.
Taguchi K, Matsumoto T, Kamata K, Kobayashi T. G protein-coupled receptor kinase 2, with β-arrestin 2, impairs insulin-induced Akt/endothelial nitric oxide synthase signaling in ob/ob mouse aorta. Diabetes. 2012a;61:1978–85.
Liu S, Premont RT, Kontos CD, Zhu S, Rockey DC. A crucial role for GRK2 in regulation of endothelial cell nitric oxide synthase function in portal hypertension. Nat Med. 2005;11:952–8.
Brinks H, Das A, Koch WJ. A role for GRK2 in myocardial ischemic injury: indicators of a potential future therapy and diagnostic. Future Cardiol. 2011;7:547–56.
Dorn GW 2nd. GRK mythology: G-protein receptor kinases in cardiovascular disease. J Mol Med (Berl). 2009;87:455–63.
Penela P, Murga C, Ribas C, Lafarga V, Mayor F Jr.. The complex G protein-coupled receptor kinase 2 (GRK2) interactome unveils new physiopathological targets. Br J Pharmacol. 2010;160:821–32.
Taguchi K, Matsumoto T, Kamata K, Kobayashi T. Inhibitor of G protein-coupled receptor kinase 2 normalizes vascular endothelial function in type 2 diabetic mice by improving β-arrestin 2 translocation and ameliorating Akt/eNOS signal dysfunction. Endocrinology. 2012b;153:2985–96.
Gurevich EV, Tesmer JJ, Mushegian A, Gurevich VV. G protein-coupled receptor kinases: more than just kinases and not only for GPCRs. Pharmacol Ther. 2012;133:40–69.
Rupp H, Maisch B, Brilla CG. Drug withdrawal and rebound hypertension: differential action of the central antihypertensive drugs moxonidine and clonidine. Cardiovasc Drugs Ther. 1996;10:251–62.
Kobayashi T, Taguchi K, Yasuhiro T, Matsumoto T, Kamata K. Impairment of PI3-K/Akt pathway underlies attenuated endothelial function in aorta of type 2 diabetic mouse model. Hypertension. 2004;44:956–62.
Ikeda LS, Harm SC, Arcuri KE, Goldberg AI, Sweet CS. Comparative antihypertensive effects of losartan 50 mg and losartan 50 mg titrated to 100 mg in patients with essential hypertension. Blood Press. 1997;6:35–43.
Naruse T, Ishii R, Tagawa T. Absence of tolerance to hypotensive effects of clonidine in spontaneously hypertensive rats. Jpn J Pharmacol. 1995;67:407–10.
Matsumoto T, Kobayashi T, Kamata K. Mechanisms underlying lysophosphatidylcholine-induced potentiation of vascular contractions in the Otsuka Long-Evans Tokushima Fatty (OLETF) rat aorta. Br J Pharmacol. 2006;149:931–41.
Matsumoto T, Kobayashi T, Ishida K, Taguchi K, Kamata K. Enhancement of mesenteric artery contraction to 5-HT depends on Rho kinase and Src kinase pathways in the ob/ob mouse model of type 2 diabetes. Br J Pharmacol. 2010;160:1092–104.
Ishida K, Matsumoto T, Taguchi K, Kamata K, Kobayashi T. Pravastatin normalizes endothelium-derived contracting factor-mediated response via suppression of Rho-kinase signalling in mesenteric artery from aged type 2 diabetic rat. Acta Physiol (Oxf). 2012;205:255–65.
Ishida K, Matsumoto T, Taguchi K, Kamata K, Kobayashi T. Mechanisms underlying reduced P2Y(1)-receptor-mediated relaxation in superior mesenteric arteries from long-term streptozotocin-induced diabetic rats. Acta Physiol (Oxf). 2013;207:130–41.
Ishida K, Taguchi K, Hida M, Watanabe S, Kawano K, Matsumoto T, Hattori Y, Kobayashi T. Circulating microparticles from diabetic rats impair endothelial function and regulate endothelial protein expression. Acta Physiol (Oxf). 2016;216:211–20.
Harvey AN, Nguyen K, Lymperopoulos A. GRK2 and beta-arrestins in cardiovascular disease: established and emerging possibilities for therapeutic targeting. Curr Mol Pharmacol. 2011
Lymperopoulos A. GRK2 and β-arrestins in cardiovascular disease: something old, something new. Am J Cardiovasc Dis. 2011;1:126–37.
Lucas E, Jurado-Pueyo M, Fortuño MA, Fernández-Veledo S, Vila-Bedmar R, Jiménez-Borreguero LJ, Lazcano JJ, Gao E, Gómez-Ambrosi J, Frühbeck G, Koch WJ, Díez J, Mayor F Jr., Murga C. Downregulation of G protein-coupled receptor kinase 2 levels enhances cardiac insulin sensitivity and switches on cardioprotective gene expression patterns. Biochim Biophys Acta. 2014;1842:2448–56.
Potenza MA, Marasciulo FL, Tarquinio M, Quon MJ, Montagnani M. Treatment of spontaneously hypertensive rats with rosiglitazone and/or enalapril restores balance between vasodilator and vasoconstrictor actions of insulin with simultaneous improvement in hypertension and insulin resistance. Diabetes. 2006;55:3594–603.
Goyal BR, Mesariya P, Goyal RK, Mehta AA. Effect of telmisartan on cardiovascular complications associated with streptozotocin diabetic rats. Mol Cell Biochem. 2008;314:123–31.
Cheang WS, Tian XY, Wong WT, Huang Y. The peroxisome proliferator-activated receptors in cardiovascular diseases: experimental benefits and clinical challenges. Br J Pharmacol. 2015;172:5512–22.
Pontes Andersen CC, Flyvbijerg A, Buschard K, Holmstrup P. Relationship between periodontitis and diabetes: lessons from rodent studies. J Periodontol. 2007;78:1264–75.
Srinivasan K, Ramarao P. Animal models in type 2 diabetes research: an overview. Indian J Med Res. 2007;125:451–72.
Masiello P, Broca C, Gross R, Roye M, Manteghetti M, Hillaire-Buys D, Novelli M, Ribes G. Experimental NIDDM: development of a new model in adult rats administered streptozotocin and nicotinamide. Diabetes. 1998;47:224–9.
Herrlich S, Spieth S, Messner S, Zengerle R. Osmotic micropumps for drug delivery. Adv Drug Deliv Rev. 2012;64:1617–27.
Iino M, Furugori T, Mori T, Moriyama S, Fukuzawa A, Shibano T. Rational design and evaluation of new lead compound structures for selective betaARK1 inhibitors. J Med Chem. 2002;45:2150–9.
Kobayashi T, Taguchi K, Nemoto S, Nogami T, Matsumoto T, Kamata K. Activation of the PDK-1/Akt/eNOS pathway involved in aortic endothelial function differs between hyperinsulinemic and insulin-deficient diabetic rats. Am J Physiol Heart Circ Physiol. 2009;297:H1767–H1775.
Fleming I, Busse R. Molecular mechanisms involved in the regulation of the endothelial nitric oxide synthase. Am J Physiol Regul Integr Comp Physiol. 2003;284:R1–R12.
Mount PF, Kemp BE, Power DA. Regulation of endothelial and myocardial NO synthesis by multi-site eNOS phosphorylation. J Mol Cell Cardiol. 2007;42:271–9.
Coffer PJ, Jin J, Woodgett JR. Protein kinase B (c-Akt): a multifunctional mediator of phosphatidylinositol 3-kinase activation. Biochem J. 1998;335:1–13.
Vanhaesebroeck B, Alessi DR. The PI3K-PDK1 connection: more than just a road to PKB. Biochem J. 2000;346:561–76.
Dimmeler S, Fleming I, Fisslthaler B, Hermann C, Busse R, Zeiher AM. Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature. 1999;399:601–5.
Fleming I, Busse R. Signal transduction of eNOS activation. Cardiovasc Res. 1999;43:532–41.
Daiber A, Steven S, Weber A, Shuvaev VV, Muzykantov VR, Laher I, Li H, Lamas S, Münzel T. Targeting vascular (endothelial) dysfunction. Br J Pharmacol. 2017;174:1591–619.
The present work was supported, in part, by JSPS KAKENHI (Grant Numbers JP15K07975 and JP17K08318). We thank Mari Hida for technical support. We also thank Enago for the English language review.
Conflict of interest
The authors declare that they have no conflict of interest.
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Journal of Endocrinology (2019)
Hypertension Research (2018)