MAPK signaling pathway regulates cerebrovascular receptor expression in human cerebral arteries

BMC Neuroscience

Lars Edvinsson¹  Hans Säveland²  Ola Nilsson²  Elisabeth Nilsson¹  Roya Waldsee¹  Sajedeh Eftekhari¹  Saema Ansar¹ 

[1] Department of Clinical Sciences, Division of Experimental Vascular Research, Lund University, Lund, Sweden;Department of Neurosurgery, Lund University Hospital, Lund, Sweden

关键词: Mitogen activated protein kinase;    Thromboxane receptors;    5-hydroxytryptamine receptors;    Angiotensin receptors;    Endothelin receptors;    Human cerebral arteries;   

Others  :  1140526
DOI  :  10.1186/1471-2202-14-12

 received in 2012-06-29, accepted in 2012-12-26,  发布年份 2013

【 摘 要 】

Background

Cerebral ischemia results in enhanced expression of contractile cerebrovascular receptors, such as endothelin type B (ET_B), 5-hydroxytryptamine type 1B (5-HT_1B), angiotensin II type 1 (AT₁) and thromboxane (TP) receptors in the cerebral arteries within the ischemic area. The receptor upregulation occurs via activation of the mitogen-activated protein kinases (MAPK) pathway. Previous studies have shown that inhibitors of the MAPK pathway diminished the ischemic area and contractile cerebrovascular receptors after experimental cerebral ischemia. The aim of this study was to examine if the upregulation of contractile cerebrovascular receptors after 48 h of organ culture of human cerebral arteries involves MAPK pathways and if it can be prevented by a MEK1/2 inhibitor. Human cerebral arteries were obtained from patients undergoing intracranial tumor surgery. The vessels were divided into ring segments and incubated for 48 h in the presence or absence of the specific MEK1/2 inhibitor U0126. The vessels were then examined by using in vitro pharmacological methods and protein immunohistochemistry.

Results

After organ culture of the cerebral arteries the contractile responses to endothelin (ET)-1, angiotensin (Ang) II and thromboxane (TP) were enhanced in comparison with fresh human arteries. However, 5-carboxamidotryptamine (5-CT) induced decreased contractile responses after organ culture as compared to fresh arteries. Incubation with U0126 diminished the maximum contraction elicited by application of ET-1, Ang II and U46619 in human cerebral arteries. In addition, the MEK1/2 inhibitor decreased the contractile response to 5-CT. Immunohistochemistry revealed that organ culture resulted in increased expression of endothelin ET_A, endothelin ET_B angiotensin AT_2, 5-hydroxytryptamine 5-HT_1B and thromboxane A2 receptors, and elevated levels of activated pERK1/2, all localized to the smooth muscle cells of the cerebral arteries. Co-incubation with U0126 normalized these proteins.

Conclusion

The study demonstrated that there is a clear association between human cerebrovascular receptor upregulation via transcription involving activation of the MAPK pathway after organ culture. Inhibition of the MAPK pathways attenuated the vasoconstriction mediated by ET_, AT and TP receptors in human cerebral arteries and the enhanced expression of their receptors. The results indicate that MAPK inhibition might be a novel target for treatment of cerebrovascular disorders.

【 授权许可】

   
2013 Ansar et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]O'Collins VE, Macleod MR, Donnan GA, Horky LL, van der Worp BH, Howells DW: 1,026 experimental treatments in acute stroke. Ann Neurol 2006, 59(3):467-477.
• [2]Edvinsson L: Cerebrovascular endothelin receptor upregulation in cerebral ischemia. Curr Vasc Pharmacol 2009, 7(1):26-33.
• [3]Viossat I, Duverger D, Chapelat M, Pirotzky E, Chabrier PE, Braquet P: Elevated tissue endothelin content during focal cerebral ischemia in the rat. J Cardiovasc Pharmacol 1993, 22(Suppl 8):S306-309.
• [4]Lampl Y, Fleminger G, Gilad R, Galron R, Sarova-Pinhas I, Sokolovsky M: Endothelin in cerebrospinal fluid and plasma of patients in the early stage of ischemic stroke. Stroke 1997, 28(10):1951-1955.
• [5]Zervas NT: Vasospasm: an update. Clin Neurosurg 1979, 26:643-656.
• [6]Zervas NT, Lavyne MH, Negoro M: Neurotransmitters and the normal and ischemic cerebral circulation. N Engl J Med 1975, 293(16):812-816.
• [7]Saavedra JM, Benicky J, Zhou J: Mechanisms of the Anti-Ischemic Effect of Angiotensin II AT(1) Receptor Antagonists in the Brain. Cell Mol Neurobiol 2006, 26(7–8):1099-1111.
• [8]Koudstaal PJ, Ciabattoni G, van Gijn J, Nieuwenhuis HK, de Groot PG, Sixma JJ, Patrono C: Increased thromboxane biosynthesis in patients with acute cerebral ischemia. Stroke 1993, 24(2):219-223.
• [9]Saloheimo P, Juvela S, Riutta A, Pyhtinen J, Hillbom M: Thromboxane and prostacyclin biosynthesis in patients with acute spontaneous intracerebral hemorrhage. Thromb Res 2005, 115(5):367-373.
• [10]Furuta H, Guo DF, Inagami T: Molecular cloning and sequencing of the gene encoding human angiotensin II type 1 receptor. Biochem Biophys Res Commun 1992, 183(1):8-13.
• [11]Masaki T, Vane JR, Vanhoutte PM: International Union of Pharmacology nomenclature of endothelin receptors. Pharmacol Rev 1994, 46(2):137-142.
• [12]Zimmermann M, Seifert V: Endothelin and subarachnoid hemorrhage: an overview. Neurosurgery 1998, 43(4):863-875. discussion 875-866
• [13]Timmermans PB, Chiu AT, Herblin WF, Wong PC, Smith RD: Angiotensin II receptor subtypes. Am J Hypertens 1992, 5(6 Pt 1):406-410.
• [14]Chiu AT, Herblin WF, McCall DE, Ardecky RJ, Carini DJ, Duncia JV, Pease LJ, Wong PC, Wexler RR, Johnson AL: Identification of angiotensin II receptor subtypes. Biochem Biophys Res Commun 1989, 165(1):196-203.
• [15]Coyle AT, Miggin SM, Kinsella BT: Characterization of the 5' untranslated region of alpha and beta isoforms of the human thromboxane A2 receptor (TP). Differential promoter utilization by the TP isoforms. Eur J Biochem 2002, 269(16):4058-4073.
• [16]Beg SA, Hansen-Schwartz JA, Vikman PJ, Xu CB, Edvinsson LI: ERK1/2 inhibition attenuates cerebral blood flow reduction and abolishes ET(B) and 5-HT(1B) receptor upregulation after subarachnoid hemorrhage in rat. J Cereb Blood Flow Metab 2006, 26(6):846-856.
• [17]Beg SS, Hansen-Schwartz JA, Vikman PJ, Xu CB, Edvinsson LI: Protein kinase C inhibition prevents upregulation of vascular ET(B) and 5-HT(1B) receptors and reverses cerebral blood flow reduction after subarachnoid haemorrhage in rats. J Cereb Blood Flow Metab 2007, 27(1):21-32.
• [18]Vikman P, Beg S, Khurana T, Hansen-Schwartz J, Edvinsson L: Gene expression and molecular changes in cerebral arteries following subarachnoid hemorrhage in the rat. J Neurosurg 2006, 105(3):438-444.
• [19]Hansen-Schwartz J, Hoel NL, Zhou M, Xu CB, Svendgaard NA, Edvinsson L: Subarachnoid hemorrhage enhances endothelin receptor expression and function in rat cerebral arteries. Neurosurgery 2003, 52(5):1188-1194. 1194-1185
• [20]Hansen-Schwartz J, Hoel NL, Xu CB, Svendgaard NA, Edvinsson L: Subarachnoid hemorrhage-induced upregulation of the 5-HT1B receptor in cerebral arteries in rats. J Neurosurg 2003, 99(1):115-120.
• [21]Ansar S, Vikman P, Nielsen M, Edvinsson L: Cerebrovascular ETB, 5-HT1B, and AT1 receptor upregulation correlates with reduction in regional CBF after subarachnoid hemorrhage. Am J Physiol Heart Circ Physiol 2007, 293(6):H3750-3758.
• [22]Stenman E, Edvinsson L: Cerebral ischemia enhances vascular angiotensin AT1 receptor-mediated contraction in rats. Stroke 2004, 35(4):970-974.
• [23]Stenman E, Malmsjo M, Uddman E, Gido G, Wieloch T, Edvinsson L: Cerebral ischemia upregulates vascular endothelin ET(B) receptors in rat. Stroke 2002, 33(9):2311-2316.
• [24]Ansar S, Larsen C, Maddahi A, Edvinsson L: Subarachnoid hemorrhage induces enhanced expression of thromboxane A2 receptors in rat cerebral arteries. Brain Res 2010, 1316:163-172.
• [25]Ansar S, Edvinsson L: Subtype activation and interaction of protein kinase C and mitogen-activated protein kinase controlling receptor expression in cerebral arteries and microvessels after subarachnoid hemorrhage. Stroke 2008, 39(1):185-190.
• [26]Namura S, Iihara K, Takami S, Nagata I, Kikuchi H, Matsushita K, Moskowitz MA, Bonventre JV, Alessandrini A: Intravenous administration of MEK inhibitor U0126 affords brain protection against forebrain ischemia and focal cerebral ischemia. Proc Natl Acad Sci USA 2001, 98(20):11569-11574.
• [27]Henriksson M, Stenman E, Vikman P, Edvinsson L: MEK1/2 inhibition attenuates vascular ETA and ETB receptor alterations after cerebral ischaemia. Exp Brain Res 2007, 178(4):470-476.
• [28]Irving EA, Bamford M: Role of mitogen- and stress-activated kinases in ischemic injury. J Cereb Blood Flow Metab 2002, 22(6):631-647.
• [29]Sandhu H, Ansar S, Edvinsson L: Comparison of MEK/ERK pathway inhibitors on the upregulation of vascular G-protein coupled receptors in rat cerebral arteries. Eur J Pharmacol 2010, 644(1–3):128-137.
• [30]Ansar S, Maddahi A, Edvinsson L: Inhibition of cerebrovascular raf activation attenuates cerebral blood flow and prevents upregulation of contractile receptors after subarachnoid hemorrhage. BMC Neurosci 2011, 12:107. BioMed Central Full Text
• [31]Adner M, Cantera L, Ehlert F, Nilsson L, Edvinsson L: Plasticity of contractile endothelin-B receptors in human arteries after organ culture. Br J Pharmacol 1996, 119(6):1159-1166.
• [32]Henriksson M, Xu CB, Edvinsson L: Importance of ERK1/2 in upregulation of endothelin type B receptors in cerebral arteries. Br J Pharmacol 2004, 142(7):1155-1161.
• [33]Mulvany MJ, Halpern W: Contractile properties of small arterial resistance vessels in spontaneously hypertensive and normotensive rats. Circ Res 1977, 41(1):19-26.
• [34]Hogestatt ED, Andersson KE, Edvinsson L: Mechanical properties of rat cerebral arteries as studied by a sensitive device for recording of mechanical activity in isolated small blood vessels. Acta Physiol Scand 1983, 117(1):49-61.
• [35]Hansen-Schwartz J, Nordstrom CH, Edvinsson L: Human endothelin subtype A receptor enhancement during tissue culture via de novo transcription. Neurosurgery 2002, 50(1):127-133. discussion 133-125
• [36]Edvinsson LI, Povlsen GK: Vascular plasticity in cerebrovascular disorders. J Cereb Blood Flow Metab 2011, 31(7):1554-1571.
• [37]Povlsen GK, Waldsee R, Ahnstedt H, Kristiansen KA, Johansen FF, Edvinsson L: In vivo experimental stroke and in vitro organ culture induce similar changes in vasoconstrictor receptors and intracellular calcium handling in rat cerebral arteries. Exp Brain Res 2012, 219(4):507-520.
• [38]Vikman P, Edvinsson L: Gene expression profiling in the human middle cerebral artery after cerebral ischemia. Eur J Neurol 2006, 13(12):1324-1332.
• [39]Johansson S, Povlsen GK, Edvinsson L: Expressional changes in cerebrovascular receptors after experimental transient forebrain ischemia. PLoS One 2012, 7(7):e41852.
• [40]Prehn JH, Welsch M, Backhauss C, Nuglisch J, Ausmeier F, Karkoutly C, Krieglstein J: Effects of serotonergic drugs in experimental brain ischemia: evidence for a protective role of serotonin in cerebral ischemia. Brain Res 1993, 630(1–2):10-20.
• [41]Guilbert F, Lainee P, Dubreuil B, McCort G, O'Connor SE, Janiak P, Herbert JM: Serotonin aggravates exercise-induced cardiac ischemia in the dog: effect of serotonin receptor antagonists. Eur J Pharmacol 2004, 497(1):55-63.
• [42]Maddahi A, Edvinsson L: Enhanced expressions of microvascular smooth muscle receptors after focal cerebral ischemia occur via the MAPK MEK/ERK pathway. BMC Neurosci 2008, 9:85. BioMed Central Full Text
• [43]Zubkov AY, Nanda A, Zhang JH: Signal transduction pathways in cerebral vasospasm. Pathophysiology 2003, 9(2):47-61.
• [44]Vajkoczy P, Meyer B, Weidauer S, Raabe A, Thome C, Ringel F, Breu V, Schmiedek P: Clazosentan (AXV-034343), a selective endothelin A receptor antagonist, in the prevention of cerebral vasospasm following severe aneurysmal subarachnoid hemorrhage: results of a randomized, double-blind, placebo-controlled, multicenter phase IIa study. J Neurosurg 2005, 103(1):9-17.