title: Regulatory mechanism of endothelin receptor B in the cerebral arteries after focal cerebral ischemia.
abstract: OBJECTIVE
  Increased expression of endothelin receptor type B (ETBR), a vasoactive receptor, has recently been implied in the reduced cerebral blood flow and exacerbated neuronal damage after ischemia-reperfusion (I/R). The study explores the regulatory mechanisms of ETBR to identify drug targets to restore normal cerebral artery contractile function as part of successful neuroprotective therapy.
  
  
  METHODS
  We have employed in vitro methods on human and rat cerebral arteries to study the regulatory mechanisms and the efficacy of target selective inhibitor, Mithramycin A (MitA), to block the ETBR mediated contractile properties. Later, middle cerebral artery occluded (MCAO) rats were used to substantiate the observations. Quantative PCR, immunohistochemistry, western blot and wire myograph methods were employed to study the expression and contractile properties of cerebral arteries.
  
  
  RESULTS
  Increased expression of specificity protein (Sp1) was observed in human and rat cerebral arteries after organ culture, strongly correlating with the ETBR upregulation. Similar observations were made in MCAO rats. Treatment with MitA, a Sp1 specific inhibitor, significantly downregulated the ETBR mRNA and protein levels. It also significantly reduced the ETBR mediated cerebrovascular contractility. Detailed analysis indicated that ERK1/2 mediated phosphorylation of Sp1 might be essential for ETBR transcription.
  
  
  CONCLUSIONS
  Transcription factor Sp1 regulates the ETBR mediated vasoconstriction in focal cerebral ischemia via MEK-ERK signaling, which is also conserved in humans. The results show that MitA can effectively be used to block ETBR mediated vasoconstriction as a supplement to an existing ischemic stroke therapy.
authors.affiliation: Department of Clinical Experimental Research, Glostrup research institute, University of Copenhagen, Glostrup, Denmark.
authors.forename: Anne-Sofie
authors.initials: AS
authors.lastname: Grell
authors.affiliation: Division of Experimental Vascular Research, Department of Clinical Sciences, Solvegatan 17, Lund University, Lund, Sweden.
authors.forename: Rushani
authors.initials: R
authors.lastname: Thigarajah
authors.affiliation: Department of Clinical Experimental Research, Glostrup research institute, University of Copenhagen, Glostrup, Denmark; Division of Experimental Vascular Research, Department of Clinical Sciences, Solvegatan 17, Lund University, Lund, Sweden.
authors.forename: Lars
authors.initials: L
authors.lastname: Edvinsson
authors.affiliation: Department of Clinical Experimental Research, Glostrup research institute, University of Copenhagen, Glostrup, Denmark.
authors.forename: Ajoy Kumar
authors.initials: AK
authors.lastname: Samraj
mesh_terms: D000818:Animals; D002545:Brain Ischemia / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D002536:Cerebral Arteries / Q000187:drug effects / Q000473:pathology; D002560:Cerebrovascular Circulation / Q000187:drug effects* / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D009924:Organ Culture Techniques; D008926:Plicamycin / Q000008:administration & dosage / Q000031:analogs & derivatives; D051381:Rats; D044023:Receptor, Endothelin B; D017466:Receptors, Endothelin / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics; D016329:Sp1 Transcription Factor / Q000037:antagonists & inhibitors; D014661:Vasoconstriction / Q000187:drug effects*
chemical_list: C000588596:EDNRB protein, human; D044023:Receptor, Endothelin B; D017466:Receptors, Endothelin; D016329:Sp1 Transcription Factor; C066851:mithramycin A; D008926:Plicamycin