title: The PI3K-Akt-eNOS pathway is involved in aortic hyporeactivity to Phenylephrine associated with late pregnancy in spontaneously hypertensive rats.
abstract: OBJECTIVE
  This study aimed to evaluate the effects of Wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt), on aortic hyporeactivity to Phenylephrine (Phe) and nitric oxide bioavailability associated with pregnancy in hypertensive rats.
  
  
  METHODS
  The intact aortic rings of pregnant and non-pregnant Wistar or spontaneously hypertensive rats (SHRs) were stimulated with Phe (1nmol/L to 10mmol/L) before and after incubation with Wortmannin (10nmol/L, 30min). Western blot experiments analyzed the expression of phosphorylated PI3K [p85-PI3K], Akt [p-Akt (Ser 473)] and eNOS [p-eNOS (Ser 1177)] in aorta homogenates of pregnant and non-pregnant Wistar rats or SHRs. The effect of Wortmannin (10nmol/L) on the cytosolic concentrations of nitric oxide (NO; measured using 4,5-diaminofluorescein diacetate [DAF-2DA], 10mmol/L), Ca(2+) (using Fluo 3-AM, 5μmol/L) and reactive oxygen species (ROS; using dihydroethidium [DHE], 2.5mmol/L) were measured fluorimetrically in freshly isolated endothelial cells.
  
  
  RESULTS
  Wortmannin increases the reactivity of the aorta to Phe and decreases NO concentrations in the aortic endothelial cells of pregnant Wistar rats and SHR.
  
  
  CONCLUSIONS
  The PI3/AKT/endothelial nitric oxide synthase (eNOS) pathway contributes to aortic hyporeactivity to Phenylephrine associated with pregnancy in normo- and hypertensive rats.
authors.affiliation: Programa de Pós Graduação Multicêntrico em Ciências Fisiológicas, SBFis, Brazil; Department of Basic Sciences, School of Dentistry of Araçatuba, UNESP - Univ Estadual Paulista, Araçatuba, São Paulo, Brazil.
authors.forename: Dalise
authors.initials: D
authors.lastname: Zancheta
authors.affiliation: Programa de Pós Graduação Multicêntrico em Ciências Fisiológicas, SBFis, Brazil; Department of Basic Sciences, School of Dentistry of Araçatuba, UNESP - Univ Estadual Paulista, Araçatuba, São Paulo, Brazil.
authors.forename: Jéssica A
authors.initials: JA
authors.lastname: Troiano
authors.affiliation: Programa de Pós Graduação Multicêntrico em Ciências Fisiológicas, SBFis, Brazil; Department of Basic Sciences, School of Dentistry of Araçatuba, UNESP - Univ Estadual Paulista, Araçatuba, São Paulo, Brazil.
authors.forename: Simone R
authors.initials: SR
authors.lastname: Potje
authors.affiliation: Department of Basic Sciences, School of Dentistry of Araçatuba, UNESP - Univ Estadual Paulista, Araçatuba, São Paulo, Brazil; School of Pharmacy of Araçatuba, UNIP - Univ Paulista, Araçatuba, São Paulo, Brazil.
authors.forename: Priscila
authors.initials: P
authors.lastname: Cavalari
authors.affiliation: Programa de Pós Graduação Multicêntrico em Ciências Fisiológicas, SBFis, Brazil; Department of Basic Sciences, School of Dentistry of Araçatuba, UNESP - Univ Estadual Paulista, Araçatuba, São Paulo, Brazil.
authors.forename: Doris H
authors.initials: DH
authors.lastname: Sumida
authors.affiliation: Programa de Pós Graduação Multicêntrico em Ciências Fisiológicas, SBFis, Brazil; Department of Basic Sciences, School of Dentistry of Araçatuba, UNESP - Univ Estadual Paulista, Araçatuba, São Paulo, Brazil. Electronic address: crisant@foa.unesp.br.
authors.forename: Cristina
authors.initials: C
authors.lastname: Antoniali
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology*; D015153:Blotting, Western; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D010656:Phenylephrine / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011247:Pregnancy; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D051381:Rats; D011918:Rats, Inbred SHR; D017208:Rats, Wistar; D014662:Vasoconstrictor Agents / Q000494:pharmacology*; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D047428:Protein Kinase Inhibitors; D014662:Vasoconstrictor Agents; D010656:Phenylephrine; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D000077191:Wortmannin
keywords: Aorta; Nitric oxide; Phenylephrine; Pregnancy; Spontaneously hypertensive rats; Wortmannin