title: Change of concept about the regulation of angiotensin II-induced monocyte chemoattractant protein-1 production in human endothelial cells.
abstract: OBJECTIVE
  Some intriguing clinical observations about the anti-inflammatory effects of angiotensin type 1 (AT1) receptor blockers and angiotensin converting enzyme inhibitors in cardiovascular patients brought us to study the signalling pathways which lead to angiotensin II (ANG)-induced monocyte chemoattractant protein-1 (MCP-1) production in human endothelial cells.
  
  
  METHODS
  MCP-1 production in human umbilical vein endothelial cells (HUVECs) under treatments with ANG, AT1 and angiotensin type 2 (AT2) receptor blockers and pravastatin was measured by ELISA. The expression of AT1 and AT2 receptors and NADPH oxidase catalytic subunits (NOX 1-5) was analysed at mRNA and protein levels. Nuclear factor-kappa B (NF-κB) activation was studied by p65 subunit translocation to the cellular nucleus. Cell viability was tested by the MTT method. Nox4 subcellular distribution was analysed by subcellular protein fractionation and by immunoprecipitation followed by matrix-assisted laser desorption/ionization mass spectrometry analysis.
  
  
  RESULTS
  ANG-induced MCP-1 production was mediated by AT2 receptor, but not AT1 receptor in HUVECs in culture, which in turn activated NF-κB, promoting p65 subunit translocation to the nucleus. Reactive oxygen species produced by NADPH oxidase participated in this activation, mainly by the Nox4 subunit, ubiquitously expressed in all the compartments of HUVECs. Pravastatin inhibited ANG-induced MCP-1 production.
  
  
  CONCLUSIONS
  Our results support that ANG-induced MCP-1 production in HUVECs is mediated by AT2 instead AT1 receptor activation, which in turn activates NF-κB involving reactive oxygen species produced by the NADPH oxidase complex. Statins can also block ANG-induced MCP-1 production, probably by their inhibitory effects on NADPH oxidase activity.
authors.affiliation: Servicio de Cardiología, Complejo Hospitalario Universitario de Santiago de Compostela and Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela 15706, Spain.
authors.forename: M I
authors.initials: MI
authors.lastname: Castiñeiras-Landeira
authors.affiliation: Servicio de Cardiología, Complejo Hospitalario Universitario de Santiago de Compostela and Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela 15706, Spain.
authors.forename: B K
authors.initials: BK
authors.lastname: Rodiño-Janeiro
authors.affiliation: Servicio de Cardiología, Complejo Hospitalario Universitario de Santiago de Compostela and Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela 15706, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Paradela-Dobarro
authors.affiliation: Servicio de Cardiología, Complejo Hospitalario Universitario de Santiago de Compostela and Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela 15706, Spain.
authors.forename: A L
authors.initials: AL
authors.lastname: Batista-Oliveira
authors.affiliation: Servicio de Cardiología, Complejo Hospitalario Universitario de Santiago de Compostela and Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela 15706, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Raposeiras-Roubín
authors.affiliation: Departamento de Enfermería, Universidad de Santiago de Compostela, Santiago de Compostela 15782, Spain.
authors.forename: M
authors.initials: M
authors.lastname: González-Peteiro
authors.affiliation: Servicio de Cardiología, Complejo Hospitalario Universitario de Santiago de Compostela and Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela 15706, Spain; Departamento de Medicina, Universidad de Santiago de Compostela, Santiago de Compostela 15782, Spain.
authors.forename: J R
authors.initials: JR
authors.lastname: González-Juanatey
authors.affiliation: Servicio de Cardiología, Complejo Hospitalario Universitario de Santiago de Compostela and Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela 15706, Spain. Electronic address: ezequiel.alvarez.castro@gmail.com.
authors.forename: E
authors.initials: E
authors.lastname: Álvarez
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology*; D047228:Angiotensin II Type 1 Receptor Blockers / Q000494:pharmacology*; D057912:Angiotensin II Type 2 Receptor Blockers / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D018932:Chemokine CCL2 / Q000096:biosynthesis*; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007150:Immunohistochemistry; D009249:NADP / Q000235:genetics; D021122:Protein Subunits; D044140:Receptor, Angiotensin, Type 1 / Q000235:genetics; D044139:Receptor, Angiotensin, Type 2 / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013481:Superoxides / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism
chemical_list: D047228:Angiotensin II Type 1 Receptor Blockers; D057912:Angiotensin II Type 2 Receptor Blockers; C111835:CCL2 protein, human; D018932:Chemokine CCL2; D021122:Protein Subunits; D044140:Receptor, Angiotensin, Type 1; D044139:Receptor, Angiotensin, Type 2; D051996:Transcription Factor RelA; D013481:Superoxides; D000804:Angiotensin II; D009249:NADP
keywords: Angiotensin II; Angiotensin receptors; Atherogenesis; Human endothelial cells; Monocyte chemoattractant protein-1