title: Upregulation of S1P1 and Rac1 receptors in the pulmonary vasculature of nitrofen-induced congenital diaphragmatic hernia.
abstract: OBJECTIVE
  Sphingolipids play a crucial role in pulmonary development. The sphingosine kinase 1 (SphK1) modulates the synthesis of sphingolipid sphingosine-1-phosphate (S1P). S1P regulates cell proliferation and angiogenesis via different receptors, S1P1, S1P2 and S1P3, which all influence the expression of Ras-related C3 botulinum toxin substrate 1 (Rac1). We designed this study to test the hypothesis that the S1P/Rac1 pathway is altered in the nitrofen-induced CDH model.
  
  
  METHODS
  Pregnant rats received nitrofen or vehicle on D9. On D21, fetuses were killed and divided into nitrofen and control group (n = 12). QRT-PCR, western blotting and confocal-immunofluorescence microscopy were performed to reveal pulmonary gene and protein expression levels of SphK1, S1P1, S1P2, S1P3 and Rac1.
  
  
  RESULTS
  Pulmonary gene expression of S1P1 and Rac1 was significantly increased in the CDH group compared to controls, whereas S1P2 and S1P3 expression was decreased. These results were confirmed by western blotting and confocal microscopy. SphK1 expression was not found to be altered.
  
  
  CONCLUSIONS
  The increased expression of S1P1 and Rac1 in the pulmonary vasculature of nitrofen-induced CDH lungs suggests that S1P1 and Rac1 are important mediators of PH in this model.
authors.affiliation: National Children's Research Centre, Our Lady's Children's Hospital, Crumlin, Dublin, Ireland.
authors.forename: Julia
authors.initials: J
authors.lastname: Zimmer
authors.affiliation: National Children's Research Centre, Our Lady's Children's Hospital, Crumlin, Dublin, Ireland.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Takahashi
authors.affiliation: National Children's Research Centre, Our Lady's Children's Hospital, Crumlin, Dublin, Ireland.|School of Medicine and Medical Science and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Dublin, Ireland.
authors.forename: Johannes W
authors.initials: JW
authors.lastname: Duess
authors.affiliation: National Children's Research Centre, Our Lady's Children's Hospital, Crumlin, Dublin, Ireland.|Department of Pediatric Surgery, Hannover Medical School, Hannover, Germany.
authors.forename: Alejandro D
authors.initials: AD
authors.lastname: Hofmann
authors.affiliation: National Children's Research Centre, Our Lady's Children's Hospital, Crumlin, Dublin, Ireland. prem.puri@ucd.ie.|School of Medicine and Medical Science and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Dublin, Ireland. prem.puri@ucd.ie.
authors.forename: Prem
authors.initials: P
authors.lastname: Puri
mesh_terms: D000818:Animals; D015153:Blotting, Western; D004195:Disease Models, Animal; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D065630:Hernias, Diaphragmatic, Congenital / Q000235:genetics*; D008168:Lung / Q000098:blood supply*; D010647:Phenyl Ethers; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D049349:Receptors, Lysosphingolipid / Q000235:genetics*; D000081025:Sphingosine-1-Phosphate Receptors; D015854:Up-Regulation / Q000235:genetics*; D020830:rac1 GTP-Binding Protein / Q000235:genetics*
chemical_list: D010647:Phenyl Ethers; D049349:Receptors, Lysosphingolipid; D000081025:Sphingosine-1-Phosphate Receptors; C570053:sphingosine-1-phosphate receptor-2, rat; C499134:Rac1 protein, rat; D020830:rac1 GTP-Binding Protein; C007350:nitrofen
keywords: Congenital diaphragmatic hernia; Nitrofen; Pulmonary hypertension; Rac1; Sphingosine kinase; Sphingosine-1-phosphate