title: Increased pulmonary RhoA expression in the nitrofen-induced congenital diaphragmatic hernia rat model.
abstract: OBJECTIVE
  Persistent pulmonary hypertension remains a major cause of mortality and morbidity in cases of congenital diaphragmatic hernia (CDH). Recently, RhoA/Rho-kinase-mediated vasoconstriction has been reported to be important in the pathogenesis of pulmonary hypertension (PH). Several recent reports have described that fasudil, a potent Rho-kinase inhibitor and vasodilator, could represent a potential therapeutic option for PH. We designed this study to investigate the hypothesis that the expression level of RhoA is increased in the nitrofen-induced CDH rat model. The expression level of Wnt11, an activator of RhoA, was also evaluated.
  
  
  METHODS
  Pregnant rats were treated with or without nitrofen on gestational day 9 (D9). Fetuses were sacrificed on D17, D19 and D21 and were divided into control and CDH groups. Quantitative real-time polymerase chain reaction was performed to determine the pulmonary gene expression levels of both Wnt11 and RhoA. An immunofluorescence study was also performed to evaluate the expression and localization of RhoA.
  
  
  RESULTS
  The relative mRNA expression levels of pulmonary Wnt11 and RhoA on D21 were significantly increased in the CDH group compared with the control group (p=0.016 and p=0.008, respectively). The immunofluorescence study confirmed the overexpression of RhoA in the pulmonary vessels of CDH rats on D21.
  
  
  CONCLUSIONS
  Our results provide evidence that the RhoA/Rho-kinase-mediated pathway is involved in the pathogenesis of PH in the nitrofen-induced CDH rat model. Our data also suggest that the fasudil, a Rho-kinase inhibitor, could represent a therapeutic option for the treatment of PH in CDH.
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan. Electronic address: hajimeta@md.tsukuba.ac.jp.
authors.forename: Hajime
authors.initials: H
authors.lastname: Takayasu
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Kouji
authors.initials: K
authors.lastname: Masumoto
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Koki
authors.initials: K
authors.lastname: Hagiwara
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Takato
authors.initials: T
authors.lastname: Sasaki
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Ono
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Jimbo
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Uesugi
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Chikashi
authors.initials: C
authors.lastname: Gotoh
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Urita
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Toko
authors.initials: T
authors.lastname: Shinkai
authors.affiliation: Department of Pediatric Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1, Tennoudai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Tanaka
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D065630:Hernias, Diaphragmatic, Congenital / Q000139:chemically induced / Q000235:genetics* / Q000378:metabolism; D008168:Lung / Q000378:metabolism*; D010647:Phenyl Ethers / Q000633:toxicity; D011247:Pregnancy; D011270:Pregnancy, Animal; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D020742:rhoA GTP-Binding Protein / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D010647:Phenyl Ethers; D012333:RNA, Messenger; D020742:rhoA GTP-Binding Protein; C007350:nitrofen
keywords: Congenital diaphragmatic hernia; Nitrofen; Pulmonary hypertension; RhoA/Rho-kinase; Vasoconstrictor