title: Murine prolylcarboxypeptidase depletion induces vascular dysfunction with hypertension and faster arterial thrombosis.
abstract: Prolylcarboxypeptidase (PRCP) activates prekallikrein to plasma kallikrein, leading to bradykinin liberation, and degrades angiotensin II. We now identify PRCP as a regulator of blood vessel homeostasis. β-Galactosidase staining in PRCP(gt/gt) mice reveals expression in kidney and vasculature. Invasive telemetric monitorings show that PRCP(gt/gt) mice have significantly elevated blood pressure. PRCP(gt/gt) mice demonstrate shorter carotid artery occlusion times in 2 models, and their plasmas have increased thrombin generation times. Pharmacologic inhibition of PRCP with Z-Pro-Prolinal or plasma kallikrein with soybean trypsin inhibitor, Pro-Phe-Arg-chloromethylketone or PKSI 527 also shortens carotid artery occlusion times. Aortic and renal tissues have uncoupled eNOS and increased reactive oxygen species (ROS) in PRCP(gt/gt) mice as detected by dihydroethidium or Amplex Red fluorescence or lucigenin luminescence. The importance of ROS is evidenced by the fact that treatment of PRCP(gt/gt) mice with antioxidants (mitoTEMPO, apocynin, Tempol) abrogates the hypertensive, prothrombotic phenotype. Mechanistically, our studies reveal that PRCP(gt/gt) aortas express reduced levels of Kruppel-like factors 2 and 4, thrombomodulin, and eNOS mRNA, suggesting endothelial cell dysfunction. Further, PRCP siRNA treatment of endothelial cells shows increased ROS and uncoupled eNOS and decreased protein C activation because of thrombomodulin inactivation. Collectively, our studies identify PRCP as a novel regulator of vascular ROS and homeostasis.
authors.affiliation: Department of Medicine, Hematology and Oncology Division, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106-7284, USA.
authors.forename: Gregory N
authors.initials: GN
authors.lastname: Adams
authors.forename: Gretchen A
authors.initials: GA
authors.lastname: LaRusch
authors.forename: Evi
authors.initials: E
authors.lastname: Stavrou
authors.forename: Yihua
authors.initials: Y
authors.lastname: Zhou
authors.forename: Marvin T
authors.initials: MT
authors.lastname: Nieman
authors.forename: Gretta H
authors.initials: GH
authors.lastname: Jacobs
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Cui
authors.forename: Yuan
authors.initials: Y
authors.lastname: Lu
authors.forename: Mukesh K
authors.initials: MK
authors.lastname: Jain
authors.forename: Fakhri
authors.initials: F
authors.lastname: Mahdi
authors.forename: Zia
authors.initials: Z
authors.lastname: Shariat-Madar
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Okada
authors.forename: Louis G
authors.initials: LG
authors.lastname: D'Alecy
authors.forename: Alvin H
authors.initials: AH
authors.lastname: Schmaier
mesh_terms: D000818:Animals; D001808:Blood Vessels / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D002268:Carboxypeptidases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D002341:Carotid Artery Thrombosis / Q000150:complications / Q000235:genetics*; D002478:Cells, Cultured; D055785:Gene Knockdown Techniques; D006801:Humans; D006973:Hypertension / Q000150:complications / Q000235:genetics* / Q000503:physiopathology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D034622:RNA Interference / Q000502:physiology*; D034741:RNA, Small Interfering / Q000494:pharmacology; D013918:Thrombin Time; D013997:Time Factors; D014652:Vascular Diseases / Q000150:complications / Q000235:genetics* / Q000503:physiopathology
chemical_list: D034741:RNA, Small Interfering; D002268:Carboxypeptidases; C022448:lysosomal Pro-X carboxypeptidase