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Age-related alterations in the level and expression pattern of Von Willebrand Factor (VWF)

  • Author / Creator
    Abdualla, Radya Y
  • Von Willebrand Factor (VWF) is a multimeric adhesive glycoprotein that is exclusively expressed in endothelial cells (EC) and megakaryocytes. This procoagulant protein is involved in maintaining primary hemostasis and thrombus formation. However, there are pathological and physiological conditions that alter the circulating levels and pattern of VWF expression in vasculature. Unregulated increase in VWF levels may contribute to elevate the incidence of thrombosis. Since increased thrombogenicity is observed with aging, we explored whether aging is associated with alterations in the level and/or pattern of VWF expression. This study compared circulating VWF levels in the blood of young and aged mice and rats, using ELISA. Additionally, immunofluorescence confocal microscopy, Western blot analyses, and RT-PCR analyses were used to determine VWF expression pattern, cellular protein and mRNA levels. The expression pattern of the VWF protein was determined in combination with the endothelial marker CD31 and micro vessels marker (Isolectin-GS-IB4), in various organs of young and aged mice. Furthermore, to explore the functional consequences of altered VWF expression with regard to platelet aggregation, which is a major contributing factor to thrombus formation, CD41 a marker of activated platelets was used for immunofluorescence confocal microscopy. The results of these analyses demonstrated that circulating VWF protein levels were higher in blood of aged rats and mice compared to young. Also with age VWF expression at mRNA levels were significantly increased in livers, brains, and lungs, but not in hearts and kidneys. Consistent with the mRNA analyses, the cellular protein levels were determined in lungs and livers and shown to be significantly increased in aged compared to young mice. Moreover, the endothelial staining for VWF was observed significantly in microvessels of brains, lungs, and livers of aged but not young mice, demonstrating a phenotypic shift of microvascular endothelial cells of these organs with aging towards a procoagulant state with de novo expression of VWF. Additionally, activated platelet aggregates and occluded vessels were significantly increased in livers, brains, and lungs, but not in hearts and kidneys of aged mice compared to young, consistently with the mRNA and protein levels, and redistribution patterns of VWF expression. These results demonstrate that with aging, VWF levels are increased in circulation, and vasculature of distinct organs exhibit an altered VWF expression pattern. Specifically, in liver, lung, and brain, but not heart and kidney, with aging increased VWF levels and de novo expression in microvascular endothelial cells are observed. Furthermore, the increased VWF levels specifically in these organs were concomitant with detection of significantly increased platelet aggregate formation and occluded vessels. Overexpression of VWF in circulation and microvasculature vessels of distinct organs as a result of aging may contribute to vascular diseases such as thrombosis.

  • Subjects / Keywords
  • Graduation date
    2017-06:Spring 2017
  • Type of Item
    Thesis
  • Degree
    Master of Science
  • DOI
    https://doi.org/10.7939/R3TB0Z71Z
  • License
    This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.
  • Language
    English
  • Institution
    University of Alberta
  • Degree level
    Master's
  • Department
    • Department of Medicine
  • Supervisor / co-supervisor and their department(s)
    • Dr. Nadia, Jahroudi (Department of Medicine)
  • Examining committee members and their departments
    • Dr. Gina, Rayat (Department of Surgery).
    • Dr. Fakhreddin, Jamali (Faculty of Pharmacy)
    • Dr.Stephane, Bourque (Department of Pharmacology)