Document Detail


Protective effects of ezrin on cold storage preservation injury in the pig kidney proximal tubular epithelial cell line (LLC-PK1).
MedLine Citation:
PMID:  19461485     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Renal damage caused by cold preservation and warm reperfusion has been well documented and involves tissue edema, cell swelling, ATP depletion, calcium toxicity, and oxidative stress. However, more common proximal mechanisms have not been identified, which may limit the development of effective clinical treatment strategies. Previous work indicates that many cytoskeletal structures are affected by cold preservation and reperfusion, including membrane-rich ezrin-associated complexes. The aim of this study was to investigate whether the sublamellar cytoskeletal protein ezrin is causally involved in cold preservation injury in renal tubule epithelial cells. METHODS: We created a stably transfected cell line (LLC-EZ) using the pig kidney proximal tubular epithelial cell line (LLC-PK1), which constitutively overexpresses wild-type ezrin. These cells were cold stored in University of Wisconsin Solution and reperfused in vitro to model renal tubule preservation injury, which was assessed by biochemical, metabolic, functional, and structural endpoints. RESULTS: Overexpression of ezrin increased cell viability (lactate dehydrogenase release), mitochondrial activity (ATP synthesis, dehydrogenase activity, and inner mitochondrial membrane potential), and protected the structure of cell membrane microvilli and mitochondria after cold storage preservation injury. Reperfusion-induced apoptosis was also significantly reduced in LLC-EZ cells overexpressing ezrin. CONCLUSIONS: Enhanced ezrin expression protects tubule epithelial cells from cold storage preservation injury, possibly by membrane or mitochondrial mechanisms.
Authors:
Tao Tian; Susanne L Lindell; Scott C Henderson; Martin J Mangino
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Transplantation     Volume:  87     ISSN:  1534-6080     ISO Abbreviation:  Transplantation     Publication Date:  2009 May 
Date Detail:
Created Date:  2009-05-22     Completed Date:  2009-07-08     Revised Date:  2010-09-27    
Medline Journal Info:
Nlm Unique ID:  0132144     Medline TA:  Transplantation     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1488-96     Citation Subset:  IM    
Affiliation:
Department of Surgery, Virginia Commonwealth University, Medical College of Virginia Campus, Richmond, VA 23298, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Culture Techniques / methods
Cell Membrane / physiology,  ultrastructure
Cold Temperature
Cytoskeletal Proteins / genetics,  therapeutic use*
DNA, Complementary / genetics
Epithelial Cells / cytology*
Kidney
Kidney Tubules, Proximal / cytology*
Mice
Mitochondria / physiology,  ultrastructure
Reperfusion Injury / prevention & control*
Swine
Tissue Preservation / methods*
Transfection
Grant Support
ID/Acronym/Agency:
5P30NS04746/NS/NINDS NIH HHS; DK63309/DK/NIDDK NIH HHS; P30 CA016059-239011/CA/NCI NIH HHS; P30 CA16059/CA/NCI NIH HHS; R44 DK066654-03/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Cytoskeletal Proteins; 0/DNA, Complementary; 0/ezrin
Comments/Corrections

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