Identification of thrombospondin 1 (TSP-1) as a novel mediator of cell injury in kidney ischemia

J Clin Invest. 2005 Dec;115(12):3451-9. doi: 10.1172/JCI25461. Epub 2005 Nov 17.

Abstract

Thrombospondin 1 (TSP-1) is a matricellular protein that inhibits angiogenesis and causes apoptosis in vivo and in vitro in several cancerous cells and tissues. Here we identify TSP-1 as the molecule with the highest induction level at 3 hours of IR injury in rat and mouse kidneys subjected to ischemia/reperfusion (IR) injury using the DNA microarray approach. Northern hybridizations demonstrated that TSP-1 expression was undetectable at baseline, induced at 3 and 12 hours, and returned to baseline levels at 48 hours of reperfusion. Immunocytochemical staining identified the injured proximal tubules as the predominant sites of expression of TSP-1 in IR injury and showed colocalization of TSP-1 with activated caspase-3. Addition of purified TSP-1 to normal kidney proximal tubule cells or cells subjected to ATP depletion in vitro induced injury as demonstrated by cytochrome c immunocytochemical staining and caspase-3 activity. The deleterious role of TSP-1 in ischemic kidney injury was demonstrated directly in TSP-1 null mice, which showed significant protection against IR injury-induced renal failure and tubular damage. We propose that TSP-1 is a novel regulator of ischemic damage in the kidney and may play an important role in the pathophysiology of ischemic kidney failure.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adenosine Triphosphate / chemistry
  • Animals
  • Binding Sites
  • Blotting, Northern
  • Blotting, Western
  • CD36 Antigens / biosynthesis
  • Caspase 3
  • Caspases / metabolism
  • Colorimetry
  • Cytochromes c / metabolism
  • DNA, Complementary / metabolism
  • Enzyme Activation
  • Gene Deletion
  • Immunoblotting
  • Immunohistochemistry
  • In Situ Nick-End Labeling
  • Ischemia / pathology*
  • Kidney / metabolism
  • Kidney / pathology
  • Kidney Tubules / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Nucleic Acid Hybridization
  • Oligonucleotide Array Sequence Analysis
  • RNA / chemistry
  • RNA / metabolism
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Reperfusion Injury / pathology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Thrombospondin 1 / metabolism
  • Thrombospondin 1 / physiology*
  • Time Factors

Substances

  • CD36 Antigens
  • DNA, Complementary
  • RNA, Messenger
  • Thrombospondin 1
  • RNA
  • Adenosine Triphosphate
  • Cytochromes c
  • Casp3 protein, mouse
  • Casp3 protein, rat
  • Caspase 3
  • Caspases