Thalidomide inhibits inflammatory and angiogenic activation of human intestinal microvascular endothelial cells (HIMEC). Am J Physiol Gastrointest Liver Physiol 2010 Feb;298(2):G167-76
Date
11/21/2009Pubmed ID
19926820Pubmed Central ID
PMC2822501DOI
10.1152/ajpgi.00385.2009Scopus ID
2-s2.0-76749086170 (requires institutional sign-in at Scopus site) 32 CitationsAbstract
The glutamic acid derivative thalidomide is a transcriptional inhibitor of TNF-alpha but is also known to affect human blood vessels, which may underlie its teratogenicity. Thalidomide has been used in the treatment of refractory Crohn's disease (CD), but the therapeutic mechanism is not defined. We examined the effect of thalidomide on primary cultures of human intestinal microvascular endothelial cells (HIMEC), the relevant endothelial cell population in inflammatory bowel disease (IBD), to determine its effect on endothelial activation, leukocyte interaction, and VEGF-induced angiogenesis. HIMEC cultures were pretreated with thalidomide before activation with either TNF-alpha/LPS or VEGF. A low-shear-stress flow adhesion assay with either U-937 or whole blood was used to assess HIMEC activation following TNF-alpha/LPS, and a Wright's stain identified adherent leukocytes. Expression of cell adhesion molecules (E-selectin, intercellular adhesion molecule-1, vascular cell adhesion molecule-1) was assessed using radioimmunoassay. Effects of thalidomide on NF-kappaB activation, cyclooxygenase (COX)-2, and inducible nitric oxide synthase (iNOS) expression in TNF-alpha/LPS-activated HIMEC were determined by RT-PCR and Western blotting. Thalidomide blocked adhesion of both U-937 and whole blood leukocytes by 50% in HIMEC, inhibiting binding of all classes of leukocytes. Thalidomide also blocked NF-kappaB and cell adhesion molecule expression in HIMEC. In marked contrast, thalidomide did not affect either iNOS or COX-2 expression, two key molecules that play a role in the downregulation of HIMEC activation. VEGF-induced HIMEC transmigration, growth, proliferation, tube formation, and Akt phosphorylation were significantly inhibited by thalidomide. In summary, thalidomide exerted a potent effect on HIMEC growth and activation, suggesting that it may also function via an endothelial mechanism in the treatment of CD.
Author List
Rafiee P, Stein DJ, Nelson VM, Otterson MF, Shaker R, Binion DGAuthors
Mary F. Otterson MD Professor in the Surgery department at Medical College of WisconsinReza Shaker MD Assoc Provost, Sr Assoc Dean, Ctr Dir, Chief, Prof in the Medicine department at Medical College of Wisconsin
Daniel J. Stein MD Director, Associate Professor in the Medicine department at Medical College of Wisconsin
MESH terms used to index this publication - Major topics in bold
Angiogenesis InhibitorsCell Adhesion
Cell Division
Cell Movement
Cells, Cultured
Crohn Disease
Cyclooxygenase 2
E-Selectin
Endothelial Cells
Humans
Intercellular Adhesion Molecule-1
Intestines
Leukocytes
Lipopolysaccharides
Microvessels
NF-kappa B
Neovascularization, Pathologic
Nitric Oxide Synthase Type II
Phosphorylation
Proto-Oncogene Proteins c-akt
Thalidomide
Tumor Necrosis Factor-alpha
Vascular Cell Adhesion Molecule-1
Vascular Endothelial Growth Factor A