Regulation and distribution of MAdCAM-1 in endothelial cells in vitro.
Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is a 60-kDa endothelial cell adhesion glycoprotein that regulates lymphocyte trafficking to Peyer's patches and lymph nodes. Although it is widely agreed that MAdCAM-1 induction is involved in chronic gut inflammation, few studies have investigated regulation of MAdCAM-1 expression. We used two endothelial lines [bEND.3 (brain) and SVEC (high endothelium)] to study the signal paths that regulate MAdCAM-1 expression in response to tumor necrosis factor (TNF)-alpha using RT-PCR, blotting, adhesion, and immunofluorescence. TNF-alpha induced both MAdCAM-1 mRNA and protein in a dose- and time-dependent manner. This induction was tyrosine kinase (TK), p42/44, p38 mitogen-activated protein kinase (MAPK), and nuclear factor (NF)-kappa B/poly-ADP ribose polymerase (PARP) dependent. Because MAdCAM-1 is regulated via MAPKs, we examined mitogen/extracellular signal-regulated kinase (MEK)-1/2 activation in SVEC. We found that MEK-1/2 is activated by TNF-alpha within minutes and is dependent on TK and p42/44 MAPKs. Similarly, TNF-alpha activated NF-kappa B through TK, p42/44, p38 MAPKs, and PARP pathways in SVEC cells. MAdCAM-1 was also shown to be frequently distributed to endothelial junctions both in vitro and in vivo. Cytokines like TNF-alpha stimulate MAdCAM-1 in high endothelium via TK, p38, p42/22 MAPKs, and NF-kappa B/PARP. MAdCAM-1 expression requires NF-kappa B translocation through both direct p42/44 and indirect p38 MAPK pathways in high endothelial cells.
Oshima
T
T
Department of Molecular and Cellular Physiology, Louisianna State University Health Sciences Center, Shreveport, Louisiana, 71130-3932, USA.
Pavlick
K P
KP
Laroux
F S
FS
Verma
S K
SK
Jordan
P
P
Grisham
M B
MB
Williams
L
L
Alexander
J S
JS
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D053119:Benzophenanthridines; D001578:Benzopyrans / Q000494:pharmacology; D002227:Carbazoles; D015815:Cell Adhesion Molecules; D002461:Cell Line, Transformed; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004727:Endothelium / Q000737:chemistry / Q000166:cytology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D019833:Genistein / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D007136:Immunoglobulins / Q000032:analysis* / Q000235:genetics*; D066298:In Vitro Techniques; D007211:Indoles; D015212:Inflammatory Bowel Diseases / Q000503:physiopathology; D007413:Intestinal Mucosa / Q000737:chemistry* / Q000166:cytology*; D007546:Isoquinolines / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009088:Mucoproteins / Q000032:analysis* / Q000235:genetics*; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D010617:Phenanthridines / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000470:Alkaloids; D053119:Benzophenanthridines; D001578:Benzopyrans; D002227:Carbazoles; D015815:Cell Adhesion Molecules; D015853:Cysteine Proteinase Inhibitors; D004791:Enzyme Inhibitors; D005419:Flavonoids; C417259:GPI 6150; D007093:Imidazoles; D007136:Immunoglobulins; D007211:Indoles; D007546:Isoquinolines; D007976:Leupeptins; C489082:Madcam1 protein, mouse; D009088:Mucoproteins; D016328:NF-kappa B; D010617:Phenanthridines; D011725:Pyridines; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; C073601:KT 5823; D019833:Genistein; C016299:chelerythrine; C090942:4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one