Hyperthermia attenuates TNF-alpha-induced up regulation of endothelial cell adhesion molecules in human arterial endothelial cells.
OBJECTIVE
The activation of NF-kappaB induces production of inflammatory cytokines and up regulation of endothelial cell adhesion molecules (ECAM). ECAM (e.g., E-selectin, VCAM-1 and ICAM-1) associates to the recruitment of leukocytes into tissue exposed to inflammatory situation. In this study, we investigated the effects of hyperthermia on the activation of NF-kappaB and the up regulation of E-selectin and VCAM-1 in human endothelial cells stimulated by TNF-alpha.


METHODS
Human arterial endothelial cells (HAEC) were pretreated with hyperthermia for 60 min at 42 degrees C, followed by incubation at 37 degrees C in a passively cooled incubator, before TNF-alpha stimulation. To assess the effects of hyperthermia on TNF-alpha-induced up regulation of ECAM and TNF-alpha-induced activation of NF-kappaB, we measured ECAM by ELISA, and evaluated the activation of NF-kappaB by Western blotting after TNF-alpha stimulation. The accumulation of HO-1, Hsp70 and IkappaBalpha in hyperthermia-treated HAEC was also assessed by Western blotting. To investigate the role of Hsp70, we treated HAEC with geranylgeranylacetone (GGA, Hsp70 inducer) 2 h before hyperthermia, and then measured ECAM in TNF-alpha-stimulated HAEC by ELISA.


RESULTS
Pretreatment of hyperthermia reduced TNF-alpha-induced up regulation of E-selectin and VCAM-1. In addition, accumulation of Hsp70, HO-1 and IkappaBalpha protein were up-regulated after hyperthermia. Furthermore, Western blotting analysis revealed that pretreatment of hyperthermia attenuated TNF-alpha-induced translocation of p65 into the nuclei of HAEC. Moreover, GGA enhanced Hsp70 accumulation induced by hyperthermia. Hyperthermia pretreatment combined with GGA induced further inhibition of TNF-alpha-induced up regulation of ECAM when compared with hyperthermia alone.


CONCLUSIONS
Pretreatment of hyperthermia blocks TNF-alpha-induced NF-kappaB activation, resulting in the inhibition of ECAM up regulation in HAEC.
Nakabe
Nami
N
Inflammation and Immunology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Kokura
Satoshi
S
Shimozawa
Makoto
M
Katada
Kazuhiro
K
Sakamoto
Naoyuki
N
Ishikawa
Takeshi
T
Handa
Osamu
O
Takagi
Tomohisa
T
Naito
Yuji
Y
Yoshida
Norimasa
N
Yoshikawa
Toshikazu
T
D001158:Arteries / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D004224:Diterpenes / Q000494:pharmacology; D019040:E-Selectin / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005334:Fever / Q000503:physiopathology*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D051547:Heme Oxygenase-1 / Q000378:metabolism; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000502:physiology*; D051996:Transcription Factor RelA / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*; D015854:Up-Regulation / Q000502:physiology; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism*
D004224:Diterpenes; D019040:E-Selectin; D018840:HSP70 Heat-Shock Proteins; D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D000072000:NF-KappaB Inhibitor alpha; C496541:HMOX1 protein, human; D051547:Heme Oxygenase-1; C031049:geranylgeranylacetone