Hyperthermia attenuates TNF-alpha-induced up regulation of endothelial cell adhesion molecules in human arterial endothelial cells.

Abstract

BACKGROUND AND AIM:

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.

CONCLUSION:

Pretreatment of hyperthermia blocks TNF-alpha-induced NF-kappaB activation, resulting in the inhibition of ECAM up regulation in HAEC.

• Authors: Nami Nakabe, Satoshi Kokura, Makoto Shimozawa, Kazuhiro Katada, Naoyuki Sakamoto, Takeshi Ishikawa, Osamu Handa, Tomohisa Takagi, Yuji Naito, Norimasa Yoshida, Toshikazu Yoshikawa
• Journal: International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group (Int J Hyperthermia) Vol. 23 Issue 3 Pg. 217-24 (May 2007) ISSN: 0265-6736 [Print] England
• PMID: 17523016 (Publication Type: Journal Article)

Chemical References

• Diterpenes
• E-Selectin
• HSP70 Heat-Shock Proteins
• I-kappa B Proteins
• NF-kappa B
• NFKBIA protein, human
• Transcription Factor RelA
• Tumor Necrosis Factor-alpha
• Vascular Cell Adhesion Molecule-1
• NF-KappaB Inhibitor alpha
• HMOX1 protein, human
• Heme Oxygenase-1
• geranylgeranylacetone

Topics

• Arteries (cytology, drug effects, metabolism)
• Cells, Cultured
• Diterpenes (pharmacology)
• E-Selectin (metabolism)
• Endothelium, Vascular (cytology, drug effects, metabolism)
• Fever (physiopathology)
• HSP70 Heat-Shock Proteins (metabolism)
• Heme Oxygenase-1 (metabolism)
• Humans
• I-kappa B Proteins (metabolism)
• NF-KappaB Inhibitor alpha
• NF-kappa B (physiology)
• Transcription Factor RelA (metabolism)
• Tumor Necrosis Factor-alpha (physiology)
• Up-Regulation (physiology)
• Vascular Cell Adhesion Molecule-1 (metabolism)