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. 2007 May;23(3):217-24.
doi: 10.1080/02656730601143295.

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

Nami Nakabe  1 , Satoshi Kokura, Makoto Shimozawa, Kazuhiro Katada, Naoyuki Sakamoto, Takeshi Ishikawa, Osamu Handa, Tomohisa Takagi, Yuji Naito, Norimasa Yoshida, Toshikazu Yoshikawa
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Free article

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

Nami Nakabe et al. Int J Hyperthermia. 2007 May.
Free article

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.

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