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. 2006 Nov 13;12(1):25–38. doi: 10.2478/s11658-006-0052-0

Induction of heme oxygenase-1 and heat shock protein 70 in rat hepatocytes: The role of calcium signaling

Malte Silomon 1, Inge Bauer 1,✉, Michael Bauer 1, Julia Nolting 1, Markus Paxian 1, Hauke Rensing 1
PMCID: PMC6275859  PMID: 17103089

Abstract

Stress response genes including heat shock proteins are induced under a variety of conditions to confer cellular protection. This study investigated the role of calcium signaling in the induction of two stress response genes, heme oxygenase-1/hsp32 and hsp70, in isolated rat hepatocytes. Both genes were induced by cellular glutathione depletion. This induction could be inhibited by BAPTA-AM. Culturing in a calcium-free medium prevented the induction of hsp70 gene expression after glutathione depletion without affecting heme oxygenase-1 gene expression. Thapsigargin increased the gene expression of heme oxygenase-1 but not that of hsp70. Thapsigargin-induced heme oxygenase-1 induction was completely inhibited by BAPTA-AM. Incubation with the Ca2+-ionophore A23187 augmented heme oxygenase-1 (two-fold) and hsp70 (5.2-fold) mRNA levels. Our data suggests a significant role of Ca2+-dependent pathways in the induction of the two stress genes. An increase in the cytoplasmic Ca2+ activity seems to play a key role in the cascade of signaling leading to the induction of the two genes. However, the source of Ca2+ that fluxes into the cytoplasm seems to be different. Our data provides evidence for a compartmentalization of calcium fluxes, i.e. the Ca2+ flux from intracellular stores (e.g. the endoplasmic reticulum) plays a major role in the induction of heme oxygenase-1. By contrast, Ca2+ flux from the extracellular medium seems to be a mechanism initiating the cellular signaling cascade leading to hsp70 gene induction.

Key words: Stress response genes, Hepatocytes, Calcium, Gene expression

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Abbreviations used

AP-1

activator protein-1

BAPTA-AM

(1,2-bis(o-amino-phenoxy)ethane-N,N,N′,N′-tetra (acetoxymethyl)esther

DMSO

dimethylsulfoxide

EDTA

ethylene-diamine-tetraacetic-acid

ER

endoplasmic reticulum

GSH

glutathione

HO-1

heme oxygenase-1

hsp

heat shock protein

KHB

Krebs-Henseleit buffer

PBS

phosphate buffered saline

P/BSO

2,6 dimethyl-2,5 heptadien-4-on (phorone)/buthionine sulfoximine

Tg

thapsigargin

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