Attenuating effects of heat shock against TGF-beta1-induced apoptosis in cultured rat hepatocytes.
Heat shock proteins (HSPs) induction confers protection against diverse forms of cellular injury. However, the mechanism by which HSPs exert cytoprotective effects remains unclear. Treatment of rat hepatocyte with transforming growth factor-beta1 (TGF-beta1) induces growth arrest followed by extensive cell death by apoptosis. In this study, the effects of preexposure to heat on TGF-beta1-induced apoptosis of cultured hepatocytes were examined. Treatment of hepatocytes for 24 h with TGF-beta1 resulted in significant apoptotic cell death, as demonstrated by DNA fragmentation, caspase activation, and hypodiploid DNA peak. Moreover, TGF-beta1-induced cell death was accompanied by an enhanced generation of reactive oxygen species and a loss of the mitochondrial membrane potential. These effects were attenuated when the hepatocytes were subjected to 43 degrees C for 20 min prior to the cytokine stimulation. The enhancement in HSP70 expression at mRNA and protein levels induced by heat preexposure was accompanied by an increase in mRNA levels of intracellular antioxidant enzymes. Heat treatment also prevented TGF-beta1-induced activation of nuclear factor kappa B (NF-kappaB) by preventing the degradation of the inhibitory protein kappa Balpha (IkappaBalpha). In conclusion, these data indicate that in the mechanism by which a mild heat pretreatment increases the resistance of hepatocytes to TGF-beta1-induced apoptotic cell death, the upregulation of catalase expression and a decrease in ROS generation are involved.
Instituto de Bioqui;mica (Centro Mixto CSIC-UCM), Facultad de Farmacia, Madrid, Spain.
Carmen
C
Díez-Fernández
David
D
Andrés
María
M
Cascales
D000818:Animals; D017209:Apoptosis; D015152:Blotting, Northern; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002374:Catalase / Q000378:metabolism; D002478:Cells, Cultured; D053938:DNA Fragmentation; D004587:Electrophoresis, Agar Gel; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology*; D006358:Hot Temperature; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D015854:Up-Regulation
D017382:Reactive Oxygen Species; D016212:Transforming Growth Factor beta; D002374:Catalase; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases