Differential Cell Sensitivity between OTA and LPS upon Releasing TNF-α

Toxins

Lauy Al-Anati²  Ebtisam Essid²  Ulla Stenius¹  Knut Beuerlein³  Klaus Schuh² 

[1] Institute of Environmental Medicine, Karolinska Institutet, S-17177 Stockholm, SwedenInstitute of Pharmacology and Toxicology, College of Veterinary Medicine, Justus Liebig University Giessen. Frankfurter Street 107, D-35392 Giessen, Germany;;Rudolf-Buchheim-Institute of Pharmacology, College of Medicine, Justus Liebig University Giessen, Frankfurter Street 107, D-35392 Giessen, Germany

关键词: ochratoxin A;    lipopolysaccharide;    tumor necrosis factor α;    Kupffer cells;    macrophages;    rat liver sinusoidal endothelial cells;    HepG2 cells;    rat hepatocytes;   

DOI  :  10.3390/toxins2061279

来源: mdpi

【 摘 要 】

The release of tumor necrosis factor α (TNF-α) by ochratoxin A (OTA) was studied in various macrophage and non-macrophage cell lines and compared with E. coli lipopolysaccharide (LPS) as a standard TNF-α release agent. Cells were exposed either to 0, 2.5 or 12.5 µmol/L OTA, or to 0.1 µg/mL LPS, for up to 24 h. OTA at 2.5 µmol/L and LPS at 0.1 µg/mL were not toxic to the tested cells as indicated by viability markers. TNF-α was detected in the incubated cell medium of rat Kupffer cells, peritoneal rat macrophages, and the mouse monocyte macrophage cell line J774A.1: TNF-α concentrations were 1,000 pg/mL, 1,560 pg/mL, and 650 pg/mL, respectively, for 2.5 µmol/L OTA exposure and 3,000 pg/mL, 2,600 pg/mL, and 2,115 pg/mL, respectively, for LPS exposure. Rat liver sinusoidal endothelial cells, rat hepatocytes, human HepG2 cells, and mouse L929 cells lacked any cytokine response to OTA, but showed a significant release of TNF-α after LPS exposure, with the exception of HepG2 cells. In non-responsive cell lines, OTA lacked both any activation of NF-κB or the translocation of activated NF-κB to the cell nucleus, i.e., in mouse L929 cells. In J774A.1 cells, OTA mediated TNF-α release via the pRaf/MEK 1/2-NF-κB and p38-NF-κB pathways, whereas LPS used pRaf/MEK 1/2–NF-κB, but not p38-NF-κB pathways. In contrast, in L929 cells, LPS used other pathways to activate NF-κB. Our data indicate that only macrophages and macrophage derived cells respond to OTA and are considered as sources for TNF-α release upon OTA exposure.

【 授权许可】

CC BY   
© 2010 by the authors; licensee MDPI, Basel, Switzerland