Gliotoxin causes apoptosis and necrosis of rat Kupffer cells in vitro and in vivo in the absence of oxidative stress: exacerbation by caspase and serine protease inhibition.
OBJECTIVE
A potential application of gliotoxin therapy for liver fibrosis was suggested by its apoptotic effect on fibrogenic activated stellate cells. We investigated if gliotoxin exerts similar effects on hepatic macrophage Kupffer cells.


METHODS
Effects of gliotoxin on Kupffer cells isolated from the normal liver and in vivo following its administration to CCl(4)-induced cirrhotic rats were studied.


RESULTS
Gliotoxin caused apoptosis of cultured Kupffer cells, the effect being apparent at 0.3 microM concentration within 1h; longer incubation caused necrosis. This effect was associated with mitochondrial cytochrome c release, caspase-3 activation and ATP depletion. Interestingly, inhibition of caspase-3 and serine proteases accelerated and augmented gliotoxin-induced cell death via necrosis. Gliotoxin stimulated nuclear translocation of NFkappaB, and phosphorylation of p38, ERK1/2 and JNK MAP kinases, but these signaling molecules were not involved in gliotoxin-induced death of Kupffer cells. In vivo administration of gliotoxin to cirrhotic rats caused apoptosis of Kupffer cells, stellate cells and hepatocytes. In control rats, the effect was minimal on the nonparenchymal cells and not apparent on hepatocytes.


CONCLUSIONS
In the fibrotic liver, gliotoxin nonspecifically causes death of hepatic cell types. Modification of gliotoxin molecule may be necessary for selective targeting and elimination of activated stellate cells.
Anselmi
Kristin
K
Thomas E. Starzl Transplantation Institute, Department of Surgery, and VA medical Center, University of Pittsburgh, E-1518 BST, 200 Lothrop Street, Pittsburgh, PA 15213, USA.
Stolz
Donna B
DB
Nalesnik
Michael
M
Watkins
Simon C
SC
Kamath
Ravindra
R
Gandhi
Chandrashekhar R
CR
D000818:Animals; D017209:Apoptosis; D002251:Carbon Tetrachloride / Q000633:toxicity; D053148:Caspase 3 / Q000502:physiology*; D061945:Caspase Inhibitors; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism; D005912:Gliotoxin / Q000633:toxicity*; D022781:Hepatocytes / Q000187:drug effects* / Q000473:pathology; D007728:Kupffer Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology*; D008106:Liver Cirrhosis, Experimental / Q000139:chemically induced / Q000473:pathology; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009336:Necrosis; D018384:Oxidative Stress; D010766:Phosphorylation / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D012697:Serine Endopeptidases / Q000187:drug effects / Q000502:physiology*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology
D061945:Caspase Inhibitors; D016328:NF-kappa B; D015842:Serine Proteinase Inhibitors; D005912:Gliotoxin; D045304:Cytochromes c; D002251:Carbon Tetrachloride; D020929:Mitogen-Activated Protein Kinase Kinases; D012697:Serine Endopeptidases; D053148:Caspase 3