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Use of Precision‐Cut Liver Slices To Evaluate The Protective Effects Of Betaine In LPS‐Induced Liver Injury
Author(s) -
Kharbanda Kusum,
Cannella John J,
Ward Brian W,
Todero Sandra L,
Tuma Dean J.
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.981.1
Subject(s) - betaine , cytokine , intracellular , liver injury , chemistry , downregulation and upregulation , tumor necrosis factor alpha , viability assay , pharmacology , apoptosis , kupffer cell , caspase 3 , programmed cell death , biochemistry , biology , immunology , gene
LPS plays an important role in the pathogenesis of alcoholic liver injury via its effect on Kupffer cell‐mediated inflammatory responses. Consequently, released cytokines can activate specific intracellular pathways in hepatocytes that influence cell fate in different manners, e.g., proapoptotic signals via the caspase cascade, but also survival pathways. We have begun to utilize the precision cut liver slice model as a tool to dissect these interactions as well as to evaluate the potential protective effects of therapeutic agents. Here, chow‐fed rat liver slices were prepared in accordance with the manufacturer's instructions (Vitron Inc, Tucson, AZ) and exposed to LPS in the presence and absence of 5mM betaine. Slices were harvested after 24h for hepatocellular caspase‐3 activity measurements. Media was analyzed for LDH leakage and cytokine release (TNF and IL6). The slices exposed to media alone exhibited low caspase‐3 activity and retained viability as judged by minimal leakage of LDH. Following exposure to LPS, an induction of caspase‐3 and an increase in cytokine release (p<0.05) was observed. Betaine treatment significantly attenuated LPS‐induced caspase‐3 activation (p<0.05), but had no effect on cytokine levels. This data suggests that the protective effects of betaine on LPS are down‐stream of TNF and IL6 and may occur via upregulation of survival signals in hepatocytes.