Alterations induced on cytoskeleton by Escherichia coli endotoxin in different types of rat liver cell cultures.
Endotoxins (lipopolysaccharide, LPS) from Gram-negative bacteria are considered as the agents responsible for the induction of endotoxic shock, producing severe cellular metabolic dishomeostasis. Cytotoxic lesions, as well as functional and metabolic disturbances, occur mainly in the liver, which is one of the target organs and exerts an LPS clearance function. In an attempt to approach the molecular basis of endotoxic shock, and to propose an experimental model, we have focused this study on cytoskeleton (microtubules and microfilaments) alterations induced by different doses of endotoxin in different target liver cells. Microfilaments and microtubules were studied by immunofluorescence and different microscopy techniques (optic fluorescence microscopy and confocal laser scanning microscopy) in order to improve the cytoskeleton study resolution. Parenchymal and sinusoidal cell morphology, severely damaged by the LPS action, is related to a disturbance on the cytoskeletal organisation, even more evident in a particular proliferating rat liver cell culture. The most relevant changes are seen in the microtubule patterns in all liver cells tested, which could be related, depending on cell type, either to a direct LPS action or to [Ca+2]i dishomeostasis as well as free radical and cytokine (IL-1beta and TNF-alpha) production. Due to their features, proliferating rat liver cell cultures are used as a sensitive cell model to understand the effect of LPS on cytoskeleton organisation.
Pagani
R
R
Department of Biochemistry and Molecular Biology I, Universidad Complutense, Madrid, Spain. rpaganib@quim.ucm.es
Portolés
M T
MT
De La Viña
S
S
Melzner
I
I
Vergani
G
G
D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism*; D004731:Endotoxins / Q000494:pharmacology*; D004926:Escherichia coli / Q000378:metabolism*; D007150:Immunohistochemistry; D008070:Lipopolysaccharides / Q000378:metabolism / Q000494:pharmacology*; D008099:Liver / Q000187:drug effects; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D051381:Rats; D013997:Time Factors
D004731:Endotoxins; D008070:Lipopolysaccharides