Colchicine protects mice from the lethal effect of an agonistic anti-Fas antibody
Guoping Feng, Neil Kaplowitz
Guoping Feng, Neil Kaplowitz
Published February 1, 2000
Citation Information: J Clin Invest. 2000;105(3):329-339. https://doi.org/10.1172/JCI7398.
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Article

Colchicine protects mice from the lethal effect of an agonistic anti-Fas antibody

Abstract

The aim of this study was to determine whether colchicine, which has been reported to protect against various hepatotoxic insults, influences the susceptibility of mice to the agonistic anti-Fas antibody, Jo2. All mice that were pretreated with colchicine (2 mg/kg) survived the lethal challenge of intraperitoneal administration of 10 μg of Jo2, whereas all control mice pretreated with γ-lumicolchicine succumbed to the challenge. Twelve micrograms of Jo2 killed less than half of colchicine-pretreated mice and its lethal effects were delayed relative to control mice, which all died within 8 hours. Other microtubule-disrupting agents such as Taxol, vinblastine, and nocodazole also improved the survival of mice treated with the lethal dose of Jo2. Histologic examination showed that colchicine protected against Jo2-induced fulminant liver injury, and TUNEL assay demonstrated that colchicine protected against massive apoptosis of hepatocytes. Hepatocytes isolated from colchicine-pretreated mice exhibited decreased susceptibility to Jo2-induced apoptosis. In addition, colchicine pretreatment reduced surface expression of Fas and decreased Jo2- and TNF-α–induced apoptosis of cultured hepatocytes in the presence of actinomycin D, but did not affect the susceptibility of cultured sinusoidal endothelial cells to Jo2-induced apoptosis. Remarkably, Fas and TNF receptor-1 mRNA and intracellular protein levels increased after colchicine treatment, indicating that colchicine protects against death ligand–induced apoptosis in the liver by decreasing death-receptor targeting to the cell surface.

Authors

Guoping Feng, Neil Kaplowitz

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Figure 6

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(a) FACS® analysis of surface Fas density of hepatocytes. Hepatocytes we...
(a) FACS® analysis of surface Fas density of hepatocytes. Hepatocytes were isolated 24 hours after mice were pretreated in vivo with colchicine or γ-lumicolchicine. Hepatocytes were stained with 2 mg/mL PE-labeled Jo2 antibody and were assessed for their surface Fas density by FACS® analysis as described in Methods. The solid line represents colchicine-pretreated mice; the dotted line represents γ-lumicolchicine-pretreated mice. (b) Confocal microscopic analysis of Fas distribution. Hepatocytes were isolated 24 hours after mice were pretreated in vivo with colchicine or γ-lumicolchicine. Hepatocytes were stained using PE-labeled Jo2 antibody as described in Methods. At left, hepatocytes from γ-lumicolchicine-pretreated mice show prominent plasma membrane distribution of Fas. At right, hepatocytes from colchicine-pretreated mice show redistribution of Fas to inside the cells, with decreased plasma membrane staining. (c) Effect of colchicine on Fas and TNFR-1 protein of LPM fraction. LPM preparation and Western analysis were performed as described in Methods. For LPM preparation, LPM recovery and enrichment were estimated by marker enzymes; similar results were obtained for colchicine-pretreated samples and γ-lumicolchicine-pretreated samples. Results for 2 independent experiments are shown. The density of the bands was estimated by PhosphorImager (Molecular Dynamics, Sunnyvale, California, USA) using the ImageQuant program, revealing that as a result of colchicine pretreatment, both Fas and TNFR-1 doubled in homogenates and decreased by 50% in LPM.