Activation of caspases in intestinal villus epithelial cells of normal and nematode infected rats.

Abstract:

BACKGROUND:Small intestinal epithelial cells (IEC) show apoptosis in physiological turnover of cells and in certain inflammatory diseases. AIMS:To investigate the role of caspases in the progression of IEC apoptosis in vivo. METHODS:IEC were separated along the villus-crypt axis from the jejunum of normal and Nippostrongylus brasiliensis infected rats at 4 degrees C. Caspases were examined by a fluorometric assay method, histochemistry, and immunoblotting. RESULTS:Villus cell rich IEC from normal rats exhibited a high level of caspase-3-like activity whereas activities of caspase-1, -8, and -9 were negligible. Immunoblotting analysis of villus cell rich IEC revealed partial cleavage of procaspase-3 into a 17 kDa molecule as well as cleavage of a caspase-3 substrate, poly(ADP-ribose) polymerase (PARP), whereas in crypt cell rich IEC, caspase-3 cleavage was less significant. Caspase-3 activity was also observed histochemically in villus epithelium on frozen sections of the normal small intestine. IEC prepared at 4 degrees C did not reveal nuclear degradation whereas subsequent incubation in a suspension at 37 degrees C induced intense nuclear degradation within one hour in accordance with increases in active caspase-3. This apoptosis was partially suppressed by the caspase inhibitor Z-VAD-fmk. Nematode infected animals showed villus atrophy together with significant increases in levels of caspase-3 in IEC but not of caspase-1, -8, or -9. CONCLUSION:Caspase-3 may have an important role in the physiological replacement of IEC as well as in progression of IEC apoptosis induced by nematode infection.

journal_name

Gut

journal_title

Gut

authors

Hyoh Y,Ishizaka S,Horii T,Fujiwara A,Tegoshi T,Yamada M,Arizono N

doi

10.1136/gut.50.1.71

keywords:

subject

Has Abstract

pub_date

2002-01-01 00:00:00

pages

71-7

issue

1

eissn

0017-5749

issn

1468-3288

journal_volume

50

pub_type

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