Activation of caspases in intestinal villus epithelial cells of normal and nematode infected rats.
BACKGROUND
Small intestinal epithelial cells (IEC) show apoptosis in physiological turnover of cells and in certain inflammatory diseases.


OBJECTIVE
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.


CONCLUSIONS
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.
Hyoh
Y
Y
Department of Medical Zoology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Ishizaka
S
S
Horii
T
T
Fujiwara
A
A
Tegoshi
T
T
Yamada
M
M
Arizono
N
N
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D015153:Blotting, Western; D001973:Bromodeoxyuridine / Q000378:metabolism; D020169:Caspases / Q000502:physiology*; D002478:Cells, Cultured; D004587:Electrophoresis, Agar Gel; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000201:enzymology*; D015200:Fluoroimmunoassay / Q000379:methods; D007421:Intestine, Small / Q000166:cytology / Q000201:enzymology*; D008297:Male; D008856:Microscopy, Fluorescence; D009559:Nippostrongylus; D051381:Rats; D011914:Rats, Inbred BN; D017206:Strongylida Infections / Q000201:enzymology*
D020169:Caspases; D001973:Bromodeoxyuridine