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1 Departments of Medicine and Surgery, Rhode Island Hospital and Brown University School of Medicine, Providence, Rhode Island 02903; and 2 Digestive Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892
Human gallbladders with cholesterol stones
exhibit impaired muscle contraction induced by agonists that act on
transmembrane receptors, increased membrane cholesterol content, and
abnormal cholesterol-to-phospholipid ratio compared with those with
pigment stones. The present study was designed to investigate the
functions of the CCK receptor of gallbladder muscle membranes by
radioreceptor assay and cross-linking.
125I-labeled CCK-8 binding was
time-dependent, competitive, and specific. Scatchard analysis showed
that the maximum specific binding
(Bmax) was significantly
decreased in cholesterol compared with pigment stone gallbladders (0.18 ± 0.07 vs. 0.38 ± 0.05 pmol/mg protein, P < 0.05). In contrast, the affinity
for CCK was higher in cholesterol than pigment stone gallbladders (0.18 ± 0.06 vs. 1.2 ± 0.23 nM). Similar results were observed in
binding studies with the CCK-A receptor antagonist
[3H]L-364,718.
Cross-linking and saturation binding studies also showed significantly
less CCK binding in gallbladders with cholesterol stones. These
abnormalities were reversible after incubation with cholesterol-free
liposomes. The Bmax increased
(P < 0.01) and the dissociation
constant decreased (P < 0.001) after
incubation with cholesterol-free liposomes. In conclusion, human
gallbladders with cholesterol stones have impaired CCK receptor binding
compared with those with pigment stones. These changes are reversed by removal of the excess membrane cholesterol. These receptor alterations may contribute to the defective contractility of the gallbladder muscle
in patients with cholesterol stones.
smooth muscle; cholecystokinin receptors; liposomes
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