Current evidence indicates that the very first step in the formation of cholesterol gallstone is nucleation of cholesterol from an aqueous phase (bile) into a solid crystalline phase (cholesterol monohydrate crystals). It is now realized that cholesterol, lipid with extremely poor water solubility, is solubilized in bile in two different forms - micelles of bile salts, phospholipids and cholesterol, and vesicles of phospholipids and cholesterol. Moreover, vesicular cholesterol is the component of cholesterol which is responsible for nucleation. This finding will significantly modify thinking about both the definition of """"""""cholesterol supersaturation"""""""" as well as the mechanism of biliary cholesterol secretion and solubilization. The factors controlling the dynamic interchange between the two different forms of cholesterol solubilization and the factors leading to the nucleation and precipitation of cholesterol remain unresolved. The present experiments are designed to address these issues. Prairie dogs fed cholesterol rapidly form cholesterol gallstones. This model is recognized to be, and is accepted as, an excellent model of experimental cholelithiasis. Using this model, cholesterol in bile will be separated into the micellar and vesicular forms by column chromatography and further characterized by quasi-elastic light scattering spectroscopy (QLS) to determine the size distribution of micellar and vesicular cholesterol. The quantity of biliary cholesterol and the proportion of micellar vesicular cholesterol will also be determined as a function of dietary cholesterol loading and hepatic bile salt secretory rate. Because of the new insight offered by the vesicular mode of cholesterol transport, the phase boundary and equilibria of bile salt-phospholipid-cholesterol systems will have to be extended beyond the mixed micellar tri-ordinates. Model bile systems will be used to define the phase boundaries of micelles and vesicles. QLS and small angle neutron scattering (SANS) will be used to characterize mixed micelles, vesicles, and to document the kinetics of interchange between them. Gallstones are a common problem; the affliction involves approximately 10% of men and 20% of women in the age group 55-64. The magnitude of the problem will only increase as the population ages. The results of the proposed study will yield new and important information that will lead to a better understanding of the information of gallstones, and also have an important impact on its treatment and prevention.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK041678-03
Application #
3242510
Study Section
General Medicine A Subcommittee 2 (GMA)
Project Start
1990-08-10
Project End
1993-07-31
Budget Start
1992-08-01
Budget End
1993-07-31
Support Year
3
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Washington
Department
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
Ko, Cynthia W; Schulte, Scott J; Lee, Sum P (2005) Biliary sludge is formed by modification of hepatic bile by the gallbladder mucosa. Clin Gastroenterol Hepatol 3:672-8
Haigh, W G; Lee, S P (2001) Identification of oxysterols in human bile and pigment gallstones. Gastroenterology 121:118-23
Wrenn, S P; Gudheti, M; Veleva, A N et al. (2001) Characterization of model bile using fluorescence energy transfer from dehydroergosterol to dansylated lecithin. J Lipid Res 42:923-34
Ko, C W; Sekijima, J H; Lee, S P (1999) Biliary sludge. Ann Intern Med 130:301-11
Wrenn, S P; Kaler, E W; Lee, S P (1999) A fluorescence energy transfer study of lecithin-cholesterol vesicles in the presence of phospholipase C. J Lipid Res 40:1483-94
Ko, C W; Lee, S P (1999) Gallstone formation. Local factors. Gastroenterol Clin North Am 28:99-115
Ko, C W; Kowdley, K V; Haigh, W G et al. (1998) Biliary lipid composition after liver transplantation: effect of allograft function and cyclosporine. Liver Transpl Surg 4:258-64
Klinkspoor, J H; Yoshida, T; Lee, S P (1998) Bile salts stimulate mucin secretion by cultured dog gallbladder epithelial cells independent of their detergent effect. Biochem J 332 ( Pt 1):257-62
Luk, A S; Kaler, E W; Lee, S P (1998) Protein lipid interaction in bile: effects of biliary proteins on the stability of cholesterol-lecithin vesicles. Biochim Biophys Acta 1390:282-92
Luk, A S; Kaler, E W; Lee, S P (1997) Structural mechanisms of bile salt-induced growth of small unilamellar cholesterol-lecithin vesicles. Biochemistry 36:5633-44

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