Studies on the Pathogenesis of Pigment Gallstones in Hemolytic Anemia

Trotman, Bruce W.; Bernstein, Seldon E.; Bove, Kevin E.; Wirt, G.

doi:10.1172/jci109793

Cited by 60 publications

(23 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The reported mean values for UCB when using TLC range from 14 to 33 pmol/l in control biles and from 26 to 181 pmol/l in pigment lithogenic biles (25,29,30). In contrast, our values range from 1 to 7pmol/l in controls and from 1 to 38 pmol/l in gallstone patients.…”

Section: Discussioncontrasting

confidence: 66%

High-Performance Liquid Chromatography of Bilirubin Conjugates in Bile: Effect ofβ-Glucuronidase on the Bile Pigments

Skar

Saxerholt²

1989

Scandinavian Journal of Gastroenterology

View full text Add to dashboard Cite

A simple, specific, and technically easy high-performance liquid chromatography (HPLC) method for the separation and quantification of unconjugated bilirubin, bilirubin monoglucoside-monoglucuronide, bilirubin diglucuronide, and bilirubin monoglucuronide has been developed. The method was used to determine the bilirubin compounds of bile obtained endoscopically from the common bile duct in 43 patients with gallstone disease and in 6 subjects without gallstones or liver disease. The bile samples were also assessed for the presence of beta-glucuronidase-producing bacteria. The amount of unconjugated bilirubin was significantly higher (p less than 0.01) in bile containing bacteria producing beta-glucuronidase than in bile without such bacterial strains. In six 'normal' bile samples the following quantities of bilirubin conjugates and unconjugated bilirubin were found (median and range): bilirubin monoglucoside-monoglucuronide (mixed conjugate), 61 (27-80) mumol/l; bilirubin diglucuronide, 632 (512-861) mumol/l; bilirubin monoglucuronide, 113 (70-175) mumol/l; and unconjugated bilirubin, 3 (1-7) mumol/l. These results are in good agreement with those obtained with other HPLC methods. The concentrations of unconjugated bilirubin were lower than those found when using conventional diazo methods and thin-layer chromatography. HPLC proved to be a useful tool in gallstone pathogenesis studies. Our results support bacterial glucuronidase as a possible pathogenic factor in pigment gallstone disease.

show abstract

Section: Discussioncontrasting

confidence: 66%

High-Performance Liquid Chromatography of Bilirubin Conjugates in Bile: Effect ofβ-Glucuronidase on the Bile Pigments

Skar

Saxerholt²

1989

Scandinavian Journal of Gastroenterology

View full text Add to dashboard Cite

show abstract

“…As we previously reported, the nb/nb mouse with hereditary hemolytic anemia spontaneously forms black pigment gallstones (1). By infrared spectroscopy the composition of mouse hemolysis-induced black pigment gallstones was similar to those from humans with hemolysis and both were nearly identical to synthesized calcium bilirubinate (1).…”

Section: Fig 3 Distribution Of Intact Conjugated and Unconjugated Tsupporting

confidence: 64%

“…In order to study the pathogenesis of black pigment gallstones we have used the nblnb mouse with hereditary hemolytic anemia as an animal model of hemolysisinduced gallstone disease (1). These animals spontaneously form black pigment gallstones that are indistinguishable from those of humans with hemolysis (1).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Monoconjugated bilirubin is a major component of hemolysis-induced gallstones in mice

1988

Self Cite

View full text Add to dashboard Cite

The role of bilirubin conjugates in the formation of pigment gallstones is not known. In this study, we completely solubilized and then analyzed by high-performance liquid chromatography specimens of black pigment gallstones from eight nb/nb mice with hereditary hemolytic anemia. Each dried gallstone specimen of about 200 micrograms was dissolved in 5 ml of dimethyl sulfoxide/0.15 M HCI/50 mM disodium-EDTA (8:1:1 by volume) at room temperature. Stone dissolution was complete by 30 min as monitored by the A456 and direct observation, and no oxidative products of bilirubin were observed in the visible spectrum, 350 to 750 nm. By high-performance liquid chromatography, the intact tetrapyrroles were separated as diconjugated and monoconjugated bilirubins; unconjugated bilirubin was resolved as XIII, IX and III alpha-isomers. The isocratic solvent system used was 0.1 M di-n-dodecylamine acetate/0.1 M di-n-octylamine acetate (4:1, v/v) in methanol, pH 7.4, at a flow of 1 ml per min. Diconjugated bilirubin accounted for 6.0 +/- 2.4 molar % (mean +/- S.E.), monoconjugated bilirubin for 37.4 +/- 8.4% and unconjugated bilirubin for 56.3 +/- 8.9% of the solubilized pigments. The IX alpha-isomer represented 96 +/- 1.9% of the unconjugated bilirubin. The presence of bilirubin conjugates in gallstones was confirmed by ethylanthranilate diazotization: the conjugated azodipyrrole in stone had the same retention time as that of conjugated azodipyrrole from rat and mouse bile. A majority of the bilirubin conjugates was sensitive to beta-glucuronidase of liver origin, indicating that the C-1 glucuronide ester was present.(ABSTRACT TRUNCATED AT 250 WORDS)

show abstract

“…5). It has been demonstrated that, in the black pigment gallstones of the hemolytic nb/nb mouse model (61), Ca salts of bilirubin monoconjugates also separate from solution in addition to Ca(HUCB) 2 . The experimental solubility products (dimensionless) for Ca(HUCB) 2 and CaUCB in 50 mM Na taurocholate are reported to be 6.5 ϫ 10 Ϫ13 and 1.8 ϫ 10 Ϫ8 , respectively (9).…”

Section: Discussionmentioning

confidence: 99%

Pathophysiological preconditions promoting mixed “black” pigment plus cholesterol gallstones in a ΔF508 mouse model of cystic fibrosis

Freudenberg

Leonard

Liu

et al. 2010

American Journal of Physiology-Gastrointestinal and Liver Physi

View full text Add to dashboard Cite

Gallstones are frequent in patients with cystic fibrosis (CF). These stones are generally "black" pigment (i.e., Ca bilirubinate) with an appreciable cholesterol admixture. The pathophysiology and molecular mechanisms for this "mixed" gallstone in CF are unknown. Here we investigate in a CF mouse model with no overt liver or gallbladder disease whether pathophysiological changes in the physical chemistry of gallbladder bile might predict the occurrence of "mixed" cholelithiasis. Employing a DeltaF508 mouse model with documented increased fecal bile acid loss and induced enterohepatic cycling of bilirubin (Am J Physiol Gastrointest Liver Physiol 294: G1411-G1420, 2008), we assessed gallbladder bile chemistry, morphology, and microscopy in CF and wild-type mice, with focus on the concentrations and compositions of the common biliary lipids, bilirubins, Ca(2+), and pH. Our results demonstrate that gallbladder bile of CF mice contains significantly higher levels of all bilirubin conjugates and unconjugated bilirubin with lower gallbladder bile pH values. Significant elevations in Ca bilirubinate ion products in bile of CF mice increase the likelihood of supersaturating bile and forming black pigment gallstones. The risk of potential pigment cholelithogenesis is coupled with higher cholesterol saturations and bile salt hydrophobicity indexes, consistent with a proclivity to cholesterol phase separation during pigment gallstone formation. This is an initial step toward unraveling the molecular basis of CF gallstone disease and constitutes a framework for investigating animal models of CF with more severe biliary disease, as well as the human disease.

show abstract

Studies on the Pathogenesis of Pigment Gallstones in Hemolytic Anemia

Abstract: A B S T R A C T The pathogenesis of hemolysis-in

Cited by 60 publications

References 17 publications

High-Performance Liquid Chromatography of Bilirubin Conjugates in Bile: Effect ofβ-Glucuronidase on the Bile Pigments

High-Performance Liquid Chromatography of Bilirubin Conjugates in Bile: Effect ofβ-Glucuronidase on the Bile Pigments

Monoconjugated bilirubin is a major component of hemolysis-induced gallstones in mice

Pathophysiological preconditions promoting mixed “black” pigment plus cholesterol gallstones in a ΔF508 mouse model of cystic fibrosis

Contact Info

Product

Resources

About