Visualization of the transport of primary and secondary bile acids across liver tissue in rats: in vivo study with fluorescent bile acids.
OBJECTIVE
Lysyl fluorescein conjugated bile acid analogues (LFCBAA) closely parallel their natural counterparts. To assess LFCBAA as a tool for the visualization of bile acid transport within liver tissue.


METHODS
Wistar rats were administered physiological concentrations of the primary bile acid analogue cholyllysyl fluoroscein (CLF) and of the secondary bile acid analogue lithocholyllysyl fluorescein (LLF) and serial liver biopsies were taken at fixed intervals. Both compounds were also injected retrogradely into the biliary tree. Frozen sections were examined by fluorescence microscopy.


RESULTS
Both CLF and LLF were rapidly taken up from sinusoidal blood but differed significantly in their hepatic handling. CLF was rapidly transported into bile, whereas LLF transport was slower and produced significantly more bile duct fluorescence. LLF clearance showed a lobular gradient with last remaining bile acid being confined largely to zone 3. Both compounds were avidly taken up by cholangiocytes after injection intravenously or retrogradely into the biliary tree.


CONCLUSIONS
Visualization of LFCBAA by fluorescence microscopy may yield further information regarding hepatobiliary bile acid localization during studies of physiological and pathological mechanisms involved in transport of bile acids. The presence of both compounds within cholangiocytes strongly suggests that they may undergo a degree of chole-hepatic recirculation.
Milkiewicz
P
P
Liver Research Laboratories, Queen Elizabeth Hospital, Edgbaston, Birmingham, UK. p.milkiewicz@bham.ac.uk
Mills
C O
CO
Hubscher
S G
SG
Cardenas
R
R
Cardenas
T
T
Williams
A
A
Elias
E
E
D000818:Animals; D001692:Biological Transport; D001706:Biopsy; D002793:Cholic Acids / Q000493:pharmacokinetics*; D005452:Fluoresceins / Q000493:pharmacokinetics*; D005456:Fluorescent Dyes / Q000493:pharmacokinetics*; D008095:Lithocholic Acid / Q000031:analogs & derivatives* / Q000493:pharmacokinetics*; D008099:Liver / Q000378:metabolism*; D008239:Lysine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics*; D008297:Male; D008856:Microscopy, Fluorescence; D051381:Rats; D017208:Rats, Wistar
D002793:Cholic Acids; D005452:Fluoresceins; D005456:Fluorescent Dyes; C072313:cholyl-lysylfluorescein; C109091:lithocholyl-lysyl-fluorescein; D008095:Lithocholic Acid; D008239:Lysine