Different pathways of canalicular secretion of sulfated and non-sulfated fluorescent bile acids: a study in isolated hepatocyte couplets and TR- rats.
OBJECTIVE
Fluorescent bile acids have proved useful for characterizing bile salt transport mechanisms. The aim of this study was to further validate the use of lysyl-fluorescein conjugated bile acid analogues as surrogate bile acids.


METHODS
We analyzed biliary excretion kinetics of cholyl lysyl fluorescein (CLF), lithocholyl lysyl fluorescein (LLF) and sulfo-lithocholyl lysyl fluorescein (sLLF), both in the isolated rat hepatocyte couplet model and in TR- rats with a selective canalicular transport defect of non-bile acid organic anions.


RESULTS
CLF and LLF, which like their natural nonsulfated bile acid congeners are expected to be handled by the canalicular bile salt export pump, were transferred into the bile canaliculus much faster than sLLF, a putative substrate for the canalicular multispecific organic anion transporter in both the in vivo and the in vitro models employed. The contention that different transport systems are involved in sulfated and non-sulfated lysyl fluorescein conjugated bile acids biliary excretion was supported further by studies using TR- rats, in which the cumulative biliary excretion of sLLF was reduced to 6% as compared with that of normal Wistar rats, in good agreement with values for its naturally-occurring radiolabeled parent compound sulfoglycolithocholate. In contrast, CLF and LLF were reduced to 66% and 52%, similar values to these for their congeners, [14C] glycocholate and [14C] lithocholate.


CONCLUSIONS
The close similarity in behavior of lysyl fluorescein conjugated bile acids to that of their naturally-occurring parent compounds in these different models gives support for both sulfated and nonsulfated lysyl fluorescein conjugated bile acids as substitute molecules for studies of bile acid transport.
Mills
C O
CO
Liver Unit, Queen Elizabeth Hospital, Edgbaston, Birmingham, UK.
Milkiewicz
P
P
Müller
M
M
Roma
M G
MG
Havinga
R
R
Coleman
R
R
Kuipers
F
F
Jansen
P L
PL
Elias
E
E
D000818:Animals; D027321:Anion Transport Proteins; D001647:Bile Acids and Salts / Q000378:metabolism*; D001648:Bile Canaliculi / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D002793:Cholic Acids; D003287:Contrast Media; D005452:Fluoresceins; D005453:Fluorescence; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D051381:Rats; D011922:Rats, Mutant Strains / Q000378:metabolism; D017208:Rats, Wistar; D013431:Sulfates / Q000378:metabolism*
D027321:Anion Transport Proteins; D001647:Bile Acids and Salts; D002352:Carrier Proteins; D002793:Cholic Acids; D003287:Contrast Media; D005452:Fluoresceins; D013431:Sulfates; C072313:cholyl-lysylfluorescein