Role of hepatocytes in direct clearance of lipopolysaccharide in rats.
OBJECTIVE
The liver is the clearance organ for lipopolysaccharide (LPS). The aim of this study was to investigate the biliary excretion of LPS using fluorescein isothiocyanate (FITC)-labeled LPS.


METHODS
After FITC-LPS was injected intravenously into rats, the cellular localization of fluorescence in the liver was examined and the biliary excretion of fluorescence was measured. The effects of gadolinium chloride, a blocker of Kupffer cells, and colchicine, an inhibitor of microtubules, on the biliary excretion of fluorescence was investigated, and bile was analyzed using high-performance liquid chromatography.


RESULTS
Laser scanning confocal microscopy showed that fluorescence was taken up by hepatocytes 5 minutes after injection of FITC-LPS into the portal vein. When FITC-LPS was injected into the portal vein, fluorescence was rapidly secreted into bile, peaking at 20 minutes, and 25.1% of the injected dose appeared in bile within 60 minutes. When the same dose of FITC-LPS was injected into the tail vein, 15.8% appeared in bile within 60 minutes. Chromatography showed that FITC-LPS was excreted into bile in an unchanged form over a period of 20 minutes after injection. Colchicine significantly reduced the biliary excretion of fluorescence, but gadolinium chloride had no effect.


CONCLUSIONS
LPS was directly and effectively processed by hepatocytes and secreted into the bile canalicular system via a microtubule-dependent vesicular pathway.
D000818:Animals; D001646:Bile / Q000187:drug effects / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003078:Colchicine / Q000494:pharmacology; D016650:Fluorescein-5-isothiocyanate; D005682:Gadolinium / Q000494:pharmacology; D007728:Kupffer Cells / Q000187:drug effects; D008070:Lipopolysaccharides / Q000378:metabolism*; D008099:Liver / Q000166:cytology*; D008297:Male; D018613:Microscopy, Confocal; D008870:Microtubules / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar
D008070:Lipopolysaccharides; D005682:Gadolinium; D016650:Fluorescein-5-isothiocyanate; C038958:gadolinium chloride; D003078:Colchicine