Role of hepatic transporters in prevention of bile acid toxicity after partial hepatectomy in mice

The enterohepatic recirculation of bile acids (BAs) is important in several physiological processes. Although there has been considerable research on liver regeneration after two-thirds partial hepatectomy (PHx), little is known about how the liver protects itself against BA toxicity during regeneration. In this study, various BAs in plasma and liver, the composition of micelle-forming bile constituents, as well as gene expression of the main hepatobiliary transporters were quantified in sham-operated and PHx mice 24 and 48 h after surgery. PHx did not influence the hepatic concentrations of taurine-conjugated BAs (T-BA) but increased the concentration of glycine-conjugated (G-BA) and unconjugated BAs. Total BA excretion (μg·min−1·g liver wt−1) increased 2.4-fold and was accompanied by a 55% increase in bile flow after PHx. The plasma concentrations of T-BAs (402-fold), G-BAs (17-fold), and unconjugated BAs (500-fold) increased. The mRNA and protein levels of the BA uptake transporter Ntcp were unchanged after PHx, whereas the canalicular Bsep protein increased twofold at 48 h. The basolateral efflux transporter Mrp3 was induced at the mRNA (2.6-fold) and protein (3.1-fold) levels after PHx, which may contribute to elevated plasma BA and bilirubin levels. Biliary phospholipid excretion was nearly doubled in PHx mice, most likely owing to increased mRNA expression of the phospholipid transporter, Mdr2. In conclusion, the remnant liver after PHx excretes 2.5-fold more BAs and three times more phospholipids per gram liver than the sham-operated mouse liver. Upregulation of phospholipid transport may be important in protecting the biliary tract from BA toxicity during PHx.

Endogenous neurotensin facilitates enterohepatic bile acid circulation by enhancing intestinal uptake in rats

Initial studies on the digestive hormone neurotensin (NT) showing that intestinal NT mRNA expression and blood levels were altered in rats fed chow containing bile acid (BA) and the BA chelator cholestyramine led us to investigate the role of NT in the enterohepatic circulation of BA. In fasted, anesthetized rats with common bile ducts cannulated for bile collection, intravenous NT infusion (10 pmol · kg−1 · min−1) enhanced BA output relative to control over 3 h in animals administered donor bile into the duodenum (30 μl/min). This suggested that the effect of NT was on the return of BA from the intestine to the liver, which is rate determining in the normal process. In rats prepared as described above and administered [3H]taurocholate ([3H]TC; 5 mM, 1 ml) duodenally, NT infusion (3–10 pmol · kg−1 · min−1) increased the [3H]TC recovery rate in bile approximately twofold, whereas sulfated CCK-8 (12–50 pmol · kg−1 · min−1) had no effect. To investigate the roles of endogenous NT and CCK, we administered [3H]TC into the rat duodenum or lower jejunum and tested the effect of the NT antagonist SR-48692 (2 nmol · kg−1 · min−1) or CCK-A antagonist lorglumide (100 nmol · kg−1 · min−1). SR-48692 reduced the [3H]TC recovery rate by ≅50% and ≅24% in the duodenum and jejunum, respectively, whereas lorglumide had no effect. These results suggest that NT or a similar peptide is an endogenous regulator of enterohepatic BA cycling, which acts by enhancing BA uptake in the intestine.

BENZYL ADENINE UPTAKE AND METABOLISM DURING PETUNIA SHOOT ORGANOGENESIS

The uptake and metabolism of exogenous tritium-labelled benzyl adenine was studied during the shoot induction period of petunia leaf explants in tissue culture. Transfer experiments with Petunia `MD1' leaf explants (1 cm2) on MS media with 2.2 uM BA show that 27% and 100% of the leaf explants are committed to shoot induction on days 6 and 10, respectively. To study BA uptake and metabolism, leaf explants were placed on media containing tritium-labled BA for 1, 3, 6 and 10 days. BA was taken up from the media on days 1-6. BA metabolizes were analyzed using HPLC, a UV absorbancc detector and enzymatic techniques. Metabolizes produced include: BA, BAdo, BA 7G, BA 9G, BAdoMP, BAdoDP, BAdoTP and 3 unidentified compounds. BA and BAdo were detected on days 1 and 3 but not during day 6-10, the time of shoot induction. The pool of ribotide metabolites decreased from days 1 to 10, from 26.5% of all metabolites to 1.6%. Glucosylated compounds, BA 7G and BA 9G, increased continuously from 24.9% to 69.8% between days 1 and 10. An unidentified compound C increased from 13% on day 3 to 24.8% on day 10. In separate experiments, BA uptake and metabolism were compared in two Petunia hybrida lines, St40 and TLV1, with different shoot organogenic responses in tissue culture. These data show interesting patterns of BA metabolism in relationship to shoot induction and organogenesis.