Bilirubin kinetics in intact rats and isolated perfused liver. Evidence for hepatic deconjugation of bilirubin glucuronides.

Gollan, John L.; Hammaker, Lydia; Ličko, Vojtech; Schmid, Rudi

doi:10.1172/jci110111

Cited by 61 publications

(22 citation statements)

References 44 publications

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“…These oxygen consumption values were lower than those for mammals given in the literature (Tavoloni et al, 1978;Gollan et al, 1981 but were similar to other values in fish described by different authors (Reed et al, 1982;Blanco, 1994).…”

Section: Validation Assayssupporting

confidence: 89%

“…Two different oxygenation systems have traditionally been used for the perfusion medium: a multi-bulb glass oxygenator (Miller et al, 1951;Schimassek, 1963;Hems et al, 1966) and a membrane oxygenator Gollan et al, 1981 Even though both methods have been described in the literature as providing sufficient amounts of oxygen to cover the liver requirements, we chose to use the former because under our experimental conditions, the amount of oxygen in the perfusion medium was significantly higher (P < 0.05) when the multibulb glass oxygenator was used. Moreover, the amount of oxygen consumed by the liver was significantly higher (P < 0.001) for this system.…”

Section: Sample Collectingmentioning

confidence: 99%

“…Another criterion frequently used for validating this kind of technique is LDH activity (Rasenack et al, 1980;Gollan et al, 1981;Guaitani et al, 1983 (Sugano et al, 1978;Nashat et al, 1985;Styne and Vierling, 1986 When the influence of fasting on glucose production or liberation was studied, a change in the metabolic capacity of the liver was observed. For the fish that had undergone 96 h of fasting, the glucose production was significantly higher (P < 0.05) than for those undergoing 24 and 48 h of fasting for every 15 min interval period.…”

Section: Validation Assaysmentioning

confidence: 99%

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Perfusion of trout liver in situ. Description and validation of the technique

Cascales

Pérez‐Llamas²,

Marín³

et al. 1997

Reprod. Nutr. Dev.

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Summary ― The aim of this study was to develop a method for perfusing isolated trout livers that would make it possible to study hepatic metabolism in the whole organ. Rainbow trout (Oncorhynchus mykiss Walbaum) subjected to different fasting periods of 24, 48 or 96 h were used in all the experiments. A non-recirculating system was applied at a flow rate of 0.2 mL/min. The perfusion medium was a specific saline solution for salmonids oxygenated by a multibulb glass oxygenator. Viability assays included the measurement of oxygen consumption, lactate dehydrogenase activity and liver metabolic capacity. In addition, a histological study was carried out. Our results showed that the metabolic capacity of the liver survived throughout the perfusion process and that the functioning of this organ changed depending on the length of the fasting period to which the animal had been submitted. The method described here was shown to be suitable for studying the

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Section: Validation Assayssupporting

confidence: 89%

Section: Sample Collectingmentioning

confidence: 99%

Section: Validation Assaysmentioning

confidence: 99%

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Perfusion of trout liver in situ. Description and validation of the technique

Cascales

Pérez‐Llamas²,

Marín³

et al. 1997

Reprod. Nutr. Dev.

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“…Organic solvents often are used for extraction, with the assumption that interfering compounds such as heme are not extracted (10,19). This assumption is not strictly correct.…”

Section: Extraction Of Bilirubinmentioning

confidence: 99%

Clearance of Bilirubin from Rat Brain after Reversible Osmotic Opening of the Blood-Brain Barrier

1985

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ABSTRACT. Utilizing multicomponent spectrophotometry, we assayed the bilirubin content of rat cerebral hemispheres. With this assay, we determined the clearance of bilirubin from the rat brain following reversible, osmotic opening of the blood-brain barrier. Clearance was rapid, with a half-time of 1.7 h. This half-time was the same as that for clearance of bilirubin from the serum, suggesting that brain bilirubin was removed by transport or diffusion back into the general circulation. Hyperbilirubinemia is likely the most common medical diagnosis in the newborn. Physicians treat neonatal jaundice in order to prevent the neurotoxicity associated with brain uptake of bilirubin (kernicterus). Research on the cerebral transport and toxicity of bilirubin continues, but the results remain unclear (1). Uncertainty extends to clinical practice as well. For example, the preceding edition of Hospital Care of Newborn Infants, published by the American Academy of Pediatrics, included guidelines for exchange transfusion (2). The current edition omits guidelines, noting a diversity of approaches among physicians (3). In part, this diversity results from uncertainty about how bilirubin enters the brain and the anatomic and biochemical loci of bilirubin toxicity.The blood-brain bamer normally restricts exchange of watersoluble substances and proteins between blood and brain, but is damaged in diseases such as hypertension, by ischemia or trauma. Experimentally, the blood-brain bamer can be unilaterally and reversibly opened without causing brain damage by infusing a hypertonic solution of arabinose into one of the carotid arteries (4). Permeability returns to normal within about I h after arabinose infusion (5). We have previously shown that circulating albumin-bound bilirubin enters the side of the rat brain receiving the arabinose (4, 6). Furthermore, the regional cerebral distribution of albumin mimicks that seen in human kernicterus, providing a potential model for the human disease.We designed this study to determine the time-course of bilirubin entry into and removal from the brain after osmotic opening of the blood-brain bamer. For the study, we also developed rapid, simple techniques for the extraction and quantitation of bilirubin in brain tissue.

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“…Isolated perfused rat livers maintain physiological rates of bile salt uptake and secretion for up to 3 hours, but bile flow decreases thereafter. 5 Isolated rat hepatocytes exhibit profound decreases in rates of bile salt uptake over the first 72 hours of primary culture. [6][7][8] Bile salt transport may be virtually absent in cultured hepatocyte tumor cell lines.…”

mentioning

confidence: 99%

Enhanced Na+-dependent bile salt uptake by WIF-B cells, a rat hepatoma hybrid cell line, following growth in the presence of a physiological bile salt

et al. 1998

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Western blots immunostained for the basolateral Na ؉ -dependent plasma membrane protein, ntcp, revealed the appropriate Ϸ50-kd band in control and TC-grown cells, and confocal immunofluorescence microscopy demonstrated staining along the basolateral plasma membrane. Northern blots hybridized with a cDNA probe directed against ntcp indicated a modest TC-induced increase in mRNA levels. Reverse-transcriptase polymerase chain reaction (RT-PCR) using RNA isolated from WIF-B cells and oligonucleotide primers specific for rat ntcp or human NTCP transcripts revealed only the presence of the rat ntcp transcript. We conclude that bile salts, at concentrations normally found in mammalian portal blood, may be capable of promoting enhanced hepatocellular bile salt uptake via an increase in basolateral Na ؉ -dependent plasma membrane transport capacity. (HEPATOLOGY 1998;27:191-199.)The mammalian liver is continually exposed to low concentrations of bile salts present in portal and peripheral blood. Hepatocytes avidly take up bile salts across the basolateral plasma membrane and rapidly secrete them into bile; less than 5% of secreted bile salts are derived from de novo synthesis within the hepatocyte. 1 In rats, the postprandial bile salt concentration in the portal blood is Ϸ60 µmol/L, with taurocholate (TC) comprising up to half of the bile salt species. 2 In humans, the bile salt load reaching the liver fluctuates considerably over a 24-hour period, with portal vein plasma concentrations ranging from Ϸ14 µmol/L in the fasting state to Ϸ43 µmol/L in the postprandial state. 3 Hepatocytes are thus exposed routinely to changes in rates of bile salt delivery, and therefore must respond with variable rates of bile salt uptake across the basolateral plasma membrane. High plasma bile salt concentrations (up to 1 mmol/L) are associated with down-regulation of basolateral bile salt uptake into hepatocytes. 4 It is unclear, however, whether plasma bile salt concentrations within physiological ranges may promote bile salt uptake into hepatocytes.Difficulties are encountered when examining the physiological role of bile salts in intact animals, because many variables change simultaneously when conducting in vivo studies. A major experimental problem also is encountered when addressing normal bile salt regulation of hepatic physiology in vitro, because of the difficulty in maintaining liver preparations with stable rates of basolateral bile salt uptake. Isolated perfused rat livers maintain physiological rates of bile salt uptake and secretion for up to 3 hours, but bile flow decreases thereafter. 5 Isolated rat hepatocytes exhibit profound decreases in rates of bile salt uptake over the first 72 hours of primary culture. [6][7][8] Bile salt transport may be virtually absent in cultured hepatocyte tumor cell lines. 9 Recently, an in vitro polarized hepatocyte cell system, WIF-B cells, has been reported by Hubbard et al. 10,11 as a refinement of an existing rat hepatoma-human fibroblast cell line developed by Cassio et al. 12 WIF-B cell...

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Bilirubin kinetics in intact rats and isolated perfused liver. Evidence for hepatic deconjugation of bilirubin glucuronides.

Cited by 61 publications

References 44 publications

Perfusion of trout liver in situ. Description and validation of the technique

Perfusion of trout liver in situ. Description and validation of the technique

Clearance of Bilirubin from Rat Brain after Reversible Osmotic Opening of the Blood-Brain Barrier

Enhanced Na+-dependent bile salt uptake by WIF-B cells, a rat hepatoma hybrid cell line, following growth in the presence of a physiological bile salt

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