Synthesis Of Retinol-binding Protein Research Articles

Development of a Compartmental Model to Investigate the Influence of Inflammation on Predictions of Vitamin A Total Body Stores by Retinol Isotope Dilution in Theoretical Humans

BackgroundInflammation, both acute and chronic, is associated with reductions in the synthesis of retinol-binding protein (RBP) and the concentration of retinol in plasma. Consequently, it is currently recommended that the retinol isotope dilution (RID) method not be used to estimate vitamin A total body stores (TBS) in subjects with inflammation. ObjectivesTo apply compartmental analysis to study the effects of inflammation on hepatic apo-RBP input, plasma retinol pool size, and RID-predicted TBS in theoretical subjects with known steady state values for these parameters. MethodsWe selected 6 previously generated hypothetical subjects who ingested a dose of stable isotope–labeled vitamin A (day 0). Starting with each subject’s published steady state model for retinol tracer kinetics, we developed a parallel model for unlabeled retinol and RBP that incorporated links between these entities and tied liver retinol secretion to RBP availability. Then we perturbed the steady state model by initiating chronic or acute inflammation on day 0 or acute inflammation on day 3 or 9 and simulating results for RBP, plasma retinol, and TBS. ResultsChronic inflammation led to substantial reductions in RID-predicted TBS for at least 2 weeks after tracer administration. In contrast, acute inflammation induced on day 0 or 3 resulted in less dramatic impacts on TBS (37% or 20% reduction, respectively, from steady state levels, with levels rebounding by 14 days). When inflammation was induced 9 days after administration of the tracer, the effects on predicted TBS were minimal. Overall, for acute inflammation initiated at 0, 3, or 9 days, accurate predictions of TBS were obtained by 2 weeks. ConclusionsCompartmental analysis can be applied in the novel way described here to study the influence of perturbations such as inflammation on predictions of vitamin A status using RID. Such an approach has potential value for studying other perturbations and different nutrients.

Vitamin A and retinol-binding protein deficiency among chronic liver disease patients

ObjectiveVitamin A deficiency (VAD) is associated with the progression of chronic liver disease (CLD). The aim in this study was to assess levels of serum retinol and retinol-binding protein (RBP) as well as liver vitamin A stores in the presence of liver cirrhosis and hepatocellular carcinoma. MethodsWe ascertained the serum retinol and RBP levels of randomly selected CLD patients divided into two groups, one given 1500 UI (n = 89) and the other receiving 2500 UI (n = 89) doses of retinyl palmitate for the relative dose response test. Blood samples were collected in a fasting state and 5 and 7 h after supplementation. ResultsThe prevalence of VAD was 62.4%. There was a progressive drop in serum retinol (P < 0.001) and RBP (P = 0.002) according to the severity of the liver disease, and a greater prevalence of severe VAD was noted in cirrhosis Child & Pugh C (52.8%). Fifty percent of the patients presented a low availability of RBP relative to retinol concentration, and there was no peak in RBP levels regardless of the dose of retinyl palmitate administered. ConclusionsOur findings suggest serum retinol and RBP are relevant as indicators of vitamin A nutritional status in the presence of CLD. Liver vitamin A store cannot be evaluated using the RDR test because CLD causes a reduction in RBP synthesis and interferes with the mobilization of endogenous vitamin A. Considering how the patients already showed a drop in RBP relative to retinol concentrations, it is reasonable to assume vitamin A supplementation may trigger harmful effects in CLD patients.

High levels of vitamin E affect retinol binding protein but not CYP26A1 in liver and hepatocytes from laying hens

This study was conducted to determine the influence of vitamin E on the retinol binding protein (RBP) and cytochrome p450 family 26 subfamily A polypeptide 1 (CYP26A1), which are specific transporters and catabolic enzymes of vitamin A, respectively. In the in vivo experiments, a total of 450 laying hens was fed 5 levels of vitamin E (0, 20, 80, 320, and 1,280 IU/kg of feed) supplementation. For the in vitro assays, hepatocytes from laying hens were cultured in 4 levels of α-tocopherol (0, 10, 50, and 100 μM). High dietary vitamin E increased the concentration of vitamin A in liver (P < 0.05). The RBP and its mRNA expression in liver and hepatocytes were markedly inhibited by dietary vitamin E (320 and 1,280 IU/kg) and α-tocopherol (100 μM) in culture medium (P < 0.05). However, CYP26A1 and its mRNA expression were not affected by vitamin E in both liver and hepatocytes (P > 0.05). The results indicate that excessive vitamin E could increase the concentration of vitamin A in liver by inhibiting RBP synthesis in hepatocytes.

DEFISIENSI VITAMIN A DAN ZINC SEBAGAIFAKTOR RISIKO TERJADINYA STUNTING PADA BALITA DI NUSA TENGGARA BARAT

Children of 1 - 5 years old often face nutrition problems such as undernourishment, vitamin A and zinc deficiency, anemia, stunting and low mental development index (MDI). Vitamin A deficiency can cause growth disorder and declining zinc transport and mobilization in the heart meanwhile zinc is needed in retinol binding protein synthesis. If the deficiency lasts for long it can cause growth disorder as manifested in stunting incidence. To study vitamin A and zinc deficiency as risk factors for the incidence of stunting among underfives at Nusa Tenggara Barat. The study was analytic observational with cross sectional design. Subject of the study were 327 underfives of 6 - 59 months at the Province of Nusa Tenggara Barat. Nutrition status was assessed through measurement of anthopometry, retinol serum level using HPLC, zinc serum level using AAS and hemoglobin using hemoCue. Data of individual characteristics and rearing pattern were obtained through interview and nutrient intake were measured using recall 2 x 24 hours. Data analysis used bivariate technique for variable related to stunting, chi square test for category data, independent t-test for ratio and logistic regression test to measure risk of some variables simultaneously related to incidence of stunting. The result of bivariate analysis showed that was difference in age between stunted and normal underfives (p<0.05) . In the group of underfives that were no longer breastfed there was difference in intake of zinc between stunted and normal underfives (p<0.05). The result of multivariate analysis showed that underfives that were no longer breastfed had risk 2 times greater for being stunted after the control of age, status of vitamin A and zinc. The effect is modified by age and breast-feeding. There was no evidence that showed that vitamin A and zinc deficiency among underfives were risk factors for the incidence of stunting. Variable that was individually as well as simultaneously significant for the incidence of stunting was breastfeeding status. Underfives that were not breastfed had risk 2 times greater for being stunted than those that were breastfed, and the effect is modified by age and breast-feeding. Keywords: Stunting, Vitamin A, Zinc, Retinol Serum, Zinc Serum, Hemoglobin, Breastfeeding Status

Expression of Genes Associated with Retinoid Metabolism in the Trout Ovarian Follicle1

In vertebrates producing oviparous eggs, retinoids and their precursor molecules need to be deposited in oocytes during vitellogenesis. While most studies focus on the transport of retinoids and carotenoids formed outside the fish ovary and their deposition within the developing oocyte, recent investigations in mammalian species suggest the ovary is an important site for retinoid and carotenoid metabolism. Therefore, we investigated the expression of six genes (bcmo1, bcdo2, rbp1, lrat, rbp4, and stra6) associated with retinoids and carotenoids in juvenile and adult trout ovaries. Except for bcdo2, these genes were expressed in the ovary. Expression of stra6 was detected in the ovary but not in the liver. Gene expression levels of bcmo1 and stra6 were significantly higher in juvenile ovaries, in contrast to those of rbp1, rbp4, and lrat, which were similar in all tested ovarian stages. The mean values of the relative mRNA levels of the tested genes differed between the ovary and the liver. Gene transcripts of rbp4 and bcmo1 were identified by in situ hybridization in the theca layer, and all five genes were expressed in the granulosa, stromal cells, and only the early vitellogenic oocyte. The occurrence of retinol-binding protein in the theca and granulosa cells and within oocytes at all developmental stages was revealed by immunocytochemistry. These results indicate that ovarian cells express genes putatively associated with cleavage of beta-carotene, storage and mobilization of retinyl-esters, and of retinol-binding protein synthesis, suggesting a novel pathway for providing retinoids and carotenoids to developing fish ovarian follicles.

Accumulation of retinol in the liver after prolonged hyporetinolemia in the vitamin A-sufficient rat

We assessed the effects of prolonged reduction of plasma retinol concentrations (hyporetinolemia) on the distribution of tissue vitamin A (VA) and of its active compounds using a model of continuous recombinant human interleukin-6 (rhIL-6) infusion via osmotic minipumps in VA-sufficient male rats. Plasma retinol and retinol-binding protein (RBP) concentrations remained decreased and lower in rhIL-6-treated rats compared with controls from 7.5 h throughout 7 days of infusion (P < 0.001). This reduction was accompanied by a 68% increase in hepatic retinol concentration by 7 days (P < 0.05). Hepatic and renal retinyl palmitate and retinoic acid concentrations did not change, and renal megalin content remained unchanged; hepatic RBP concentrations were 41% lower in rhIL-6-treated rats compared with controls (P < 0.05). These results indicate that instead of being lost, retinol accumulated in the liver during inflammation and that hyporetinolemia was attributable to a decrease in the availability of hepatic RBP. A plausible consequence of the effect of rhIL-6-induced hyporetinolemia is that by 7 days tissues that are dependent on plasma retinol may become deprived of VA. These results have important implications in understanding the mechanism by which measles infection induces hyporetinolemia and VA deficiency of extrahepatic tissues.

Increased Vitamin A and E Levels in Adult Cystic Fibrosis Patients after Lung Transplantation

Most patients with cystic fibrosis (CF) and pancreatic insufficiency require supplementation with fat-soluble vitamins to maintain normal serum levels. Even with supplementation, toxicity is rare. We evaluated serum vitamin A and E levels in 23 adult patients with CF who underwent double lung transplantation. Twenty-one of the subjects were pancreatic insufficient. Fifteen subjects had serum vitamin levels before and after transplant. The median time posttransplantation for these subjects was 9 months. Mean serum vitamin A and E levels were significantly higher posttransplantation (P<0.0001, P<0.001, respectively). Eight subjects who only had posttransplant vitamin levels also had abnormally high vitamin A levels. Although the etiology of this novel finding is unclear, possibilities include altered absorption, drug interactions, impaired retinol metabolism, or increased hepatic synthesis of retinol binding protein.

Expression of retinol-binding protein and cellular retinol-binding protein in the bovine ovary.

Retinol (vitamin A) is essential for reproduction, and retinoids have been suggested to play a role in ovarian steroidogenesis, oocyte maturation, and early embryonic development. Retinol is transported systemically and intercellularly by retinol-binding protein (RBP). Within the cell, cellular retinol-binding protein (CRBP) functions in retinol accumulation and metabolism. Since the actions of retinoids are mediated, in part, by retinoid-binding proteins, the objective of this study was to investigate cell-specific expression of RBP and CRBP in the bovine ovary. Immunocytochemical analysis (ICC) localized RBP to the thecal and granulosa cell layers of antral and preantral follicles with the most intense staining in the cells of large, healthy follicles. The tunica adventitia of arterial blood vessels also exhibited RBP staining. Immunostaining of CRBP was most intense in the granulosa cells of preantral follicles and present, but diminished, in thecal and granulosa cells of antral follicles. Within the corpus luteum, both proteins were observed in large luteal cells, but only RBP was observed in small luteal cells. Northern blot analysis demonstrated that thecal and granulosa cells from antral follicles and luteal tissue expressed RBP and CRBP mRNA. Synthesis and secretion of RBP by thecal cells, granulosa cells, and luteal cells were demonstrated by immune-complex precipitation of radiolabeled RBP from the medium of cultured cells or explants, followed by SDS-PAGE and fluorography. Follicular fluid was collected from small (<5 mm) and large (8-14 mm) follicles, pooled according to follicular size, and analyzed for retinol, RBP, estradiol-17beta, and progesterone. Concentrations of retinol, RBP, and estradiol were greater in the fluid of large follicles. Results demonstrate retinoid-binding protein expression by bovine ovaries and provide physical evidence that supports the concept that retinoids play a role in ovarian function.

Plasma Retinol Binding Protein for Monitoring the Acetaminophen-induced Hepatotoxicity

Retinol-binding protein (RBP) is a specific transport protein which carries retinol in the circulation. RBP concentration in plasma and liver of rats following a large dose of acetaminophen (APAP) intraperitoneally was examined. The RBP concentration in plasma decreased significantly at 12 hr after the APAP administration, while the plasma albumin concentration was affected a little. Western blot and northern blot analyses showed marked changes in RBP but not in albumin. Thus, RBP was suggested to be more sensitive for the acute drug-induced hepatotoxicity than albumin. The decrease of RBP concentration in plasma was suggested to be caused by the dysfunction of RBP synthesis in the liver.

Retinol mobilization from cultured rat hepatic stellate cells does not require retinol binding protein synthesis and secretion

Retinol mobilization from retinyl esters stores of hepatic stellate cells (HSCs) is a key step in the regulation of mammalian retinol homeostasis, but the precise mechanisms of such a mobilization are still poorly understood. Using primary cultures of HSCs, we first demonstrated that HSCs expressed immunoreactivity against retinol-binding-protein (RBP) when cultured in a medium containing RBP but were unable to synthesize RBP transcripts and proteins. Using pulse and chase-type experiments, we demonstrated that radioactive retinol was released in culture medium without binding proteins. Inhibition of protein secretion by brefeldin A did not modify quantitatively retinol release. This data ruled out, for the first time, the direct involvement of RBP in retinol mobilization from HSCs. Moreover, HSCs co-cultured with primary isolated hepatocytes displayed an increase of retinol transfer from HSCs to hepatocytes when they established direct physical contacts, as compared with co-cultures without contact. Based on this latter data, a mechanism of retinol mobilization from HSCs via the hepatocytes using retinol transfer through cellular membranes is proposed.

Developmental expression, tissue distribution and hormonal regulation of fish (Sparus aurata) serum retinol-binding protein

Retinol-binding protein (RBP) is the specific carrier of retinol in vertebrates and forms a 1:1 complex with transthyretin (TTR). A cDNA encoding serum RBP was cloned from liver and 7-day larvae of the marine fish Sparus aurata. The mature protein is 176 amino acids long and shows sequence identity of 77–78%, 56%, 63% and 62% with rainbow trout, Xenopus, chicken and human RBP, respectively. Northern blot analysis of hepatic RBP revealed two transcripts: a major one of approximately 1.4–1.5 kb and a minor of approximately 0.7 kb. Distribution of RBP mRNA in various tissues was studied by RT-PCR and showed high expression in liver and skin, and low expression in brain, kidney and gill filament (20–35% of the level in liver). RBP expression in intestine, pyloric caeca, muscle and pituitary was estimated to be approximately 7–14% of the level in liver. The ontogeny of RBP expression in S. aurata was examined in unfertilized eggs, embryos and larvae by using RT-PCR followed by hybridization with a specific probe. RBP transcript was found in all larval stages studied. Very low levels of RBP mRNA were detected in unfertilized eggs and in embryos 8 h after fertilization with a gradual increase at 12 h and 15–16 h post-fertilization. A single injection of estradiol-17β to S. aurata immature, bisexual fish or to adult males reduced steady-state levels of hepatic RBP by 37 and 25%, respectively. The same treatment induced vitellogenin expression. The present data suggest that in fish, liver is the main site of RBP synthesis, but that RBP may have an important function in fish skin. RBP is expressed early in embryonic development and in fish its expression can be down regulated by estrogen.

Cooperation of liver cells in health and disease.

Cooperation of liver cells in health and disease.

The Metabolic Availability of Vitamin A Is Decreased at the Onset of Diabetes in BB Rats

Streptozotocin (STZ)-induced diabetic rats have been associated with an impaired metabolic availability of vitamin A (retinol). This study was undertaken to investigate whether Biobreeding (BB) rats, in which diabetes mellitus resembling human type I diabetes develops spontaneously, respond the same way at the onset of diabetes. Weaning diabetes-prone (BBdp) and normal (BBn) BB rats consumed NIH-07 nonpurified diet ad libitum until 120 d of age. Plasma and hepatic concentrations of retinol and its carriers, retinol-binding protein (RBP) and transthyretin (TTR) were lower in diabetic BB (BBd) rats than in BBn rats. In parallel with RBP, the abundance of mRNA was lower in the liver of BBd rats. Furthermore, the status of zinc, an important factor for the synthesis of RBP, was also disturbed in BBd rats, as indicated by lower circulatory levels and greater urinary excretion. To determine whether the biochemical evidence of vitamin A deficiency in BBd rats could be reversed, BBdp rats were fed a diet supplemented with vitamin A either alone or in combination with zinc. None of these treatments increased plasma vitamin A concentration. The hepatic abundance of RBP mRNA was significantly greater, whereas circulatory RBP concentrations were unaffected by vitamin A plus zinc supplementation. Overall, these results suggest that impaired metabolic availability of vitamin A, possibly caused by its decreased transport from hepatic stores, is another metabolic derangement associated with type I diabetes.

MMH cells: An in vitro model for the study of retinol-binding protein secretion regulated by retinol.

The untransformed stable cell line Met murine hepatocytes (MMH), generated from liver explants of transgenic mice expressing a constitutively active truncated form of the human hepatocyte growth factor receptor (cyto-Met), represents an innovative tool for in vitro studies of liver function. In the present report, we show that the MMH-D3 line isolated from the liver of a 3-day-old mouse is a useful model to investigate the regulation of the synthesis and secretion of retinol-binding protein (RBP) by retinol (vitamin A alcohol). Experiments with Northern blot hybridization, metabolic labeling of cellular proteins followed by immunoprecipitation, and Western blot analysis demonstrated that, similarly to the in vivo situation, in MMH-D3 cells the presence of retinol does not affect transcriptional and translational rate of the RBP gene but is essential for regulating the secretion rate of the protein. Unlike HepG2 human hepatocarcinoma cells used thus far in studies of retinoid metabolism, including the synthesis and secretion of RBP, vitamin A deficiency causes, in MMH-D3 cells, the inhibiton of RBP secretion and the protein accumulation in the cell, whereas retinol repletion promptly results in RBP secretion. This model will be very useful in future studies on vitamin A distribution in the organism.

Steroid regulation of retinol-binding protein in the ovine oviduct.

Two studies were conducted to identify retinol-binding protein (RBP) expression in the ovine oviduct and to determine the role of ovarian steroids in its regulation. Ewes were salpingectomized on Days 1, 5, or 10 of their respective estrous cycles, and oviducts were homogenized for RNA analysis, fixed for immunocytochemistry (ICC), or cultured for 24 h for protein analysis. ICC localized RBP to the epithelium of all oviducts. RBP synthesis was demonstrated by immunoprecipitation of radiolabeled RBP from the medium of oviductal explant cultures. Explant culture medium from oviducts harvested on Day 1 contained significantly more RBP than medium from oviducts collected on Days 5 or 10. Slot-blot analysis demonstrated that steady-state RBP mRNA levels were significantly higher on Day 1 than Day 5 or 10. In the second experiment, ovariectomized ewes were treated with estradiol-17beta (E2), progesterone (P4), E2+P4 (E2+P4), or vehicle control, and oviducts were analyzed as above. P4 alone or in combination with E2 significantly reduced steady-state RBP mRNA levels compared to those in E2-treated animals. Oviductal explants from E2- and E2+P4-treated animals released 3- to 5-fold more RBP into the medium than control and P4 treatments as determined by ELISA. RBP synthesis of metabolically labeled RBP was increased by E2 and E2+P4 treatments. This study demonstrates that P4 applied on an estradiol background negatively regulates RBP gene expression in the oviduct whereas estradiol appears to stimulate RBP synthesis and secretion.

Acute Inflammation Induces Hyporetinemia and Modifies the Plasma and Tissue Response to Vitamin A Supplementation in Marginally Vitamin A–Deficient Rats

Plasma retinol is reduced during numerous infections, and inflammation alters the hepatic synthesis of retinol-binding protein (RBP). In this study, we have investigated the effects of endotoxin-induced inflammation on vitamin A (VA) supplementation in a rat model of marginal VA deficiency. Marginally VA-deficient rats received an intraperitoneal dose of lipopolysaccharide (LPS, n = 14) or saline (n = 10); 6 h later, six LPS + VA and six saline + VA rats received 7.1 μmol VA orally. Twenty-four hours after endotoxin administration, rats with inflammation (LPS) had lower plasma retinol, RBP, and hepatic RBP than saline rats (37, 31 and 44%, respectively, P < 0.05). Inflammation did not affect VA concentrations in liver and perirenal adipose tissue, although kidney VA was reduced relative to saline rats. However, urinary VA was not detected. Eighteen hours after VA supplementation, inflammation reduced the plasma unesterified retinol response (P < 0.05) in LPS + VA relative to saline + VA rats, although total VA increased as a result of the presence of retinyl esters in LPS + VA rats. Hepatic esterified retinol concentration was reduced (P < 0.01) in LPS + VA compared with saline + VA rats; however, hepatic unesterified retinol did not differ. Renal total retinol increased in VA-supplemented rats, but urinary retinol excretion, when observed, was low, independently of inflammation. These findings indicate that inflammation-induced hyporetinemia does not necessarily imply a loss of VA, but rather represents a redistribution of tissue VA brought about by a reduced hepatic synthesis of RBP. Practical implications from these collective results are to recommend the determination of both unesterified and esterified retinol to fully assess the plasma response to VA supplementation and to caution the use of VA assessment methodologies that depend on the hepatic synthesis of RBP during acute inflammation.

Bovine endometrial retinol-binding protein secretion, messenger ribonucleic acid expression, and cellular localization during the estrous cycle and early pregnancy.

Retinol-binding protein (RBP) mRNA was localized to luminal and glandular epithelial cells of bovine endometrium by in situ hybridization. Relative levels of endometrial RBP mRNA expression during the estrous cycle and early pregnancy were determined by quantitative slot blot analysis and contrasted with uterine luminal concentrations of RBP. Expression of mRNA was moderate at Day 1 after estrus, declined at Day 5, reached lowest levels by Day 10, rose significantly through Days 15-17, and peaked at Day 20. RBP mRNA expression in pregnant animals was similar to that in cyclic animals on Day 15, doubled between Days 17 and 20, remained constant through Day 22, and rose slightly thereafter. Luminal RBP concentrations of cyclic cows, as determined by ELISA, decreased from Day 1 through Day 10, rose dramatically on Day 15, then declined through Day 20. Concentrations of RBP in uterine flushes from pregnant animals were similar to those of cyclic cows on Day 15 but remained relatively constant through Day 17. It is concluded that 1) RBP synthesis occurs in the luminal and glandular epithelial cells, 2) RBP transcription and secretion are correlated with each other, and 3) ovarian steroids, possibly in conjunction with uterine concentrations of their receptors, modulate uterine RBP expression.

A retinol-binding protein receptor-mediated mechanism for uptake of vitamin A to postimplantation rat embryos.

The major form of dietary vitamin A in blood is retinol bound to retinol-binding protein (RBP), but it is not clear how maternal retinol reaches the embryo. We demonstrate here that uptake of radiolabeled retinol from holo-RBP to the visceral yolk sac of cultured rat embryos exhibits receptor-mediated characteristics (Km = 4.1 x 10(-7) M, Vmax = 3.0 x 10(-12) mol/sec per gram yolk sac protein) and also demonstrate the presence in the yolk sac endoderm of a protein previously described as an RBP receptor. Furthermore, transport of radiolabeled retinol from the yolk sac to the embryo proper can be markedly decreased by inhibition of visceral yolk sac RBP synthesis. Thus embryonic accumulation of retinol depends on a relay of retinol from maternal RBP to RBP synthesized in the yolk sac, via interaction with a visceral yolk sac RBP receptor.

Retinyl Ester Hydrolysis and Retinol Efflux from BFC-1β Adipocytes

Adipose tissue is an important storage depot for retinol, but there are no data regarding retinol mobilization from adipose stores. To address this, dibutyryl cAMP was provided to murine BFC-1beta adipocytes and its effects on retinol efflux assessed. High performance liquid chromatography analysis of retinol and retinyl esters in adipocytes and media indicated that cAMP stimulated, in a time- and dose-dependent manner, retinol accumulation in the culture media and decreased cellular retinyl ester concentrations. Study of adipocyte retinol-binding protein synthesis and secretion indicated that cAMP-stimulated retinol efflux into the media did not result from increased retinol-retinol-binding protein secretion but was dependent on the presence of fetal bovine serum in the culture media. Since our data suggested that retinyl esters can be hydrolyzed by a cAMP-dependent enzyme like hormone-sensitive lipase (HSL), in separate studies, we purified a HSL-containing fraction from BFC-1beta adipocytes and demonstrated that it catalyzed retinyl palmitate hydrolysis. Homogenates of Chinese hamster ovary cells overexpressing HSL catalyzed retinyl palmitate hydrolysis in a time-, protein-, and substrate-dependent manner, with an apparent Km for retinyl palmitate of 161 microM, whereas homogenates from control Chinese hamster ovary cells did not.

Impaired synthesis of retinol-binding protein and transthyretin in rat liver with bile duct obstruction.

To gain further insight into the protein metabolism in bile duct-obstruction, we examined the synthesis of retinol-binding protein (RBP) and transthyretin (TTR) in rats with common bile duct-ligation. In these rats, liver and plasma levels of RBP and TTR decreased markedly, whereas liver retinoid contents remained unchanged. Although there appeared no decrease in the total amount of RBP or TTR mRNA expressed in the liver, the subcellular distribution of these mRNAs changed from the membrane-bound polysome fraction to the membrane-unbound polysome fraction. This abnormal distribution recovered rapidly after biliary drainage, resulting in the subsequent recovery of the plasma RBP and TTR levels. These observations suggest that cholestasis inhibits the synthesis and secretion of RBP and TTR by disrupting the binding of their mRNAs to membrane-bound polysomes. Plasma levels of RBP and TTR might be sensitive indicators of the recovery of protein synthesis after biliary drainage in patients with obstructive biliary disorders.