Hepatic Retinol Concentrations Research Articles

Nanoparticles Loaded with a Carotenoid-Rich Extract from Cantaloupe Melon Improved Hepatic Retinol Levels in a Diet-Induced Obesity Preclinical Model.

The study evaluated the effect of the carotenoid-rich extract from cantaloupe melon (CE) nanoencapsulated in porcine gelatin (EPG) on hepatic retinol concentration and liver damage scores in Wistar rats with obesity induced by high glycemic index and high glycemic load diet (HGLI diet). For 17 days, animals were fed the HGLI diet. They were divided into three groups and treated for 10 days [HGLI diet + water, HGLI diet + CE (12.5 mg/kg), and HGLI diet + EPG (50 mg/kg)]. The groups were evaluated for dietary intake, retinol, weight variation, hematological parameters, fasting glucose, lipid profile, hepatic retinol concentration, AST/ALT ratio, FIB-4 (Fibrosis-4 Index for Liver Fibrosis), and APRI (AST to Platelet Ratio Index) scores to evaluate the effects on the liver. Animals treated with EPG showed a lower dietary intake (p < 0.05). No significant weight change was detected in the evaluated groups (p > 0.05). The EPG-treated group had significantly higher concentrations (p < 0.05) of hepatic retinol [266 (45) μg/g] than the untreated group [186 (23.8) μg/g] and the one treated with CE [175 (8.08) μg/g]. Liver damage assessment scores did not show significant differences, but the lowest means were observed in the group treated with EPG. The nanoencapsulation of the extract rich in beta-carotene promoted reduced food consumption and increased hepatic retinol without causing significant changes in liver damage scores. Thus, EPG is a candidate for future clinical studies to evaluate the beneficial effects of treating diseases involving vitamin A deficiencies.

β-Cryptoxanthin Attenuates Cigarette-Smoke-Induced Lung Lesions in the Absence of Carotenoid Cleavage Enzymes (BCO1/BCO2) in Mice.

High dietary intake of β-cryptoxanthin (BCX, an oxygenated provitamin A carotenoid) is associated with a lower risk of lung disease in smokers. BCX can be cleaved by β-carotene-15,15'-oxygenase (BCO1) and β-carotene-9',10'-oxygenase (BCO2) to produce retinol and apo-10'-carotenoids. We investigated whether BCX has protective effects against cigarette smoke (CS)-induced lung injury, dependent or independent of BCO1/BCO2 and their metabolites. Both BCO1-/-/BCO2-/- double knockout mice (DKO) and wild type (WT) littermates were supplemented with BCX 14 days and then exposed to CS for an additional 14 days. CS exposure significantly induced macrophage and neutrophil infiltration in the lung tissues of mice, regardless of genotypes, compared to the non-exposed littermates. BCX treatment significantly inhibited CS-induced inflammatory cell infiltration, hyperplasia in the bronchial epithelium, and enlarged alveolar airspaces in both WT and DKO mice, regardless of sex. The protective effects of BCX were associated with lower expression of IL-6, TNF-α, and matrix metalloproteinases-2 and -9. BCX treatment led to a significant increase in hepatic BCX levels in DKO mice, but not in WT mice, which had significant increase in hepatic retinol concentration. No apo-10'-carotenoids were detected in any of the groups. In vitro BCX, at comparable doses of 3-OH-β-apo-10'-carotenal, was effective at inhibiting the lipopolysaccharide-induced inflammatory response in a human bronchial epithelial cell line. These data indicate that BCX can serve as an effective protective agent against CS-induced lung lesions in the absence of carotenoid cleavage enzymes.

Vitamin A deficiency during the perinatal period induces changes in vitamin A metabolism in the offspring. The regulation of intestinal vitamin A metabolism via ISX occurs only in male rats severely vitamin A-deficient.

1) To test the hypothesis of the existence of a perinatal vitamin A (VA) programming of VA metabolism and to better understand the intestinal regulation of VA metabolism. Offspring from rats reared on a control (C) or a VA-deficient (D) diet from 6weeks before mating until offspring weaning, i.e., 7weeks after mating, were themselves reared on a C or D diet for 19weeks, resulting in the following groups: C-C (parents fed C-offspring fed C), D-C, C-D and D-D. VA concentrations were measured in plasma and liver. β-Carotene bioavailability and its intestinal conversion rate to VA, as well as vitamin D and E bioavailability, were assessed after gavages with these vitamins. Expression of genes involved in VA metabolism and transport was measured in intestine and liver. C-D and D-D had no detectable retinyl esters in their liver. Retinolemia, hepatic retinol concentrations and postprandial plasma retinol response to β-carotene gavage were higher in D-C than in C-C. Intestinal expression of Isx was abolished in C-D and D-D and this was concomitant with a higher expression of Bco1, Scarb1, Cd36 and Lrat in males receiving a D diet as compared to those receiving a C diet. β-Carotene, vitamin D and E bio-availabilities were lower in offspring receiving a D diet as compared to those receiving a C diet. A VA-deficient diet during the perinatal period modifies the metabolism of this vitamin in the offspring. Isx-mediated regulation of Bco1 and Scarb1 expression exists only in males severely deficient in this vitamin. Severe VA deficiency impairs β-carotene and vitamin D and E bioavailability.

High dietary fat–induced obesity in Wistar rats and type 2 diabetes in nonobese Goto-Kakizaki rats differentially affect retinol binding protein 4 expression and vitamin A metabolism

Obesity is a major risk factor for type 2 diabetes, which is caused mainly by insulin resistance. Retinol binding protein 4 (RBP4) is the only specific transport protein for retinol in the serum. RBP4 level is increased in the diabetic state and high-fat condition, indicating that retinol metabolism may be affected under these conditions. However, the precise effect of diabetes and high fat–induced obesity on retinol metabolism is unknown. In this study, we examined differences in retinol metabolite levels in rat models of diet-induced obesity and type 2 diabetes (Goto-Kakizaki [GK] rat). Four-week-old male Wistar and GK rats were given either a control diet (AIN-93G) or a high-fat diet (HFD, 40% fat kJ). After 15 weeks of feeding, the RBP4 levels increased by 2-fold in the serum of GK rats but not HFD-fed rats. The hepatic retinol concentration of HFD-fed rats was approximately 50% that of the controls (P < .01). In contrast, the renal retinol concentrations of GK rats increased by 70% (P < .01). However, expression of RARβ in the kidney, which was induced in a retinoic acid–dependent manner, was downregulated by 90% (P < .01) in GK rats. In conclusion, diabetes and obesity affected retinol metabolism differently, and the effects were different in different peripheral tissues. The impact of HFD may be limited to the storage of hepatic vitamin A as retinyl palmitate. In particular, our data indicate that renal retinoic acid production might represent an important target for the treatment of type 2 diabetes mellitus.

Vitamin A Deficiency and Hepatic Retinol Levels in Sea Otters, Enhydra lutris

Vitamin A deficiency has rarely been reported in captive or free-ranging wildlife species. Necropsy findings in two captively housed southern sea otters (Enhydra lutris nereis) included irregular thickening of the calvaria characterized by diffuse hyperostoses on the internal surface. One animal also had moderate squamous metaplasia of the seromucinous glands of the nose. There was no measurable retinol in the liver of either sea otter. For comparison, hepatic retinol concentration was determined for 23 deceased free-ranging southern and northern (Enhydra lutris kenyoni) sea otters from California and Alaska. Free-ranging otters were found to have similar hepatic retinol concentrations (316 +/- 245 mg/kg wet weight) regardless of their location and subspecies. All of these values were significantly higher than the levels in the affected animals. Consumption of a diet with very low vitamin A concentrations and noncompliance in daily supplementation are hypothesized as the causes of vitamin A deficiency in these two sea otters.

Altered hepatic retinol and CYP26 levels in adult European common frogs (Rana temporaria) exposed to p,p′-DDE

The effects of environmental contamination on amphibians are of particular concern because there are reports of declining numbers of species and individuals in most parts of the world during the last 50 years. During the last decade there has been increased focus on the role of persistent organic pollutants as retinoid (vitamin A) disrupters, and their effects on development, growth and sexual differentiation. To study the effects of p,p'-DDE, one of the most persistent metabolites of the pesticide DDT, on retinol homeostasis, we subcutaneously exposed adult male European common frogs (Rana temporaria) to different doses of p,p'-DDE (0, 0.01, 0.1, 1 and 10 mg/kg body mass) and studied the effect of a short term exposure (14 days) on hepatic retinoid levels and CYP26 gene and protein expression. Hepatic retinol concentrations, CYP26 gene and protein levels were analysed using HPLC, quantitative RT-PCR and indirect ELISA, respectively. Our results showed a significant p,p'-DDE dose-specific increase in the hepatic retinol concentration. CYP26 gene and protein expression were reduced in an apparent p,p'-DDE dose-specific manner. The results suggest that p,p'-DDE may interfere with the hepatic metabolism of retinol in adult frogs by decreasing CYP26 expression patterns.

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.

Dose-responsive alteration in hepatic lipid peroxidation and retinol metabolism with increasing dietary beta-carotene in iron deficient rats.

Phosphatidylcholine hydroperoxide (PCOOH) levels are increased in the iron-deficient rat liver. We investigated the antioxidative effect of dietary beta-carotene and altered retinol metabolism in iron-deficient rats. Experiment 1: Male Wistar-strain rats were divided into six groups and fed a control diet, an iron-deficient diet, and iron-deficient diets with four different levels of dietary beta-carotene. The PCOOH concentration in the iron-deficient rat liver was decreased by supplementation with dietary beta-carotene. However, the beta-carotene dose response was not related to antioxidative potency. Hepatic and plasma beta-carotene concentrations were increased by iron deficiency. The hepatic retinol concentration was increased while the plasma retinol concentration was decreased in iron-deficient rats. Experiment 2: Male Wistar-strain rats were divided into two groups, with one group receiving a control diet with beta-carotene and the other an iron-deficient diet with beta-carotene. Intestinal iron was decreased and intestinal beta-carotene was unchanged in iron-deficient rats. The intestinal beta-carotene conversion ratio and beta-carotene cleavage enzyme activity were decreased in iron-deficient rats. Dietary beta-carotene played the role of an antioxidant in hepatic lipid peroxidation in the iron-deficient state, but there was no dose dependency. Moreover, intestinal beta-carotene cleavage and hepatic retinol release appear to be altered in iron-deficient rats.

Beta-carotene, retinol and alpha-tocopherol status in calves fed the first colostrum at 0-2, 6-7, 12-13 or 24-25 hours after birth.

Effects on beta-carotene, retinol and alpha-tocopherol status of feeding 1st colostrum at 0-2, 6-7, 12-13 and 24-25 h after birth were studied in calves. beta-carotene, retinol and alpha-tocopherol concentrations decreased in colostrum during the first 2.5 d of lactation. Plasma beta-carotene, retinol and alpha-tocopherol concentrations in newborn calves were very low. Plasma beta-carotene concentrations increased up to d 3 after the 1st meal and during the 1st month were higher in calves fed 1st colostrum at < 6-7 h than at > 12-13 h after birth. Plasma retinol concentrations increased up to d 5 after the 1st meal and were higher during the 1st month in calves fed 1st colostrum at < 12-13 h than at > 24-25 h after birth, whereas hepatic concentrations increased up to d 5 independent of time of 1st colostrum feeding. Plasma alpha-tocopherol concentrations increased after the 1st meal except in calves fed 1st colostrum at 24-25 h after birth and were higher during the 1st month in calves fed the 1st colostrum at 6-7 h than at 24-25 h after birth. In conclusion, delaying 1st colostrum intake by more than 12-13 h after birth impaired the plasma beta-carotene, retinol and alpha-tocopherol status during the 1st month of life, but did not negatively influence hepatic retinol concentrations.

Reproductive success and biochemical effects in tree swallows(Tachycineta bicolor)exposed to chlorinated hydrocarbon contaminants in wetlands of the great lakes and st. lawrence river basin, USA and Canada

AbstractReproduction and biochemical indicators (vitamin A, ethoxyresorufin‐O‐deethylase [EROD], highly carboxylated porphyrins) of contaminant effects, and concentrations of organochlorine residues were measured in tree swallows from wetlands within the watersheds of the Great Lakes and the St. Lawrence River, USA and Canada. To assess the vitamin A sources in the tree swallow diets, insects were collected, by ligature, from nestlings at four of these sites. Diet samples were analyzed for retinol and α‐ and β‐carotene. Marked differences were found in chlorinated hydrocarbon residues in eggs and nestlings among sites; however, we did not find any significant differences in hatching or fledging success. The maximum polychlorinated biphenyl (PCB) concentration found was 11.1 μg/g in eggs from Akwesasne Reserve, New York, USA, on the St. Lawrence River, whereas the highest p, p′‐DDE concentration of 2.57 μg/g was found in eggs from Mud Creek, Ontario, Canada, in the Lake Erie watershed. Concentrations of other organochlorine pesticides and chlorobenzenes were low and not variable among sites. Significant differences in hepatic EROD activity, retinol and retinyl palmitate, and uroporphyrin in nestling birds existed among sites. EthoxyresorufinO‐deethylase activity was highest and retinol and retinyl palmitate were lowest at Cornwall Island, Ontario, Canada, in the St. Lawrence River, whereas porphyrins were highest at Toronto and Hamilton harbors, Ontario, Canada, in Lake Ontario. EthoxyresorufinO‐deethylase induction was significantly and negatively correlated with nestling hepatic retinol concentrations, whereas uroporphyrin concentrations were positively correlated with PCB congener 118 in nestlings. In contrast, few differences occurred in vitamin A concentrations in diet samples among sites. Together, these findings suggest that depressed hepatic vitamin A and elevated highly carboxylated porphyrin concentrations are strongly associated with sites with the highest organochlorine exposure in tree swallows.

Alterations in plasma and hepatic retinoid levels in flounder ( Platichthys flesus) after chronic exposure to contaminated harbour sludge in a mesocosm study

Flounder ( Platichthys flesus) were exposed to polluted harbour sludge or to sludge wash-off in mesocosms for 3 years. At the end of the study, flounder from all mesocosms were sacrificed, and plasma and liver were isolated for determination of retinoid levels. In addition, effects of harbour sludge on liver tumour development and skin infections, on spermatogenesis and testicular steroidogenesis, on plasma steroid levels and on induction of the CYP1A enzyme system were determined in flounder. Furthermore, levels of heavy metals and PHAHs were determined in both flounder and sediment. Here, we present the effects on retinoid levels in flounder. Data on other parameters are presented elsewhere. Retinol concentrations in flounder plasma and liver were significantly reduced in mesocosms containing polluted harbour sludge and sludge wash-off as compared to the control mesocosm. Retinoid stores in the liver, measured as the concentration of retinyl palmitate, were also reduced significantly in the mesocosm containing sludge wash-off as compared to the control mesocosm but not in the directly polluted mesocosm. A negative non-linear association was found between hepatic retinol concentrations and CYP1A protein levels ( r 2=0.69; P<0.05), suggesting the involvement of PHAH and PAH inducible enzymes. The present study clearly indicates that long-term exposure to environmental contaminants in complex matrices decreases retinoid levels in flounder.

Fetal, Neonatal, and Long-Term Alterations in Hepatic Retinoid Levels Following Maternal Polychlorinated Biphenyl Exposure in Rats

This study was undertaken to investigate the effects of perinatal polychlorinated biphenyl (PCB; Aroclor 1254) exposure on hepatic and plasma retinoid levels in fetal rats, their dams, and neonatal and adult offspring. Pregnant Wistar rats were treated with 0, 5, or 25 mg Aroclor 1254/kg body wt from Days 10 to 16 of gestation. Hepatic retinoid (retinol, retinyl palmitate, and retinyl stearate) levels were determined in fetuses and dams from Day 20 of gestation, in male and female neonates 21 days postpartum, and in young adult offspring 90 days after birth. Retinol levels were determined in fetal and maternal plasma (Gestation Day 20) and plasma from the offspring 21 and 90 days after birth. Maternal and fetal plasma retinol levels were decreased by 35 and 38% on Day 20 of gestation following exposure to the highest dose of Aroclor 1254. Male, but not female, neonatal plasma retinol levels were significantly decreased (23%) in the high-dose group. No effects of PCB treatment were seen on plasma retinol levels in the offspring 90 days after birth. Only slight reductions in fetal and maternal hepatic retinol and retinyl palmitate concentrations were observed after prenatal PCB exposure. Male neonatal hepatic retinyl palmitate levels were reduced by 25 and 50% in the 5 and 25-mg Aroclor 1254/kg dose groups, respectively, while female neonatal hepatic retinyl palmitate levels were significantly reduced only in the high-dose group. Ninety days after birth, male hepatic retinyl palmitate levels were only slightly reduced in the highest dose group; however, hepatic retinol concentrations were significantly reduced by 50% in both PCB treatment groups. Female adult offspring exhibited significant reductions in hepatic retinyl palmitate levels (25%) in both PCB treatment groups, while hepatic retinol levels exhibited an unusual increase of more than 100% of controls in the low-dose group, while levels in the high-dose group were similar to controls. This study demonstrates that even a relatively low maternal dose of Aroclor 1254 results in long-term alterations in retinoid status of the offspring in the rat.

Abnormal antioxidant defence in some tissues of congenitally obese mice.

The concentration of lipoperoxides (estimated as thiobarbituric acid-reactive material) and some components of the antioxidant defence system have been compared in various tissues of lean and congenitally obese mice. NADPH-stimulated lipoperoxide generation in vitro was significantly higher in microsomes (microsomal fractions) prepared from obese hepatic tissue than lean. Plasma, liver and brain lipoperoxide concentration was significantly higher in obese mice. In blood derived from obese mice the concentration of non-enzymic antioxidants including caeruloplasmin and vitamin A was higher, but hepatic retinol concentration was lower in these animals. In all the tissues assayed the glutathione peroxidase activity against H2O2 was less than its activity against cumene hydroperoxide. Assayed with either substrate, glutathione peroxidase activity was significantly higher in the brain and blood of obese mice than their lean counterparts. Conversely, liver glutathione peroxidase was decreased in obese animals, representing 43% of the activity of the lean-mouse liver enzyme against H2O2 and 81% of the cumene hydroperoxide-reducing activity. The liver of obese mice had significantly less, and the kidneys more, oxidized glutathione than the corresponding tissues of lean mice. Further investigations on hepatic tissue indicated that glutathione reductase activity was lower in the obese animals, but there was no significant difference between glucose-6-phosphate dehydrogenase activity in obese and lean mice.