Ectopic expression of RBP4 impairs the insulin pathway and inguinal fat deposition in mice

A large body of evidence has linked retinol-binding protein 4 (RBP4) to systemic insulin resistance, but little is known about its function in fat deposition. This study aimed to confirm the involvement of RBP4 in inguinal fat deposition and insulin by intraperitoneal injection of adenovirus-mediated RBP4 to mice. Intraperitoneal injection of adenoviral vectors was validated as an efficient gene manipulation tool for over-expressing recombinant proteins in vivo. Ectopic expression of RBP4 decelerated inguinal fat deposition by decreasing the size of adipocytes. Moreover, the introduction of exogenous RBP4 blunted the response of inguinal adipocytes to insulin signals. These findings suggest that RBP4 impaired in vivo adipogenesis, partly through the repression of the insulin pathway

Changes on metabolic parameters induced by acute cannabinoid administration (CBD, THC) in a rat experimental model of nutritional vitamin A deficiency / Cambios en parámetros metabólicos inducidos por la administración aguda de canabinoides (CBD, THC) en un modelo experimental de rata deficiente en vitamina A inducido por la dieta

INTRODUCTION: Vitamin A deficiency can result from malnutrition, malabsorption of vitamin A, impaired vitamin metabolism associated with liver disease, or chronic debilitating diseases like HIV infection or cancer. BACKGROUND & AIMS: Cannabis administration has been described as a palliative symptom management therapy in such pathological stages. Therefore, this research aimed to study the effects of acute administration of cannabidiol (CBD) or thetrahydrocannabinol (THC) on the levels of retinol in plasma and in the liver, and biochemical parameters related to lipid and glucose metabolism (cholesterolaemia, triglyceridemia and glycemia) in a rat experimental model of vitamin A deficiency. METHODS: The experimental animal model of Vitamin A deficiency was developed during a 50-day experimental period in which rats consumed a vitamin A-free diet. Cannabidiol (10 mg/kg body weight) or thetrahydrocannabinol (5 mg/kg body weight) were administered intraperitoneally 2 hours prior to sacrifice of the animals. Results: The nutritional deficiency caused a significant decrease in plasmatic and liver contents of retinol and biochemical parameters of glycemic, lipidic, and mineral metabolism. Acute intraperitoneal administration of Cannabidiol and thetrahydrocannabinol did not improve the indices of vitamin A status in either control or vitamin A-deficient rats. However, it had a significant effect on specific biochemical parameters such as glucose, triglycerides, and cholesterol. CONCLUSION: Under our experimental conditions, the reported effects of cannabinoid administration on certain signs of nutritional vitamin A deficiency appeared to be mediated through mechanisms other than changes in retinol metabolism or its mobilization after the acute administration of such compounds (AU)

Introducción: La deficiencia en vitamina A está asociada a la malnutrición, malabsorción de este nutriente, metabolismo alterado de vitaminas por enfermedad hepática, o enfermedades crónicas debilitantes como VIH, cáncer o infección. La administración de cannabis ha sido descrita como una terapia eficaz en el tratamiento sintomático de determinadas manifestaciones de la deficiencia nutricional en vitamina A y de diversas enfermedades crónicas debilitantes. Objetivos: El objetivo de este trabajo era estudiar el efecto de la administración de tetrahidrocannabinol (THC) y cannabidiol (CBD) sobre las concentraciones plasmáticas y hepáticas de retinol y sobre parámetros bioquímicos relacionados con el metabolismo glucídico y lipídico (colesterolemia, trigliceridemia, glucemia) en un modelo experimental de rata deficiente en vitamina A. Métodos: El modelo experimental de deficiencia en vitamina A se desarrolló durante un periodo experimental de 50 días en los que las ratas consumieron una dieta libre en vitamina A. La administración de tetrahidrocannabinol (THC) (10 mg/kg peso corporal) y cannabidiol (CBD) (5 mg/kg peso corporal) se llevo a cabo por vía intraperitoneal 2 horas antes del sacrificio de los animales al final del periodo experimental. Resultados: La deficiencia nutricional en vitamina A causó un descenso significativo en el contenido plasmático y hepático de retinol y en parámetros bioquímicos de metabolismo glucídico, lipídico y mineral. La administración intraperitoneal aguda de tetrahidrocannabinol y cannabidiol no mejoró los índices de estado nutricional de vitamina A en ratas deficientes o control. Sin embargo, tuvo un efecto significativo sobre parámetros bioquímicos específicos como la glucemia, colesterolemia y trigliceridemia. Conclusión: Bajo nuestras condiciones experimentales, el efecto de la administración de cannabinoides sobre determinadas manifestaciones de la deficiencia en vitamina A parece estar mediada por mecanismos no relacionados con cambios en el metabolismo de retinol o su movilización tras la administración aguda de los compuestos cannabinoides ensayados (AU)

Evidence for a specific cell membrane retinol-binding protein transport mechanism in a human keratinocyte line.

The epidermis is highly sensitive to retinoids, and vitamin A (retinol) is a critical factor in the regulation of skin cell differentiation and proliferation. Despite extensive knowledge of retinoid-mediated gene transcription effects on epidermal cells and evidence for retinoid-mediated suppression of carcinogenesis in skin, basic transport events, especially cellular uptake, of this vitamin remain poorly understood and controversial. Herein, evidence is presented for receptor-mediated uptake of retinol-binding protein, RBP, the specific circulatory vitamin A carrier, in the A431 human epidermal cell line. Cellular RBP uptake was significantly inhibited by anti-RBP IgG. Addition of transthyretin (TTR), a circulatory protein that can interact with RBP, to the internalization assay also significantly reduced RBP uptake to 49.4+/-4.6% (+/- SEM) of control values (p<0.01). RBP uptake was impaired by sucrose, a known inhibitor of early endocytosis, but not significantly affected by a disruptor of later trafficking events, chlorpromazine. Binding analysis indicated saturable RBP binding to the cell surface and a total of about 94,000 binding sites/cell. Based on dissociation constants, two RBP binding sites were detected with a 50-fold affinity difference: 0.7 and 35.0 nM, with 12,000 and 82,000 receptors/cell, respectively. These results indicate that high affinity RBP receptors capable of internalizing RBP independently of TTR exist in these malignant keratinocytes, and that TTR influences binding of RBP to its putative receptor(s). Overall, the data establish membrane transport parameters for RBP, and provide a basis for examining modulation of vitamin A endocytosis that may accompany changes in proliferation or differentiation state of epidermal cells.

Carboxyl ester lipase overexpression in rat hepatoma cells and CEL deficiency in mice have no impact on hepatic uptake or metabolism of chylomicron-retinyl ester.

To study the role of carboxyl ester lipase (CEL) in hepatic retinoid (vitamin A) metabolism, we investigated uptake and hydrolysis of chylomicron (CM)-retinyl esters (RE) by rat hepatoma (McArdle-RH7777) cells stably transfected with a rat CEL cDNA. We also studied tissue uptake of CM-RE in CEL-deficient mice generated by targeted disruption of the CEL gene. CEL-transfected cells secreted active enzyme into the medium. However, both control and CEL-transfected cells accumulated exogenously added CM-RE or CM remnant (CMR)-derived RE in equal amounts. Serum clearance of intravenously injected CM-RE and cholesteryl ester were not different between wild-type and CEL-deficient mice. Also, the uptake of the two compounds by the liver and other tissues did not differ. These data indicate that the lack of CEL expression does not affect the uptake of dietary CM-RE by the liver or other tissues. Moreover, the percentage of retinol formed in the liver after CM-RE uptake, the levels of retinol and retinol-binding protein in serum, and retinoid levels in various tissues did not differ, indicating that CEL deficiency does not affect hepatic retinoid metabolism and retinoid distribution throughout the body. Surprisingly, in both pancreas and liver of wild-type, heterozygous, and homozygous CEL-deficient mice, the levels of bile salt-dependent retinyl ester hydrolase (REH) activity were similar. This indicates that in the mouse pancreas and liver an REH enzyme activity, active in the presence of bile salt and distinct from CEL, is present, compatible with the results from our accompanying paper that the intestinal processing and absorption of RE were unimpaired in CEL-deficient mice.

Lipoprotein lipase expression level influences tissue clearance of chylomicron retinyl ester.

Approximately 25% of postprandial retinoid is cleared from the circulation by extrahepatic tissues. Little is known about physiologic factors important to this uptake. We hypothesized that lipoprotein lipase (LpL) contributes to extrahepatic clearance of chylomicron vitamin A. To investigate this, [3H]retinyl ester-containing rat mesenteric chylomicrons were injected intravenously into induced mutant mice and nutritionally manipulated rats. The tissue sites of uptake of 3H label by wild type mice and LpL-null mice overexpressing human LpL in muscle indicate that LpL expression does influence accumulation of chylomicron retinoid. Skeletal muscle from mice overexpressing human LpL accumulated 1.7- to 2.4-fold more 3H label than wild type. Moreover, heart tissue from mice overexpresssing human LpL, but lacking mouse LpL, accumulated less than half of the 3H-label taken up by wild type heart. Fasting and heparin injection, two factors that increase LpL activity in skeletal muscle, increased uptake of chylomicron [3H] retinoid by rat skeletal muscle. Using [3H]retinyl palmitate and its non-hydrolyzable analog retinyl [14C]hexadecyl ether incorporated into Intralipid emulsions, the importance of retinyl ester hydrolysis in this process was assessed. We observed that 3H label was taken up to a greater extent than 14C label by rat skeletal muscle, suggesting that retinoid uptake requires hydrolysis. In summary, for each of our experiments, the level of lipoprotein lipase expression in skeletal muscle, heart, and/or adipose tissue influenced the amount of [3H]retinoid taken up from chylomicrons and/or their remnants.

Tissue distribution of the receptor for plasma retinol-binding protein.

The tissue distribution of the retinol-binding-protein receptor has been studied by using a cell-free binding assay. High binding activity was found in placenta, retina pigment epithelial cells, bone marrow and kidneys. Specific binding activity was also found in the small intestines, spleen and liver, and to a lesser extent in lung. Scatchard analysis revealed that the difference in binding activity was due to variations in receptor level and not affinity changes. When the kidneys were separated into cortex and medulla we found that almost all the specific binding activity present in kidneys was recovered in the cortex. The choroid plexus, an important site in the delivery of nutrients to the cerebrospinal fluid, expressed very high binding activity. The pineal gland, which has been shown to store vitamin A, also showed high binding activity. Testes from immature animals showed higher binding activity than testes from mature rabbits. Cultured undifferentiated kidney keratinocytes showed about 40 times higher binding activity than differentiated cells. Skin fibroblasts demonstrated no binding activity. In conclusion, the data presented in this report show that the level of the retinol-binding-protein receptor varies considerably between cell types. The observed tissue distribution of the receptor agrees well with the present knowledge on retinol function and metabolism by various cells.

[Usefulness of vitamin A binding protein as a marker for capillary endothelial permeability].

We performed a preliminary study to assess the usefulness of Vitamin A binding protein (VABP) as a gamma-camera marker for capillary endothelial permeability. We used a guinea pig model of endotoxin (LPS) induced acute lung injury. We calculated the concentration ratio of either 125I-albumin or 125I-VABP in lung tissue to that in plasma (tissue plasma ratio; T/P) as a parameter of capillary endothelial permeability. 99mTc-diethylene triamine pentaacetic acid (DTPA) was used as a marker for pulmonary interstitial volume. We estimated wet to dry lung weight ratio as a parameter of lung water accumulation (W/D). LPS increased the T/P of 125I-albumin and W/D, suggesting the development of permeability edema. The T/P for 125I-VABP was also increased, indicating that 125I-VABP can be used to detect elevated capillary endothelial permeability. In both groups, LPS and saline, the T/P was higher for 125I-VABP than for 125I-albumin. These data suggest that the pulmonary capillary endothelium is more permeable to VABP than albumin.