Studies on metabolism of vitamin A. Absorption of retinal (vitamin A aldehyde) in rats.
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ABSTRACT:
SUBMITTER: Deshmukh DS
PROVIDER: S-EPMC1207050 | biostudies-other | 1965 Aug
REPOSITORIES: biostudies-other
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Project description:Purpose:Diabetic retinopathy (DR) is the most common complication of diabetes involving microvasculature and neuronal alterations in the retina. Previously, we reported that vitamin B12 deficiency could be an independent risk factor for DR in humans. However, the effect of vitamin B12 supplementation in experimental DR is unknown. Thus, in this study, we investigated the impact of dietary supplementation of vitamin B12 on retinal changes in diabetic rats. Methods:Diabetes was induced in 2-month-old Sprague-Dawley rats and maintained for 4 months. One group of diabetic rats were fed normal levels of vitamin B12, and one group double the quantity of vitamin B12 (50 µg/kg diet). Vitamin B12 and homocysteine levels in the plasma were analyzed with radioimmunoassay (RIA) and high-performance liquid chromatography (HPLC), respectively. At the end of 4 months of experimentation, the eyeballs were collected. Retinal changes were analyzed with hematoxylin and eosin (H&E) staining, immunoblotting, and immunofluorescence methods. Results:Dietary supplementation of vitamin B12 had no effect on food intake, bodyweight, fasting blood glucose, and plasma homocysteine levels in the diabetic rats. However, vitamin B12 supplementation prevented loss of rhodopsin, and overexpression of VEGF, and completely prevented overexpression of HIF1?, GFAP, and endoplasmic reticulum (ER) stress markers (GRP78, ATF6?, XBP1, CHOP, and caspase 12) in the diabetic rat retina. Further, vitamin B12 ameliorated apoptosis in the retina as shown with terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) and prevented retinal thinning. Conclusions:Vitamin B12 supplementation of diabetic rats appeared to be beneficial by circumventing retinal hypoxia, VEGF overexpression, and ER stress-mediated cell death in the retina. The present study adds another potential therapeutic strategy of vitamin B12 in diabetes.
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Project description:1. Female rats, raised to maturity on a vitamin A-deficient diet supplemented with retinoate as the sole source of vitamin A, conceived when allowed to mate with normal males, but the conception resulted in foetal resorption, beginning from day 14 or 15 of pregnancy. 2. Daily injections of pregnenolone or oestradiol-17beta, but not of progesterone, were fully effective, and transplantation of pituitary homografts under kidney capsules was 80% effective, in preventing resorption. 3. Unilateral ovariectomy, leading to compensatory hypertrophy of the remaining ovary, resulted in significantly fewer corpora lutea and in lower weight of the hypertrophied ovary of the retinoate-treated rats, as compared with the corresponding retinyl acetate-fed ones. 4. The activity of the enzyme system Delta(5)-3beta-hydroxy steroid dehydrogenase was significantly less in the ovaries of the retinoate-fed rats that were subjected to unilateral ovariectomy or were made pregnant, when compared with that of the corresponding controls. Also, the ovaries of the retinoate-treated rats were insensitive to both exogenous and endogenous gonadotrophin stimulus. 5. These results indicate that one of the reasons for foetal resorption in the retinoate-fed rats might be inadequate synthesis of steroid hormones such as pregnenolone and oestrogen.
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Project description:1. Mucosal cells of the small intestine obtained from rats deprived of vitamin D or given excessive amounts of the vitamin accumulated significantly more calcium than did cells from control animals. 2. Mucosal cells from vitamin D-deficient rats released less calcium than did cells from normal or hypervitaminotic D animals. 3. Studies in vivo showed that the transfer of (45)Ca from the intestine to the blood was delayed in vitamin D deficiency, but was accelerated in hypervitaminosis D. 4. The findings support the thesis that vitamin D is involved in the release of calcium rather than in its uptake by mucosal cells. 5. Further evidence is presented suggesting that uptake of calcium by intestinal mucosal cells at 0 degrees is primarily passive, whereas at 38 degrees uptake and release are effected by an active process that depends on energy derived from glycolytic activity.
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Project description:ScopeBlueberries are rich sources of bioactive polyphenols that may provide health benefits when consumed regularly, leading to their increased marketing as dietary supplements. However, the metabolic changes associated with consuming concentrated doses of purified polyphenols, as may be present in dietary supplements, are unknown, especially when considering the colonic metabolites formed. This study aimed to evaluate the pharmacokinetics of high doses of purified blueberry polyphenols.Methods and results5-month old, ovariectomized Sprague-Dawley rats are acutely dosed with purified blueberry polyphenols (0, 75, 350, and 1000 mg total polyphenols per kg body weight (bw)) and 45 Ca to measure calcium absorption. Blood and urine are collected for 48 h after dosing and phenolic metabolites measured via ultra high-pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The most prominent metabolites are colonically generated cinnamic and hippuric acids. Smaller amounts of other phenolic acids, flavonols, and anthocyanins are also detected. Most metabolites follow a dose-response relationship, though several show saturated absorption. Maximal metabolite concentrations are reached within 12 h for a majority of compounds measured, while some (e.g., hippuric acid) peaked up to 24 h post-dosing. Calcium absorption is significantly increased in the highest dose group (p = 0.03).ConclusionThese results indicate that increased doses of blueberry polyphenols induce changes in intestinal phenolic metabolism and increase calcium absorption.
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Project description:Vitamin A (VA) plays critical roles in prenatal and postnatal development; however, limited information is available regarding maternal VA metabolism during pregnancy and lactation. We investigated the impact of pregnancy and lactation on VA metabolism and kinetics in rats, hypothesizing that changes in physiological status would naturally perturb whole-body VA kinetics. Eight-week old female rats (n = 10) fed an AIN-93G diet received an oral tracer dose of 3H-labeled retinol to initiate the kinetic study. On d 21 after dosing, six female rats were mated. Serial blood samples were collected from each female rat at selected times after dose administration until d 14 of lactation. Model-based compartmental analysis was applied to the plasma tracer data to develop VA kinetic models. Our compartmental model revealed that pregnancy resulted in a gradual increase in hepatic VA mobilization, presumably to support different stages of fetal development. Additionally, the model indicates that during lactation, VA derived from dietary intake was the primary source of VA delivered to the mammary gland for milk VA secretion. During pregnancy and lactation in rats with an adequate VA intake and previous VA storage, the internal redistribution of VA and increased uptake from diet supported the maintenance of VA homeostasis.
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Project description:Consumption of polyphenols and dietary fiber as part of a normal diet is beneficial to human health. In this study, we examined whether different amounts of dietary soluble fiber (pectin) affect the absorption and metabolism of polyphenols from blackcurrant and green tea in rats. After 28 days, the rats fed blackcurrant and green tea with pectin (4 or 8%) had significantly lower body weight gain and food intake compared to the rats fed a control diet. Rats fed a blackcurrant and green tea diet with 8% pectin had significantly higher fecal nitrogen output and lower protein digestibility. No polyphenols were observed in the urine, feces and plasma of rats fed the control diet. Parent catechins and flavonols were absent in urine obtained from all diet groups. Gallocatechin glucuronide was only observed in the plasma of rats fed the blackcurrant and green tea diet without pectin. Meanwhile, epicatechin and catechin gallate were present in the feces of rats fed a blackcurrant and green tea diet with and without 4% pectin. Pectin (4 or 8%) added to the blackcurrant and green tea diet increased the plasma antioxidant capacity in rats. Inclusion of pectin in the diet altered the host absorption and metabolism of polyphenols from blackcurrant and green tea.
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Project description:Selenite is an inorganic form of selenium that has a cytotoxic effect against several human cancer cell lines: one or more selenite metabolites are considered to be responsible for its toxicity. X-ray absorption spectroscopy was used to monitor Se speciation in A549 human lung cancer cells incubated with selenite over 72 h. As anticipated, selenodiglutathione and elemental Se both comprised a large proportion of Se in the cells between 4 and 72 h after treatment, which is in accordance with the reductive metabolism of selenite in the presence of glutathione and glutathione reductase/NADPH system. Selenocystine was also present in the cells but was only detected as a significant component between 24 and 48 h concomitant with a decrease in the proportion of selenocysteine and the viability of the cells. The change in speciation from the selenol, selenocysteine, to the diselenide, selenocystine, is indicative of a change in the redox status of the cells to a more oxidizing environment, likely brought about by metabolites of selenite. X-ray fluorescence microscopy of single cells treated with selenite for 24 h revealed a punctate distribution of Se in the cytoplasm. The accumulation of Se was associated with a greater than 2-fold increase in Cu, which was colocalized with Se. Selenium K-edge extended X-ray absorption fine structure (EXAFS) spectroscopy revealed Se-Se and Se-S bonding, but not Se-Cu bonding, despite the spatial association of Se and Cu. Microprobe X-ray absorption near-edge structure spectroscopy (μ-XANES) showed that the highly localized Se species was mostly elemental Se.
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Project description:1. The enzymic activation of sulphate by various tissue extracts of vitamin A-deficient rats and their pair-fed controls was studied. 2. Vitamin A deficiency does not impair the enzymic activity in liver, colon and brain. However, a significant decrease in activity was observed in epiphyseal cartilage.
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Project description:Although currently available treatment options for age-related macular degeneration (AMD) are limited, particularly for atrophic AMD, the identification of predisposing genetic variations has informed clinical studies addressing therapeutic options such as complement inhibitors and anti-inflammatory agents. To lower risk of early AMD, recommended lifestyle interventions such as the avoidance of smoking and the intake of low glycemic antioxidant-rich diets have largely followed from the identification of nongenetic modifiable factors. On the other hand, the challenge of understanding the complex relationship between aging and cumulative damage leading to AMD has fueled investigations of the visual cycle adducts that accumulate in retinal pigment epithelial (RPE) cells and are a hallmark of aging retina. These studies have revealed properties of these compounds that provide insights into processes that may compromise RPE and could contribute to disease mechanisms in AMD. This work has also led to the design of targeted therapeutics that are currently under investigation.
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