Vitamin A deficiency modifies lipid metabolism in rat liver

Liver fatty acid metabolism of male rats fed on a vitamin A-deficient diet for 3 months from 21 d of age was evaluated. Vitamin A restriction produced subclinical plasma and negligible liver retinol concentrations, compared with the control group receiving the same diet with 4000 IU vitamin A (8 mg retinol as retinyl palmitate)/kg diet. Vitamin A deficiency induced a hypolipidaemic effect by decreasing serum triacylglycerol, cholesterol and HDL-cholesterol levels. The decrease of liver total phospholipid was associated with low phosphatidylcholine synthesis observed by lower [14C]choline incorporation into phosphatidylcholine, compared with control. Also, liver fatty acid synthesis decreased, as was indicated by activity and mRNA expression of acetyl-CoA carboxylase (ACC), and incorporation of [14C]acetate into saponified lipids. A decrease of the PPARalpha mRNA expression was observed. Liver mitochondria of vitamin A-deficient rats showed a lower total phospholipid concentration coinciding with a decrease of the cardiolipin proportion, without changes in the other phospholipid fractions determined. The mitochondria fatty acid oxidation increased by 30 % of the control value and it was attributed to a high activity and mRNA expression of carnitine palmitoyltransferase-I (CPT-I). An increase in serum beta-hydroxybutyrate levels was observed in vitamin A-deficient rats. Vitamin A deficiency alters the mitochondria lipid composition and also enhances fatty acid oxidation by modifying the production of malonyl-CoA, the endogenous inhibitor of CPT-I, due to decreased activity of liver ACC. The incorporation of vitamin A into the diet of vitamin A-deficient rats reverted all the changes observed.     

Some effects of vitamin A deficiency on the isolated rat lung alveolar type II cell.

Alveolar Type II cells were isolated from control and vitamin A deficient rats and allowed to form a monolayer in plastic dishes for 16-18 hours. The vitamin A content (retinol plus retinyl palmitate) of deficient cells was 50-75% less than in control cells on a per mg protein basis. Isolated Type II cells took up [3H]-retinol, synthesized [3H]-retinyl palmitate, and after 4 hours, 24% of the radioactivity in the Type II cells was [3H]-retinoic acid. Deficiency did not appear to alter retinoic acid synthesis. Phosphatidylcholine (PC) and disaturated phosphatidylcholine (DSPC) synthesis, were slightly less in deficient cells compared to control (95 and 85% respectively). In addition, 10(-6) M and 10(-5) M retinoic acid in the reaction media stimulated both PC and DSPC synthesis by 120-140% in control cells. The stimulating effect of retinoic acid was present in deficient cells as well, but less pronounced (120% with 10(-5) M). Vitamin A deficient Type II cells also had less basal levels of both tissue transglutaminase and epidermal transglutaminase activity than control cells.     

Effects of vitamin A deficiency on mitochondrial function in rat liver and heart

The aim of this study was to investigate comparative effects of vitamin A deficiency on respiratory activity and structural integrity in liver and heart mitochondria. Male rats were fed a liquid control diet (control rats) or a liquid vitamin A-deficient diet (vitamin A-deficient rats) for 50 days. One group of vitamin-A deficient rats was refed a control diet for 15 days (vitamin A-recovered rats). To assess the respiratory function of mitochondria the contents of coenzyme Q (ubiquinone, CoQ), cytochrome c and the activities of the whole electron transport chain and of each of its respiratory complexes were evaluated. Chronic vitamin A deficiency promoted a significant increase in the endogenous coenzyme Q content in liver and heart mitochondria when compared with control values. Vitamin A deficiency induced a decrease in the activity of complex I (NADH-CoQ reductase) and complex II (succinate-CoQ reductase) and in the levels of complex I and cytochrome c in heart mitochondria. However, NADH and succinate oxidation rates were maintained at the control levels due to an increase in the CoQ content in accordance with the kinetic behaviour of CoQ as an homogeneous pool. On the contrary, the high CoQ content did not affect the electron-transfer rate in liver mitochondria, whose integrity was preserved from the deleterious effects of the vitamin A deficiency. Ultrastructural assessment of liver and heart showed that vitamin A deficiency did not induce appreciable alterations in the morphology of their mitochondria. After refeeding the control diet, serum retinol, liver and heart CoQ content and the activity of complex I and complex II in heart mitochondria returned to normality. However, the activities of both whole electron transfer chain and complex I in liver were increased over the control values. The interrelationships between physiological antioxidants in biological membranes and the beneficial effects of their administration in mitochondrial diseases are discussed.     

Vitamin A deficiency impairs fetal islet development and causes subsequent glucose intolerance in adult rats

To determine the role of vitamin A in fetal islet development, beta- and alpha-cell mass, apoptosis, and alpha- and beta-cell replication were measured in rats using a model of marginal vitamin A deficiency. Female rats before and during pregnancy and their offspring postweaning were fed a diet containing retinol as retinyl palmitate at a low marginal (LM, 0.25 mg/kg diet) or a sufficient (SUFF, 4.0 mg/kg diet) level. Fetal islet size, replication, apoptosis, and offspring glucose tolerance were examined. Both beta-cell area and number per islet were reduced approximately 50% in fetuses from dams fed an LM vitamin A diet compared with those from dams fed the SUFF vitamin A diet. The alpha-cell area and number per fetal islet were not affected by vitamin A deficiency. Apoptosis was not increased. The percentage of newly replicated beta-cells in the LM fetal pancreas was 42% less than that of SUFF offspring, but alpha-cell replication was not affected. To determine whether this decrease in beta-cell area affected adult glucose tolerance and insulin secretion, 65-d-old offspring were subject to glucose tolerance tests. LM rats had a 55% lower plasma insulin level and a 76% higher serum glucose than SUFF rats. The same pattern could be seen in 35-d-old rats. These findings show that vitamin A deficiency decreases beta-cell mass and this reduction can be attributed to a reduced rate of fetal beta-cell replication in LM offspring. This may contribute to impaired glucose tolerance later in adult life.     

Vitamin A deficiency increases hepcidin expression and oxidative stress in rat

ObjectiveThe interaction between vitamin A and iron status has been widely reported; however, the exact mechanism involved in this interaction has not been well characterized. The present study investigated the mechanism involved in tissue iron accumulation and changes in the oxidative status in vitamin A–deficient rats.MethodsRats were treated with a control diet, a vitamin A–deficient diet, or a vitamin A/iron-deficient diet for 57 d. The animals were sacrificed; the blood, liver, and spleen were collected for biochemical analysis. Analysis of variance or Mann-Whitney tests were used to compare groups and Pearson's or Spearman's tests to investigate the bivariate correlation.ResultsVitamin A deficiency increased liver hepcidin mRNA and iron spleen concentrations; however, iron deficiency in vitamin A–deficient rats deeply inhibits liver hepcidin mRNA expression and significantly increases divalent metal transporter-1 mRNA levels. Liver ferroportin and hereditary hemochromatosis gene mRNA levels did not change in either treatment. In the vitamin A–deficient groups, liver carbonyl protein increased, whereas catalase and glutathione S-transferase activities decreased, suggesting that vitamin A protects the liver against protein oxidation. A significant positive correlation was found between lipid oxidative damage and iron concentration in the liver and spleen (r = 0.611, P = 0.007; r = 0.558, P = 0.025, respectively).ConclusionThese results suggest that vitamin A maintains iron homeostasis by the modulation of liver hepcidin expression. The increase of lipid peroxidation in vitamin A deficiency seems to be iron dependent, whereas protein oxidation is not.     

Vitamin B-6 deficiency impairs interleukin 2 production and lymphocyte proliferation in elderly adults

The effect of vitamin B-6 deficiency on immune response was studied in eight healthy elderly adults. The protocol consisted of a 5-d baseline (BL) period; a vitamin B-6-depletion period of less than or equal to 20 d; three stages of vitamin B-6-repletion, each lasting 21 d; and a 4-d final phase. The amounts of vitamin B-6 ingested during the different phases of the study were 3.00, 15.00, 22.50, and 33.75 micrograms.kg body wt-1.d-1, respectively. During the final phase the subjects ingested 50 mg vitamin B-6/d. Fasting blood was collected at the end of each period. Vitamin B-6 depletion significantly decreased percentage and total number of lymphocytes, mitogenic responses of peripheral blood lymphocytes to T- and B-cell mitogens, and interleukin 2 production. These indices returned to BL values after the third vitamin B-6-repletion period, when the total vitamin B-6 intakes were 1.90 +/- 0.18 mg/d for women and 2.88 +/- 0.17 mg/d for men. Vitamin B-6 deficiency impairs in vitro indices of cell-mediated immunity in healthy elderly adults. This impairment is reversible by vitamin B-6 repletion.     

Vitamin E deficiency and prostaglandin synthesis in rat testes

A group of 8 weanling rats was fed a vitamin E(VE) deficient diet (D) and a control group was fed a VE sufficient diet (C) for 19 weeks. Levels of docosapentaenoic acid (22:5w6) in testis parenchyma lipids were significantly lower (p<0.05) in group D compared with group C. This decrease in 22:5w6, however, was not accompanied by the documented decrease in total testis weight of the VE deficient group. These results suggest that the 22:5w6 level in testis parenchyma lipids may be a sensitive indicator of VE deficiency in rats. There was no difference in the arachidonic acid (AA) level of testis parenchyma lipids between two groups. The tunica was the major site of prostaglandin (PG) synthesis in the testicular tissue and prostacyclin was the major PG synthesized from exogenous AA in the tunica. Both PGE₂ and 6-keto-PGF_1α derived from endogenous AA in tunica homogenate affer 30 min incubation appear to be greater in the control group than in the deficient group, although the difference was not statistically significant. These results imply that increased lipid peroxidation in VE deficiency can result in greater inactivation of PGH synthetase and thus can reduce the synthesis of PGs during the incubation.     

Vitamin A deficiency in patients with hepatitis C virus-related chronic liver disease

Hepatitis C virus (HCV) infection is associated with oxidative stress and vitamin A possesses antioxidant activity. The objective of the present study was to investigate vitamin A nutritional status in chronic hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC), according to biochemical, functional and dietetic indicators correlating these findings with liver function, liver damage and death. Vitamin A nutritional status was analysed by serum retinol levels, dietetic indicators and functional indicators. A total of 140 patients with HCV-related liver disease were enrolled. Vitamin A deficiency was detected in 54·3 % of all patients, and there was a progressive drop in serum retinol levels from chronic hepatitis C patients towards cirrhosis and HCC patients. Increased total bilirubin, liver transaminases and prothrombin time, presence of hepatic encephalopathy and ascites were related to reduced serum retinol levels, and values ≤ 0·78 μmol/l of serum retinol were associated with liver-related death. A high prevalence of inadequate intake of vitamin A was observed in all stages of chronic liver disease. The functional indicator was not an adequate parameter for evaluating the vitamin A nutritional status. Therefore, serum retinol concentration is related to severity of the disease, liver complications and mortality. The effectiveness of nutritional counselling and measures of intervention in this group in improving vitamin A nutritional status should be examined further in a controlled study.     

Biotin deficiency inhibits heme synthesis and impairs mitochondria in human lung fibroblasts

Four of the 5 biotin-dependent carboxylases (BDC) are in the mitochondria. BDC replace intermediates in the Krebs [tricarboxylic acid (TCA)] cycle that are regularly removed for the synthesis of key metabolites such as heme or amino acids. Heme, unlike amino acids, is not recycled to regenerate these intermediates, is not utilized from the diet, and must be synthesized in situ. We studied whether biotin deficiency (BD) lowers heme synthesis and whether mitochondria would be disrupted. Biotin-deficient medium was prepared by using bovine serum stripped of biotin with charcoal/dextran or avidin. Biotin-deficient primary human lung fibroblasts (IMR90) lost their BDC and senesced before biotin-sufficient cells. BD caused heme deficiency; there was a decrease in heme content and heme synthesis, and biotin-deficient cells selectively lost mitochondrial complex IV, which contains heme-a. Loss of complex IV, which is part of the electron transport chain, triggered oxidant release and oxidative damage, hallmarks of heme deficiency. Restoring biotin to the biotin-deficient medium prevented the above changes. Old cells were more susceptible to biotin shortage than young cells. These findings highlight the biochemical connection among biotin, heme, and iron metabolism, and the mitochondria, due to the role of biotin in maintaining the biochemical integrity of the TCA cycle. The findings are discussed in relation to aging and birth defects in humans.     

Vitamin A and lung cancer

In a dozen case-control and cohort studies, high intake of fruits and vegetables containing carotenoids has been associated with a reduced risk of lung cancer. In contrast, little relation has been found between intake of preformed vitamin A and this disease. Although initial studies suggested that persons with lower levels of serum retinol have higher future rates of lung cancer, this idea was not confirmed in subsequent investigations. Prediagnostic levels of beta-carotene in blood, however, have been inversely related with risk of lung cancer. Available data thus strongly support the hypothesis that dietary carotenoids reduce the risk of lung cancer, but the data are also compatible with the possibility that some other factor in these foods is responsible for the lower risk. Even if ultimately shown to be casual, the relation between diet and lung cancer is modest compared with the deleterious effect of cigarette smoking.     

Iron deficiency impairs protein synthesis in immune tissues of rat pups

To determine if decreased protein synthesis is a factor in reduced immunocompetence of iron deficiency, RNA, DNA and in vitro protein synthesis were measured. Rats were fed diets containing 6 (severe anemia), 11 (moderate anemia) or 250 (iron sufficient) mg iron/kg diet throughout gestation and lactation. On d 2 of lactation, litters were adjusted to contain six pups. On d 12 of lactation, two pups from each litter were immunized with sheep red blood cells (SRBC) and on d 17, tissues were removed for the determination of protein synthesis and evaluation of RNA and DNA contents. In the moderately iron-deficient pups, protein synthesis was lower (30%) in spleen than that in iron-sufficient pups. Protein synthesis in liver and thymus was not changed by moderate iron deficiency. In spleen, liver and thymus, protein synthesis in severely iron-deficient pups was less than half that of iron-sufficient pups. Protein synthesis in the spleen of the moderately iron-deficient group was higher after immunization with SRBC than in iron-sufficient controls, whereas the severely iron-deficient pups failed to respond. Impaired protein synthesis may be the mechanism responsible for compromised ability to produce antibody in iron deficiency.     

Vitamin A deficiency and the metabolism of glycosaminoglycans and ascorbic acid in the rat.

Weanling rats were fed diets with and without the addition of retinyl palmitate at 6,500 units/kg. The supplemented groups were fed either ad libitum or food was restricted daily to that amount consumed by the group of rats receiving the unsupplemented diet. After a 10 week experimental period, signs of vitamin A deficiency were observed (growth plateau, xerophthalmia) and liver values as retinol were only 1% of control values. Relative to the two control groups, vitamin A deficiency resulted in approximately 30% lower liver, 50% lower blood and 40% lower urinary ascorbic acid. Vitamin A deficiency did not appear to result in significant and direct impairment of GAG sulfate metabolism. Although the total amount of GAG in rat skin was increased, the composition of GAG fractions did not appear to be altered by vitamin A deficiency. Studies regarding the incorporation and disappearance of 35S-sulfate in vivo into GAG fractions obtained from skin indicated no serious impairment in GAG turnover with vitamin A deficiency. Twenty-four hour urine samples were also collected for estimation of 35SO4 excreted in GAG and non-GAG fractions. Likewise, little change was observed with respect to radioactivity associated with sulfate fractions excreted in urine. Although many previous studies have directly linked vitamin A with sulfation of GAG, the results reported here suggest that if there is an alteration in GAG sulfate metabolism, it is probably an indirect consequence of vitamin A status.     

Vitamin A deficiency alters the structure and collagen IV composition of rat renal basement membranes

Retinoids can modulate the expression of extracellular matrix (ECM) proteins with variable results depending on other contributing factors. Because changes in these proteins may alter the composition and impair the function of specialized ECM structures such as basement membranes (BMs), we studied the effects of vitamin A deficiency on renal BMs during the growing period. Newborn male rats were fed a vitamin A-deficient (VAD) diet for 50 d. The ultrastructure of renal BMs was analyzed by electron microscopy. Total collagen IV, the different alpha(IV) chains, matrix degrading metalloproteinases (MMP), and tissue inhibitors of metalloproteinases (TIMP) were quantified by immunocytochemistry and/or Western blotting. Tumor necrosis factor-alpha and interleukin-1beta were measured by ELISA. Semiquantitative RT-PCR was used for determining the steady-state levels for each alpha(IV) chain mRNA. VAD renal BMs showed an irregular thickening, particularly tubular BM. The total collagen IV content was increased, but there was a differential expression of the collagen IV chains. The protein amounts for alpha1(IV), alpha4(IV), and alpha5(IV) were similarly increased, whereas alpha2(IV) and alpha3(IV) were decreased. The levels of mRNA for each collagen IV chain changed in parallel with those of the corresponding protein. Both MMP2 and MMP9 were diminished, but no change was detected in TIMP1 or TIMP2. Our data indicate that nutritional VAD leads to alterations in the structure of renal BMs and to quantitative and qualitative variations in its collagen IV composition. These changes may be a factor predisposing to or resulting in kidney malfunction and renal disease.     

Effect of zinc and copper deficiency on microsomal NADPH-dependent active oxygen generation in rat lung and liver

Both dietary zinc and copper deficiencies can cause lipid peroxidation in microsomes in rats. The cytochrome P-450 enzyme system can generate active oxygens by uncoupling of the P-450-oxy complex in the catalytic cycle and/or the electron transfer mediated by the NADPH-cytochrome P-450 reductase. The effects of dietary zinc and copper deficiencies on NADPH-dependent H2O2 generation, the catalytic activity of the cytochrome P-450 enzyme with aminopyrine as the substrate and the activity of NADPH-cytochrome P-450 reductase were determined. Zinc deficiency caused increased H2O2 production, increased NADPH-cytochrome P-450 reductase activity, decreased aminopyrine demethylation and two- and fivefold increases in iron concentration in lung and liver microsomes, respectively, compared to Zn-adequate, ad libitum--fed controls. Active oxygen generation by uncoupling of the cytochrome P-450 enzyme system and accumulation of iron are thus possible mechanisms by which zinc deficiency causes microsomal lipid peroxidation. Copper deficiency did not affect H2O2 production; however, it caused two- and fourfold increases in iron concentration in lung and liver microsomes, respectively, compared to Cu-adequate, ad libitum--fed controls. The mechanism by which cooper deficiency causes microsomal lipid peroxidation is still unknown but could be related to the observed accumulation of iron.     

Vitamin A analogs as tests for liver vitamin A status in the rat

Two chlorinated retinol analogs (Ro 11-0503 and Ro 11-8284) were assayed in rat serum and correlated to retinol in liver. Rats were fed a retinol-free diet to deplete their liver stores, then repleted with 0.5, 1.0, 3.0, and 10.0 mg retinyl palmitate/kg diet. Rats were given intraperitoneal injections of the analog then killed after an additional week on the diets. Analogs were measured by HPLC. The relative abundance of both analogs in serum was inversely correlated with the amount of retinyl palmitate in the liver. Serum analog concentrations may be useful as indications of liver retinol stores.