Effects of ursodeoxycholic acid in combination with vitamin E on adipokines and apoptosis in patients with nonalcoholic steatohepatitis

BACKGROUND/AIMS: Adipokines and hepatocellular apoptosis participate in the pathogenesis of nonalcoholic steatohepatitis (NASH). In a randomized trial ursodeoxycholic acid (UDCA) with vitamin E (VitE) improved serum aminotransferases and hepatic histology. The present work evaluates the effect of this combination on adipokines and hepatocellular apoptosis. METHODS: Circulating levels of adiponectin, resistin, leptin, interleukin (IL)-6, IL-8, retinol binding protein-4, monocyte chemoattractant protein-1 and tumour necrosis factor-alpha were measured by enzyme-linked immunoassays at the beginning and after 2 years of treatment with either UDCA+VitE, UDCA+placebo (P) or P+P. Apoptosis was assessed by immunohistochemistry for activated caspase-3 and circulating levels of apoptosis-associated cytokeratin 18 fragments (M30). RESULTS: Levels of adiponectin increased in patients treated with UDCA+VitE, whereas they decreased in the two other groups (P<0.04) and correlated with the improvement of liver steatosis (P<0.04). M30 levels worsened in the P/P group and improved in the other two groups. They correlated with hepatocellular apoptosis (P<0.02) and steatosis (P<0.02) as well as negatively with adiponectin levels (P<0.04). CONCLUSIONS: UDCA+VitE improves not only aminotransferase levels and liver histology of patients with NASH, but also decreases hepatocellular apoptosis and restores circulating levels of adiponectin. These results suggest that the UDCA+VitE combination has metabolic effects in addition to its beneficial cytoprotective properties.

Metabolomics reveals a novel vitamin E metabolite and attenuated vitamin E metabolism upon PXR activation

Pregnane X receptor (PXR) is an important nuclear receptor xenosensor that regulates the expression of metabolic enzymes and transporters involved in the metabolism of xenobiotics and endobiotics. In this study, ultra-performance liquid chromatography (UPLC) coupled with electrospray time-of-flight mass spectrometry (TOFMS), revealed altered urinary metabolomes in both Pxr-null and wild-type mice treated with the mouse PXR activator pregnenolone 16alpha-carbonitrile (PCN). Multivariate data analysis revealed that PCN significantly attenuated the urinary vitamin E metabolite alpha-carboxyethyl hydroxychroman (CEHC) glucuronide together with a novel metabolite in wild-type but not Pxr-null mice. Deconjugation experiments with beta-glucuronidase and beta-glucosidase suggested that the novel urinary metabolite was gamma-CEHC beta-D-glucoside (Glc). The identity of gamma-CEHC Glc was confirmed by chemical synthesis and by comparing tandem mass fragmentation of the urinary metabolite with the authentic standard. The lower urinary CEHC was likely due to PXR-mediated repression of hepatic sterol carrier protein 2 involved in peroxisomal beta-oxidation of branched-chain fatty acids (BCFA). Using a combination of metabolomic analysis and a genetically modified mouse model, this study revealed that activation of PXR results in attenuated levels of the two vitamin E conjugates, and identification of a novel vitamin E metabolite, gamma-CEHC Glc. Activation of PXR results in attenuated levels of the two vitamin E conjugates that may be useful as biomarkers of PXR activation.

A paradoxical rise of neonatal seizures after treatment with vitamin B6

We report the case of a newborn with intractable epileptic seizures developing a paradoxical rise of seizure frequency and electroencephalogram alterations after administration of vitamin B6. We have been unable to determine the aetiology of this disorder. In a newborn presenting with drug-resistant epileptic seizures, the first therapeutic option remains the application of intravenous pyridoxine, but the physician should be aware of the risk of an increase in seizure frequency.

Experimental folate and vitamin B12 deficiency does not alter bone quality in rats

Hyperhomocysteinemia (HHCY) has been linked to fragility fractures and osteoporosis. Folate and vitamin B(12) deficiencies are among the main causes of HHCY. However, the impact of these vitamins on bone health has been poorly studied. This study analyzed the effect of folate and vitamin B(12) deficiency on bone in rats. We used two groups of rats: a control group (Co, n = 10) and a vitamin-deficient group (VitDef, n = 10). VitDef animals were fed for 12 wk with a folate- and vitamin B(12)-free diet. Co animals received an equicaloric control diet. Tissue and plasma concentrations of homocysteine (HCY), S-adenosyl-homocysteine (SAH), and S-adenosyl-methionine (SAM) were measured. Bone quality was assessed by biomechanical testing (maximum force of an axial compression test; F(max)), histomorphometry (bone area/total area; B.Ar./T.Ar.], and the measurement of biochemical bone turnover markers (osteocalcin, collagen I C-terminal cross-laps [CTX]). VitDef animals developed significant HHCY (Co versus VitDef: 6.8 +/- 2.7 versus 61.1 +/- 12.8 microM, p < 0.001) that was accompanied by a high plasma concentration of SAH (Co versus VitDef: 24.1 +/- 5.9 versus 86.4 +/- 44.3 nM, p < 0.001). However, bone tissue concentrations of HCY, SAH, and SAM were similar in the two groups. Fmax, B.Ar./T.Ar., OC, and CTX did not differ between VitDef and Co animals, indicating that bone quality was not affected. Folate and vitamin B(12) deficiency induces distinct HHCY but has no effect on bone health in otherwise healthy adult rats. The unchanged HCY metabolism in bone is the most probable explanation for the missing effect of the vitamin-free diet on bone.

Effect of disodium monofluorophosphate, calcium and vitamin D supplementation on bone mineral density in patients chronically treated with glucocorticosteroids: a prospective, randomized, double-blind study

To study the effect of fluoride on bone mineral density (BMD) in patients treated chronically with glucocorticosteroids, 15 subjects (renal grafted, n = 12; skin disease, n = 1; broncho pulmonary disorder, n = 1; Crohn's disease, n = 1) were prospectively studied in a double-blinded manner and randomly allocated either to group 1 (n = 8) receiving 13.2 mg/day fluoride given as disodium monofluorophosphate (MFP) supplemented with calcium (1,000 mg/day) and 25-hydroxyvitamin D (calcifediol) (50 micrograms/day), or to group 2 (n = 7) receiving Cas+ calcifediol alone. An additional group of 14 renal transplant patients treated chronically with glucocorticosteroids but exempt of specific therapeutic intervention for bone disease was set up as historical controls. BMD was measured by dual-energy X-ray absorptiometry (DXA, Hologic QDR 1000) performed at months 0, 6 and 12 for groups 1 and 2 (lumbar spine, total upper femur, diaphysis and epiphysis of distal tibia), or 11-31 months apart with calculation of linear yearly changes for the historical cohort. Lumbar BMD tended to rise in groups 1 and 2, and to fall in group 3, the change reaching statistical significance (p < 0.05) in group 1, thus leading to a significant difference between groups 1 and 3 (p < 0.05). At upper femur, tibial diaphysis and tibial epiphysis, no significant change in BMD occurred in any of the groups. In conclusion, lumbar BMD rises more after a mild dosis of fluoride given as MFP and combined to calcium and calcifediol than on Ca+ calcifediol alone, without changes in BMD at the upper femur or distal tibia.

Effects of mild vitamin a deficiency on lung maturation in newborn rats: a morphometric and morphologic study

OBJECTIVE To evaluate the effects of a 60% vitamin A deficiency (VAD) on the two postnatal stages of lung development: alveolarization and microvascular maturation. Lungs from deficient rats were compared to age-matched controls. STUDY DESIGN Starting at 3 weeks before mating, female rats were maintained under a diet lacking vitamin A. Due to the slow depletion of the vitamin A liver stores the pregnant rats carried to term and delivered pups under mild VAD conditions. Mothers and offspring were then kept under the same diet what resulted in a mean reduction of vitamin A plasma concentration of about 60% vs. controls during the whole experimental period. Pups were sacrificed on days 4, 10 and 21 and their lungs fixed and analyzed by means of a combined morphologic and morphometric investigation at light and electron microscopic levels. RESULTS During the whole experiment, body weights of VAD animals were lower than controls with a significant decrease on day 10. On days 4, 10 and 21 the pulmonary structure was in a comparable gross morphologic state in both groups. Despite this morphologic normality, quantitative alterations in some functional parameters could be detected. On day 4, lung volume and the volume and surface area of air spaces were decreased, while the arithmetic mean barrier thickness and type 2 pneumocyte volume were increased in the VAD group. On day 21, some changes were again manifest mainly consisting in an augmentation of the vascularization and a decrease in interstitial volume in deficient animals. CONCLUSIONS Mild VAD causes no gross disturbances in the postnatal phases of lung development in rats. However, a body weight-related transient retardation of lung maturation was detectable in the first postnatal week. At 3 weeks, the VAD lungs showed a more mature vascular system substantiated by an increase in volume of both capillary volume and the large non-parenchymal vessels. In view of these quantitative alterations, we suspect that mild VAD deregulates the normal phases of body and lung growth, but does not induce serious functional impairments.

Effect of Parenteral Vitamin E and/or A on Reproductive Performance of Ewes Mated on Pasture or in Drylot

Forty-four Hampshire (H) and 41 Suffolk (S) ewes were allotted within breed to one of four treatment groups (VitA, VitE, VitAE, and Control) to evaluate the effect of supplemental vitamin E and A on reproductive performance of ewes mated on pasture or in drylot. Beginning two weeks before the mating period, ewes received 0 or 300 IU of vitamin E every 14 days and 0 or 250,000 IU of vitamin A every 28 days. Hampshire ewes remained on pasture during the mating period, whereas S ewes were moved to drylot. Treatment did not affect ovulation rate (OR), embryonic loss (EL), fetal loss (FL) or litter size (LS) of H ewes. Embryonic loss was higher (P<.05) in the H yearlings and two-year olds than in older ewes. Litter size was lower (P<.01) for H yearlings compared with other age groups. Suffolk ewes in the VitE group exhibited a lower (P<.01) OR than S ewes in other treatment groups, but no effect of treatment was observed for EL, FL, or LS. Although S yearling ewes did not differ from ewes of other age groups for OR and EL, they did produce fewer (P<.05) lambs. Analysis of serum samples revealed that H yearling ewes exhibited lower (P<.05) serum a-tocopherol levels than older H ewes. In addition, H ewes had a higher (P<.05) serum a-tocopherol level than S ewes at the beginning of study when they were managed as one group. Even though age and breed influenced certain reproductive parameters, results of this study indicate little effect of supplemental vitamin E and A on the overall reproductive efficiency of ewes mated on pasture or in drylot.

Effect of Supplemental Vitamin E and A on Reproductive Performance and Serological Profiles of Ewes Managed in Drylot

Forty Hampshire and 40 Suffolk ewes were allotted to one of four groups (VitA, VitE, VitA&E, Control) in a 2 x 2 factorial treatment arrangement to evaluate the effect of supplemental vitamin E (0 or 300 IU) and vitamin A (0 or 250,000 IU) on reproductive performance. Laparoscopy and ultrasonography were used to measure ovulation rate, embryonic loss, and fetal loss. Serum profiles of a-tocopherol (vitamin E) and retinol (vitamin A) also were monitored. There were no differences (P>.05) among treatment groups in any reproductive trait. Suffolk ewes exhibited a higher (P<.02) ovulation rate than Hampshire ewes, and yearling ewes incurred higher (P<.001) embryonic loss than other age groups, resulting in a lower (P<.001) litter size. Serum levels of a-tocopherol were higher (P<.05) for Hampshire than for Suffolk ewes and were lower (P<.001) in yearling ewes versus ewes two years of age and older. Serum levels of a-tocopherol declined (P<.01) throughout the study in VitA and Control ewes but remained unchanged in VitE and VitA&E ewes. Serum level of retinol remained unchanged in VitA ewes, whereas the level increased (P<.01) initially in VitE, VitA&E, and Control ewes before declining toward initial levels. Correlations were detected between ovulation rate and the change of pre-mating a-tocopherol serum level (r=-.29; P<.02), the change in pre-mating retinol serum level (r=-.50; P<.02) and the interval from vitamin A injection (r=-.60; P<.05). These data indicate significant influences of breed, age, and treatment on a-tocopherol and retinol serum levels in ewes and suggest that the timing of vitamin A administration may influence ovulation rate; however, vitamin supplementation, administered at random stages of the estrous cycle, was unable to alter flock reproductive performance.