Hepatic Levels Of Vitamin Research Articles

NFR2 EXPRESSION IN APAP ACUTE INTOXICATION

Acetaminophen (APAP) overdose promotes the formation of NAPQI, the depletion of liver glutathione and oxidative stress. N‐acetilcysteine (NAC) is a cysteine donator that is a precursor for the synthesis of GSH. Nrf2 is a transcription factor, important in antioxidant response pathway, promoting the expression of phase II enzyme and the regulation of liver GSH. Male C57 mices, were fasted for 16h prior to the application of APAP (250mg/kg). The treatment with NAC (204mg/kg) occurred 1h after APAP challenge. The collection of liver tissue occurred 24h after treatment. Analysis: Histopathology by score of necrosis fields; hepatic levels of GSH by spectrophotometry and hepatic levels of Vitamin E (VE) by HPLC; the gene expression of NFR2 by RT‐qPCR (Taqman probes). Statistic Analysis: Student. t test (Graphpad Prism 5.0). The group (ANAC) that received treatment with NAC 1h after APAP challenge, showed a reduction of 28% of necrosis (p <0.05). ANAC GSH levels were 1.5 higher than the APAP levels (p <0.05), although VE presents no differences between the groups. The Nfr2 gene expression fold change had almost doubled compared with APAP group (p <0.05). The results show that after the treatment with NAC, the increase of NFR2 gene expression promotes the increase of GSH levels, saving the reduction of VE and consequently, the oxidative stress, protecting the liver from necrosis. Support by FAPESP (2011/12749–0 & 2011/17616–9)

Effect of α-tocopherol on carbon tetrachloride intoxication in the rat liver

Carbon tetrachloride (1 ml/kg body weight as a 1:1 mixture of CCl(4) and mineral oil) was orally administered to rats. After 12 h, the activity of plasma ALT (alanine aminotransferase) was significantly higher than that of the control group, and plasma ALT and AST (aspartate aminotransferase) activities significantly increased 24 h after CCl(4) administration. These results indicated that the necrotic process had initiated at about 12 h and developed thereafter. After 6-24 h of CCl(4) administration, the hepatic level of vitamin C, the most sensitive indicator of oxidative stress, decreased significantly, indicating that oxidative stress was significantly enhanced 6 h after CCl(4) intoxication and thereafter. Oral administration of vitamin E (1 ml/kg body weight as a 1:1 mixture of alpha-tocopherol and mineral oil) 12 h before CCl(4) administration caused a significant elevation of liver vitamin E level and ameliorated liver necrosis 24 h after CCl(4) intoxication based on plasma AST and ALT. Vitamin E also significantly restored the hepatic vitamin C concentration 12 and 24 h after CCl(4) intoxication, demonstrating that vitamin E functioned as an antioxidant. The liver vitamin E concentration was not changed by vitamin E supplementation to rats that did not receive CCl(4). This result indicated that vitamin E accumulated in the damaged liver. The activation of JNK, ERK1/2 and p38 MAPK took place 1.5 h after CCl(4) administration. Co-administration of alpha-tocopherol with CCl(4) did not affect these early changes in MAPKs.

Effect of dimethyl sulphoxide on oxidative stress, activation of mitogen activated protein kinase and necrosis caused by thioacetamide in the rat liver

Thioacetamide (400 mg/kg body weight, i.p.) was administered to rats. After 12 h the activity of plasma glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) was significantly higher than that of the control group, and after 24 h plasma GOT and GPT activities strongly increased. These results indicated that the necrotic process was initiated at about 12 h and developed thereafter. By co-administration of dimethyl sulphoxide (DMSO, 18 and 1 h before, and 8 h after administration of thioacetamide: each time, 2.5 ml/kg body weight, p.o.), plasma GOT and GPT were significantly decreased and were even comparable to the control group, showing that DMSO totally prevented the necrotic action of thioacetamide. After 12 and 24 h of thioacetamide administration, the hepatic level of vitamin C, the most sensitive chemical indicator of oxidative stress, decreased significantly, indicating that oxidative stress was significantly enhanced 12 h after thioacetamide intoxication and thereafter. DMSO totally restored the liver vitamin C level, demonstrating that DMSO effectively ameliorated the oxidative stress caused by thioacetamide, resulting in the prevention of necrosis of the liver. Phosphorylated c-Jun NH2-terminal kinase (JNK) significantly increased transiently 12 h after treatment with thioacetamide. These results indicated that oxidative stress and the activation of JNK took place almost simultaneously. Phosphorylated extracellular signal-related kinase (ERK) 2 was significantly increased 6–12 h after thioacetamide injection. Phosphorylated p38 MAPK (mitogen activated protein kinase) was significantly decreased 24 h after administration of thioacetamide. DMSO treatment inhibited the change of these MAPKs by thioacetamide, corresponding with the prevention of the liver necrosis as well as the attenuation of oxidative stress.

Activation of mitogen activated protein kinase (MAPK) during carbon tetrachloride intoxication in the rat liver

Carbon tetrachloride (CCl(4): 4 ml/kg body weight as a 1:1 mixture of CCl(4) and mineral oil) was orally administered to rats. After 12 h the activity of plasma AST (aspartate aminotransferase) and ALT (alanine aminotransferase) was significantly higher than that of the control group and plasma AST and ALT activities increased thereafter. These results indicated that the necrotic process was active at about 12 h and developed thereafter. After 2-24 h of CCl(4) administration, the hepatic level of vitamin C, the most sensitive indicator of oxidative stress, decreased significantly, indicating that oxidative stress was significantly enhanced as early as 2 h after CCl(4) intoxication and thereafter. Phosphorylated JNK (c-Jun NH(2)-terminal kinase) and phospho-ERK1/2 (extracellular signal-regulated kinase1/2) were significantly increased transiently 1-3 h after treatment with CCl(4), while phosphorylated p38 decreased significantly 1-24 h after CCl(4) treatment. These results indicated that the change in MAPKs (mitogen activated protein kinases) slightly preceded that in vitamin C, the most sensitive chemical indicator of oxidative stress.

Inhibitory Effect of Dimethyl Sulfoxide (DMSO) on Necrosis and Oxidative Stress Caused by D-Galactosamine in the Rat Liver

D-Galactosamine (D-Galn: 300 mg/kg) was intraperitoneally administered to rats. After 6 h the activity of plasma GOT and GPT was significantly higher than that of the control group and plasma GOT and GPT activities increased thereafter. These results indicated that the necrotic process was initiated at about 6 h and developed thereafter. With coadministration of DMSO (1 h before administration of D-Galn: 2.5 mL/kg, oral), plasma GOT and GPT were significantly lower, showing that DMSO inhibited the necrotic action of D-Galn. After 6-24 h of D-Galn administration, the hepatic level of vitamin C, the most sensitive indicator of oxidative stress, decreased significantly, indicating that oxidative stress was significantly enhanced 6 h after D-Galn intoxication and thereafter. DMSO significantly restored the liver vitamin C level 24 h after D-Galn injection, demonstrating that DMSO effectively ameliorated the oxidative stress caused by D-Galn, resulting in the prevention of necrosis of the liver. Phosphorylated JNK and phospho-ERK were significantly increased transiently 6-12 h after treatment with D-Galn. These results indicated that oxidative stress and the activation of JNK took place almost simultaneously. Phosphorylated p38 MAPK was not changed and DMSO treatment did not affect the change of these MAPKs by D-Galn.

The mode of delivery of parenteral multivitamins influences nutrient handling in an animal model of total parenteral nutrition

Very low birthweight preterm infants receive early total parenteral nutrition (TPN) to optimize protein balance. Adding multivitamins (MVP) to the lipid emulsion (MVP+LIP) rather than to the amino acid+dextrose moiety of TPN (AA+MVP) limits the effects of light exposure on lipid peroxidation and vitamin loss. Compare the effects of the mode of delivery of MVP on nutrient handling and indices of oxidant stress. Three-day old guinea pig pups were assigned to TPN containing MVP+amino acids+dextrose+heparin and electrolytes, with lipids provided separately (AA+MVP). Solutions were light exposed (LE, n = 8) or light protected (LP, n = 9). In a further group (n = 7), MVP was co-administered with the lipid moiety and light exposed (LIP+MVP). Variables measured in urine (creatinine, nitrogen, vitamin C) and in liver (protein, glutathione, isoprostane, vitamins A, E, C) were compared by ANOVA. Urinary nitrogen and vitamin C were higher (P<0.05) during LE, while hepatic levels of vitamin C were higher (P<0.05) with LIP+MVP. These results were not related to total peroxide levels in TPN or to markers of oxidant stress. CONCLUSION. Co-administration of MVP with lipid or light protected amino acids offers comparable beneficial effects on nitrogen and vitamin C metabolism.

Serum and hepatic vitamin E assessment in cirrhotics before transplantation.

Hepatic vitamin E may have a protective effect against hepatocyte injury; therefore, vitamin E replacement or supplementation may be beneficial in patients with cirrhosis. However, serum vitamin E may not correlate with hepatic vitamin E stores, making decisions regarding treatment difficult based on serum levels alone. The specific aims of this study were to determine hepatic concentrations of vitamin E and to determine whether serum levels of vitamin E correlate with hepatic vitamin E stores in cirrhotics. A prospective study of cirrhotics undergoing orthotopic liver transplantation (OLT) was completed. Serum and hepatic levels of vitamin E were measured by high-performance liquid chromatography. Statistical analysis was performed using rank sum tests and Spearman's rank correlation coefficient. Fifty cirrhotics (33 males, 17 females; mean age of 53 years) were studied. The control group (25 males, 25 females; mean age of 47 years) consisted of the liver donors. The median serum levels of vitamin E in controls and cirrhotics were 5.95 and 7.8 mg/L, respectively (p = .009). The median hepatic levels (0.10 mg/g) in the control and cirrhotic groups were similar (p = .037). There was a significant correlation between serum and hepatic vitamin E levels in cirrhotics (R = 0.335; p = .017). A positive correlation exists between serum and hepatic concentrations of vitamin E in cirrhotics, therefore making serum vitamin E levels a useful reference for treatment using exogenous vitamin E.

Nutrition Therapy in Liver Disease

The following is an excellent review by Drs Patton and Aranda-Michel regarding the etiology and nutrition therapy of liver disease. As the authors clearly point out, malnutrition is a significant problem in end-stage liver disease, resulting in increased morbidity and mortality. Unfortunately, there are few data to suggest that aggressive nutritional supplementation (ie, oral, enteral, or parenteral) improves clinical outcome in those patients with end-stage cirrhosis. It has been my clinical observation that clinical improvement in terms of function and quality of life is marginal at least. I suspect this may be the result of a sick liver not being able to appropriately process nutrients. It is fascinating to observe that usually within 4 months after hepatic transplantation, these patients begin to regain muscle mass and function and exhibit an improved quality of life. I therefore believe that the treatment of choice of the malnutrition in this group of patients is hepatic transplantation and not prolonged enteral or parenteral support. In regards to the causes of malnutrition, the authors mention that early satiety and subsequent decreased food intake may be the result of gastroparesis or delayed gastric emptying secondary to the ascites. We have recently reported that gastric emptying is unchanged after large-volume paracentesis in cirrhotics. I suspect the problem of early satiety experienced by cirrhotics with ascites is the result of reduced gastric accommodation or relaxation and not necessarily delayed gastric emptying. Studies evaluating gastric accommodation in cirrhotics with ascites are currently underway. In terms of enteral access for feeding, I would agree with the authors that a soft small-bore feeding tube (8 to 10 French) is usually well tolerated for a short period, ie, 4 to 8 weeks. I would also caution against the use of a gastrostomy tube because the risk of leakage and peritonitis is significantly increased in cirrhotics with ascites. I would agree with the authors that the use of total parenteral nutrition (TPN) should be limited because of the increased incidence of catheter infection. If TPN is given and the patient has cholestatic liver disease, ie, elevated bilirubin, copper and manganese should be held because these are excreted through the hepatic circulation. In terms of dietary changes or supplements, I would point out that supplying protein from a vegetable source may be better than an animal source because fewer aromatic amino acids are produced from vegetable sources. Careful attention should also be given to excess vitamin A intake because high hepatic levels of vitamin A may worsen hepatic function. We have recently reported that serum vitamin A levels may not correlate with hepatic tissue levels. Therefore, caution should be given for vitamin A replacement based on serum vitamin A levels alone. I agree with the authors that future randomized studies are needed to evaluate clinical outcome of aggressive nutrition support in end-stage cirrhotics, specifically, those on the waiting list for hepatic transplantation. These studies will be limited by lack of specific markers of nutritional response and duration of therapy required.

Nutritional assessment of serum and hepatic vitamin A levels in patients with cirrhosis.

Serum vitamin A (retinol) levels may not correlate with hepatic vitamin A stores in patients with cirrhosis; thus, supplementation of vitamin A based on serum levels may have a detrimental effect. Our aim was to determine whether serum levels correlate with hepatic stores in cirrhotic patients. A prospective study of patients with cirrhosis undergoing orthotopic liver transplantation was completed. Serum and hepatic levels of vitamin A were measured by high-performance liquid chromatography. Statistical analysis was performed using rank sum tests and Spearman rank correlation coefficients. Fifty cirrhotic patients (33 men and 17 women, mean age 53 years) were compared with a control group (25 men and 25 women, mean age 47 years) of liver donors. Median serum levels of retinol were 259 microg/L in controls and 166 microg/L in cirrhotic patients (p < .001). Median hepatic levels of retinol were 25 microg/g in controls and 27.5 +/- g/g in cirrhotic patients (p not significant). Total hepatic vitamin A levels (retinol plus retinyl esters) were 471 microg/g in controls and 244 microg/g in cirrhotic patients (p = .028). Serum retinol did not correlate with total hepatic vitamin A stores in cirrhotic patients (rs = .10, p = .332). Serum retinol and total hepatic vitamin A stores are lower in cirrhotic patients than in controls. However, because levels of serum retinol do not correlate with hepatic vitamin A levels, the decision to prescribe vitamin A replacement for patients with cirrhosis should not be made solely on the basis of serum retinol levels.

Vitamins A1and A2in hepatic tissue and subcellular fractions in mink feeding on fish‐based diets and exposed to Aroclor 1242

Two-month-old female mink were fed diets based on either Baltic herring (Clupea harengus membras) or freshwater smelt (Osmerus eperlanus) for 21 weeks. A portion of the smelt-fed mink were exposed orally to polychlorinated biphenyls (PCBs), Aroclor 1242 (1 mg/d). Retinol (vitamin A1), 3,4-didehydroretinol (vitamin A2), and their different fatty acyl esters were studied in hepatic tissue, microsomes, and cytosol by argentated reversed-phase high-performance liquid chromatography. As a result of Aroclor exposure, concentrations of the fatty acyl esters of vitamins A1 and A2 were about one-tenth and those of unesterified A2 one-fourth those of the control levels. In the fatty acyl esters, percentages of stearates (A1--18:0 and A2--18:0) increased at the expense of the other fatty acyl esters. The Aroclor exposure decreased concentrations of alcoholic and esterified forms of the A2 analog more than those of the corresponding A1 analog. In microsomes. Aroclor decreased the alcoholic and esterified vitamin analogs to the same extent (to 9-17%). In the cytosol compared to the control, the concentrations of the vitamin esters fell below 10%, but the alcoholic analogs remained at 30 to 40%. Despite equal dietary supply, in mink fed on Baltic herring, the hepatic levels of vitamin A, were only about one-third of the values found in the smelt-fed mink. The organochlorines also altered hepatic lipid composition and impaired breeding and kit growth. In the kits of the females fed on Baltic herring, blood hemoglobin was decreased.

Nutritional assessment of serum and hepatic vitamin A levels in patients with cirrhosis

BACKGROUND Serum vitamin A (retinol) levels may not correlate with hepatic vitamin A stores in patients with cirrhosis; thus, supplementation of vitamin A based on serum levels may have a detrimental effect. Our aim was to determine whether serum levels correlate with hepatic stores in cirrhotic patients. METHODS A prospective study of patients with cirrhosis undergoing orthotopic liver transplantation was completed. Serum and hepatic levels of vitamin A were measured by high-performance liquid chromatography. Statistical analysis was performed using rank sum tests and Spearman rank correlation coefficients. RESULTS Fifty cirrhotic patients (33 men and 17 women, mean age 53 years) were compared with a control group (25 men and 25 women, mean age 47 years) of liver donors. Median serum levels of retinol were 259 microg/L in controls and 166 microg/L in cirrhotic patients (p < .001). Median hepatic levels of retinol were 25 microg/g in controls and 27.5 +/- g/g in cirrhotic patients (p not significant). Total hepatic vitamin A levels (retinol plus retinyl esters) were 471 microg/g in controls and 244 microg/g in cirrhotic patients (p = .028). Serum retinol did not correlate with total hepatic vitamin A stores in cirrhotic patients (rs = .10, p = .332). CONCLUSIONS Serum retinol and total hepatic vitamin A stores are lower in cirrhotic patients than in controls. However, because levels of serum retinol do not correlate with hepatic vitamin A levels, the decision to prescribe vitamin A replacement for patients with cirrhosis should not be made solely on the basis of serum retinol levels.

VITAMINS A1, A2, AND E IN MINKS EXPOSED TO POLYCHLORINATED BIPHENYLS (AROCLOR 1242®) AND COPPER, VIA DIET BASED ON FRESHWATER OR MARINE FISH

Minks (Mustela vison) fed diets based on either freshwater fish or marine fish were exposed to 1 mg of polychlorinated biphenyls (PCBs) (Aroclor 1242) daily for 28 d. To minks on the freshwater diet, copper was also given with or without PCBs. The marine diet included more vitamin A, and E than the freshwater diet. The authors studied how the exposures affected levels of vitamins A{sub 1}, A{sub 2}, and E in liver and adipose tissues and levels of vitamins A{sub 1} and A{sub 2} in plasma. In females and males on the freshwater diet, the hepatic level of vitamin A{sub 2} was decreased because of the PCBs, and in these males the hepatic levels of vitamin E also decreased. Interestingly, with coexposure to PCBs and copper, the vitamin levels were, in general, close to the control values. In adipose tissues also, the PCBs induced significant changes in the concentrations of vitamins A{sub 1} and A{sub 2}. In plasma, vitamins A{sub 1} and A{sub 2} were decreased in all patterns of exposure and on both diets. However, plasma thyroxine was slightly increased, a finding opposite to that reported previously in rodent studies. The results suggest that, in mink, diet greatlymore » modulates the responses to PCBs, which may also differ in males and females. Furthermore, vitamins A{sub 1} and A{sub 2} may not be metabolized equally during PCB administration.« less

Low doses of eicosapentaenoic acid, docosahexaenoic acid, and hypolipidemic eicosapentaenoic acid derivatives have no effect on lipid peroxidation in plasma.

It was of interest to investigate the influence of both high doses of eicosapentaenoic acid (EPA) and low doses of 2- or 3-methylated EPA on the antioxidant status, as they all cause hypolipidemia, but the dose required is quite different. We fed low doses (250 mg/d/kg body wt) of different EPA derivatives or high doses (1500 mg/d/kg body wt) of EPA and DHA to rats for 5 and 7 d, respectively. The most potent hypolipidemic EPA derivative, 2,2-dimethyl-EPA, did not change the malondialdehyde content in liver or plasma. Plasma vitamin E decreased only after supplementation of those EPA derivatives that caused the greatest increase in the fatty acyl-CoA oxidase activity. Fatty acyl-CoA oxidase activity increased after administration of both EPA and DHA at high doses. High doses of EPA and DHA decreased plasma vitamin E content, whereas only DHA elevated lipid peroxidation. In liver, however, both EPA and DHA increased lipid peroxidation, but the hepatic level of vitamin E was unchanged. The glutathione-requiring enzymes and the glutathione level were unaffected, and no significant changes in the activities of xanthine oxidase and superoxide dismutase were observed in either low- or high-dose experiments. In conclusion, increased peroxisomal beta-oxidation in combination with high amounts of polyunsaturated fatty acids caused elevated lipid peroxidation. At low doses of polyunsaturated fatty acids, lipid peroxidation was unchanged, in spite of increased peroxisomal beta-oxidation, indicating that polyunsaturation is the most important factor for lipid peroxidation.

The effects of running stress on plasma vitamin A levels in rats

The plasma concentrations of vitamin A, zinc and proteins and the hepatic level of vitamin A were determined in rats subjected to running as a model for stress and which were receiving standard or vitamin-A free diets. All rats showed a decrease in plasma vitamin A with running compared with non-running control animals. Hepatic levels of vitamin A were higher in these two test groups than in their weight- and age-matched non-running controls. The data support that running, like other forms of stress, decreases plasma vitamin A, consistent with the retention of vitamin A in the liver.

Vitamin E protection against nitrosamine-induced esophageal tumor incidence in mice immunocompromised by retroviral infection.

Retrovirally induced immunosuppression may elevate the incidence of chemically induced cancers. A proposed hypothesis to explain this relationship is the increased free radical activity observed during retroviral infection and carcinogen activation. We previously found that vitamin E retarded growth of esophageal tumors accompanied by reductions of free radical products. This study investigated the contribution that retroviral immunosuppression has on esophageal cancer induced by the carcinogen N-nitrosomethylbenzylamine (NMBzA), and the response that increased levels of dietary vitamin E has on this induced carcinogenesis. Female C57BL/6 mice received NMBzA or vehicle (corn oil) i.p. weekly for 3 weeks. Then some of the mice were infected with LP-BM5 murine retrovirus and fed diets containing 30 IU vitamin E or 172 IU vitamin E/kg of diet. As an assessment of free radical activity, exhaled ethane was measured prior to killing the animals at 26 weeks. Esophagi from the various mice groups were assessed for size and frequency of tumors. Livers homogenates were analyzed for vitamins A and E, lipid fluorescence, conjugated dienes and malondialdehyde. Hepatic levels of vitamin A and E were decreased (P < 0.05) and indices of lipid peroxidation were greater (P < 0.05) in NMBzA-treated mice relative to controls. Lipid peroxidation and serum transaminases (ALT and AST) were greatest in mice given NMBzA and infected with the retroviruses. Incidence of esophageal tumors were also greatest in the NMBzA-treated, immunocompromised animals. Mice fed vitamin E-supplemented diets showed increased (P < 0.05) hepatic concentrations of vitamin E and vitamin A, decreased activities of serum transaminases, decreased indices of lipid peroxidation, and decreased size and frequency of esophageal tumors in both the immunocompromised and non-immunocompromised mice. These results suggest that vitamin E plays an antioxidant function that retards the incidence of esophageal cancers in immunocompromised and non-immunocompromised animals.

Glutathione depletion: Its effects on other antioxidant systems and hepatocellular damage

1. The mechanisms of the liver damage produced by three glutathione (GSH)-depleting agents, bromobenzene, allyl alcohol and diethyl maleate, were investigated. 2. With each toxin liver necrosis was accompanied by lipid peroxidation that developed only after severe depletion of GSH. 3. Changes in antioxidant systems by alpha-tocopherol (vitamin E) and ascorbic acid were studied. A decrease in the hepatic level of vitamin E, and a change in the redox state of vitamin C (increase in oxidized over reduced form) were evident whenever extensive lipid peroxidation developed. However, in the case of bromobenzene intoxication these alterations preceded lipid peroxidation, and may be an index of oxidative stress leading to subsequent membrane damage. 4. Experiments carried out with vitamin E-deficient or supplemented diets indicated that pathological phenomena occurring as a consequence of GSH depletion depend on hepatic levels of vitamin E. In vitamin E-deficient animals, lipid peroxidation and liver necrosis appeared earlier than in animals fed the control diet. In animals fed a vitamin E-supplemented diet, bromobenzene and allyl alcohol had only limited toxicity, and diethyl maleate none, in spite of similar hepatic GSH depletion. Thus, vitamin E may largely modulate the expression of toxicity by GSH-depleting agents.

Lipid peroxidation and antioxidant systems in the liver injury produced by glutathione depleting agents

The mechanisms of the liver damage produced by three glutathione (GSH) depleting agents, bromobenzene, allyl alcohol and diethylmaleate, was investigated. The change in the antioxidant systems represented by α-tocopherol (vitamin E) and ascorbic acid were studied under conditions of severe GSH depletion. With each toxin liver necrosis was accompanied by lipid peroxidation that developed only after severe depletion of GSH. The hepatic level of vitamin E was decreased whenever extensive lipid peroxidation developed. In the case of bromobenzene intoxication, vitamin E decreased before the onset of lipid peroxidation. Changes in levels of the ascorbic and dehydroascorbic acid indicated a redox cycling of vitamin C with the oxidative stress induced by all the three agents. Such a change of the redox state of vitamin C (increase of the oxidized over the reduced form) may be an index of oxidative stress preceding lipid peroxidation in the case of bromobenzene. In the other cases, such a change is likely to be a consequence of lipid peroxidation. Experiments carried out with vitamin E deficient or supplemented diets indicated that the pathological phenomena occurring as a consequence of GSH depletion depend on hepatic levels of vitamin E. In vitamin E deficient animals, lipid peroxidation and liver necrosis appeared earlier than in animals fed the control diet. Animals fed a vitamin E supplemented diet had an hepatic vitamin E level double that obtained with a commercial pellet diet. In such animals, bromobenzene and allyl alcohol had only limited toxicity and diethylmaleate none in spite of comparable hepatic GSH depletion. Thus, vitamin E may largely modulate the expression of the toxicity by GSH depleting agents.

Perturbations in cerebral oxygen radical formation and membrane order following vitamin E deficiency

The effects of dietary vitamin E deficiency on mouse cerebral membrane order and oxygen reactive species were studied. Quantitation of vitamin E levels in several brain regions showed greatest deficiencies in striatum and cerebellum, followed by substantia nigra, and cortex. Vitamin E deficiency increased central-core membrane order in cerebral P2 fraction, but was without effect in the superficial hydrophilic membrane domain. Oxygen radical formation was studied using the probe 2',7'-dichlorofluorescein diacetate. Basal generation rates of oxygen reactive species were 2.5-fold higher when compared to control animals. While hepatic levels of vitamin E are much more reduced than brain levels, in deficient mice, the rate of oxygen radical formation in the liver was unaltered. This implies an special susceptibility of the brain to deficiency of this lipophilic antioxidant vitamin. Data demonstrate that endogenous levels of free radical scavengers, such as vitamin E, may play an important role in maintaining basal oxygen radical levels and membrane integrity. The dietary vitamin E depletion paradigm suggests that a relation exists between elevated levels of oxygen radicals and more rigid hydrophobic central-cores in cerebral membranes, effects that may play a role in mechanisms underlying the neuropathologic lesions observed following vitamin E deficiency.

Vitamin A levels in regenerating rat liver. Effects on microsomal drug metabolism.

After being maintained on a vitamin A-deficient or complete diet for a period of five weeks, male Sprague-Dawley rats were subjected to a two-thirds partial hepatectomy (PH) or sham operation. The vitamin A content of the liver of vitamin A-deficient, PH rats was below the limit of detection (less than 1 microgram/g liver). Rats fed the control diet and subjected to PH had hepatic levels of vitamin A that were 37% and 49% lower 48 and 72 hours after surgery, respectively, when compared with sham-operated controls. Hepatic microsomal cytochrome P-450 levels were significantly reduced in PH rats fed the complete diet 48 hours after PH and in PH rats fed either the deficient or complete diet 72 hours after. Vitamin A deficiency alone significantly reduced cytochrome P-450 levels. A combination of vitamin A deficiency and PH had the most dramatic effect on cytochrome P-450 and aminopyrine N-demethylase, which reduced the activity to approximately 50% of the activities found in the sham-operated control group. PH resulted in the greatest reduction in the rate of disappearance of benzo[a]pyrene in the presence of liver microsomes prepared from vitamin A-deficient rats.

Effect of vitamin A deficiency on the pulmonary and hepatic drug-metabolizing enzymes in rat.

The effect of vitamin A deficiency on the drug-metabolizing enzyme system of the lung and liver was analyzed in rats fed diets with or without vitamin A for 5-6 weeks. The hepatic level of vitamin A was significantly reduced in vitamin A deficient animals. The hepatic cytochrome P-450 and b5 contents and activity of benzo(a)pyrene hydroxylase was significantly reduced in deficient animals. Contrary to this, pulmonary cytochrome P-450 and b5 contents were above the control values. No alteration in pulmonary benzo(a)pyrene hydroxylase was noted. The uridine diphosphate-glucuronosyltransferase activity of digitonin-treated microsomal membranes was below the control values both in lung and liver. However, the native uridine diphosphate-glucuronosyltransferase activity remained unchanged in the liver and was below control values in the lung.