Comparative antioxidant effectiveness of dietary beta-carotene, vitamin E, selenium and coenzyme Q10 in rat erythrocytes and plasma

Five groups of five weanling rats were each fed a Torula yeast-based diet either unsupplemented or supplemented with 30 mg beta-carotene/kg, 30 IU vitamin E/kg, 1 mg selenium/kg or 30 mg coenzyme Q10/kg. Elevated levels of plasma aspartate aminotransferase and alanine aminotransferase are sensitive indicators of liver damage. The former enzyme was lower (P less than 0.01) in the vitamin E-, selenium- and beta-carotene-supplemented groups than in the unsupplemented control group, and the latter enzyme was lower in the vitamin E- and selenium-supplemented groups, suggesting a relatively equal effectiveness of these three antioxidants against liver damage. Erythrocytes were tested for protection against uninduced oxidative damage or that induced by 1 mmol/L bromotrichloromethane (BrCl3C) by measuring thiobarbituric acid-reactive substances (TBARS), hemoglobin, hemolysis, protein precipitation, alanine release and several enzyme activities. In untreated erythrocytes, selenium, beta-carotene and coenzyme Q10 exhibited protection by lowering (P less than 0.05) TBARS and alanine release, but only vitamin E protected against hemolysis. In BrCl3C-treated erythrocytes, vitamin E, selenium and beta-carotene protected by decreasing (P less than 0.05) protein precipitation, whereas selenium and beta-carotene decreased alanine release. The results of this study suggested that, in a manner analogous to vitamin E and selenium, beta-carotene and coenzyme Q10 function as antioxygenic nutrients.     

Dietary coenzyme Q10 and vitamin E alter the status of these compounds in rat tissues and mitochondria

Vitamin E (VE) and coenzyme Q (CQ) are essential for maintaining functions and integrity of mitochondria, and high concentrations of these compounds are found in their inner membranes. This study was conducted to examine the interaction between exogenously administered CQ10 and VE in rats. Male Sprague-Dawley rats (12 mo old) were fed a basal diet (10 IU VE or 6.7 mg RRR-alpha-tocopherol equivalent) supplemented with either 0 or 500 mg CQ10, and 0, 100 or 1310 IU VE/kg diet for 14 or 28 d. Liver, spleen, heart, kidney, skeletal muscle, brain and serum were analyzed for the levels of CQ10, CQ9 and VE. CQ10 supplementation significantly (P: < 0.05) increased CQ10 concentration in the liver and spleen (total and mitochondria) and serum, but not in other organs. Interestingly, rats supplemented with CQ10 plus 100 IU VE/kg diet had significantly higher CQ10 levels in the liver and spleen, whereas those supplemented with CQ10 plus 1310 IU VE/kg diet had lower levels, compared with those supplemented with CQ10 alone. As expected, dietary VE increased VE content in all of the organs analyzed in a dose-dependent manner. However, rats fed the basal diet supplemented with CQ10 had significantly higher VE levels in liver (total and mitochondria) than those not receiving CQ10 supplementation. CQ9 levels were higher in the liver and spleen, lower in skeletal muscle and unaltered in brain, serum, heart and kidney of rats supplemented with CQ10 compared with the controls. These data provide direct evidence for an interactive effect between exogenously administered VE and CQ10 in terms of tissue uptake and retention, and for a sparing effect of CQ10 on VE. Data also suggest that dietary VE plays a key role in determining tissue retention of exogenous CQ10.     

A significant correlation between the plasma levels of coenzyme Q10 and vitamin B-6 and a reduced risk of coronary artery disease

Coronary artery disease (CAD) is the leading cause of death worldwide. The purpose of this study was to investigate the relationship between plasma levels of coenzyme Q10 and vitamin B-6 and the risk of CAD. Patients with at least 50% stenosis of one major coronary artery identified by cardiac catheterization were assigned to the case group (n = 45). The control group (n = 89) comprised healthy individuals with normal blood biochemistry. The plasma concentrations of coenzyme Q10 and vitamin B-6 (pyridoxal 5’-phosphate) and the lipid profiles of the participants were measured. Subjects with CAD had significantly lower plasma levels of coenzyme Q10 and vitamin B-6 compared to the control group. The plasma coenzyme Q10 concentration (β = 1.06, P = .02) and the ratio of coenzyme Q10 to total cholesterol (β = .28, P = .01) were positively correlated with vitamin B-6 status. Subjects with higher coenzyme Q10 concentration (≥516.0 nmol/L) had a significantly lower risk of CAD, even after adjusting for the risk factors for CAD. Subjects with higher pyridoxal 5’-phosphate concentration (≥59.7 nmol/L) also had a significantly lower risk of CAD, but the relationship lost its statistical significance after adjusting for the risk factors of CAD. There was a significant correlation between the plasma levels of coenzyme Q10 and vitamin B-6 and a reduced risk of CAD. Further study is needed to examine the benefits of administering coenzyme Q10 in combination with vitamin B-6 to CAD patients, especially those with low coenzyme Q10 level.     

Oxidative stress in mouse plasma and lungs induced by cigarette smoke and lipopolysaccharide

Short-term exposure to cigarette smoke (CS) or lipopolysaccharide (LPS) leads to acute lung inflammation through oxidant-antioxidant imbalance. We studied the response in mice exposed to smoke or LPS during five consecutive days, as measured by superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities, as well as lipid peroxidation and nitric oxide levels in bronchoalveolar lavage fluid (BALF), lung homogenates, and plasma. Control mice were exposed to ambient air. Exposure to CS or LPS led to a similar influx of alveolar macrophages and neutrophils into the BALF; however, hydroxyproline levels were increased only in the CS group (p<0.001); SOD activity was increased in the BALF (p<0.001) and lung homogenates (p<0.05) of the CS group but was decreased in the BALF (p<0.05), lung homogenates (p<0.05) and plasma (p<0.01) of the LPS group. CAT activity was increased in the BALF (p<0.01), lung homogenates (p<0.001) and plasma (p<0.05) of the CS group but decreased in the BALF (p<0.001) and plasma (p<0.05) of the LPS group. GPx activity was reduced in the BALF (p<0.01) and plasma (p<0.01) of both the CS and LPS groups. Lipid peroxidation was increased in the BALF (p<0.001) and lung homogenates (p<0.001) of the CS group. Finally, the levels of nitrite were reduced in the CS (p<0.01) and LPS (p<0.001) groups. Our data show that the activity profiles of enzymes contributing to oxidant-antioxidant imbalance in the lungs differ depending on the inflammatory stimulus, and that SOD, CAT and GPx may be useful markers of oxidative stress in acute lung inflammation induced by exposure to CS.     

Body fat deposition: effects of dietary fat and two exercise protocols.

We evaluated the effects of two levels of dietary fat (0 and 20g beef tallow/100g diet) and two treadmill exercise protocols (low-intensity, high-intensity) on fat deposition in rats.

Male Wistar rats (n = 50) remained sedentary or were forced to run 840 meters/day, 5 days/week, on a rodent treadmill. Those on the high-intensity protocol covered this distance in less time (38 min) than those on the low-intensity program (60 min). Responses to high-fat (HF) and low-fat (LF) diets were compared within each exercise group for this 8-week study.

High fat feeding, as a single factor, did not affect energy intake, carcass fat, intramuscular fat, or fat associated with any tissue studied. The HF diet also did not affect responses to either exercise protocol. The high-intensity-exercised animals had less carcass fat (LF: 21% less; HF: 33% less), smaller omental fat pads (LF: 20% less; HF: 37% less), and retroperitoneal fat pads (LF: 19% less; HF: 38% less), and lower serum triglyceride levels (LF: 26% less; HF: 41% less) than sedentary rats. Those differences were less marked for the low-intensity-exercise rat. Neither mode of exercise or diet affected lipid concentrations in hindlimb muscles, livers, hearts, or kidneys.

Exercised animals generally had less fat deposition than sedentary rats but this was more pronounced for high-intensity than low-intensity-exercised rats, and was not affected by feeding a 20% beef tallow diet.     

Lung eicosanoid synthesis is affected by age, dietary fat and vitamin E.

The effect of age, dietary fat type and all-rac-alpha-tocopheryl acetate (vitamin E) supplementation on ex vivo synthesis of lung eicosanoids was measured in C57BL/6NIA mice using a 2 (age) x 3 (fat) x 3 (vitamin E) factorial design. Young (3-mo-old) and old (24-mo-old) mice were fed a semipurified diet containing 5% (by wt) corn oil, coconut oil or fish oil supplemented with 30, 100 or 500 mg vitamin E/kg for 4 wk. Ex vivo synthesis of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1 alpha (PGI2) were measured by RIA in lung homogenates. Old mice had significantly higher concentrations of TXB2 and PGI2 than did young mice, resulting in a significant increase in the TXB2:PGI2 ratio with aging. Young and old mice fed fish oil had significantly lower concentrations of PGI2 and TXB2 than those fed corn oil or coconut oil. The degree of reduction varied according to age and vitamin E status. Old mice fed fish oil and 30 mg vitamin E/kg diet had the lowest plasma vitamin E concentration and the highest TXB2:PGI2 ratio. The TXB2:PGI2 ratio was significantly reduced in old mice fed coconut oil or fish oil by vitamin E supplementation. Vitamin E supplementation (100 mg/kg) significantly increased PGI2 concentration in young mice fed coconut oil. Thus, significant changes in the capacity of lung to synthesize eicosanoids occur with age and are influenced by dietary fat type and vitamin E. J. Nutr.     

Oxidative stress in abetalipoproteinemia patients receiving long-term vitamin E and vitamin A supplementation

BACKGROUND: Patients with abetalipoproteinemia develop progressive ataxic neuropathy and retinopathy that are thought to be due, in part, to oxidative damage resulting from deficiencies of vitamins E and A. OBJECTIVE: The goal was to determine the degree of oxidative stress in abetalipoproteinemia patients who had received vitamin E (100 mg/kg) and vitamin A (10 000-15 000 IU/d) since infancy. DESIGN: Ten patients aged 3-25 y were studied. Assessed were plasma carbonyl concentrations as a marker of oxidative damage to proteins; total plasma oxidizability, which was used to evaluate the susceptibility of plasma lipoproteins to oxidation; and cyclic voltammetry, which represents the overall reducing and antioxidant capacity stemming from low-molecular-weight antioxidants in plasma. RESULTS: Concentrations of plasma carbonyls did not differ significantly between patients and control subjects ( +/- SE: 0.5670 +/- 0.031 and 0.5039 +/- 0.0134 nmol/mg protein, respectively). The lag phase of plasma oxidizability was 28.03 +/- 3.16 min in the patients and 24.0 +/- 2.79 min in healthy subjects in whom oxidizability of isolated HDL was measured (NS). Cyclic voltammetry showed a peak potential of 330 +/- 8.3 mV in all samples studied, denoting that the same antioxidants were present in the plasma of the patients and the control subjects. The anodic current of the samples, a measure of the concentration of hydrophilic low-molecular-weight antioxidants, was 5.227 +/- 0.25 and 5.38 +/- 0.20 micro A in the patients and the control subjects, respectively (NS). CONCLUSION: Enhanced oxidative stress is not apparent in the plasma of abetalipoproteinemia patients receiving long-term supplementation with vitamins E and A.     

Cardiorespiratory fitness modifies the association between dietary fat intake and plasma fatty acids

OBJECTIVES: To investigate the relation between (1) cardiorespiratory fitness and plasma saturated, monounsaturated and polyunsaturated fatty acids and (2) the interactions between cardiorespiratory fitness, dietary fat intake and plasma fatty acid composition. DESIGN: Cross-sectional analysis. SETTING AND SUBJECTS: The subjects were randomly selected, 127 middle-aged Finnish men participating in the DNASCO exercise intervention study. INTERVENTIONS: Cardiorespiratory fitness was determined spiroergometrically, dietary intake of macro- and micronutrients by 4-day food records and plasma fatty acids by gas chromatography. The subjects were divided into tertiles of aerobic fitness. RESULTS: Differences between fitness tertiles were not observed for dietary intake of total fat, and saturated, monounsaturated or polyunsaturated fatty acids (percent of total energy). In contrast, plasma saturated fatty acids were significantly lower (P <0.01) and polyunsaturated fatty acids significantly higher (P <0.05) in the highest fitness tertile compared to the lowest tertile. Dietary saturated fat intake was positively associated with plasma saturated fatty acids (r=0.342; P <0.05) and inversely with plasma polyunsaturated fatty acids (r=-0.453; P <0.01) only in the lowest fitness tertile. In addition, a positive correlation between body mass index and plasma saturated fatty acids (r=0.516; P <0.01) as well as a negative correlation between body mass index and plasma polyunsaturated fatty acids (r=-0.516; P <0.01) was observed in the lowest tertile solely. CONCLUSION: Different levels in cardiorespiratory fitness are associated with different levels in plasma saturated and polyunsaturated fatty acids and lead to modifications in the association between dietary and plasma fatty acids. These findings can perhaps be explained by a reduced hepatic fatty acid and lipoprotein synthesis as well as by an enhanced muscular lipid utilization, which are commonly seen in those who are physically active and who exhibit a higher level of fitness.     

Body weight and depot fat changes as influenced by exercise and dietary fat sources in adult BHE rats.

Adult male BHE rats were fed diets containing 15% of either corn oil (CO) or medium chain triglycerides (MCT) as the dietary source of fat. Further, rats were allowed to remain sedentary or were forced to exercise by swimming for 1 hour daily, for 3 weeks, followed by swimming for 2 hours daily for 3 weeks. The exercise for 3 weeks caused significant reductions in average body weight gains. After 6 weeks of exercise the lipid content of the adipose cells was reduced by about 50%. Fat cell numbers were not changed by either fat source or exercise, but fat cell size was significantly reduced after swimming daily for 6 weeks.     

Modulation of the exercise and retirement effects by dietary fat intake in hamsters

An experiment was conducted to determine the effects and interactions of exercise-retirement and dietary fat intake on body composition and hepatic lipogenic enzyme activities in hamsters. Forty-eight adult female hamsters were randomly allotted to eight groups of six each for a 40-d experiment. Exercise was in the form of voluntary wheel-running. Four groups served as either exercise or sedentary controls and were fed either a low or a high fat diet for 40 d, in a factorial fashion. Another four groups had access to exercise for 32 d and were then retired for 8 d. Of these four groups, two were fed either the low or high fat diet for the entire 40-d period; the other two were changed to the other diet on d 32. Results showed that compared to sedentary hamsters, exercise hamsters had greater body weight gain but less body fat content and hepatic lipogenic enzyme activities. Under both sedentary and exercise conditions, high fat-fed hamsters had lower hepatic lipogenic enzyme activities and less body fat content than low fat-fed hamsters. Upon retirement, high fat feeding led to a faster increase in body fat especially in previously high fat-fed hamsters. Results of this study suggest that dietary fat intake can significantly modulate the exercise and retirement effects with respect to body weight gain and body fat content.     

液质联用法同时测定保健食品中维生素E、维生素E乙酸酯和辅酶Q10

目的采用高效液相色谱-串联质谱法(HPLC-MS/MS),建立保健食品中维生素E、维生素E乙酸酯和辅酶Q10的检测方法。方法样品经过乙腈超声提取(50℃),采用HPLC-MS/MS法进行检测。色谱条件:采用Agilent Eclipse-plus C18(2.1 mm×50 mm,1.8μm)色谱柱,流动相为乙醇-甲醇(含0.1%甲酸)溶液,梯度洗脱,流速为0.2 ml/min,柱温为35℃。质谱条件:电喷雾离子源(ESI+),干燥气温度为250℃,鞘气流速为11 L/min,源电压为4 kV,检测方式为多重反应监测(MRM)模式。结果维生素E、维生素E乙酸酯和辅酶Q10在相应的浓度范围内与峰面积都呈良好线性关系(r>0.99),加样回收率为78.1%~96.3%,精密度RSD<6%,方法检出限在0.008μg/g^0.15μg/g。结论此方法测定快速准确、灵敏度高,专属性强,可用于保健食品中维生素E、维生素E乙酸酯和辅酶Q10的检测。     

Effect of dietary fat and vitamin E on mouse lung lipids.

To examine the effect of dietary fat on lung lipids, male weanling mice (CD-1 strain) were fed purified diets containing 5% stripped lard or corn oil and kept in chambers supplied with air filtered free of airborne bacteria. Vitamin E was fed at 0, 10.5 or 105 mg dl-alpha-tocopheryl acetate/kg diet. Dietary fat and vitamin E (0 or 10.5 mg/kg) had no significant effects on the lung levels of triacylglycerol (TG) or phospholipid (PL) molecular species through 4 weeks of intake. Alterations in lung fatty acid composition were followed through 6 weeks of intake at 0, 10.5 and 105 mg vitamin E/kg diet. Vitamin E, at all levels of supplementation, had no significant effect on mouse lung fatty acid composition. Saturated fatty acids of the lung also showed little alteration by diet, but feeding the lard diet significantly elevated oleic and palmitoleic acids. In mice fed the corn oil diet the levels of linoleic acid (18:2) were twice those of lard-fed mice, and arachidonic acid (20:4) was elevated by 15.8%. The diet elevated the mean peroxidizability index (PI) on lung tissue in corn oil-fed mice.     

Influence of dietary fat on beta-carotene absorption and bioconversion into vitamin A

Dietary fat facilitates the utilization of carotenoids and, based on serum beta-carotene or retinol responses following ingestion of meals containing carotene and fat sources, it has been reported that the amount of fat required in a meal may be minimal (approximately 3-5 g). However, the dietary fat requirement for optimal carotene utilization in humans cannot be fully ascertained without longer-term dose-response studies that measure the changes in vitamin A body stores in response to varying levels of dietary fat. In humans, vitamin A body stores can be determined by use of stable isotope-dilution methods. Animal studies have shown that although the level of dietary fat has no effect on serum vitamin A concentrations of animals fed beta-carotene, higher liver vitamin A concentrations were found in those that ingested higher fat levels. Other factors that might influence the relationship of fat intake and beta-carotene utilization include the type of fat ingested, physicochemical properties of the carotenoid source, amount of carotene ingested, whether fat and beta-carotene sources are provided in the same meal, the presence of helminthic infections, age, and vitamin A status.     

Role of vitamin E and oxidative stress in exercise.

Reactive oxygen species (ROS) play an important role as mediators of skeletal muscle damage and inflammation after strenuous exercise. These ROS arise largely from increases in mitochondrial oxygen consumption and electron transport flux. Bouts of intense exercise are associated with increases in lipid peroxidation, generating malondialdehyde and F(2alpha)-isoprostanes, and the release of muscle enzymes like lactate dehydrogenase and creatine kinase. Dietary and enzymatic antioxidant defenses appear to play a protective role in muscle cells by reducing associated oxidative damage to lipids, nucleic acids, and protein. However, studies of the use of dietary antioxidants like vitamin E to reduce exercise-induced muscle injury have met with mixed success. The equivocal nature of these results appear to reflect a diversity of factors including the antioxidant(s) tested, the nature and timing of the exercise, the age and fitness of the subjects, and the methodology for assessing oxidative stress.     

No effect of adult dietary fat on tumors induced prenatally by diethylstilbestrol.

Strain CD-1 female mice exposed prenatally to diethylstilbestrol (DES) or vehicle were placed on semipurified diets containing 2.6%, 10%, 20%, or 29% fat by weight at four weeks of age. These mice were used as a breeding colony for a few weeks and then maintained to terminal illness on the semipurified diets. Females exposed prenatally to DES developed mammary tumors, pituitary tumors, and glandular tumors of the reproductive tract. There was no significant difference in tumor frequency between low- and high-fat dietary groups. Fewer tumors appeared in the vehicle-exposed mice, as expected, and their frequency did not differ between the dietary groups. Pregnancy reduced tumor frequency in DES-exposed mice, but the incidence of pregnancy was not significantly different between low- and high-fat dietary groups. In the adult the failure of a high-fat diet to increase the frequency of reproductive system tumors induced prenatally is in marked contrast to the effectiveness of high-fat diets in promoting mammary tumors induced by carcinogens given to rats postnatally. This difference is critical in the interpretation of epidemiological studies. The relationship of dietary fat to reproductive system cancer in human populations was reviewed in comparison with these two animal models. The epidemiological literature was found to be more consistent with the animal model, showing high sensitivity to dietary fat prenatally but no significant sensitivity at the adult stage of life.     

The influence of dietary levels of vitamin A and fat on colon cancer

Rats fed diets high (24%) or low (5%) in fat were given dietary levels of vitamin A (retinyl acetate) ranging from 0.3 to 30 micrograms/g food. The lowest tumor incidence was in the group fed diets high in vitamin A and low in fat. When the diet was high in fat and low in vitamin A, tumor incidence and frequency were significantly increased over that in rats fed the high-fat diet with normal levels of vitamin A (10 micrograms/g feed). However, even with a high level of fat in the diet, raising the level of vitamin A above 10 micrograms/g feed had no further beneficial effect. Thus, although there was a significant interaction between vitamin A and fat, it is the latter that appears to require the most attention, once the vitamin A intake is adequate. These data support the view that we should set as a goal an adequate, diversified diet that is low in fat but that an excessive intake of vitamins such as vitamin A that are toxic should be avoided.     

Breast cancer and dietary and plasma concentrations of carotenoids and vitamin A

A case-control study of breast cancer was conducted in Buffalo. Participants completed a food frequency questionnaire and donated a fasting blood sample before definitive workup for breast masses. Dietary and plasma concentrations of carotenoids and retinol for 83 women found to have breast cancer were compared with those of 113 women found to be free of breast cancer (control subjects). There were no case-control differences in dietary estimates of vitamin A intake or in plasma alpha-carotene and lycopene. However, subjects with breast cancer had lower concentrations of plasma beta-carotene than did control subjects (P = 0.02). There was no overall association between plasma retinol and breast cancer but a positive relationship was observed between retinol and breast cancer in the subgroup with low beta-carotene values. These results suggest that low plasma beta-carotene is associated with increased risk of breast cancer. Other studies will need to determine whether low carotene concentrations are a subtle effect of the disease or might be causally related to breast cancer.