Coenzyme Q differentially modulates phospholipid hydroperoxide glutathione peroxidase gene expression and free radicals production in malignant and non-malignant prostate cells.

The aim of this study was to investigate the role of coenzyme Q on the mRNA abundance of PHGPx and the reactive oxygen species (ROS) production in two different cell lines from human prostate, a line of non cancer cells (PNT2) and a line of cancer cells (PC3). Results showed that malignant cells markedly differ in their response to coenzyme Q compared to non-malignant cells, with no changes in PHGPx expression and greater ROS production. Furthermore coenzyme Q supplementation significantly lowered cell growth of the PC3 cancer line without affecting the PNT2. If these results are confirmed with additional experiments, it could represent a novel and interesting approach on the biomedical use of coenzyme Q10 in cancer therapy.

Dietary fat type and regular exercise affect mitochondrial composition and function depending on specific tissue in the rat.

Physical exercise and fatty acids have been studied in relation to mitochondrial composition and function in rat liver, heart, and skeletal muscle. Male rats were divided into two groups according to dietary fat type (virgin olive and sunflower oils). One-half of the animals from each group were subjected to a submaximal exercise for 8 weeks; the other half acted as sedentary controls. Coenzyme Q, cytochromes b, c + c1, a + a3 concentrations, and the activity of cytochrome c oxidase were determined. Regular exercise increased (P < 0.05) the concentration of the above-mentioned elements and the activity of the cytochrome c oxidase by roughly 50% in liver and skeletal muscle. In contrast, physical exercise decreased (P < 0.05) cytochrome c oxidase activity in the heart (in micromol/min/g, from 8.4+/-0.1 to 4.9+/-0.1 in virgin olive oil group and from 9.7+/-0.1 to 6.7+/-0.2 in sunflower oil animals). Dietary fat type raised the levels of coenzyme Q, cytochromes, and cytochrome c oxidase activity in skeletal muscle (P < 0.05) among the rats fed sunflower oil. In conclusion, dietary fat type, regular exercise, and the specific tissue modulate composition and function of rat mitochondria.

Olive oil supplemented with vitamin E affects mitochondrial coenzyme Q levels in liver of rats after an oxidative stress induced by adriamycin.

In this study we have evaluated the supplementation of olive oil with vitamin E on coenzyme Q concentration and lipid peroxidation in rat liver mitochondrial membranes. Four groups of rats were fed on virgin olive, olive plus 200 mg/kg of vitamin E or sunflower oils as lipid dietary source. To provoke an oxidative stress rats were administered intraperitoneally 10 mg/kg/day of adriamycin the last two days of the experiment. Animals fed on olive oil plus vitamin E had significantly higher coenzyme Q and vitamin E levels but a lower mitochondrial hydroperoxide concentration than rats fed on olive oil. Retinol levels were not affected, by either different diets or adriamycin treatment. In conclusion, an increase in coenzyme Q and alpha-tocopherol in these membranes can be a basis for protection against oxidation and improvement in antioxidant capacity.

Virgin olive oil and coenzyme Q10 protect heart mitochondria from peroxidative damage during aging.

The mitochondrial theory of aging suggests that this phenomenon is the consequence of random somatic mutations in mitochondrial DNA, induced by long-term exposure to free radical attack. There are two potential dietary means of delaying the effects of free radicals on cellular aging, i.e., enrichment of mitochondrial membranes with monounsaturated fatty acids and supplementation with antioxidants. We have performed a preliminary study on male rats, 6 or 12 month old, fed with diets differing in the nature of the fat (virgin olive oil or sunflower oil) and/or with antioxidant supplementation (coenzyme Q10), analysing hydroperoxide and coenzyme Q9 and Q10 in heart mitochondria. Preliminary results allow us to conclude that the CoQ10 dietetic supplementation as well as the enrichment of the cellular membranes with monounsaturated fatty acids, successfully protect mitochondrial membranes from aged rats against the free radical insult.

Physical exercise affects the lipid profile of mitochondrial membranes in rats fed with virgin olive oil or sunflower oil.

The effects of physical exercise on the lipid profile in mitochondrial membranes of liver and skeletal muscle were examined in rats fed with virgin olive oil or sunflower oil. Thirty male Wistar rats, 21 d old, were randomly assigned to four groups according to fat ingestion and physical activity over an 8-week period. For each type of oil, one group acted as a control group while rats from the other were trained to run for 40 min daily on a horizontal treadmill, at a speed of 35 m/min. The results show that diet affected the fatty acid profile of the mitochondrial membranes from skeletal muscle and liver. Physical exercise also modified the fatty acid profile of the mitochondrial membranes. Total monounsaturated fatty acids decreased (P < 0.001) in liver mitochondria of exercised animals. Total polyunsaturated fatty acids in mitochondrial membranes of liver increased (P < 0.005) after exercise but those in mitochondrial membranes of skeletal muscle decreased (P < 0.05). These changes due to the exercise may arise via several mechanisms, e.g. fluidity regulation; changes in the eicosanoid metabolism; differences in the availability or oxidation rate of the different fatty acids.

Relative importance of the saponified and unsaponified fractions of dietary olive oil on mitochondrial lipid peroxidation in rabbit heart.

The effects of four edible oils on lipid peroxidation have been investigated in rabbit heart mitochondrial membranes. The experimental oils (olive oil from the variety "picual", washed olive oil from the variety "picual", olive oil from the variety "arbequina" and high-oleic sunflower oil) had a similar fatty-acid composition, but differed in their unsaponified fraction (polyphenols, tocopherols, and others). The lowest hydroperoxide levels were found with picual and washed picual. No differences in mitochondrial membrane thiobarbituric acid reactive substance (TBARS), alpha-tocopherol concentrations and cytosolic antioxidant enzymes (superoxide dismutase, glutathione peroxidase, glutathione reductase and catalase) were found, whereas the CoQ10 content correlated inversely with hydroperoxide levels in all groups. These results suggest that mitochondrial membranes with high levels of monounsaturated fatty acids generate low levels of lipid peroxidation. Moreover, the saponified fraction of the experimental diets proved more important in preventing lipid peroxidation than the unsaponified fraction. Lastly, coenzyme Q may help to prevent peroxidative stress damage in rabbit heart mitochondria.

Vitamin E supplementation increases the stability and the in vivo antioxidant capacity of refined olive oil.

Two experiments were carried out to investigate if the supplementation with vitamin E affects refined olive oil response to oxidation regarding the stability of the oil and the protection in vivo against lipid peroxidation in rats after its intake in comparison with other edible oils. In experiment 1, samples of virgin olive oil, refined olive oil, refined olive oil supplemented by us with 200 mg/kg vitamin E, and sunflower oil were collected before and after a 60 min frying process. After frying, refined olive oil supplemented with vitamin E compared with the non-supplemented refined olive oil had a higher concentration of alpha-tocopherol (240.34+/-6.07 mg/kg vs. 131.94+/-8.14 mg/kg), more resistance against oxidation (19.01+/-1.88% vs. 10.6+/-2.08%) and less polar components (4.2+/-0.06% vs. 5.45+/-0.22%). In experiment 2, 24 male Wistar rats, divided into 4 groups, were fed on diets based on the same unfried oils (8% w/w) as in experiment 1, for 4 weeks. Two days prior to the end of the experiment, the rats were intraperitoneally administered with adriamycin (10 mg/kg/ day) to provoke an oxidative stress. The rats fed on refined olive oil plus vitamin E compared to the rats fed on non-supplemented refined olive oil had lower hydroperoxides concentrations (26.8+/-2.6 nmol/mg vs. 35.6+/-2.49 nmol/mg) higher coenzyme Q levels (128.1+/-11.97 pmol/mg vs. 81.25+/-9.25 pmol/mg) and higher alpha-tocopherol values (1.23+/-0.04 mmol/mg vs. 0.93+/-0.06 mmol/mg) in microsomes of liver. In conclusion, the supplementation of refined olive oil with 200 mg/kg of vitamin E increases the stability of this oil under pro-oxidant conditions, and its intake decreases the oxidative damage generated by adriamycin in rats.

Tissue specific interactions of exercise, dietary fatty acids, and vitamin E in lipid peroxidation.

Both physical exercise and ingestion of polyunsaturated fatty acids that play an essential role in free radical-mediated damages cause lipid peroxidation. The intake of specific fatty acids can modulate the membrane susceptibility to lipid peroxidation. Data confirmed that liver, skeletal muscle, and heart have different capabilities to adapt their membrane composition to dietary fatty acids, the heart being the most resistant to changes. Such specificity affects membrane hydroperoxide levels that depend on the type of dietary fats and the rate of fatty acid incorporation into the membrane. Sedentary rats fed a monounsaturated fatty acid-rich diet (virgin olive oil) showed a higher protection of their mitochondrial membranes against peroxidation than sedentary rats fed a polyunsaturated fatty acid-rich diet (sunflower oil). Rats subjected to training showed higher hydroperoxide contents than sedentary animals, and exhaustive effort enhanced the aforementioned results as well as in vitro peroxidation with a free radical inducer. This study suggests that peroxide levels first depend on tissue, then on diet and lastly on exercise, both in liver and muscle but not in heart. Finally, it appears that alpha-tocopherol is a less relevant protective agent against lipid peroxidation than monounsaturated fatty acids.

Effects of the amount and type of dietary fat on exocrine pancreatic secretion in dogs after different periods of adaptation.

Mongrel dogs were fed, from weaning to 6 months of age, on one of two 9% lipid diets that differed only in the type of fat content (sunflower oil or virgin olive oil) to study their effects on exocrine pancreatic secretion, in the basal period and in response to food. In addition, the results were compared with those obtained in a previous work performed by us on dogs adapted for 8 months to diets containing a higher (15%) amount of the same dietary fats to further evaluate the influence of the amount of dietary fat and the length of the adaptation period. The results from the present study show that both the volume and bicarbonate secreted in the absence of stimuli are unaffected by the quality of dietary fat. In contrast, in response to food, the pancreatic juice flow and the bicarbonate output were significantly higher in the group of animals given the sunflower oil diet. The differences seem to be related with the oleic acid content in the diets and the effectiveness of this fatty acid in triggering the release of inhibitory peptides such as pancreatic polypeptide and peptide YY. The comparison between the results from our present and previous studies supports the afore-mentioned hypothesis and confirms the existence of a clear influence of the amount and type of dietary fat, especially the oleic acid content, upon the pancreatic response to food, without ruling out a role for the duration of the adaptation period.

Coenzyme Q content depends upon oxidative stress and dietary fat unsaturation.

The presence of Coenzyme Q (CoQ) in food, its role in cellular bioenergetics and antioxidant protection and the key role played by dietary fatty acids on membrane structure support the interest for a wide research concerning the relationship between dietary fats, CoQ content and biochemical behaviour. Several models of peroxidative stress 'in vivo' have been extensively investigated in our laboratory, with particular regards to the influence of dietary fat upon mitochondrial CoQ levels. First studies showed that the unsaturation degree of dietary fat leads to different CoQ9 and CoQ10 mitochondrial contents. The highest levels were found using polyunsaturated fat. A significant CoQ9 decrease after adriamycin peroxidative induction was found when dietary fat was polyunsaturated; on the contrary, a light increase was found in the case of monounsaturated fat. Another example of oxidative stress is that produced by food frying. The results obtained were in some cases similar to those of the previous experimental design: in fact monounsaturated dietary fats increased CoQ mitochondrial contents, whereas the polyunsaturated ones decreased CoQ levels. Finally, the combined effect of physical exercise and dietary fats on tissue and plasma CoQ levels has been studied. CoQ levels did not change during aerobic performances when dietary fat was monounsaturated whereas light increases were detected in the case of polyunsaturated fats. On the contrary, in anaerobic conditions, CoQ levels clearly increased with monounsaturated fats and no alterations were found in the case of polyunsaturated ones.

Long-term effects on lipid metabolism in miniature swine (Sus scrofa) of diets enriched in saturated, monounsaturated and polyunsaturated (N-6 and N-3) fatty acids.

In this paper we study the effect of long-term adaptation (twelve months) of lipidic parameters of miniature swine to diets enriched in saturated (lard; L-group), monounsaturated (olive-oil; O-group) and polyunsaturated (sunflower and fish-oil; S- and F-groups respectively). The experimental group with the highest level of total cholesterol, free cholesterol and phospholipids was the S-group. This group had as well levels of HDL-C and LDL-C significantly higher when compared with the remaining groups. The L-group had the lowest value of HDL-C. In spite of that, the index of artherogenicity (HDL/LDL+VLDL) was significantly higher in the L-group, followed by the O-, F- and S-group respectively. On the other hand, after 12 months adaptation we observed that the fatty acid composition of serum lipids clearly reflects the quality of the dietary fats. The O-group had significantly higher serum oleic acid levels than all the other groups, and its content in saturated fatty acids was the lowest. The same happens with red blood cell (Rbcs) membranes fatty acids but the effect is less marked. Membranes of the L-group were found to have the highest saturation index (SI) in Rbcs membranes, while the F-group had the highest unsaturation index (UI), followed by the O-group. Taken together, our findings show that the diet enriched in olive oil produces a lipid pattern intermediate between that obtained with the fish-oil-diet and the sunflower-oil-diet but with the advantage of Rbcs membranes with a lower amount of PUFAS. As it is known, membranes with high percentages of PUFAS are more accessible for peroxidation, and the degree of peroxidation of lipids is directly related with changes in the membranes functionality.

Effects of adaptation to diets enriched with saturated, monounsaturated and polyunsaturated fats on lipid and serum fatty acid levels in miniature swine (Sus scrofa).

A total of 37 miniature swine (Sus scrofa) was fed with three diets of 9% fat content, differing only in the quality of the fat source (sunflower oil, olive oil and lard fat). Three groups of animals were fed for a 3 month period, and three other groups were fed for a 12 month period. After the two experimental periods, the influence of the diet on serum lipids and serum fatty acids was studied. In the short term, the serum cholesterol level was higher in the lard group, but with time, it undergoes a significant increase in the sunflower group, due to increases both in HDL-C and in LDL-C. In the olive oil group, LDL-C hardly varies with time, while HDL-C tends to decrease. In the lard group, LDL-C increases and HDL-C decreases. The atherogenic index is, in the long term, lower in the lard group and higher in the polyunsaturated fat group. The fatty acid composition of serum lipids in the long term shows a greater saturation index (SI) for the sunflower group and lard group, an increase in monounsaturated fatty acids in the olive oil group, and a higher percentage of linoleic acid in the sunflower group.

Peroxidative extent and coenzyme Q levels in the rat: influence of physical training and dietary fats.

Sport practice is widely recognized as capable of producing peroxidative damages, even of severe intensity. Dietary manipulations can also modify membrane susceptibility to peroxidation. In previous experiments we found that, while dietary virgin olive oil successfully protects mitochondrial and microsomal membranes from endogenous, xenobiotics-induced peroxidation, dietary polyunsaturated oils lead to increased peroxidative levels. In the latter conditions, cell machinery tries to counteract the structural and functional changes which have occurred, by modulating enzyme activities and concentrations, by increasing biosynthesis of coenzyme Q and by mobilizing cholesterol. In the present study we hypothesized that combining these two aspects could give useful information on the membrane response to peroxidation phenomena that daily occur throughout the lifespan. Rats fed different dietary oils as only fat source underwent a carefully designed training program and were killed at different times following acute or chronic exercise. Results show that peroxidation related to chronic training and to an acute bout of exercise sum up with peroxidative effects induced by dietary factors. The above mentioned phenomena occurred simultaneously with increased tissue levels of coenzyme Q, possibly triggered within a physiological reactive antioxidant strategy.

Effect of dietary fat composition on biliary cholesterol saturation index in dogs.

In dogs, the effect of dietary fat on bile cholesterol saturation index and the evolution of molar percentages of biliary lipids have been studied in both fasting and postprandial periods after a long-term adaptation period to diets which only differ in their lipidic source (olive oil and sunflower oil). It has been observed that for similar bile cholesterol saturation indexes in both groups, dietary fat altered differently biliary lipid composition through a double mechanism which involved bile acids and phospholipids. Dietary fat is postulated to affect differently the lipidic composition of bile as well as the biliary tree motility both during interdigestive and postprandial periods.

[Cytochrome oxidase induction following oxidative stress produced by adriamycin in the heart of rats fed olive oil]. / Induzione della citocromo ossidasi in seguito a stress ossidativo prodotto da adriamicina in cuore di ratti alimentati con olio di oliva.

Cytochrome oxidase is extremely susceptible to modifications that could occur at membrane level such as, for example, the deep influence that cardiolipin has on its maximal activity. Moreover, even the different ratio of fatty acid species in cardiolipin molecule could affect cytochrome oxidase properties. The influence of dietary fat and of endogenous lipid peroxidation with regard to cytochrome c oxidase activity and cytochrome a+a3 concentrations in mitochondria from rat heart has been investigated. Whilst the endogenous oxidative stress led to increase the peroxidation products, detected by means of hydroperoxides and conjugated dienes determinations, the features of the lipid environment were not affected as shown by fluorescence polarization technique. However, the oxidative stress was able to induce a significant increase of cytochrome a+a3 level even if the phenomenon did not reach the same extent as in the case of previous investigations on rat liver. Moreover, the results vary with the different fat source used and this might indicate a possible involvement of different dietary fats in peroxidation mechanisms. Furthermore the data we have obtained enable us to confirm once more that an induction of the synthesis of cytochrome a+a3 might be related to an enhanced production of peroxides at membrane level.

Adaptation of biliary response to dietary olive oil and sunflower-seed oil in dogs.

The effects of adaptation to dietary fat of different degrees of unsaturation (olive oil and sunflower oil) on bile secretion were studied in dogs at rest and after food intake. The animals were prepared with a bidirectional biliary cannula and a duodenal cannula to provide bile return. The two experimental groups were fed on diets containing 150 g fat/kg in the form of either olive oil (O) or sunflower-seed oil (S). The flow-rate under resting conditions and the patterns of response to food were similar in both experimental groups, although postprandial hypersection were significantly greater in volume and more prolonged in group O. No appreciable differences in concentration and output of biliary cholesterol or phospholipids were noted between the two groups. In contrast, the concentration and output of bile acids differed significantly both at rest and after food: concentration and output of bile acids were greater at rest in group S. However, after food intake, these responses were increased only in group O. The results suggest that the type of dietary fat affects biliary response to food, probably through differences in the contribution of the gall bladder in the two experimental groups.

Mitochondrial and microsomal cholesterol mobilization after oxidative stress induced by adriamycin in rats fed with dietary olive and corn oil.

The influence of three different dietary fats (8%) and of endogenous lipid peroxidation with regard to cholesterol concentrations in liver mitochondria and microsomes and in serum has been investigated in the rat. Although the different diet fat used did not produce any effect on serum cholesterol, it was possible to show that each experimental diet differently influenced the microsomal and mitochondrial levels of cholesterol. The highest mitochondrial and microsomal cholesterol content was found in case of diet supplemented with virgin olive oil and the lowest with rectified olive oil. An endogenous oxidative stress induced by adriamycin was able to produce a clear decrease in microsomal and mitochondrial cholesterol level and a sharp increase in serum concentration in all three groups. However, dietary fats and adriamycin had no effect on the microsomal and mitochondrial membrane viscosity as detected by fluorescence polarization. These results are consistent with the hypothesis that mitochondrial and microsomal cholesterol can exchange with exogenous pools when phospholipid peroxidation occurs.

Cytochrome oxidase induction after oxidative stress induced by adriamycin in liver of rats fed with dietary olive oil.

The influence of different kinds of dietary fat (8%) and of endogenous lipid peroxidation with regard to cytochrome c oxidase activity and cytochrome a + a3 concentrations in mitochondria from rat liver has been investigated. It was possible to confirm that the dietary fat induced higher phospholipid degradation in mitochondrial membranes; moreover an endogenous oxidative stress induced by adriamycin was able to increase the peroxidative effects. We have found that the peroxidative effects could sometimes induce an apparent enhancement of cytochrome oxidase activity due to a significant increase of cytochrome a + a3 content. This finding lets us suppose that both changes in the lipid environment and some peroxidation damage could occur in the membrane as a consequence of the fat assumed. Furthermore we should suggest that an induction of the synthesis of cytochrome a + a3 might be related to an enhanced production of peroxides at membrane level.

Changes in mitochondrial and microsomal rat liver coenzyme Q9 and Q10 content induced by dietary fat and endogenous lipid peroxidation.

The influence of different kinds of dietary fat (8%) and of endogenous lipid peroxidation with regard to coenzyme Q9 (CoQ9) and coenzyme Q10 (CoQ10) concentrations in mitochondria and microsomes from rat liver has been investigated by means of an HPLC technique. Although the different diet fats used did not produce any effect on microsomes, it was possible to show that each experimental diet differently influenced the mitochondrial levels of CoQ9 and CoQ10. The highest mitochondrial CoQ content was found in case of a diet supplemented with corn oil. An endogenous oxidative stress induced by adriamycin was able to produce a sharp decrease in mitochondrial CoQ9 levels in the rats to which corn oil was administered. The results suggest that dietary fat ought to be considered when studies concerning CoQ mitochondrial levels are carried out.