E. Turchetto

Age-related changes in linoleate and α-linolenate desaturation by rat liver microsomes

The first and rate limiting step in the conversion of linoleic and alpha-linolenic acid is catalyzed by the delta - 6 - desaturase (D6D) enzyme. Rat liver microsomal D6D activity decreases on linolenic acid at a rate proportional to the animal age; on alpha-linolenic acid the decrease in D6D activity begins only later than on linoleic acid. The fatty acid composition of liver microsomes determined by gas chromatographic analysis confirms the impairment of the enzymatic activity directly measured. Our data indicate a correlation between aging and D6D activity impairment. The loss of D6D activity may be a key factor in aging through altering the eicosanoid balance.

The effect of dietary lipid changes on the fatty acid composition and function of liver, heart and brain mitochondria in the rat at different ages

A correlation between dietary lipids and cellular enzyme activities is a problem that has only been partially addressed by nutritionists. Therefore, changes in the fatty acid composition and the activities of some key metabolic enzymes (ubiquinol-2-cytochrome c reductase (EC 1.10.2.2), cytochrome oxidase (EC 1.9.3.1) and ATPase (EC 3.6.1.3)) in the mitochondria of liver, heart and brain of rats fed on diets differing extensively in their polyunsaturated fatty acid compositions have been investigated. The results showed that fatty acid compositional changes brought about by the dietary differences were associated with extensive changes in the activities of these key enzymes in the mitochondria. The extent of the influence differed considerably with the period over which the diets were fed. The role of dietary lipids to effect changes through the preservation of membrane structural integrity is discussed.

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.

Oxisterol determination in selected coffees.

The main aim of green-coffee processing techniques, such as decqffeination and roasting, is always to maintain a very high level of quality in taste and flavor, the beverages most important cliaracteristics to consumers. Oxidative alterations of coffee lipids, which can occur in roasting, exert a very marked influence on these quality traits. Determining the extent of oxidation thus can provide an indication of the products potential shelf-life and reveal traces of any newly-formed oxidative products that might prove nutritionally unsafe. Yet, while much attention has recently been focused on certain by-products induced by cholesterol oxidation and their proven toxicity as risk factors in atherosclerosis and cancer, oxidated phytosterols have largely gone unnoticed, being considered along with β-sitosterol as not very dangerous in that neither is absorbed by the intestinal tract. The present study investigates the substances derived from phytosterol oxidation (oxisterols) in samples of regular and decaffeinated commercial coffees. The findings show that oxisterols were absent in some samples and that the traces of oxidate phytosterols detected in others were well below the threshold considered as toxicologically active.

The effect of dietary polyenylphosphatidylcholine on microsomal delta-6-desaturase activity, fatty acid composition, and microviscosity in rat liver under oxidative stress

Abstract Polyenylphosphatidylcholine is a choline-glycerophospholipid containing up to 80% of total fatty acids as linoleic acid and may be an important factor in ensuring normal functioning of cell membranes. We tested the effect of a polyenylphosphatidylcholine-supplemented diet and compared it with both a trilinolein-supplemented and a laboratory chow diet on the fatty acid composition, microviscosity, and delta-6-desaturase activity of liver microsomal membranes of 12-month-old rats, in the absence or presence of oxidative stress induced by adriamycin. Polyenylphosphatidylcholine- and trilinolein-supplemented diets showed a similar increase in linoleic acid content and delta-6-desaturase activity in liver microsomes, indicating that low amounts of linoleic acid are able to partially restore the enzyme activity in old rats, independent of the source of linoleic acid. After adriamycin treatment, delta-6-desaturase activity increased in polyenylphosphatidylcholine and trilinolein groups, indicating a protective mechanism against the damage induced by polyunsaturated fatty acid peroxidation. The measurement of malondialdehyde production showed a protective effect on adriamycin-induced lipid peroxidation by polyenylphosphatidylcholine supplementation only. Microsomal membrane microviscosity did not change independent of diet and adriamycin treatment, suggesting that the response of microsomes to lipid peroxidation might be the maintenance of a given membrane order. Administration of polyenylphosphatidylcholine can prevent or minimize the liver damage induced by adriamycin treatment.

Possible implications in the use of exogenous phospholipids

Abstract The development of new techniques has shed more light on the physical state of phospholipids within the living cell. This has helped in the development of new concepts of the structural inter-relationship of phospholipids with proteins and other natural constituents and also of their functional role (e.g. enzyme modulators). On the other hand, understanding of phospholipid behaviour within the biological environment has encouraged research into the possibility of assigning to these molecules the role of carriers for exogenous or endogenous biologically active agents. Possible applications of this carrier potential in biology and medicine are discussed.

Age.related changes in essential fatty acid metabolism in cultured rat heart myocytes

We previously demonstrated that cultured neonatal rat myocytes have the capacity to desaturate/elongate essential fatty acids, alpha-linolenic acid conversion being higher than linoleic acid conversion. The whole process of highly unsaturated fatty acid formation from linoleic and alpha-linolenic acids slows with aging. In this study we grew heart myocytes in culture for different periods of time, and we observed a decrease in the desaturating/elongating activities for both substrates as the cells aged in culture. Alpha-linolenic acid conversion into highly unsaturated fatty acids was less impaired by aging than linoleic acid conversion. These modifications are correlated to the age-dependent alterations observed in the total lipid fatty acid composition, which caused a decrease in the unsaturation index. Changes in the lipid composition that occur in aging cultures parallel those reported for several tissues upon aging in the whole animal. The data herein reported may suggest the possibility of counteracting the effects of aging on lipid metabolism by supplementing cultures with appropriate amounts of highly unsaturated fatty acids.

Effect of dietary oils containing graded amounts of 18:3 n-6 and 18:4 n-3 on cell plasma membranes

Abstract Rats were fed diets containing a constant supply of lipids (10% by weight) differing from one another in the content of oleic acid (18:1 n-9), linoleic acid (18:2 n-6), and most peculiar 18-carbon polyunsaturated fatty acids, such as gamma-linolenic (18:3 n-6), alpha-linolenic (18:3 n-3), and stearidonic acid (18:4 n-3). Heart plasma membrane fatty acid composition from rats fed the different diets was affected. The diet containing the highest oleic acid content (olive oil diet) and the lowest PUFA content produced a significant decrease of linoleic acid content, while the oleic acid content was significantly lower when diets contained the lowest oleic acid percentage (black currant oil diet). Similarly, liver plasma membrane fatty acid composition was influenced by the different diets; in particular a significant higher content of the 20:4 n-6 was observed when rats were fed diets containing black currant oil (alone or into a 1:1 mixture with olive oil). Finally, the three diets tested influence only to a low level of significancy the fatty acid composition of the brain plasma membrane. Accordingly, the specific activity of 5′-nucleotidase, a typical cell plasma membrane enzyme, was not affected by diets neither in brain, nor in heart membrane preparations. On the contrary, the 5′-nucleotidase activity was highest in the membrane of liver cells from rats fed olive oil. A correlation between 5′-nucleotidase activity and membrane fatty acid composition cannot be stated in the conditions examined, however, possible biochemical mechanisms as the basis of 5′-nucleotidase behavior are discussed.

Influence of dietary fatty acids on phospholipid composition and prostaglandin E synthesis in rat kidneys

The effects of oils with different amounts of n6 and n3 fatty acid precursors and derivatives were evaluated on phospholipid composition and PGE2 synthesis of rat kidneys. Dietary lipids were: olive oil, an olive-blackcurrant-fish oil mixture and a blackcurrant-fish oil mixture. We observed in the kidneys of rats fed the blackcurrant-fish oil mixture a significant decrease in PGE2 synthesis, while arachidonate values did not show significant variations. A decrease of PGE2 synthesis could be due to competitive and inhibitory effects of fatty acids other than arachidonate, observed in the kidney phospholipid composition in our dietary conditions.

Dietary-induced changes in lipid and fatty acid composition can modify chronaxie values in the rat sciatic nerve

The present investigation aimed at clarifying the possible correlations among dietary lipids, peripheral fatty acid composition of nerve lipids and an index of the nervous tissue excitability, the chronaxie. The experiments were performed on female albino rats fed diets containing olive oil (OO) and fish oil (FO) along two generations. Total lipids fatty acid composition of the sciatic nerves from the two groups differed in the proportions of 18:1(n-9), 20:1(n-9), 22-1, 20:5(n-3) and 22:6(n-3). Also the lipid class composition showed significant differences among FO and OO specimens (free cholesterol more concentrated in the OO lipids; triacylglycerols more concentrated in the FO ones). The sciatic nerve isolated from FO rats showed a significant decrease in the chronaxie values if compared to the OO specimens. These results could follow from dietary-induced changes in the perineural permeability and/or possible modifications in the cable properties of the peripheral nerve fibers related to the myelin sheath composition.