Oleanolic acid and ursolic acid stabilize liposomal membranes

The effects of oleanolic acid (OA) and ursolic acid (UA) on the fluidity and stability of dipalmitoyl phosphatidylcholine (DPPC) liposomal membrane were monitored by measuring the fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene labeled in the liposomal membrane and the leakage of calcein from the probe-encapsulated liposomes. The experiments with the liposomes made of DPPC and OA or UA showed that OA and UA exhibited a moderate fluidity-modulating effect for the liquid-crystalline liposomal membrane, and a strong condensing effect for both crystalline and liquid-crystalline liposomal membranes. Their effects were comparable to those of cholesterol. These results suggest that their fluidity-modulating and condensing effects might have some implications in their biological functions.     

Incorporation of n-3 fatty acids into plasma lipid fractions, and erythrocyte membranes and platelets during dietary supplementation with fish, fish oil, and docosahexaenoic acid-rich oil among healthy young men

The effects of n-3 fatty acid supplementation in the form of fresh fish, fish oil, and docosahexaenoic acid (DHA) oil on the fatty acid composition of plasma lipid fractions, and platelets and erythrocyte membranes of young healthy male students were examined. Altogether 59 subjects (aged 19–32 yr, body mass index 16.8–31.3 kg/m²) were randomized into the following diet groups: (i) control group; (ii) fish diet group eating fish meals five times per week [0.38±0.04 g eicosapentaenoic acid (EPA) and 0.67±0.09 g DHA per day]; (iii) DHA oil group taking algae-derived DHA oil capsules (1.68 g/d DHA oil group taking algae-derived DHA oil capsules (1.68 g/d DHA in triglyceride form); and (iv) fish oil group (1.33 g EPA and 0.95 g DHA/d as free fatty acids) for 14 wk. The fatty acid composition of plasma lipids, platelets, and erythrocyte membranes was analyzed by gas chromatography. The subjects kept 4-d food records four times during the study to estimate the intake of nutrients. In the fish diet, in DHA oil, and in fish oil groups, the amounts of n-3 fatty acids increased and those of n-6 fatty acids decreased significantly in plasma lipid fractions and in platelets and erythrocyte membranes. A positive relationship was shown between the total n-3 polyunsaturated fatty acids (PUFA) and EPA and DHA intake and the increase in total n-3 PUFA and EPA and DHA in all lipid fractions analyzed. DHA was preferentially incorporated into phospholipid (PL) and triglyceride (TG) and there was very little uptake in cholesterol ester (CE), while EPA was preferentially incorporated into PL and CE. The proportion of EPA in plasma lipids and platelets and erythrocyte membranes increased also by DHA supplementation, and the proportion of linoleic acid increased in platelets and erythrocyte membranes in the DHA oil group as well. These results suggest retroconversion of DHA to EPA and that DHA also interferes with linoleic acid metabolism.     

Copper(II)-catalyzed lipid peroxidation in liposomes and erythrocyte membranes

Cu⁺⁺ was uniquely capable of catalyzing the peroxidation of rat erythrocyte membrane lipid in the presence of 10 mM H₂O₂, whereas several other transition metal ions were without significant effect. In contrast, peroxidation of soybean phospholipid liposomes could be catalyzed with decreasing efficiency by Co⁺⁺, Cu⁺⁺, Pb⁺⁺, or Cr⁺⁺⁺ also in the presence of H₂O₂. The effect of imidazole on Cu⁺⁺-catalyzed lipid peroxidation was stimulatory in liposomes and inhibitory in membrane preparations, whereas EDTA, histidine, citrate and alanine inhibited peroxidation in both systems. EDTA could stop the peroxidation after initiation, but catalase could not, indicating that Cu⁺⁺ alone was necessary for the propagation of the chain reaction. Competitive inhibition studies with various scavengers of hydroxyl radicals or singlet oxygen and the absence of significant reaction enhancement by D₂O indicated that neither of these reactive oxygen species was a major mediator in the Cu⁺⁺-H₂O₂ oxidative system. A copper-oxygen complex may be directly involved in the initiation of peroxidation. Normal erythrocyte membranes and phospholipid liposomes also differ in their sensitivities toward external oxidative stress. In the absence of H₂O₂, Cu⁺⁺ (0.2 mM) was capable of catalyzing lipid peroxidation in liposomes, aged erythrocyte membranes and membranes from vitamin-E-deficient rats; however, freshly prepared membranes from control rats and liposomes containing α-tocopherol required H₂O₂ greater than 2 mM for the catalytic effect of Cu⁺⁺ to be observed.     

Clusters of Phospholipid Vesicles as Platforms for Glucose Oxidase‐Catalyzed Reaction in a Bubble‐Column Bioreactor

Clusters of negatively charged liposomes encapsulated with glucose oxidase were prepared in the presence of Ca²⁺ and used to catalyze the oxidation of glucose in an external loop airlift bubble column. The clusters exhibited higher catalytic activity compared to nonclustered glucose oxidase‐containing liposomes (GOLs) when the liposome membranes were incorporated with cholesterol. The clusters were structurally altered in shear flow to give sufficient interfacial area accessible to glucose. The reactivity of GOL clusters could be modulated on the basis of lipid composition of the membranes which affected the mode of interaction among liposomes through Ca²⁺. Part of GOL clusters could be separated from the reaction mixture by centrifugation, which would be advantageous for reusing liposomal catalysts. The liposome clusters can be the platforms to regulate the catalytic performance of glucose oxidase in the airlift.     

Kinetic behaviour and stability of glucose oxidase entrapped in liposomes

Glucose oxidase (EC 1.1.3.4) was encapsulated in liposomes (prepared from phosphatidyl choline and cholesterol) by the dehydration–rehydration method. The enzymatic activities of native and liposomal glucose oxidase were followed by the amount of H₂O₂ obtained in the enzymatic β‐D ‐glucose oxidation. Some characteristics of the liposomal and free glucose oxidase were compared. The enzyme encapsulated in liposomes showed an apparent inhibition by glucose at concentrations higher than 0.28 mol dm^?3 with a substrate inhibition constant of 0.95 ± 0.12 mol dm^?3. The enzyme entrapped showed an apparent K_m value higher than that of the free enzyme. The apparent V_max of liposomal enzyme decreased by a factor of 0.35 with respect of that of the native enzyme. The optimum temperature of the free and entrapped enzymes remained similar but the liposomal enzyme showed maximal activity at a more acid pH (5.2). The thermal and proteolytic stabilities were enhanced by encapsulation in liposomes. The stabilization factors (relationship between half‐lives of entrapped form and free enzyme) at 45, 50 and 55 °C for liposomal glucose oxidase were 2.6, 1.6 and 1.6, respectively. Copyright © 2003 Society of Chemical Industry     

Dietary fat effect on incorporation and release of lipids and cholesterol by rat intestinal slices

The effect of saturated and unsaturated fats on in vitro formation and release of lipids and cholesterol from¹⁴C acetate by rat intestinal tissue was investigated. The rats were fed a basal diet enriched with either 25% corn oil or lard and then sacrificed after a 10- or 25-day feeding period. It was observed that a similar¹⁴C lipid content but a greater¹⁴C cholesterol content was found in the intestinal tissue of rats fed corn oil than in rats fed lard for 10 days. After a longer period of feeding of 25 days, the intestinal tissue¹⁴C cholesterol level was decreased in the corn oil fed rats without any significant effect on other lipids. These data suggest that corn oil in some way influences cholesterol biosynthesis depending upon its degree of unsaturation and the period of time for which it is fed. The decrease at the later time might involve some mechanism which aids in getting rid of accumulated tissue cholesterol. Less¹⁴C lipid and¹⁴C cholesterol were released by the intestinal tissue of rats fed the unsaturated fat as compared with those fed the saturated fat, suggesting a possible role in vivo in reducing blood lipids and blood cholesterol levels. Robert A. Welch Foundation.     

Studies on the hydrogen belts of membranes: II. Non-electrolyte permeability of liposomes of diester,diether, and dialkyl phosphatidylcholine and cholesterol

We have postulated the existence of lipid-lipid and protein-lipid hydrogen bonding in the hydrogen belts of membranes, i.e., the regions of hydrogen bond acceptors (carbonyl oxygens of esters and amides) and hydrogen bond donors (hydroxyls of cholesterol, sphingosine, proteins, water). To assess the possible effects of modifications of the hydrogen belts on membrane permeability, we prepared a diester phosphatidylcholine and two analogs lacking carbonyl oxygens, a diether and a dialkyl phosphatidylcholine, care being taken to synthesize lipids of identical efficient hydrophobic chain length. Relative permeation rates for glycerol and urea were determined by osmotic swelling of liposomes containing the phospholipids alone or with an equimolar quantity of cholesterol, with 4 mole % of dioleylphosphate added. The permeation rates of both solutes were similar for all three lipids, with Arrhenius activation energies ΔE^* around 16 kcal/mole. Cholesterol reduced the permeability of all three membranes. The activation energy ΔE^* of permeation did not change for diester and dialkyl phosphatidylcholine with cholesterol, but was lower by about 5 kcal/mole for the diether lipid with cholesterol. This corresponds to a reduction in the entropy of activation ΔΔS^*∼-16 cal/mole/degree. We interpret the results as supporting the hypothesis of interaction between cholesterol hydroxyl and phospholipid carbonyl.     

Changes in renal phospholipid fatty acids in diabetes mellitus: Correlation with changes in adenylate cyclase activity

Male Sprague-Dawley rats made diabetic with alloxan (37.5 mg/kg) or streptozotocin (65 mg/kg) were killed after 3–6 weeks of disease; renal tissues were studied for phospholipid content and for fatty acid composition of the phospholipids. No consistent change was noted in total phospholipid content nor in the proportion of various phospholipids in diabetics. However, diabetic animals showed a consistent reduction of arachidonic acid content in phosphatidylcholine (PC) and phosphatidylethanolamine in whole renal cortex, plasma membranes purified from renal cortex, and in isolated glomeruli. Associated with the fall in arachidonic acid was a rise in linoleic acid in the samples studied. Insulin therapy returned the fatty acid profiles to normal. These results are similar to patterns observed in other diabetic tissues and suggest that diabetes is associated with generalized changes in cell membranes. That these structural changes may have functional significance is suggested by demonstrated alterations in the temperature-dependence of adenylate cyclase in renal plasma membranes of diabetic animals. Adenylate cyclase is thought to be intimately associated with PC in plasma membranes, a phospholipid showing significant changes in fatty acid content in diabetes (unsaturation index 165±2 for normals, 147±5 for diabetics). Na⁺,K au+-ATPase which is thought to be primarily associated in vivo with phosphatidylinositol (PI), shows no change in apparent energy of activation in diabetes. The fatty acid content of PI is minimally altered in diabetes, and the unsaturation index is unchanged.     

Preparation and properties of liposomes composed of various phospholipids with different hydrophobic chains using a supercritical reverse phase evaporation method

Liposomes were prepared by the supercritical reverse phase evaporation method developed in our laboratory using various phospholipids with different hydrocarbon chains. The effects of the length of alkyl chain and number of unsaturated bonds of phospholipids on the properties of liposomal membranes were examined through trapping efficiency measurements, transmission electron microscopic observations, and osmotic response measurements. The trapping efficiency for water-soluble drugs of liposomes prepared by our method was greatly higher than that of liposomes prepared by the conventional Bangham method. Liposomes prepared using unsaturated phospholipids showed a high trapping efficiency compared with those prepared using saturated phospholipids. In addition, the trapping efficiency of liposomes prepared using 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC), a complex phospholipid with both saturated and unsaturated alkyl groups, had a value intermediate between L-alpha-dipalmitoyl-phosphtidylcholine (DPPC), a saturated phospholipid, and L-alpha-dioleoylphosphatidylcholine (DOPC), an unsaturated phospholipid. That is, the trapping efficiency of liposomes was dependent on the number of unsaturated bonds rather than the alkyl chain length of phospholipid molecule and it increased with increasing bulkiness of the molecule. The osmotic response was higher for liposomes prepared using unsaturated phospholipids than for those formed by saturated phospholipids.     

Application of experimental design to the formulation of glucose oxidase encapsulation by liposomes

Fractional factorial screening design and response surface methodology were applied to optimize the entrapment of glucose oxidase in liposomes by the dehydration–rehydration vesicle (DRV) method. Phosphatidylcholine from different sources, cholesterol:phosphatidylcholine (Ch:PC) and enzyme:lipid (E:L) ratios, buffer pH, sonication frequency and trehalose concentration were the parameters selected for this study. The type of phosphatidylcholine was found to be the most important factor followed by the trehalose concentration, Ch:PC ratio, sonication frequency and E:L ratio. The pH did not play an important role in the response. By treating liposomes with trehalose, as cryoprotectant, the activity of entrapped enzyme decreased by 16%. Two of the factors (cholesterol:phosphatidylcholine and enzyme:lipid ratios) were further studied in a 3² central composite design. The optimized liposomal formulation with an entrapment efficiency of 24% was obtained for egg yolk PC with Ch:PC and E:L ratios of 0.95 and 14.69, respectively, at pH 6 and applying a sonication frequency of 150 W. Copyright © 2004 Society of Chemical Industry     

Lipid composition of further purified bovine liver nuclear membranes

The lipid content, distribution and fatty acid composition of highly purified bovine liver nuclear membranes was determined and compared to those of microsomes prepared in parallel. Contrasted with microsomes, nuclear membranes while containing nearly the same levels of lipid had more cholesterol and total neutral lipid and less phospholipid. Phospholipid and neutral lipid patterns generally were similar for the two types of membranes. The same fatty acids, in similar proportions, were observed in respective total lipid, total polar lipid, phosphatidyl choline and phosphatidyl ethanolamine fractions of the two membrane types. The microsomal lipid fractions contained slightly greater percentages of unsaturated fatty acids. With respect to previous results from preparations contaminated with nonmenbranous nuclear material, purified fractions contained more total lipid on a protein basis and more total unsaturated fatty acids. Only minor differences in levels and distribution of phospholipids and neutral lipids were observed between the crude and highly purified fractions. Purdue University AES Journal Paper No. 4482.     

Cholesterol autoxidation in phospholipid membrane bilayers

Lipid peroxidation in unilamellar liposomes of known cholesterol-phospholipid composition was monitored under conditions of autoxidation or as induced by a superoxide radical generating system, γ-irradiation or cumene hydroperoxide. Formation of cholesterol oxidation products was indexed to the level of lipid peroxidation. The major cholesterol oxidation products identified were 7-keto-cholesterol, isomeric cholesterol 5,6-epoxides, isomeric 7-hydroperoxides and isomeric 3,7-cholestane diols. Other commonly encountered products included 3,5-cholestadiene-7-one and cholestane-3β,5α,6β-triol. Superoxide-dependent peroxidation required iron and produced a gradual increase in 7-keto-cholesterol and cholesterol epoxides. Cholesterol oxidation was greatest in liposomes containing high proportions of unsaturated phospholipid to cholesterol (4∶1 molar ratio), intermediate with low phospholipid to cholesterol ratios (2∶1) and least in liposomes prepared with dipalmitoylphosphatidylcholine and cholesterol. This relationship held regardless of the oxidizing conditions used. Cumene hydroperoxide-dependent lipid peroxidation and/or more prolonge oxidations with other oxidizing systems yielded a variety of products where cholesterol-5β,6β-epoxide, 7-ketocholesterol and the 7-hydroperoxides were most consistently elevated. Oxyradical initiation of lipid peroxidation produced a pattern of cholesterol oxidation products distinguishable from the pattern derived by cumene hydroperoxide-dependent peroxidation. Our findings indicate that cholesterol autoxidation in biological membranes is modeled by the peroxide-induced oxidation of liposomes bearing unsaturated fatty acids and suggest that a number of cholesterol oxidation products are derived from peroxide-dependent propagation reactions occurring in biomembranes.     

Uptake and metabolism of dolichol and cholesterol in perfused rat liver

The uptake of dolichol and cholesterol by perfused rat liver was studied. When these radioactive lipids were incorporated into egg phosphatidylcholine liposomes, both dolichol and cholesterol appeared initially in the supernatant and in the microsomal fraction and, later on, in the mitochondrial-lysosomal fraction. The lipids taken up were esterified to some extent, but no phosphorylation of dolichol occurred. Incorporation of dolichol and cholesterol into lipoproteins increased the efficiency of uptake, which was receptor-mediated in this case. Accumulation of these lipids occurred in lysosomes followed by a transport to the endoplasmic reticulum (ER). Both labeled dolichol and cholesterol appeared in the bile. In the case of dolichol, the majority of this radioactivity was not associated with the original substance itself, and probably represented lipid-soluble catabolites. In the case of cholesterol, most of the radioactivity was associated with bile acids. It appears that, in contrast to the receptor-mediated uptake of lipoproteins from the perfusate, the uptake of liposomal lipids involves a different mechanism. After association with the plasma membrane, the lipids enter into the cytoplasm and are transported to the ER and later to the lysosomes.     

Production of the criegee ozonide during the ozonation of 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine liposomes

It is likely that Criegee ozonides are formed in small amounts in the lungs of animals breathing ozone-containing air. This makes these compounds potential candidates to act as secondary toxins which relay the toxic effects of ozone deeper into lung tissue than ozone itself could penetrate. Therefore, we have determined the yields of Criegee ozonides from unsaturated lipids in liposomal systems as a model of the types of yields of Criegee ozonides that might be expected both in the lung lining fluid layer and in biological membranes. Ozonation of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine liposomes produced bothcis- andtrans-Criegee ozonides. These ozonides have been isolated by solid phase extraction and high-performance liquid chromatography of the ozonized lipid, and the products have been identified by two-dimensional¹H nuclear magnetic resonance. The combined yield of thecis- andtrans-Criegee ozonides is 10.7±2.8% (avg. ±SD, n=7) with small unilamellar liposomes and 10.6±2.7% (n=3) with large multilamellar liposomes. We had previously reported (Chem. Res. Toxicol. 5 505–511, 1992) that ozonation of methyl oleate in sodium dodecylsulfate micelles also produces an 11% yield of the Criegee ozonides. Thus, ozonation in a variety of models gives about 11% of the Criegee ozonide, suggesting that these products also would be formed in small but significant amounts in the lungs of animals breathing polluted air. Further research on the pharmacokinetics and possible toxicity of the Criegee ozonides of fatty acids is suggested.     

Homeostatic control of membrane fatty acid composition in the rat after dietary lipid treatment

Diets in which both the lipid content and composition (polyunsaturated to saturated fatty acid ratio) were varied were fed to rats for 20 weeks, and the effects on the tissue lipid profiles were determined. The fatty acid profile of the plasma lipids, and the phospholipid fatty acids of the mitochondrial and microsomal fractions of liver, heart, kidney and brain, as well as erythrocyte membranes were determined. Despite large differences in the level and type of lipid present in the experimental diets and in the proportion of saturated fatty acids in the plasma lipids in response to the various diets, there was little effect on the proportion of saturated to unsaturated fatty acids in the phospholipids of the various membranes examined. The major effect of altering the dietary level of polyunsaturated to saturated fatty acids was on the ratio of the ω6/ω3 series of unsaturated fatty acids in the membrane lipids. This change occurred in all tissues except the brain, in which only a small response to altered dietary lipid intake was observed. The ω6/ω3 ratio was elevated upon feeding a diet rich in ω6 polyunsaturated fatty acids, but decreased when a diet rich in saturated fatty acids was fed. The failure to significantly alter membrane lipid saturation/unsaturation in the tissues examined would suggest that a homeostatic mechanism is operative in biological membranes and may act to buffer membranes from the effects of changes in the nature of the dietary lipid intake.     

<Emphasis Type="Italic">Trans</Emphasis> fatty acids and atopic eczema/dermatitis syndrome: The relationship with a free radical <Emphasis Type="Italic">cis-trans</Emphasis> isomerization of membrane lipids

The formation of trans FA residues in membrane phospholipids may be due to a free radical-catalyzed isomerization process occurring to the cis unsaturated FA moieties. Radical stress is well documented in inflammatory processes of atopic diseases, but no data are yet available about a possible association with trans FA detected in these patients. We investigated the presence of trans lipid isomers in the erythrocyte and T-lymphocyte membranes of 26 children affected by atopic eczema/dermatitis syndrome (AEDS). Trans lipid isomers were found in both cell membranes, up to a total content of 2.7 and 4.9% of the FA composition, respectively. By using the geometrical trans lipid library derived from in vitro models of thiyl radical-catalyzed isomerization, oleic and arachidonic acid isomers were detected. The statistical significance was evaluated by comparison with an age-matched control group. These results suggest the role of an endogenous free radical isomerization path occurring to membrane unsaturated lipids, complementary to the dietary contribution, which can be involved in the lipid impairment in AEDS. This study contributes to lipidomic research regarding the double bond structure and the influence of a geometrical change of membrane lipids in physiology and diseases.     

Polymer Encapsulated Liposomes for Oral Co-Delivery of Curcumin and Hydroxytyrosol

Curcumin (Cur) is a hydrophobic polyphenol from the rhizome of Curcuma spp., while hydroxytyrosol (HT) is a water-soluble polyphenol from Olea europaea. Both show outstanding antioxidant properties but suffer from scarce bioavailability and low stability in biological fluids. In this work, the co-encapsulation of Cur and HT into liposomes was realized, and the liposomal formulation was improved using polymers to increase their survival in the gastrointestinal tract. Liposomes with different compositions were formulated: Type 1, composed of phospholipids and cholesterol; Type 2, also with a PEG coating; and Type 3 providing an additional shell of Eudragit^® S100, a gastro-resistant polymer. Samples were characterized in terms of size, morphology, ζ-potential, encapsulation efficiency, and loading capacity. All samples were subjected to a simulated in vitro digestion and their stability was investigated. The Eudragit^®S100 coating demonstrated prevention of early releases of HT in the mouth and gastric phases, while the PEG shell reduced bile salts and pancreatin effects during the intestinal digestion. In vitro antioxidant activity showed a cumulative effect for Cur and HT loaded in vesicles. Finally, liposomes with HT concentrations up to 40 μM and Cur up to 4.7 μM, alone or in combination, did not show cytotoxicity against Caco-2 cells.     

Vitamin E,cholesterol, and lipids during atherogenesis in rabbits

Rabbits were fed diets including cholesterol and 10% butterfat to determine whether polyunsaturated butter (9% 18∶2) would be less atherogenic than normal saturated butter (3% 18∶2) when fed for 12 weeks. The cholesterol diets alone, 0.5% or 2%, produced aortic plaque development, and plasma cholesterol increased 20 times, lipids increased 10 times, and vitamin E increased 5 times. The inclusion of both fat and cholesterol in the diet produced a synergistic effect, doubling these values to 40 times for cholesterol, 20 times for lipids, and 10 times for vitamin E. The higher circulating levels of cholesterol caused increased tissue levels of cholesterol. With 2% cholesterol and fat, liver and aorta cholesterol increased 10 times, heart 4 times, and muscle cholesterol 2 times. The lower 0.5% dietary cholesterol load was successful in limiting the amount of tissue cholesterol increase. Liver, aorta, heart, and muscle levels of cholesterol were only about half the concentration attained when 2% cholesterol was fed. It was concluded that there were no differences in plasma or tissue cholesterol, vitamin E, or atherosclerosis attributable to the polyunsaturated nature of the diet. The 10% butterfat diets alone, whether saturated or unsaturated, did not induce aortic plaques and did not increase blood or tissue cholesterol, lipids, or vitamin E. Our results suggest that the lipid mobilizing effect is mediated by cholesterol, probably by conversion to bile acids and a stimulation in intestinal absorption.     

Ratio of in vivo incorporation of3H arachidonic acid and14C linoleic acid into liver lipids from normal and diabetic rats

Normal and streptozotocin diabetic rats were injected via the portal vein with a labeled solution containing³H arachidonic acid and¹⁴C linoleic acid (³H/¹⁴C ratio, 0.5) during a 1 min period. Livers were quickly frozen, pulverized, and the lipids extracted and fractioned by thin layer chromatography. The incorporation of³H and¹⁴C into liver lipids was measured and the percentage distribution of radioactivity into the different lipid fractions was determined. The incorporation of¹⁴C linoleic acid and³H arachidonic acid into liver lipids is apparently reduced in rats with severe diabetes. The higher³H/^14C ratio found in the 1,2 diglycerides from diabetic rats may be explained by the apparently smaller incorporation of¹⁴C linoleic acid or by an isotopic dilution attributable to the great availability of this acid in diabetic rats. On the other hand, the higher³H/¹⁴C ratio observed in triglycerides and phospholipids from diabetic rats, due to a relatively large incorporation of³H arachidonic acid into this fraction, may be explained by the affinity of the enzymes involved in their synthesis for some 1,2-diglyceride units. Insulin was unable to correct the changes observed in the diabetic rats.     

The effect of modest vitamin E supplementation on lipid peroxidation products and other cardiovascular risk factors in diabetic patients

Among many factors, elevated lipids and lipid peroxide levels in blood are major risk factors in the development of cardiovascular disease in diabetic patients. This study has examined whether oral supplementation of vitamin E, an antioxidant, has any effect on blood lipid peroxidation products (LP) and lipid profile of diabetic patients. Thirty-five diabetics(D) were supplemented withdl-α-tocopherol (E) capsule (orally, 100 IU/d) or placebo (P) for three months in double-blind clinical trials. Plasma E was analyzed by HPLC and LP by the thiobarbituric acid-reactivity; serum lipids by auto-analyzer. Data were analyzed using pairedt-test and Wilcoxon Signed Rank Test. Vitamin E supplementation significantly lowered LP and lipid levels in diabetic patients; there were no differences in these parameters after P supplementation. There were no differences in the duration of diabetes and ages of D between P-and E-supplemented groups. This study suggests that vitamin E supplementation significantly lowers blood LP and lipid levels in diabetic patients.