V. Ruiz-Gutierrez

Comparative Effects of Feeding Different Fats on Fatty Acid Composition of Major Individual Phospholipids of Rat Hearts

Comparative effects of feeding dietary linoleic (corn oil), oleic (olive oil), alpha-linolenic (soybean oil) and polyunsaturated fatty acids (fish oil) on lipid content and fatty acid composition of major individual phospholipids of rat hearts were examined. Feeding different diets did not result in lipid accumulation in the heart. Total triglyceride, nonesterified fatty acid, cholesteryl ester and phospholipid levels of heart tissue were not affected by the type of dietary fatty acid. However, heart free cholesterol levels decreased in both animals fed the olive and the fish oil diets. The percentage of individual phospholipids, phosphatidylcholine (PC), phosphatidylethanolamine (PE) and cardiolipin (CL) did not modify by changes in the dietary fat composition. Heart tissue from animals fed on olive oil were enriched with 18:1 (n-9 + n-7) fatty acid in all phospholipid fractions. Animals fed corn oil contained higher proportions of 18:2 (n-6) for PC, PE and CL, and the ingestion of the soybean oil diet increased 18:2 (n-6) for PC and CL in the same proportion as the ingestion of the corn oil diet. The levels of 22:6 (n-3) were increased in the fish oil-fed group, accompanied by both a decrease in total (n-6) fatty acids and an increase in total (n-3) fatty acids in the three phospholipid fractions. The 20:5 (n-3) was only detected in these animals. These results show that olive oil is as effective as fish oil in reducing heart cholesterol content and support earlier works suggesting the role of fish oil in preventing cardiovascular disease.

Influence of dietary cholesterol on polyunsaturated fatty acid composition, fluidity and membrane-bound enzymes in liver microsomes of rats fed olive and fish oil

Male rats were fed diets containing olive or marine fish oils (10% w/w) with or without added cholesterol (1% w/w). After six weeks of feeding, the major fatty acid composition, fluidity, fatty acid desaturating and cholesterol biosynthesis/esterification related enzymes of liver microsomes were determined. Both olive oil and marine fish oil diets, without added cholesterol, enriched content of oleic and docosahexaenoic acids, respectively, of rat liver microsomes. The results were consistent with reduction in delta 6 and delta 5 desaturation of n-6 essential fatty acids and higher fluidity in the marine origin oil group. Inclusion of cholesterol into diets resulted in decreased membrane arachidonic acid content, with concomitant increase in linoleic acid content. Cholesterol feeding also decreased delta 6 and delta 5 desaturase activities, as well as membrane fluidity. Furthermore, the activity of acyl-CoA:cholesterol acyltransferase decreased, whereas the activity of hydroxymethylglutaryl-CoA reductase increased, in liver microsomes from both cholesterol-fat groups.

INTAKE OF OLIVE OIL CAN MODULATE THE TRANSBILAYER MOVEMENT OF HUMAN ERYTHROCYTE MEMBRANE CHOLESTEROL

Abstract. Transbilayer movement of erythrocyte membrane cholesterol is impaired in patients affected with essential hypertension. This is an inherited disorder, but environmental factors are also involved. Dietary fats might play a role in the prevention and/or treatment of such abnormality in the kinetic pools of membrane cholesterol. We tested this hypothesis by using a diet (in which 30% of the energy came from fat) rich in olive oil or in high-oleic sunflower oil (as natural sources of monounsaturated fatty acids, MUFAs) and determining their influence on the movement of cholesterol into the lipid bilayer of the erythrocyte membrane after a four-week period. We concluded that dietary olive oil is helpful in normalizing the impaired transbilayer movement of membrane cholesterol in erythrocytes of eight normocholesterolaemic and eight hypercholesterolaemic hyperten sive patients. However, the effects cannot be attributed exclusively to the content of MUFAs (mainly oleic acid) in the diet, as high-oleic sunflower oil was unable to induce favourable changes.

Changes in fatty acid desaturation in hepatic and intestinal tissues induced by intestinal resection

We have studied the activities of Δ9, Δ6 and Δ5 desaturases in rat liver and jejunum when the entero-hepatic circulation was interrupted by either 50 or 75% intestinal resection. Desaturase activities in caecal mucosa were also determined in control and operated rats. Distal small bowel resection significantly altered the activities of desaturase enzymes in liver, jejunum and caecum. Thus, the Δ9 desaturase activity was lower in hepatic microsomes from operated animals, and this decrease was not related to the extent by which the intestine was removed. However, a significant increase in both Δ6 and Δ5 desaturases was found in these animals compared to shamoperated rats, the increase in both desaturases being higher after 75% than after 50% intestinal resection. The activities of Δ9 and Δ5 desaturases were significantly increased in jejunal mucosa of resected rats. The activity of Δ6 desaturase was increased only in 50% resected-animals. An increase in Δ6 desaturase activity was observed in caecal mucosa after resection, together with a decrease in Δ9 desaturase and no change in Δ5 desaturase activities. Enzyme activities were highest in the liver relative to the activity in jejunal and caecal homogenates. In sham rats, the caecal mucosa showed higher Δ9 and Δ6 but lower Δ5 desaturase activities than in jejunal mucosa.

Increased Na-H exchanger activity in the ileal brush-border membrane of spontaneously hypertensive rats

Abstract. In the present study, we have examined the intestinal Na+ transport, through the Na+-H+ exchanger, in ileal brush-border membrane vesicles (BBMV) isolated from spontaneously hypertensive rats (SHR), and normotensive Wistar Kyoto (WKY) rats as a control group. Na+ uptake into ileal BBMV was stimulated in the presence of a proton gradient (pH 5.5 inside/pH 7.5 outside) in SHR and WKY rats, resulting in a transient accumulation (overshoot) in both groups of rats. No overshoot was observed in the absence of a pH gradient. The magnitude of the accumulation was significantly higher in SHR than in WKY rats. Uptake of Na+ at equilibrium was identical in the presence and the absence of a proton gradient and was not changed in SHR. The use of amiloride inhibited pH gradient-driven Na+ uptake in a dose-dependent manner with a Ki of 90 μM and 100 μM for SHR and WKY rats, respectively. The relationship between proton gradient-driven Na+ uptake and external Na+ concentration was saturable and conformed to Michaelis-Menten kinetics in both SHR and WKY rats. Lineweaver-Burk analysis of the pH gradient-driven Na+ uptake indicated values of Vmax that were significantly increased in SHR compared to WKY rats (11.4±0.55 nmol/mg/8 s vs. 4.96±0.78 nmol/mg/8 s for SHR and WKY rats, respectively). In contrast, similar Km values for Na+ were found between SHR and WKY rats (4.0±0.2 mM vs. 4.9±0.6 mM for SHR and WKY rats, respectively). These studies show derangement in ileal BBMV Na+ transport of SHR, which is characterized by increased Na+-H+ exchanger activity.

Changes in uptake of linoleic acid and cholesterol by jejunal sacs of rats in vitro, after distal small-bowel resection.

Both linoleic acid and cholesterol uptake were studied in small-intestinal sacs of rats in vitro after distal small-bowel resection (DSBR). The relationship between linoleic acid concentration and its absorption was non-linear at low concentrations and became linear at high concentrations in the three groups of animals. These observations indicate that a concentration-dependent dual mechanism of transport is operative in linoleic acid intestinal uptake. Experiments with rotenone and ouabain suggest that a facilitated diffusion is the predominant mechanism of absorption at low concentrations, whereas at high concentrations simple diffusion is predominant. The apparent kinetic constants of linoleic acid uptake (Kd, Kt, and Vmax) increased after DSBR. The uptake of linoleic acid is, however, influenced by the simultaneous presence of linolenic acid, the inhibition constant being decreased after the surgical operation. After the surgical operation an increase of cholesterol uptake was observed, with a parallel enhancement in the apparent mass-transfer coefficient (Kd). Taken together, these results suggest that both organ growth and changes in transport function of the enterocytes appear to be involved in the adaptive response of the bowel to intestinal resection.

Lipid composition, phospholipid profile and fatty acid of rat caecal mucosa

In this study, we examined the lipid composition of rat caecal mucosa, including the fatty acid composition of major phospholipid classes. Phospholipids accounted for 90% of the total lipid, with cholesterol, triacylglycerols, diacylglycerols, fatty acids and cholesterol ester making up the remainder. Therefore, a phospholipid to neutral lipid ration of 9:1 was found. Phosphatidylethanolamine was the predominant phospholipid, with phosphatidylcholine as the second most abundant phospholipid. Cardiolipin, phosphatidylserine, phosphatidylinositol and lysophosphatidylcholine were present in lesser amounts. Sphingomyelin and lysophosphatidylethanolamine were only detected in trace amounts. The major fatty acids present in both the lipid and all phospholipid fractions were palmitate, stearate, oleate, linoleate and arachidonate. Other fatty acids of chain length greater than C20 were only detected in phospholipid fraction and accounted for < 5% of the total fatty acids in this fraction. However, 11.10% of 22:6 (n-3) and 7.17% of 24:0 were detected in phosphatidylserine and lysophosphatidylcholine, respectively. The results are discussed in terms of their possible physiological significance.

CAECAL AND COLONIC UPTAKE OF BOTH LINOLEIC ACID AND CHOLESTEROL IN RATS FOLLOWING INTESTINAL RESECTION

Caecal and colonic uptake of both linoleic acid and cholesterol were studied in rats after distal small bowel resection (DSBR). The results showed that the surgical operation increased the caecal and colonic uptake of linoleic acid. Supplementation with linolenic acid inhibited caecal and colonic uptake of linoleic acid. Experiments carried out in the presence of rotenone and ouabain suggest that facilitated diffusion is the predominant mechanism of caecal and colonic linoleic acid absorption, at least at low concentrations. An increase in caecal and colonic uptake of cholesterol was observed after the surgical operation. The study showed that facilitated diffusion seems to be the mechanism of linoleic acid absorption in the caecum and colon, and that both organ growth and changes in transport function of the epithelial cells of caecum and colon appear to be involved in the adaptive response of the bowel to intestinal resection.

Changes in fatty acid composition of rat liver and serum induced by distal small bowel resection.

The serum lipid composition and the fatty-acid profiles of the major lipid fraction (triglycerides, esterified cholesterol, and phospholipid) of liver and serum were examined 6 weeks after both 50% and 75% distal small bowel resection (DSBR). Total serum lipid content did not modify after DSBR. Esterified cholesterol and phospholipid levels of the serum did not significantly change after the operation. However, a significant increase in both free cholesterol and triglyceride levels was observed after DSBR. Different fatty acid changes in the liver and serum lipid fractions were found after DSBR, with the greatest differences in the hepatic esterified cholesterol fraction. These results suggest that DSBR affects both the lipid composition and the fatty acid composition of major lipid fraction of liver and serum.

Effect of distal small bowel resection on ACAT activity and microsomal lipid composition in rat small intestine

1. The acyl-CoA:cholesterol acyltransferase (ACAT) activity and lipid composition of intestinal microsomal membrane were investigated 6 weeks after both 50 and 75% distal small bowel resection (DSBR). 2. No changes in both microsomal ACAT activity and cholesteryl ester levels were found, while microsomal non-esterified cholesterol content was increased after the surgical operation. 3. The total phospholipid content of the microsomes did not change as a result of DSBR. 4. The microsomal phospholipid fatty acid composition showed a significant increase in saturated fatty acids together with no changes in both total monounsaturated and total polyunsaturated fatty acids after resection. 5. An increase in the levels of linoleic acid accompanied by a decrease in arachidonic acid was found in remnant intestine of resected rats.