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PubMedDate not supplied

Effects of butter and soybean oils on solid-phase gastric emptying in patients with functional dyspepsia.

BACKGROUND: To determine whether vegetable fats cause a slower or quicker rate of gastric emptying (GE) than animal fats, we evaluated the effect of animal butter and vegetable soybean oil on solid-phase GE in patients with functional dyspepsia. METHODS: Twenty-seven patients with functional dyspepsia were enrolled in this study. Radionuclide-labeled solid meals were used to evaluate GE. A study meal was composed of 206.8 kcal to 9.2 g protein, 45 g carbohydrate, and 10 g fat (formula 1, with animal butter: 26.2% saturated palmitic acid, 29.1% unsaturated oleic acid, 3.5% linoleic acid, and 0.5% linolenic acid; formula 2, with vegetable soybean oil: 11.0% saturated palmitic acid, 23.4% unsaturated oleic acid, 53.7% linoleic acid, and 7.8% linolenic acid). Each patient received formulas 1 and 2 as study meals on separate days. GE was represented by the gastric retention ratio of the study meal at 90 min (RR90): RR90 = residual radioactivity within the region of interest (ROI) covering the entire stomach at 90 min divided by the initial radioactivity within the ROI at 0 min. RESULTS: The RR90 was 0.648 +/- 0.156 for formula 1 and 0.600 +/- 0.131 for formula 2. There was no significant difference for the RR99 between formulas 1 and 2 (paired Student's t test, p > 0.05). Of the 27 patients, 12 (44.4%) demonstrated an increased RR99 from formula 1 to formula 2, and the RR90 of remaining 15 (55.6%) patient decreased. In addition, neither the patients with increased RR90 nor those with decreased RR90 showed a difference of symptoms between the two study meals. CONCLUSION: Our data suggest that there is no difference between these two types of fat on gastric emptying.

Adult↗
PubMed1991

Fatty acids modulate excitability in guinea-pig hippocampal slices.

A variety of fatty acids produced sustained changes in excitability in the guinea-pig hippocampal slice. Although each fatty acid was unique, a general pattern was evident. During a 30-min exposure, the synaptic potential was minimally affected, although population spike amplitude showed significant increases. With wash, synaptic efficacy increased. The increase in the synaptic potential was significant with arachidonic acid (100 microM), oleic acid (100 microM), myristic acid (250 microM) and capric acid (250 microM). Also with wash, the coupling between the synaptic potential and the population spike was reduced significantly for most of the fatty acids tested: arachidonic acid (50 microM, 100 microM), linoleic acid (100 microM) oleic acid (100 microM), stearic acid (100 microM), myristic acid (250 microM) and capric acid (250 microM, 500 microM). The fatty acids may influence neuronal excitability, in part, through a direct membrane action. The observed synaptic enhancement is consistent with a role for a fatty acid in long-term potentiation. In addition, fatty acid exposure mimics the effects of X-radiation. We suggest that free radical-induced release of fatty acids contributes to electrophysiological damage in a number of pathological states.

Action Potentials↗
PubMed2000

Identification of triacylglycerol species from high-saturated sunflower (Helianthus annuus) mutants.

The triacylglycerol (TAG) composition of oils from new high-saturated sunflower lines has been studied by means of GLC. The TAG profiles have been compared with the TAG reconstruction made after lipase hydrolysis (according to the 2-random 1,3-random theory). New TAG species with asclepic (cis,Delta11-octadecenoic acid, isomer of oleic acid), araquidic, or behenic acids have been synthesized and identified in oils from mutant lines. The TAG molecular species that contain asclepic acid instead of oleic acid have a longer retention time. Because each mutant oil has a specific TAG GLC pattern, this method could be used for a more precise validation of oil type than current fatty acid methyl ester analysis. The comparison of the results obtained by GLC with the reconstruction after pancreatic lipase hydrolysis shows, in general, a good agreement between both methods. However, results shown in this paper show that this is not always the case. TAG species containing two molecules of linoleic acid show a higher presence of palmitic or stearic acid than could be expected from a random distribution. The abundance of SLL increased in proportion to the stearic acid content of the oil, and the amount of TAG species with three unsaturated fatty acids (LLL or OLL) was therefore reduced.

Chromatography, Gas↗
PubMed1995

Comparison of the modulatory influence of maize and olive oils and butter on metabolic responses to endotoxin in rats.

1. n-3 polyunsaturated fatty acids decrease responses to cytokines and inflammatory agents. The present study examines how different intakes of n-6 and n-9 fatty acids influence the metabolic response to endotoxin in Wistar rats. 2. Weanling male rats were, for 4 weeks, fed diets containing 50, 100 or 200 g/kg fat in the form of maize oil (rich in linoleic acid), butter (poor in linoleic acid, rich in oleic acid) or olive oil (adequate in linoleic acid, rich in oleic acid) or standard laboratory chow. All butter and olive oil diets included 10 g/kg maize oil, in total fat, to avoid essential fatty acid deficiency. 3. Rats subsequently received 800 micrograms/kg Escherichia coli endotoxin or sterile saline subcutaneously. Twenty-four hours after injection, the rate of tissue protein synthesis was measured in liver, lung, kidney, tibialis muscle and spleen by the 'flooding dose' method. Protein and zinc concentrations were assayed in all tissues and serum albumin and caeruloplasmin measured. 4. In animals fed chow, protein synthetic rate increased by 18%, 29% and 27% in liver, lung and kidney respectively. Tissue zinc concentrations increased by 33% in kidney, and tissue protein increased by 17%, 23% and 17% in liver, lung and kidney respectively. Serum caeruloplasmin increased by 60% and albumin concentration fell by 14%. 5. In animals consuming the 50 g/kg maize oil diet, protein synthetic rate increased by 56%, 36% and 34% in liver, lung and kidney respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1975

Acetyl-coenzyme-A carboxylase in cultured hepatocytes. Effects of exogenous fatty acids on the content, synthesis and degradation of the enzyme.

Studies were made on the content, synthesis and degradation of acetyl-coenzyme-A carboxylase in JTC-25 - P3 cells, hepatocytes which can be maintained in a protein-free and lipid-free chemically defined medium. The addition of corn oil or fatty acid to the medium resulted in a decrease in the activity level of the enzyme without impairing the viability of cells. All the fatty acids tested exhibited this effect, although linoleic acid and oleic acid were more effective than palmitic acid, stearic acid and arachidonic acid. Immunochemical titration and Ouchterlony double-diffusion analysis indicated that the decrease in the activity level of the enzyme observed in cells incubated in medium supplemented with fatty acid can be ascribed to a reduction of the quantity of the enzyme. Isotopic leucine incorporation studies with the use of immunochemical techniques demonstrated that this reduction of the enzyme content is due to a decrease in the rate of synthesis of the enzyme. The rate of degradation of the enzyme was essentially unaffected, the half-life being 25 and 28 h, respectively, in cells incubated in the presence and absence of fatty acid. It was shown that most of the isotopic fatty acid added to the medium was incorporated into cellular phospholipids, while a very small portion of it was recovered in triglyceride and nonesterified fatty acid.

Acetyl-CoA Carboxylase↗
PubMed2001

Fatty acid and triglyceride composition of milk fat from lactating Holstein cows in response to supplemental canola oil.

The objective was to determine the influence of dietary lipid on total and sn-2 fatty acid composition and triglyceride structure of milk fat in lactating Holstein cows. Five primiparous Holstein cows surgically fitted with ruminal and duodenal cannulas were used in a 4 x 5 incomplete Latin square. All cows received a basal diet. Treatments consisted of a basal diet with no supplemental canola oil (control), basal diet with canola oil added to the concentrate portion of the diet to provide 1.6% fat, basal diet with 330 g of canola oil infused directly into the rumen, and basal diet with 330 g of canola oil infused directly into the abomasum. Canola oil treatments decreased palmitic acid and increased oleic acid content of milk fat compared with the control. Stearate was higher when canola oil was rumen available compared with control and abomasal infusion. Abomasal infusion increased linoleic and linoleic acids in milk fat compared with the other treatments. The sn-2 fatty acid composition reflected total fatty acid composition. All canola oil treatments reduced palmitic acid and increased oleic acid content at the sn-2 position. Changes in sn-2 composition reflect specificity of the acyl transferases and substrate concentration. Triglyceride composition reported as carbon number was altered by canola oil. Triglycerides in carbon number C50, C52, and C54 were increased while C32, C34, and C36 were decreased.

Abomasum↗
PubMedDate not supplied

Potential interference by free fatty acids in determination of tricyclazole in brown rice by gas chromatography with flame thermionic detection.

A convenient method is described for the determination of tricyclazole in brown rice, and the interference of free fatty acids with flame thermionic detection (FTD) is reported for the first time. Brown rice is extracted with acetone, the extract is filtered, and the filtrate is evaporated. To the residue is added 10% (w/v) NaCl solution, and the mixture is extracted with ethyl acetate. The extract is charged on a Sep-Pak Plus silica cartridge. Free fatty acids are removed from the rice by washing with diethyl ether, and tricyclazole is eluted with acetone-n-hexane (1 + 1). Tricyclazole is determined on a DB-1 capillary column by gas chromatography with FTD (GC-FTD). Linoleic acid and oleic acid, which have essentially the same retention time as tricyclazole, cannot be detected by FTD. Thus, without the Sep-Pak Plus silica cleanup, the peak height of tricyclazole in the chromatogram decreased, the extent depending on the concentration of linoleic acid. n-Hexane-acetonitrile partitioning was not used for cleanup because it could not remove 50% of the free fatty acids. Recoveries (mean +/- standard deviation, n = 5) of tricyclazole from rice fortified at 2 and 0.1 ppm were 90.5 +/- 9.4% and 81.3 +/- 10.6%, respectively. The limit of quantitation was 0.05 ppm.

Acetates↗
PubMed2001

Oleic acid-induced lung injury: thin-section CT evaluation in dogs.

PURPOSE: To validate lung attenuation measurements for quantifying extravascular lung water in oleic acid-induced pulmonary edema, compare subjective assessment with attenuation measurements, and compare this permeability-type pulmonary edema with hydrostatic-type pulmonary edema. MATERIALS AND METHODS: Thin-section computed tomography (CT) and pulmonary hemodynamic examinations were performed sequentially in six dogs before and after intravenous administration of 0.08 mg of oleic acid per kilogram of body weight. Extravascular lung water and pulmonary capillary pressure were measured. Results were compared with those reported in a canine model of hydrostatic edema. RESULTS: Oleic acid induced a progressive increase in extravascular lung water without a change in capillary pressure, which indicated pure permeability-type edema. Ground-glass opacification was detected as soon as extravascular lung water increased. Lung attenuation was highly correlated to extravascular lung water (r = 0.76, P<.001), as in hydrostatic edema, but was characterized by an almost absent gravitational gradient. CONCLUSION: Thin-section CT is sensitive for early detection and quantification of oleic acid-induced pulmonary edema in a canine model. Different from early canine hydrostatic edema, which is characterized by a gravitational gradient, early oleic acid-induced pulmonary edema in a supine dog is characterized by nearly homogeneous distribution, except for ventral sparing.

Animals↗
PubMed1994

Triacylglycerol metabolism by lymphocytes and the effect of triacylglycerols on lymphocyte proliferation.

This study investigates the ability of lymphocytes to utilize fatty acids originating from triacylglycerols and the effect of triacylglycerols upon mitogen-stimulated lymphocyte proliferation. Lymphocytes isolated from rat lymph nodes, spleen, thymus and lymphatic duct had a lipoprotein lipase activity of approx. 10 units/mg of protein, indicating that the fatty acids of circulating triacylglycerols are accessible to these cells. In culture lymph node lymphocytes hydrolysed triacylglycerols added to the medium as emulsions. Both non-esterified fatty acids and free glycerol appeared in the cell culture medium, but their concentrations indicated that a high proportion of each (65-90% of fatty acids and 60-80% of glycerol) was taken up by the cells. The incorporation and fate of triacylglycerol-fatty acids was studied by culturing the cells in the presence of tri[3H]oleoylglycerol or tri[14C]inoleoylglycerol. Both fatty acids were incorporated into lymphocyte lipids in a time-dependent manner; linoleic acid was incorporated at a significantly greater rate than oleic acid. The majority of oleic acid (greater than 70%) was incorporated into cellular triacylglycerol, while less than 10% was incorporated into phospholipids. In contrast, linoleic acid incorporation into cellular triacylglycerol never exceeded 25%, while up to 45% was incorporated into phospholipids. Triacylglycerols containing polyunsaturated fatty acids inhibited concanavalin A-stimulated lymphocyte proliferation in a concentration- and time-dependent manner; triacylglycerols containing saturated fatty acids or oleic acid were not inhibitory. Such direct effects of certain triacylglycerols on lymphocyte function may explain why some clinical trials of polyunsaturated fatty acid-rich diets have been successful in improving the condition of patients suffering from inflammatory diseases.

Animals↗
PubMed1985

Inactivation of soluble 17 beta-hydroxysteroid dehydrogenase of human placenta by fatty acids.

The sensitivity of soluble, 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) of human placenta to inactivation by fatty acids was examined. Exposure to the unsaturated fatty acids oleic, arachidonic, linoleic and linolenic acid resulted in the loss of activity. Methyl and ethyl esters of oleic acid, the saturated fatty acid, stearic acid and prostaglandins E2 and F2 alpha were without effect. Inactivation by oleic acid required the fatty acid at levels above its critical micelle concentration, 50 microM, as estimated by light-scattering. Steroid substrates and inhibitors did not protect against inactivation. NAD+, NADH, NADP+ and NADPH did protect. The concentrations of NADP+, 50 microM, and NAD, 1.5 mM, necessary for complete protection were significantly greater than their respective Michaelis constants, 0.16 microM and 15.2 microM. The data suggest that soluble 17 beta-HSD can bind to fatty acid micelles and that the binding site(s) on the enzyme are at or near pyridine nucleotide binding sites.

17-Hydroxysteroid Dehydrogenases↗
PubMed1992

Effect of feeding palmitic, oleic, and linoleic acids to Japanese quail hens (Coturnix coturnix japonica). 2. Maternal diets and stage of incubation on the lipid metabolism of quail embryos.

An experiment was conducted to evaluate the effects of diets containing 3% of either palmitic acid (Diet PA), oleic acid (Diet OA), or linoleic acid (Diet LA) and three stages of development (Days 11, 13, and 15 of incubation) on 1) weights of yolk plus yolk sac membrane (TY), yolk sac membrane (YSM), yolk, liver, and extrahepatic tissue (ET); and 2) the fatty acid composition of phospholipid, triglyceride, and cholesterol ester fractions of YSM and ET of quail embryos. Embryos from birds fed Diet LA had the highest (P less than .05) weights of TY and yolk followed by those from birds fed Diet OA and Diet PA. The weight of ET was highest (P less than .05) in embryos from birds fed Diet PA followed by those from birds fed Diet LA and Diet OA. The weights of YSM and liver were not affected by the maternal diet (P greater than .05). The weight of TY decreased whereas the weights of liver and ET increased as incubation progressed (P less than .05). The weight of YSM was maximum at Day 13 of incubation. In the phospholipid, triglyceride, and cholesterol ester fractions of both YSM and ET of embryos, the fatty acid present at the highest level, except palmitic acid, was the one added to the maternal diet. In phospholipids of both YSM and ET the oleic acid content decreased and linoleic acid increased as incubation progressed. In triglycerides and cholesterol esters of both YSM and ET, the levels of palmitic acid increased, but oleic and linoleic acids decreased, with advancing embryonic development. The results of the present study indicate that embryos from birds fed Diet PA mobilize more yolk material and produce heavier ET than embryos from birds fed Diet OA and Diet LA. The fatty acid profiles of phospholipid, triglyceride, and cholesterol esters of embryonic tissues are consistently influenced by dietary fatty acid and the stage of development.

Animals↗
PubMed1992

Synthesis and secretion of lipids by long-term cultures of female rat hepatocytes.

The objective of this work was to characterize lipid metabolism in long-term cultures of adult rat hepatocytes from female rats and explore the potential use of this culture system to study the effect of hormones, drugs and toxic chemicals on it. Hepatocytes, seeded on a feeder layer of 3T3 cells, maintained for 2 weeks their typical morphology. The cultures were able to take up [14C]acetic and [14C]oleic acid from the culture medium and incorporate them into lipids. The synthesis and secretion of lipids by [14C]acetic acid-labeled cultures had a maximum value after 11 and 13 days in culture. Triacylglycerols were the main lipidic species synthesized and secreted by hepatocytes (up to 67% of the total lipids); they also synthesized and secreted phospholipids, cholesterol and cholesterol esters from [14C]acetic acid. Similarly, [14C]oleic acid-labeled cultures synthesized and secreted mostly triacylglycerols (up to 60-70% of the total lipids), but they were also able to incorporate the labeled precursor into both cellular and secreted phospholipids and cholesterol esters. The activity of glycerol-phosphate-dehydrogenase, marker enzyme of glycerolipid synthesis, decreased slightly during the culture time whereas the activity of malic enzyme, marker of fatty acid synthesis, increased. Our results show that long-term cultures of female rat hepatocytes are able to synthesize and secrete several lipids, specially triacylglycerols, from both [14C]acetic and [14C]oleic acid for at least 2 weeks and that they maintain enzyme activities related with the synthetic pathways of glycerolipids and fatty acids.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗
PubMed2002

Atrial natriuretic peptide improves pulmonary gas exchange by reducing extravascular lung water in canine model with oleic acid-induced pulmonary edema.

OBJECTIVE: The purpose of this study was to examine and compare the effects of atrial natriuretic peptide and furosemide on pulmonary gas exchange, hemodynamics, extravascular lung water, and renal function in a dog model of oleic acid-induced pulmonary edema. DESIGN: Prospective, comparable, experimental study. SETTING: Laboratory at a university hospital. SUBJECTS: Eighteen male beagle dogs were studied under mechanical ventilation with pentobarbital anesthesia. INTERVENTIONS: Oleic acid (0.08 mL/kg) was injected and allowed for 1 hr to achieve pulmonary edema with hypoxemia at Fio2 of 0.3. After lung injury, dogs were divided into three groups; control group (n = 6) receiving saline (2.5 mL/hr for 5 hrs), atrial natriuretic peptide group (n = 6) receiving atrial natriuretic peptide (1 microg x kg(-1) x min(-1) for 5 hrs), and furosemide group (n = 6) receiving furosemide (1 mg x kg(-1) x hr(-1) for 5 hrs). MEASUREMENTS AND MAIN RESULTS: Hemodynamics, arterial blood gases, extravascular lung water, and renal function were measured hourly for 7 hrs after injury. Oleic acid increased extravascular lung water and induced hypoxemia. In the atrial natriuretic peptide group, extravascular lung water was significantly (p <.05) lower and Pao2 was significantly (p <.05) higher than in the control and furosemide groups, respectively. Pulmonary hypertension induced by oleic acid was attenuated by atrial natriuretic peptide infusion but not by saline or furosemide. Increased natriuresis/diuresis did not significantly differ between the atrial natriuretic peptide and the furosemide group, whereas creatinine clearance in the atrial natriuretic peptide group was significantly higher than that in the furosemide group. CONCLUSIONS: These findings suggest that atrial natriuretic peptide improves pulmonary gas exchange by reducing extravascular lung water and pulmonary arterial pressure, possibly independently from natriuresis/diuresis in oleic acid-induced pulmonary edema.

Animals↗
PubMed1975

[Effect of unsaturated fatty acids on the carcinogenic activity of overheated sunflower seed oil].

Non-pedigree rats were injected subcutaneously either overheated oil or the oil and one of unsaturated fatty acids (linoleic, linolenic, oleic acids), or overheated oil and fresh oil. The greatest number of tumors in earlier terms occurred in the group of rats not receiving fatty acids. In simultaneous injection of overheated sunflower oil and fatty acids (oleic and linolenic), as well as fresh oil, tumors would arise considerably more rarely. Whereas, in injection of overheated sunflower oil and linoleic acid tumors did not arise at all. The mechanism of an inhibitory action of fatty acids on tumor development is discussed.

Animals↗
PubMed1996

Measuring functional residual capacity in normal and oleic acid-injured lungs.

Functional residual capacity (FRC) is an important oxygen reserve that is often depleted in acute respiratory failure. Recent interest in the mechanisms of liquid ventilation and limited experience in measuring FRC in paralyzed, mechanically ventilated, normal and lung-injured animal models have mandated development of accurate laboratory techniques. Eight sheep, from 17 to 27 kg, were anesthetized and instrumented to provide a tracheostomy, a pulmonary artery catheter, and carotid arterial line. They were randomized to two groups, one of which received 0.07 ml/kg of intravenous oleic acid to induce lung injury. Gas ventilation of both groups was identical except for respiratory rate, which was adjusted to normalize PaCO2. FRC was measured in duplicate by both helium dilution (HD) and body plethysmography (BP). When measurements were completed, the animals were euthanized and their endotracheal tubes clamped at end expiration. The lungs were then removed and their water displacement (WD) FRC values were measured. FRC was the difference between WD and tissue weight assuming 1 ml = 1g. Pearson's correlation coefficient (R(2)) was calculated. During in vitro measurement of test lungs, HD had an R(2) value of 0.99 and BP had an R(2) value of 0.98. When compared to WD, in vivo measurement of FRC by HD had an R(2) value of 0.94 while the value for BP was 0.97. In conclusion, both HD and BP are accurate methods of determining FRC in an uninjured and injured lung model when compared to postmortem WD. Documenting changes in FRC will aid in elucidating the mechanisms of alternative ventilatory techniques.

Animals↗
PubMed1981

Comparative effects of cholic, chenodeoxycholic, and ursodeoxycholic acids on micellar solubilization and intestinal absorption of cholesterol.

Cholesterol absorption was studied in mice receiving cholic, chenodeoxycholic, or ursodeoxycholic acids (0.2% of the diet) for 2 months. Cholesterol absorption was greater with cholic acid (79%) than with chenodeoxycholic acid feeding (60%) and the lowest levels were observed during ursodeoxycholic acid feeding (37%). Under the three diets, bile acid pool and bile acid secretion were not different. Biliary cholesterol secretion was increased by cholic acid. The bile acid fed represents at least 80% of total bile acids. Micellar solubilization of oleic acid and cholesterol in the presence of each tauro-conjugated bile salt (10 mM) was determined in vitro by the coprecipitation method. Whatever the pH conditions, taurochenodeoxycholate solubilized significantly more cholesterol and more oleic acid than taurocholate. Tauroursodeoxycholate had the poorest detergent properties for both lipids. The differences between the three bile salts for cholesterol solubilization were enlarged by lowering pH and by high oleic acid concentration. Therefore the decrease in cholesterol absorption observed during ursodeoxycholic acid feeding could be explained by the poor detergent properties of this bile salt species. On the other hand, there is no relationship between the detergent properties of taurochenodeoxycholate and taurocholate and their effects on cholesterol absorption in mice. These results suggest that, in this particular case, micellar solubilization is not the rate limiting step in cholesterol absorption.

Animals↗
PubMed2001

Brown and white adipose tissue lipid composition in three successive progenies of rats: effects of ethanol exposure.

The effect of ethanol exposure on the fatty acid composition of brown and white adipose tissue in three successive rat progenies at the end of an experimental period (24 weeks) was studied. Ethanol-treated rats received a standard rat chow diet and 5, 10 and 15% ethanol in the ad libitum drinking fluid over 3 successive weeks. Then a concentration of 20% ethanol was maintained for 5 additional weeks up to the end of the experimental period. The males and females in the ethanol treated group were mated to obtain the 1st generation of offspring. Then female and male rats from the 1st generation were mated to obtain the 2nd generation. Finally, males and females from the 2nd generation were mated to obtain the 3rd generation of ethanol treated rats. Another group served as control and received only water and a standard rat chow diet. The control group was handled in the same way as the other experimental groups. In the 1st and 2nd generations the percentage of stearic acid (18:0) decreased and palmitoleic (16:1n7) and oleic acid (18:1n9) increased in both adipose tissues of ethanol-treated rats with respect to control. Additionally, n-3 and n-6 series were reduced both in brown and white adipose tissues. In the 3rd generation the fatty acid composition of the white adipose tissue was similar to that of control rats. Thus, no significant difference in essential fatty acids and oleic acid (18:1n9) were found. However, the fatty acid composition of the brown adipose tissue, in the 3rd generation, was similar to that observed in the 1st and 2nd generation. Thus, a decrease in essential fatty acids and an increase in oleic acid (18:1n9) was found. This suggests adaptation to ethanol consumption during successive progenies in white adipose tissue. However, in brown adipose tissue the values indicate a triglyceride storing during the thermogenesis, which is more important to newborns.

Adaptation, Physiological↗
PubMed2004

Mechanism of oleic acid-induced gap junctional disassembly in rat cardiomyocytes.

This study investigated the mechanism of oleic acid (OA) on gap junctions and identified the protein kinase C (PKC) isoforms involved in OA-mediated gap junction disassembly in cardiomyocytes. Control cardiomyocytes showed continuous staining of the plasma membrane at cell-cell contact areas using antibodies reacting with connexin 43 (Cx43). The spontaneous contraction rate of cultured cardiomyocytes was reduced in a time-dependent manner by OA. In addition, Cx43 expression at cell-cell junction decreased, suggesting the disassembly of gap junction. Staining for PKC and PKCalpha, which were shown to colocalize with Cx43, also decreased with increased duration of OA treatment. The effects of OA on these distributional changes at cell junctions were reversed by 24 h incubation in fresh culture medium devoid of OA. Immunoprecipitation assays confirmed the biochemical binding between Cx43 and PKC/PKCalpha, and this protein interaction was not affected by OA. This may provide the basis for simultaneous detachment of Cx and PKC/PKCalpha from the cell-cell junction to the cytosol upon OA stimulation. Western blot analysis showed that OA-induced Cx43 Ser368 phosphorylation, and that this effect could be blocked by cotreatment with the general PKC inhibitor, calphostin C, the PKC inhibitor, eV1-2, or the Src kinase inhibitor, PP1, but not by the PKCalpha inhibitor, Gö6976. eV1-2 also prevented the OA-induced disassembly of gap junctions. Taken together, these data suggest that OA-induced Cx43 Ser368 phosphorylation is mediated by activation of PKC and Src kinase and might be responsible for OA-induced gap junctional disassembly.

Animals↗