Low Density Lipoprotein Triacylglycerol Research Articles

Triacylglycerol Fatty Acid Positional Distribution and Postprandial Lipid Metabolism

Triacylglycerol (TAG) structure affects the biochemical properties of fat. According to studies with animals and infants, absorption of palmitic and stearic acids is inferior from the stereospecifically numbered (sn) positions sn-1 and sn-3 compared with from the sn-2 position of TAGs, and saturated fatty acids in the sn-2 position, delay the clearance of chylomicron remnants. Medium chain fatty acids are transported both in lymph and in portal blood, lymphatic absorption being more efficient from the sn-2 position.Recently, new information has arisen on the variation in TAG composition within the postprandial period and on differences in the TAG structure-dependent formation and clearance of chylomicron and very low density lipoprotein TAGs. This article reviews the metabolism of TAG fatty acids in different sn positions.

Isolation of very low density lipoprotein phospholipids enriched in ethanolamine phospholipids from rats injected with Triton WR 1339

Phospholipids carried by very low density lipoprotein (VLDL) are hydrolysed in circulation by lipoprotein and hepatic lipases and lecithin-cholesterol acyltransferase. We have previously demonstrated [6] [J.J. Ågren, A. Ravandi, A. Kuksis, G. Steiner, Structural and compositional changes in very low density lipoprotein triacylglycerols during basal lipolysis, Eur. J. Biochem. 269 (2002) 6223–6232] that the infusion of Triton WR 1339 (TWR), which inhibits these lipases, leads in 2 h to five-fold increase in VLDL triacylglycerol concentration along with major differences in the composition of their molecular species. The present study demonstrates that the accumulation of triacylglycerols is accompanied by major changes in the content of the VLDL phospholipids, of which the most significant is the enrichment of phosphatidylethanolamine (PtdEtn). This finding coincides with the enrichment in PtdEtn demonstrated in the VLDL of a hepatocytic Golgi fraction but it had not been demonstrated that the Golgi VLDL, along with its unusual phospholipid composition, can be directly transferred to plasma. Aside from providing an easy access to nascent plasma VLDL, the TWR infusion demonstrates that lipoprotein and hepatic lipases are also responsible for the degradation of plasma VLDL PtdEtn, as independently demonstrated for plasma phosphatidylcholine. Our results indicate also, with the exception of lysophosphatidylcholine, that preferential basal hydrolysis no dot lead to major differences in molecular species composition between circulating and newly secreted VLDL phospholipids. The comparison of the molecular species composition of VLDL and liver phospholipids suggests a selective secretion of PtdEtn and sphingomyelin molecular species during VLDL secretion.

Triacylglycerol Is Synthesized in a Specific Subclass of Caveolae in Primary Adipocytes

A principal metabolic function of adipocytes is to synthesize triacylglycerol (TG) from exogenous fatty acids. The level of fatty acids has to be tightly controlled in the adipocyte, as they can act as detergents that rapidly dissolve the plasma membrane, causing cell lysis if allowed to accumulate. Fatty acids therefore have to be efficiently converted to TG and stored in the central lipid droplet. We report that in intact primary adipocytes exogenous oleic acid was taken up and directly converted to TG in the plasma membrane, in a novel subclass of caveolae that specifically contains the protein perilipin. Isolated caveolae catalyzed de novo TG synthesis from oleic acid and glycerol 3-phosphate. Electron microscopy revealed the presence of caveolin and perilipin in caveolae and in lipid-laden bulbs in the plasma membrane, and fluorescence microscopy demonstrated colocalization of fatty acids/TG with caveolin and perilipin at the plasma membrane. A second caveolae fraction was isolated, which lacked perilipin and the triacylglycerol synthesizing enzymes. Both caveolae fractions contained caveolin-1 and the insulin receptor. The findings demonstrate that specific subclasses of caveolae carry out specific functions in cell metabolism. In particular, triacylglycerol is synthesized at the site of fatty acid entry in one of these caveolae classes.

Changes in serum lipoprotein lipids and their fatty acid compositions and lipid peroxidation in growing rats fed soybean protein versus casein with or without cholesterol*1

We compared the effects of diets based on soybean protein and casein supplemented or not supplemented with 0.1% cholesterol on plasma lipoprotein lipid amounts and their fatty acid compositions, lecithin:cholesterol acyl-transferase activity, and lipid peroxidation. The composition and concentration of lipid and apolipoprotein in different lipoprotein classes, plasma LCAT activity, and lipid peroxidation were determined in rats fed 20% highly purified soybean protein or casein with or without 0.1% cholesterol for 2 mo. Soybean protein and casein diets with or without cholesterol had similar plasma total cholesterol concentrations. Soybean protein consumption diminished very low-density lipoprotein particle number, as measured by diminished contents of very low-density lipoprotein triacylglycerol, phospholipid, and apolipoprotein-B100. Lecithin:cholesterol acyl-transferase activity was not significantly modified by either protein. The soybean protein diet decreased the linoleate desaturation index (20:4[ω-6]/18:2[ω-6]) in liver and high-density lipoprotein fraction 2–3-phospholipids but enhanced red blood cell resistance against free radical attack. Addition of cholesterol to both protein diets decreased concentrations of high-density lipoprotein fraction 2–3 cholesterol. Lecithin:cholesterol acyl-transferase activity tended to be greater after cholesterol feeding, likely due to the enhanced high-density lipoprotein fraction 2–3 apolipoprotein-AI, a cofactor activator for lecithin:cholesterol acyl-transferase. Regardless of dietary protein source, cholesterol supplementation decreased the linoleate desaturation index in liver and plasma lipoprotein lipids and red blood cell resistance to free radical attack. Our results suggest that the dietary protein origin affects lipid peroxidation and polyunsaturated fatty acid biosynthesis and distribution among liver and different lipoprotein lipid classes, but plays only a minor role in the regulation of plasma and lipoprotein cholesterol concentrations. Providing dietary cholesterol (0.1%) with casein or soybean protein attenuates the effects of these proteins, with the exception of plasma cholesterol.

Changes in serum lipoprotein lipids and their fatty acid compositions and lipid peroxidation in growing rats fed soybean protein versus casein with or without cholesterol

Objective We compared the effects of diets based on soybean protein and casein supplemented or not supplemented with 0.1% cholesterol on plasma lipoprotein lipid amounts and their fatty acid compositions, lecithin:cholesterol acyl-transferase activity, and lipid peroxidation. Methods The composition and concentration of lipid and apolipoprotein in different lipoprotein classes, plasma LCAT activity, and lipid peroxidation were determined in rats fed 20% highly purified soybean protein or casein with or without 0.1% cholesterol for 2 mo. Results Soybean protein and casein diets with or without cholesterol had similar plasma total cholesterol concentrations. Soybean protein consumption diminished very low-density lipoprotein particle number, as measured by diminished contents of very low-density lipoprotein triacylglycerol, phospholipid, and apolipoprotein-B100. Lecithin:cholesterol acyl-transferase activity was not significantly modified by either protein. The soybean protein diet decreased the linoleate desaturation index (20:4[ω-6]/18:2[ω-6]) in liver and high-density lipoprotein fraction 2–3-phospholipids but enhanced red blood cell resistance against free radical attack. Addition of cholesterol to both protein diets decreased concentrations of high-density lipoprotein fraction 2–3 cholesterol. Lecithin:cholesterol acyl-transferase activity tended to be greater after cholesterol feeding, likely due to the enhanced high-density lipoprotein fraction 2–3 apolipoprotein-AI, a cofactor activator for lecithin:cholesterol acyl-transferase. Regardless of dietary protein source, cholesterol supplementation decreased the linoleate desaturation index in liver and plasma lipoprotein lipids and red blood cell resistance to free radical attack. Conclusions Our results suggest that the dietary protein origin affects lipid peroxidation and polyunsaturated fatty acid biosynthesis and distribution among liver and different lipoprotein lipid classes, but plays only a minor role in the regulation of plasma and lipoprotein cholesterol concentrations. Providing dietary cholesterol (0.1%) with casein or soybean protein attenuates the effects of these proteins, with the exception of plasma cholesterol.

Increased VLDL-TAG turnover during and after acute moderate-intensity exercise.

Breakdown rates of very low density lipoprotein triacylglycerols (VLDL-TAG) were quantified before (3 h), during (45 min), and after (3 h) moderate physical exercise at 40% VO2 max in young sedentary subjects (four male and four female, age 29.8 +/-1.6 yr, BMI 24.1 +/- 0.9 kg x m, VO2max 37.0 +/- 1.7 mL x kg x min), using boluses of H5-glycerol (100 mu mol x kg) and H2-palmitate (6.6 mu mol x kg). The catabolic rates of VLDL-TAG were calculated from the decay in the isotopic enrichment using a single-pool model. The results were compared with those obtained during 6 h of rest in five of the same volunteers. VLDL-TAG concentration remained constant throughout the study with exercise (P = NS). The fractional catabolic rate was nearly doubled from rest to exercise (P < 0.05 vs rest) and increased to an even greater extent during the early phase of recovery (P < 0.001 vs rest). During the late recovery phase, the value returned to the preexercise value (P = NS vs rest). The values for the absolute catabolic rate (kabs) followed the same trend. VLDL-TAG turnover was increased during exercise and during the early phase of recovery.

Stable isotope tracer dilution for quantifying very low-density lipoprotein-triacylglycerol kinetics in man

Background & aim: A number of approaches have been employed in the past to measure very low-density lipoprotein (VLDL) triacylglycerol (TG) kinetics in humans in vivo, varying in the selection of tracer and mode of administration. All, however, make use of labeled TG precursors and more or less complicated mathematical models to derive the kinetic parameters of interest. The aim of the present study was to develop a conceptually straightforward method, based on the traditional tracer infusion technique, for quantifying VLDL-TG production rates in man using stable isotopes. Method: Our approach involves ingestion of [U- 13C 3]glycerol to endogenously label the glycerol in VLDL-TG, plasmapheresis, isolation of the newly 13C-labeled VLDL from plasma, and administration within the next 2–3 days via a primed constant autologous reinfusion. This procedure produces enough tracer for a priming dose plus 2–3 h of infusion. In the physiological conditions examined (basal and hyperglycemic states, fat- and carbohydrate-rich diets), with almost 3-fold ranging VLDL-TG pool sizes, a steady state in plasma VLDL-TG glycerol tracer-to-tracee ratio was readily achieved within 2 h. Consequently, calculations are made according to the isotope dilution principle, thus avoiding assumptions implicit in more complicated models. Conclusion: The stable isotope VLDL-TG tracer dilution method offers an alternative and reliable tool for the determination of endogenous VLDL-TG kinetics in man under a variety of metabolic states.

Effects of Exogenous Glucagon on Lipids in Lipoproteins and Liver of Lactating Dairy Cows

Decreased concentrations of phospholipids, free cholesterol, and cholesteryl ester in plasma and liver are associated with fatty liver, a major metabolic disease of dairy cows in early lactation. The objective was to test whether daily subcutaneous injections of 7.5 and 15mg of glucagon, which can decrease concentrations of liver triacylglycerol, affect concentrations of plasma lipoprotein components and liver lipids other than triacylglycerol. Multiparous Holstein cows (n = 32) were grouped on the basis of liver triacylglycerol concentrations at d 8 postpartum into “normal” (n = 8; triacylglycerol <1% liver wet wt) and “susceptible to fatty liver” (n = 24; triacylglycerol >1% liver wet wt) cows. Susceptible cows were assigned randomly to three groups and beginning at d 8 postpartum received 0 (same for Normal cows), 2.5, or 5mg of glucagon by subcutaneous injections every 8h for 14 d. In comparison to saline injections, subcutaneous injections of glucagon either increased or tended to increase concentrations of phospholipids and free cholesterol in liver, with greater increases of the latter during ambient temperatures below 35°C. Glucagon injections decreased or tended to decrease concentrations of very low-density lipoprotein-triacylglycerol, high-density lipoprotein1-phospholipids, and high-density lipoprotein2-free cholesterol in plasma, with no changes of the latter two during ambient temperatures below 35°C. The results indicate that subcutaneously administered glucagon has only minor effects on the lipid transport in plasma of dairy cows in early lactation with more beneficial effects occurring during ambient temperatures below 35°C and, most importantly, no indications that glucagon has negative effects.

SKELETAL MUSCLE FAT AND CARBOHYDRATE METABOLISM DURING RECOVERY FROM GLYCOGEN DEPLETING EXERCISE IN HUMANS

The primary aim of the present study was to determine whether intramuscular triacylglycerol (IMTG) utilization contributed significantly to the increase in lipid oxidation during recovery from exercise, as determined from the muscle biopsy technique. In addition, we also examined the regulation of pyruvate dehydrogenase (PDHa) and changes in muscle acetyl units during an 18 h recovery period after glycogen-depleting exercise. Eight endurance-trained males completed an exhaustive bout of exercise (~90 min) on a cycle ergometer followed by ingestion of carbohydrate (CHO)-rich meals (64–70 % of energy from carbohydrate) at 1, 4 and 7 h of recovery. Duplicate muscle biopsies were obtained at exhaustion, and 3, 6 and 18 h of recovery. Despite the large intake of CHO during recovery (491 ± 28 g or 6.8 ± 0.3 g kg _1 ), respiratory exchange ratio values of 0.77 to 0.84 indicated a greater reliance on lipid as an oxidative fuel. However, there was no net IMTG utilization during recovery. IMTG content at exhaustion was 23.5 ± 3.5 mmol (kg dry wt) _1 , and remained constant at 24.6 ± 2.6, 25.7 ± 2.8 and 28.4 ± 3.0 mmol (kg dry wt) _1 after 3, 6 and 18 h of recovery. Muscle glycogen increased significantly from 37 ± 11 mmol (kg dry wt) _1 at exhaustion, to 165 ± 13, 250 ± 18, and 424 ± 22 mmol (kg dry wt) _1 at 3, 6 and 18 h of recovery, respectively. PDHa was reduced at 6 and 18 h when compared to exhaustion, but did not change during the recovery period. Acetyl-CoA, acetylcarnitine and pyruvate contents declined significantly after 3 h of recovery compared to exhaustion, and thereafter remained unchanged. We conclude that IMTG has a negligible role in contributing to the enhanced fat oxidation during recovery from exhaustive exercise. Despite the elevation of glucose and insulin following high-CHO meals during recovery, CHO oxidation and PDH activation were decreased, supporting the hypothesis that glycogen resynthesis is of high metabolic priority. Plasma fatty acids, very low density lipoprotein triacylglycerols, as well as intramuscular acetylcarnitine stores are likely to be important fuel sources for aerobic energy, particularly during the first few hours of recovery.

Skeletal muscle fat and carbohydrate metabolism during recovery from glycogen-depleting exercise in humans.

The primary aim of the present study was to determine whether intramuscular triacylglycerol (IMTG) utilization contributed significantly to the increase in lipid oxidation during recovery from exercise, as determined from the muscle biopsy technique. In addition, we also examined the regulation of pyruvate dehydrogenase (PDHa) and changes in muscle acetyl units during an 18 h recovery period after glycogen-depleting exercise. Eight endurance-trained males completed an exhaustive bout of exercise (approximately 90 min) on a cycle ergometer followed by ingestion of carbohydrate (CHO)-rich meals (64-70 % of energy from carbohydrate) at 1, 4 and 7 h of recovery. Duplicate muscle biopsies were obtained at exhaustion, and 3, 6 and 18 h of recovery. Despite the large intake of CHO during recovery (491 +/- 28 g or 6.8 +/- 0.3 g kg-1), respiratory exchange ratio values of 0.77 to 0.84 indicated a greater reliance on lipid as an oxidative fuel. However, there was no net IMTG utilization during recovery. IMTG content at exhaustion was 23.5 +/- 3.5 mmol (kg dry wt)-1, and remained constant at 24.6 +/- 2.6, 25.7 +/- 2.8 and 28.4 +/- 3.0 mmol (kg dry wt)-1 after 3, 6 and 18 h of recovery. Muscle glycogen increased significantly from 37 +/- 11 mmol (kg dry wt)-1 at exhaustion, to 165 +/- 13, 250 +/- 18, and 424 +/- 22 mmol (kg dry wt)-1 at 3, 6 and 18 h of recovery, respectively. PDHa was reduced at 6 and 18 h when compared to exhaustion, but did not change during the recovery period. Acetyl-CoA, acetylcarnitine and pyruvate contents declined significantly after 3 h of recovery compared to exhaustion, and thereafter remained unchanged. We conclude that IMTG has a negligible role in contributing to the enhanced fat oxidation during recovery from exhaustive exercise. Despite the elevation of glucose and insulin following high-CHO meals during recovery, CHO oxidation and PDH activation were decreased, supporting the hypothesis that glycogen resynthesis is of high metabolic priority. Plasma fatty acids, very low density lipoprotein triacylglycerols, as well as intramuscular acetylcarnitine stores are likely to be important fuel sources for aerobic energy, particularly during the first few hours of recovery.

Structural and compositional changes in very low density lipoprotein triacylglycerols during basal lipolysis

Triacylglycerols secreted by liver and carried by very low density lipoprotein (VLDL) are hydrolysed in circulation by lipoprotein and hepatic lipases. These enzymes have been shown to have positional and fatty acid specificity in vitro. If there were specificity in basal lipolysis in vivo, triacylglycerol compositions of circulating and newly secreted VLDL would be different. To study this we compared the composition of normal fasting VLDL triacylglycerol of Wistar rats to that obtained after blocking lipolysis by Triton WR1339, which increased plasma VLDL triacylglycerol concentration about 4.7-fold in 2 h. Analyses of molecular species of sn-1,2- and sn-2,3-diacylglycerol moieties and stereospecific triacylglycerol analysis revealed major differences between the groups in the VLDL triacylglycerol composition. In nontreated rats, the proportion of 16:0 was higher and that of 18:2n-6 lower in the sn-1 position. The proportion of 14:0 was lower in all positions and that of 18:0 was lower in the sn-1 and sn-3 positions in nontreated rats whereas the proportions of 20:4n-6, 20:5n-3, 22:5n-3 and 22:6n-3 were higher in the sn-1 and lower in the sn-2 position. These results suggest that the fatty acid of the sn-1 position is the most decisive factor in determining the sensitivity for hydrolysis of the triacylglycerol. In addition, triacylglycerol species with highly unsaturated fatty acids in the sn-2 position also favoured hydrolysis. The in vivo substrate specificity followed only partly that obtained in in vitro studies indicating that the nature of molecular association of fatty acids in natural triacylglycerol affects its susceptibility to lipolysis. To conclude, our results indicate that preferential basal lipolysis leads to major structural differences between circulating and newly secreted VLDL triacylglycerol. These differences extend beyond those anticipated from analysis of total fatty acids and constitute a previously unrecognized feature of VLDL triacylglycerol metabolism.

Mitochondrial glycerol-3-phosphate acyltransferase-deficient mice have reduced weight and liver triacylglycerol content and altered glycerolipid fatty acid composition.

Microsomal and mitochondrial isoforms of glycerol-3-phosphate acyltransferase (GPAT; E.C. 2.3.1.15) catalyze the committed step in glycerolipid synthesis. The mitochondrial isoform, mtGPAT, was believed to control the positioning of saturated fatty acids at the sn-1 position of phospholipids, and nutritional, hormonal, and overexpression studies suggested that mtGPAT activity is important for the synthesis of triacylglycerol. To determine whether these purported functions were true, we constructed mice deficient in mtGPAT. mtGPAT(-/-) mice weighed less than controls and had reduced gonadal fat pad weights and lower hepatic triacylglycerol content, plasma triacylglycerol, and very low density lipoprotein triacylglycerol secretion. As predicted, in mtGPAT(-/-) liver, the palmitate content was lower in triacylglycerol, phosphatidylcholine, and phosphatidylethanolamine. Positional analysis revealed that mtGPAT(-/-) liver phosphatidylethanolamine and phosphatidylcholine had about 21% less palmitate in the sn-1 position and 36 and 40%, respectively, more arachidonate in the sn-2 position. These data confirm the important role of mtGPAT in the synthesis of triacylglycerol, in the fatty acid content of triacylglycerol and cholesterol esters, and in the positioning of specific fatty acids, particularly palmitate and arachidonate, in phospholipids. The increase in arachidonate may be functionally significant in terms of eicosanoid production.

Fat utilization during exercise: adaptation to a fat-rich diet increases utilization of plasma fatty acids and very low density lipoprotein-triacylglycerol in humans

Fat utilization during exercise: adaptation to a fat-rich diet increases utilization of plasma fatty acids and very low density lipoprotein-triacylglycerol in humans

Fat utilization during exercise: adaptation to a fat-rich diet increases utilization of plasma fatty acids and very low density lipoprotein-triacylglycerol in humans

1. This study was carried out to test the hypothesis that the greater fat oxidation observed during exercise after adaptation to a high-fat diet is due to an increased uptake of fat originating from the bloodstream. 2. Of 13 male untrained subjects, seven consumed a fat-rich diet (62 % fat, 21 % carbohydrate) and six consumed a carbohydrate-rich diet (20 % fat, 65 % carbohydrate). After 7 weeks of training and diet, 60 min of bicycle exercise was performed at 68 +/- 1 % of maximum oxygen uptake. During exercise [1-(13)C]palmitate was infused, arterial and venous femoral blood samples were collected, and blood flow was determined by the thermodilution technique. Muscle biopsy samples were taken from the vastus lateralis muscle before and after exercise. 3. During exercise, the respiratory exchange ratio was significantly lower in subjects consuming the fat-rich diet (0.86 +/- 0.01, mean +/- S.E.M.) than in those consuming the carbohydrate-rich diet (0.93 +/- 0.02). The leg fatty acid (FA) uptake (183 +/- 37 vs. 105 +/- 28 micromol min(-1)) and very low density lipoprotein-triacylglycerol (VLDL-TG) uptake (132 +/- 26 vs. 16 +/- 21 micromol min(-1)) were both higher (each P < 0.05) in the subjects consuming the fat-rich diet. Whole-body plasma FA oxidation (determined by comparison of (13)CO(2) production and blood palmitate labelling) was 55-65 % of total lipid oxidation, and was higher after the fat-rich diet than after the carbohydrate-rich diet (13.5 +/- 1.2 vs. 8.9 +/- 1.1 micromol min(-1) kg(-1); P < 0.05). Muscle glycogen breakdown was significantly lower in the subjects taking the fat-rich diet than those taking the carbohydrate-rich diet (2.6 +/- 0.5 vs. 4.8 +/- 0.5 mmol (kg dry weight)(-1) min(-1), respectively; P < 0.05), whereas leg glucose uptake was similar (1.07 +/- 0.13 vs. 1.15 +/- 0.13 mmol min(-1)). 4. In conclusion, plasma VLDL-TG appears to be an important substrate source during aerobic exercise, and in combination with the higher plasma FA uptake it accounts for the increased fat oxidation observed during exercise after fat diet adaptation. The decreased carbohydrate oxidation was apparently due to muscle glycogen sparing and not to diminished plasma glucose uptake.

EFFECT OF ACUTE EXERCISE UPON LIPID METABOLISM IN THE LIVER

Very low density lipoprotein-triacylglycerol (VLDL-TAG) and ketone bodies secreted by the liver constitute important substrates for the exercising muscle, and the former is also important in the reposition of intramuscular TAG stores. Our aim was to investigate the effect of exercise upon fatty acid partitioning in the liver. Male Wistar rats were submitted to a single bout of exercise (running on a treadmill at 65% VO2max for 1h). The exercised (E) and control (C) rats were killed immediately after exercise and liver relative weight, fat content (petroleum ether extraction), fat uptake capacity (14C-oleate content in tissue after oral administration of labelled triolein - 2.5 μCi per rat) and VLDL-TAG secretion were evaluated. Liver weight in relation to total body mass was decreased (12.5%) in E vs C group (p < 0.05) while fat content (mg fat/g tissue) was not altered (42.9 ± 2.2 and 41.5 ± 2.5 mg fat/g tissue for E and C, respectively). The reduction of liver mass was associated with decreased glycogen content in E (58%, p < 0.01). 14C-oleate incorporation into the lipid fraction of the liver was reduced (39%, p < 0.05) in E in relation to C, while the incorporation into VLDL-TAG was increased in the exercised animals (15%, p < 0.05). The results suggest that lipid is being diverted from storage towards oxidation/secretion in the exercised animals. Sponsor: CAPES and FAPESP

Glyceroneogenesis and the Source of Glycerol for Hepatic Triacylglycerol Synthesis in Humans

Glyceroneogenesis, i.e. the synthesis of the glycerol moiety of triacylglycerol from pyruvate, has been suggested to be quantitatively important in both the liver and adipose tissue during fasting. However, the actual contribution of glyceroneogenesis to triacylglycerol synthesis has not been quantified in vivo in human studies. In the present study we have measured the contribution of glycerol and pyruvate to in vivo synthesis of hepatic triacylglycerol in nonpregnant and pregnant women after an overnight fast. Five nonpregnant women were administered [(13)C(3)]glycerol tracer as prime constant rate infusion, and the appearance of tracer in plasma glucose and triacylglycerol was quantified using gas chromatography-mass spectrometry. The contribution of pyruvate to hepatic triacylglycerol was quantified in nonpregnant and pregnant women using the deuterium labeling of body water method. The appearance of [(2)H] in hydrogens on C(1) and C(3) of triacylglycerol was measured following periodate oxidation of the glycerol isolated from hydrolyzed triacylglycerol. After a 16-h fast, approximately 6.1% of the plasma triacylglycerol pool was derived from plasma glycerol, whereas 10 to 60% was derived from pyruvate in nonpregnant women and pregnant women early in gestation. Our data suggest that glyceroneogenesis from pyruvate is quantitatively a major contributor to plasma triacylglycerol synthesis and may be important for the regulation of very low density lipoprotein triacylglycerol production. Our data also suggest that 3-glycerol phosphate is in rapid equilibrium with the triosephosphate pool, resulting in rapid labeling of the triose pool by the administered tracer glycerol. Because the rate of flux of triosephosphate to glucose during fasting far exceeds that to triacylglycerol, more glycerol ends up in glucose than in triacylglycerol. Alternatively, there may be two distinct pools of 3-glycerol phosphate in the liver, one involved in generating triosephosphate from glycerol and the other involved in glyceride-glycerol synthesis.

Utilization of very low density lipoprotein by rat heart: the effect of endotoxin.

The effect of endotoxin on myocardial utilization of very low density lipoprotein (VLDL) triacylglycerol (TAG) was studied. VLDL was prepared by rat liver perfusion and tested as substrate in the isolated working rat heart. Both liver and heart donor rats were pretreated in vivo with endotoxin or vehicle (control). VLDL-TAG synthesized by endotoxin-pretreated livers was assimilated and oxidized at an increased rate by hearts compared with control VLDL-TAG, regardless of the cardiac endotoxic status, with increased cardiac mechanical performance (cardiac output, hydraulic work). There was no change in incorporation of labeled VLDL lipids into myocardial tissue lipids. Lipoprotein lipase (LPL) activity was increased in endotoxin-pretreated hearts, and after perfusion with "endotoxic" VLDL, there was a tendency for translocation of LPL from tissue-residual to heparin-releasable compartments, but these changes were modest. Analysis of the VLDL composition showed that endotoxin-pretreated livers produced apolipoprotein (apo)-B48 VLDL with decreased particle size (and hence TAG content), but apo-B100 VLDL was unchanged. Oleate content of VLDL was increased, but there was no difference in apo-C or apo-E content. These results suggest that VLDL-TAG produced during sepsis/endotoxinemia may be destined for utilization by the heart as energy substrate. However, the mechanism for its increased efficacy is uncertain.

Effects of a prudent diet containing either lean beef and mutton or fish and skinless chicken on the plasma lipoproteins and fatty acid composition of triacylglycerol and cholesteryl ester of hypercholesterolemic subjects

In this two-phase crossover study, 39 hypercholesterolemic subjects followed a prudent diet with either lean red meat or fish and skinless chicken (treatment groups), and 13 subjects (reference group) followed their habitual diet. Fasting blood samples were analyzed for plasma total cholesterol, triacylglycerol (TAG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein one- and two-cholesterol, apolipoprotein-B, very low density lipoprotein cholesterol, and very low density lipoprotein TAG, and fatty acid composition of plasma TAG and cholesteryl ester (CE). Body mass and blood pressure were determined. Seven-day dietary records were kept once at baseline and twice during the treatment periods. Significant differences were observed in dietary intake between the baseline and treatment diets and between the two treatment diets. HDL-C ( P < 0.05) and diastolic blood pressure ( P < 0.01) were higher in patients on the red meat diet than in those on the chicken-fish diet. No other significant differences in lipoproteins were observed between the effects of the two treatment diets. The linoleic acid (%), eicosapentaenoic acid (%), and the eicosapentaenoic acid/arachidonic acid ratios in TAG and CE were higher ( P < 0.01) in subjects on the chicken-fish diet than in those on the red meat diet. In conclusion, this study showed that the effect of two lipid-lowering diets containing either lean red meat or skinless chicken and fish on the atherogenic lipoproteins did not differ significantly. A prudent diet with skinless chicken and fish, however, had a more favorable effect on the fatty acid composition of the plasma TAG and the CE than did the lean red meat diet.

Plasma lipoprotein changes in hens ( Gallus domesticus) during an induced molt

Blood plasma lipoproteins were studied during food and light deprivation or prolactin injection-induced involution of ovarian follicles (molt) of laying hens. Egg laying stopped 3 days after initiation of either treatment. Food and light-deprived hens lost 29% of initial body weight during the 10-day experiment ( P<0.05), whereas prolactin-treated hens lost 9% of body weight. Yolk-directed very low density lipoprotein (VLDLy) concentration in plasma decreased in both groups, but declined more rapidly in food and light-deprived hens. Very low density lipoprotein triacylglycerol decreased 40% in food and light-deprived hens by day 2 compared with a 13% decrease in the prolactin-treated hens. By day 5, a lipoprotein particle 21–22 nm in diameter appeared in the d=1.019–1.046 g/ml density fraction of plasma in both groups. A similar lipoprotein particle, termed HDL R, developed in overfed hens with involuting ovarian follicles. In conclusion, hens undergoing ovarian regression due to food and light deprivation, prolactin treatment or overfeeding display marked decreases in plasma yolk-directed very low density lipoproteins and the appearance of HDL R. Other lipoprotein populations varied depending on whether the hens continued to feed or not.

Metabolism of trans fatty acids by hepatocytes.

The present work was undertaken to study the metabolism of fatty acids with trans double bonds by rat hepatocytes. In liver mitochondria, elaidoyl-CoA was a poorer substrate for carnitine palmitoyltransferase I (CPT-I) than oleoyl-CoA. Likewise, incubation of hepatocytes with oleic acid produced a more pronounced stimulation of CPT-I than incubation with trans fatty acids. This was not due to a differential effect of cis and trans fatty acids on acetyl-CoA carboxylase (ACC) activity and malonyl-CoA levels. Elaidic acid was metabolized by hepatocytes at a higher rate than oleic acid. Surprisingly, compared to oleic acid, elaidic acid was a better substrate for mitochondrial and, especially, peroxisomal oxidation, but a poorer substrate for cellular and very low density lipoprotein triacylglycerol synthesis. Results thus show that trans fatty acids are preferentially oxidized by hepatic peroxisomes, and that the ACC/malonyl-CoA/CPT-I system for coordinate control of fatty acid metabolism is not responsible for the distinct hepatic utilization of cis and trans fatty acids.