Changes in blood lipids during six days of overfeeding with medium or long chain triglycerides

Although medium chain triglyceride (MCT) is less calorically dense than long chain triglyceride (LCT), it produces a greater thermic effect following ingestion. We hypothesized that the previously observed high rate of thermogenesis produced by MCT overfeeding was due to hepatic de novo synthesis of long chain fatty acids (LCFA) from the excess medium chain fatty acids (MCFA). To study this, we compared the effects of overfeeding MCT- and LCT-containing diets on blood lipid profiles. Ten in-patient, nonobese males were overfed (150% of estimated energy requirements) two formula diets for 6 days each, in a randomized crossover design. Diets differed only in the composition of the fat and contained either 40% of energy as MCT or LCT (soybean oil). The major differences between diets in the resulting pattern of blood lipids were: 1) a reduction in fasting serum total cholesterol concentrations with the LCT, but not the MCT diet; and 2) a threefold increase in fasting serum triglyceride concentrations with MCT, but not LCT, diet. Moreover, 10% of the fasting triglyceride fatty acids were medium chain and 40% were 16:0 with the MCT diet. This compared to 1% and 20% for medium chain and 16:0, respectively, with the LCT diet. In addition, there were increases in 16:1, 18:0, and 18:1 in the triglycerides during MCT feeding. The changes in fatty acids in triglycerides with MCT feeding are consistent with the hypothesis that excess dietary MCT cause a significant increase in the hepatic synthesis of these fatty acids from MCFA through de novo synthesis and/or chain elongation and desaturation. These processes could account for the higher rate of postprandial thermogenesis with MCT as compared to LCT.     

Effect of dietary medium chain triacylglycerols on plasma triacylglycerol levels in horses

The hypothesis tested was that the feeding of medium chain triacylglycerols (MCT) to horses would raise the level of plasma triacylglycerols by increasing the availability of glucose as lipogenic substrate, implying that the MCT effect would be greater with glucose in the diet instead of cellulose. A Latin square experiment was carried out with 4 horses and 4 dietary treatments. The experimental periods lasted 21 d. Blood samples were taken 16 h after feeding. The diets consisted of hay and experimental concentrates, differing in fat source (MCT or soybean oil) and carbohydrate source (corn starch plus glucose or cellulose). The dietary variables, MCT or soybean oil, provided on average 27% of total dietary net energy, while glucose plus cornstarch or cellulose provided 33%. The feeding of MCT versus soybean oil raised the level of plasma triacylglycerols significantly from 196.7 ± 30.2 to 427.3 ± 85.7 mmol/1 and that of VLDL cholesterol from 0.028 ± 0.01 to 0.069 ± 0.01 mmol/ml. As based on analysis of variance, for the four experimental diets there was no significant effect of carbohydrate source and no fat‐carbohydrate interaction. Thus, the hypothesis was rejected. When the diets contained soybean oil, cellulose versus starch plus glucose produced significantly greater increase plasma triacylglycerols. This carbohydrate effect was not seen when horses were fed the MCT diets. The experimental concentrates did not differently influence the concentrations of plasma glucose, total serum cholesterol, phospholipids, insulin, free fatty acids and the activity of post‐heparin lipoprotein lipase. We suggest that the MCT‐induced increase in plasma triacylglycerols is related to an increase in hepatic VLDL secretion, with the extra substrate for increased synthesis of triacylglycerols being the acetyl‐CoA derived from the hepatic oxidation of medium chain fatty acids.     

A high-fat diet enriched in medium chain triglycerides triggers hepatic thermogenesis and improves metabolic health in lean and obese mice

Obesity, liver steatosis and type 2 diabetes are major diseases partly imputed to energy-dense diets rich in long chain triglycerides (LCT). The search for bioactive nutrients that help to overcome metabolic diseases is a growing field. In this regard, medium chain triglycerides (MCT) were shown to promote lipid catabolism and to stimulate brown adipose tissue thermogenesis. The objective of our study was to evaluate if the replacement of LCT by MCT in high-fat diets could prevent and/or reduce metabolic disorders. For this purpose, two cohorts of C57BL/6 mice were fed during 10 weeks with three isocaloric high-fat diets with variable MCT content. Cohort A was composed of lean mice while cohort B was composed of obese, insulin resistant mice. In cohort A, replacement of LCT by MCT preserved metabolic health, in part by triggering hepatic thermogenesis. We further found that medium chain fatty acids promote thermogenesis markers within cultured hepatocytes in a FFAR1/GPR40-dependent manner. In cohort B, high-fat diets enriched in MCT promoted body fat depletion and caused metabolic health improvement, together with the induction of thermogenesis markers in the liver as well as in subcutaneous white adipose tissue. Our study supports that replacement of LCT by MCT in high-fat diets improves the metabolic features associated with obesity.     

The concentration of plasma triacylglycerols in horses fed diets containing either medium chain triacylglycerols or an isoenergetic amount of starch or cellulose

In a Latin square design, six horses were fed hay and concentrates with isoenergetic amounts of either starch, cellulose or medium chain triacylglycerols (MCT). The dietary variables provided on average 22% of total dietary net energy. Plasma triacylglycerols and other variables of lipid metabolism were determined. The experimental periods lasted 21 days. Blood samples were taken just before the morning meal and three and six hours later. The diet rich in MCT significantly raised the plasma level of triacylglycerols when compared to either the starch‐ or cellulose‐rich diet. The plasma concentrations of 3‐hydroxybutyrate, total cholesterol and phospholipids were significantly higher when the horses were fed the ration with MCT instead of either cellulose or starch. Postprandial.insulin concentrations were lowest for the MCT diet, and concentrations of free fatty acids were highest Lipoprotein lipase activity was not significantly different for the three diets. Our study does not support the idea that cellulose feeding generates sufficient acetic acid in the caecum and colon, so that it would enhance the provision of cytosolic acetyl‐CoA which in turn would stimulate hepatic fatty acid synthesis and then raise plasma triacylglycerols.     

Production of structured lipids by lipase-catalysed interesterification in an ultrafiltration membrane reactor

The application of membranes to the enzymatic production of structured lipids has been investigated using an ultrafiltration membrane reactor for a reaction between medium chain triacylglycerols (MCT) and n-3 polyunsaturated fatty acids from fish oil. Lipozyme IM was used as the biocatalyst. The incorporation of polyunsaturated fatty acids into MCT was increased by about 15% over 80 h by simultaneous separation of the released medium chain fatty acids compared to control.     

Report on dietary medium chain fatty acids (MCT) and chicken growth

From the results, obtained from the comparison between diet containing medium chain triglycerides (MCT) and the growth of chicks, emerged the importance of the qualitative composition of the diet; most of all it became evident the influence that MCT exert on the amount of energy rendered per ration consumed. Less body growth is obtained due to the adapting, probably temporary, of oxidative metabolism to the increased quantity of medium chain fatty acid consumption.     

Fasting-induced oxidative stress in very long chain acyl-CoA dehydrogenase-deficient mice

Hepatopathy and hepatomegaly as consequences of prolonged fasting or illnesses are typical clinical features of very long chain acyl-CoA dehydrogenase (VLCACD) deficiency, the most common long-chain fatty acid β-oxidation defect. Supplementation with medium-chain triglycerides (MCTs) is an important treatment measure in these defects, in order to supply sufficient energy. Little is known about the pathogenetic mechanisms leading to hepatopathy. Here, we investigated the effects of prolonged fasting and an MCT diet on liver function. Wild-type (WT) and VLCAD knockout mice were fed with either a regular long-chain triglyceride diet or an MCT diet for 5 weeks. In both groups, we determined liver and blood lipid contents under nonfasting conditions and after 24 h of fasting. Expression of genes regulating peroxisomal and microsomal oxidation pathways was analyzed by RT-PCR. In addition, glutathione peroxidase and catalase activities, as well as thiobarbituric acid reactive substances, were examined. In VLCAD knockout mice fed with a long-chain triglyceride diet, fasting is associated with excessive accumulation of liver lipids, resulting in hepatopathy and strong upregulation of peroxisomal and microsomal oxidation pathways as well as antioxidant enzyme activities and thiobarbituric acid reactive substances. These effects were even evident in nonfasted mice fed with an MCT diet, and were particularly pronounced in fasted mice fed with an MCT diet. This study strongly suggests that liver damage in fatty acid oxidation defects is attributable to oxidative stress and generation of reactive oxygen species as a result of significant fat accumulation. An MCT diet does not prevent hepatic damage during catabolism and metabolic derangement.     

Changes in carnitine octanoyltransferase activity induce alteration in fatty acid metabolism

The peroxisomal beta oxidation of very long chain fatty acids (VLCFA) leads to the formation of medium chain acyl-CoAs such as octanoyl-CoA. Today, it seems clear that the exit of shortened fatty acids produced by the peroxisomal beta oxidation requires their conversion into acyl-carnitine and the presence of the carnitine octanoyltransferase (CROT). Here, we describe the consequences of an overexpression and a knock down of the CROT gene in terms of mitochondrial and peroxisomal fatty acids metabolism in a model of hepatic cells. Our experiments showed that an increase in CROT activity induced a decrease in MCFA and VLCFA levels in the cell. These changes are accompanied by an increase in the level of mRNA encoding enzymes of the peroxisomal beta oxidation. In the same time, we did not observe any change in mitochondrial function. Conversely, a decrease in CROT activity had the opposite effect. These results suggest that CROT activity, by controlling the peroxisomal amount of medium chain acyls, may control the peroxisomal oxidative pathway.     

Melatonin attenuates monocrotaline-induced hepatic sinusoidal obstruction syndrome in rats via activation of Sirtuin-3

Melatonin possesses potent hepatoprotective properties, but it remains to be elucidated whether melatonin has a therapeutic effect on monocrotaline (MCT)-induced hepatic sinusoidal obstruction syndrome (HSOS). In this study, male Sprague Dawley rats were intraperitoneally injected with melatonin or the same volume of vehicle at 0 and 24 h after MCT intragastric administration. Next, hematoxylin–eosin staining and electron microscopy were performed to evaluate the hepatic sinusoidal injury of rats. Endothelial cell marker RECA-1 was observed by immunohistochemistry. Hepatic oxidative stress was analyzed by detecting malondialdehyde, glutathione S-transferase, and reactive oxygen species. Assessment of liver function was carried out by analysis of serum aspartate aminotransferase, alanine aminotransferase, total bilirubin, and albumin levels. Real-time polymerase chain reaction and Western blot analysis were used to identify liver Sirtuin-3 (SIRT3) and active matrix metallopeptidase 9 (MMP-9) expression. Besides, liver sinusoidal endothelial cells (LSECs) were used for the in vitro functional verification experiment. Specifically, liver histology of the melatonin-treated groups showed that the pathological damages caused by MCT were significantly attenuated, total HSOS scores were decreased, and the elevation of serum hyaluronic acid observed in the model group was also reduced. Moreover, melatonin treatment also improved the survival of rats after partial hepatectomy. Administration of melatonin ameliorated MCT-induced LSECs injury, hepatic oxidative stress, and hepatic dysfunction. Furthermore, melatonin treatment increased SIRT3 expression while attenuating MMP-9 activity in liver tissues. Cell experiment also demonstrated that SIRT3 might mediate the protective effect of melatonin on LSECs. Collectively, our study provided the potential rationale for the application of melatonin for the prevention of MCT-induced HSOS.     

Metabolic effects of glucose, medium chain triglyceride and long chain triglyceride feeding before prolonged exercise in rats

The purpose of this study was to test the hypothesis that oral ingestion of lipids could increase endurance by slowing the rate of glycogen depletion. Trained rats were killed after a 2 h run on a rodent treadmill, following an intragastric infusion of water, glucose, medium chain triglycerides (MCT) or long chain triglycerides (LCT). Glucose and triglycerides were administered in equicaloric concentrations (50 kJ). The results show that oral ingestion of lipids (MCT or LCT) did not reduce glycogen depletion in liver, heart or skeletal muscle after exercise whereas the fat diet increased muscle and heart glycogen stores in resting conditions. In contrast, glucose feeding induced a significant sparing effect on endogenous carbohydrate utilization and reduced physical exercise lipolysis. These data indicated, firstly, that enhanced lipid availability induced by a single lipid meal before exercise was not able to modify the glycogen depletion occurring after exercise and, secondly, that the glucose/fatty acid cycle was not effective in these conditions. The comparison between lipids indicated that the effect on glycogen use of MCT did not differ from that of LCT, and did not seem to be of any particular importance during physical exercise.     

Age-related changes in renal and hepatic cellular mechanisms associated with variations in rat serum thyroid hormone levels

Aging is associated with changes in thyroid gland physiology. Age-related changes in the contribution of peripheral tissues to thyroid hormone serum levels have yet to be systematically assessed. Here, we investigated age-related alterations in the contributions of the liver and kidney to thyroid hormone homeostasis using 6-, 12-, and 24-mo-old male Wistar rats. A significant and progressive decline in plasma thyroxine occurred with age, but triiodothyronine (T(3)) was decreased only at 24 mo. This was associated with an unchanged protein level of the thyroid hormone transporter monocarboxylate transporter 8 (MCT8) in the kidney and with a decreased MCT8 level in the liver at 24 mo. Hepatic type I deiodinase (D1) protein level and activity declined progressively with age. Renal D1 levels were decreased at both 12 and 24 mo but D1 activity was decreased only at 24 mo. In the liver, no changes occurred in thyroid hormone receptor (TR) TRalpha(1), whereas a progressive increase in TRbeta(1) occurred at both mRNA and total protein levels. In the kidney, both TRalpha(1) and TRbeta(1) mRNA and total protein levels were unchanged between 6 and 12 mo but increased at 24 mo. Interestingly, nuclear TRbeta1 levels were decreased in both liver and kidney at 12 and 24 mo, whereas nuclear TRalpha(1) levels were unchanged. Collectively, our data show differential age-related changes among hepatic and renal MCT8 and D1 and TR expressions, and they suggest that renal D1 activity is maintained with age to compensate for the decrease in hepatic T(3) production.     

Monocarboxylic acid transporters, MCT1 and MCT2, in cortical astrocytes in vitro and in vivo

We used sequence-specific antibodies tocharacterize two monocarboxylic acid transporters, MCT1 and MCT2, inastrocytes. Both proteins are expressed in primary cultures of corticalastrocytes, as indicated by immunoblotting and immunofluorescence. BothMCT1 and MCT2 are present in small, punctate structures in thecytoplasm and at the cell membrane. Cells showing very low levels oflabeling for glial fibrillary acidic protein (GFAP) also label moredimly for MCT2, but not for MCT1. In vivo, double-labelimmunofluorescence studies coupled with confocal microscopy indicatethat MCT1 and MCT2 are rare in astrocytes in the cortex. However, theyare specifically labeled in astrocytes of the glial limiting membraneand in white matter tracts. Both transporters are also present in themicrovasculature. Comparison of labeling for MCT1 and MCT2 with markersof the blood-brain barrier shows that the transporters are not alwayslimited to the astrocytic endfeet in vivo. Our results suggestthat the level of expression of monocarboxylic acid transporters MCT1and MCT2 by cortical astrocytes in vivo is significantly lowerthan in vitro but that astrocytes in some other regions of thebrain can express one or both proteins in significant amounts.

     

Plasma fatty acids and [13C]linoleic acid metabolism in preterm infants fed a formula with medium-chain triglycerides

Most preterm infant formulas contain medium-chain triacylglycerols (MCT), but the effects of MCT on polyunsaturated fatty acid status and metabolism are controversial. Thus, we studied the effects of MCT on linoleic acid metabolism using stable isotopes. Enterally fed preterm infants were randomized to receive for 7 days 40% of fat as MCT (n = 10) or a formula without MCT (n = 9). At study day 5, infants received orally 2 mg/kg body weight of (13)C-labeled linoleic acid. Fatty acids in plasma lipid classes and (13)C enrichment of phospholipid fatty acids were measured and tracer oxidation was monitored. Compared with the control group, the MCT group showed lower breath (13)CO(2) and higher plasma triacylglycerol contents of octanoic acid, of decanoic acid, and of total long-chain polyunsaturated fatty acids (57.1 +/- 4.4 micro mol/l vs. 37.9 +/- 4.8 micro mol/l, P < 0.01). Concentrations of several polyunsaturated fatty acids in plasma phospholipids and non esterified fatty acids were higher in the MCT group. (13)C concentrations in phospholipid n-6 fatty acids indicated no difference in the relative conversion of linoleic to arachidonic acid. We conclude that oral MCT effectively reduce polyunsaturated fatty acid and long chain polyunsaturated fatty acid oxidation in preterm infants without compromising endogenous n-6 long chain polyunsaturated fatty acid synthesis.     

Effect of pectin feeding on monocarboxylate transporters in rat adrenal gland

We have recently proved the expression and localization of seven monocarboxylate transporters (MCT1, MCT2, MCT3, MCT4, MCT5, MCT7, and MCT8) in the rat adrenal gland. So far, there are no data reporting possible regulation of any MCT isoform in the adrenal gland. Pectin is a soluble dietary fiber that is known to exert a hypocholesterolemic effect and increases the short chain fatty acids production in the large intestine. This work aimed to study the effect of pectin feeding on the expression of MCTs (MCT1–MCT5, MCT7, and MCT8) and their cellular distribution in rat adrenal gland. Western blotting demonstrated significant increase in the expression levels of MCT1, MCT2, MCT4, MCT5, and MCT7 in pectin-fed rats in comparison with the controls. Immunohistochemistry revealed extended distribution and distinctive increase in the immunoreactivities of MCT1, MCT2, MCT4, MCT5, and MCT7 in the adrenal cortical zones, besides the increase in the immunoreactive intensity of MCT5 and MCT7 in the adrenal medulla of pectin-fed versus control rats. Interestingly, zona glomerulosa which did not show any reactivity for MCT1 or MCT2 in controls, exhibited marked immunopositivities for both MCT1 and MCT2 in pectin-fed rats. MCT3 and MCT8, however, did not show significant changes in their expression levels between pectin-fed and control rats. Our data is the first to describe the up regulation of various MCTs in rat adrenal gland under the influence of pectin feeding. This up regulation might be a compensatory response to the hypocholesterolemic effect of pectin in order to maximize the intracellular availability of acetate. This article suggests that monocarboxylate transporters have an important physiological role in the regulation of adrenal hormones as well as in cholesterol homeostasis.     

Analysis of three variable number terminal repeat loci is sufficient to characterize the deoxyribonucleic acid fingerprints of a panel of human tumor cell lines

Using primers for the MCT118, YNZ22, and COL2A1 loci in polymerase chain reaction analysis we could distinguish among the approximately 20 cell lines routinely maintained in our laboratory. We also demonstrated that the cell line NB-1691 (a neuroblastoma) and its xenograft had an identical number of repeats at two loci. Rh30 (a rhabdomyosarcoma) made resistant to rapamycin was identical to its parent line and to a subline that had reverted to sensitivity after it was cultured without rapamycin in the medium.     

POLYAMINE REGULATORY PROCESSES AND OXIDATIVE STRESS IN MONOCROTALINE-TREATED PULMONARY ARTERY ENDOTHELIAL CELLS

Alterations in polyamine metabolism may be a critical mechanism of monocrotaline (MCT)-induced structural remodeling of the pulmonary vasculature. In the present study, the hypothesis that MCT, through the induction of oxidative stress, modulates cellular polyamine regulatory mechanisms which in turn might be involved in the upregulation of fibronectin production in pulmonary artery endothelial cells (PAEC) was examined. A 24-h treatment with MCT significantly increased PAEC polyamine concentrations as compared to vehicle-treated cells. In addition, exposure to MCT caused an increase in abundance of ornithine decarboxylase (ODC) mRNA, upregulation of ODC activity and enhancement of spermidine import into PAEC. Inhibition ofde novopolyamine synthesis further increased spermidine uptake in MCT-treated cells. The depletion of cellular polyamine contents through the blockade of bothde novopolyamine biosynthesis and polyamine transport prevented MCT-induced increases in the medium level of fibronectin. In addition, PAEC treatment with MCT stimulated cellular oxidative stress as determined by increased levels of thiobarbituric acid reactive substances, enhanced dichlorofluorescein fluorescence and activation of NF-KB. A co-treatment with dimethylthiourea, an oxygen radical scavenger, prevented MCT-induced increases in cellular oxidation and attenuated disturbances in polyamine metabolism. These data suggest that MCT can stimulate polyamine regulatory processes in PAEC possibly through an increase in cellular oxidative stress. The present study may have significant implication in understanding mechanisms of MCT-induced pulmonary hypertension and remodeling of pulmonary vasculature.     

Expression of monocarboxylate transporters in rat ocular tissues

The aim of the present study was to determine the distribution of monocarboxylate transporter (MCT) subtypes 1-4 in the various structures of the rat eye by using a combination of conventional and real-time RT-PCR, immunoblotting, and immunohistochemistry. Retinal samples expressed mRNAs encoding all four MCTs. MCT1 immunoreactivity was observed in photoreceptor inner segments, Müller cells, retinal capillaries, and the two plexiform layers. MCT2 labeling was concentrated in the inner and outer plexiform layers. MCT4 immunolabeling was present only in the inner retina, particularly in putative Müller cells, and the plexiform layers. No MCT3 labeling could be observed. The retinal pigment epithelium (RPE)/choroid expressed high levels of MCT1 and MCT3 mRNAs but lower levels of MCT2 and MCT4 mRNAs. MCT1 was localized to the apical and MCT3 to the basal membrane of the RPE, whereas MCT2 staining was faint. Although MCT1-MCT4 mRNAs were all detectable in iris and ciliary body samples, only MCT1 and MCT2 proteins were expressed. These were present in the iris epithelium and the nonpigmented epithelium of the ciliary processes. MCT4 was localized to the smooth muscle lining of large vessels in the iris-ciliary body and choroid. In the cornea, MCT1 and MCT2 mRNAs and proteins were detectable in the epithelium and endothelium, whereas evidence was found for the presence of MCT4 and, to a lesser extent, MCT1 in the lens epithelium. The unique distribution of MCT subtypes in the eye is indicative of the pivotal role that these transporters play in the maintenance of ocular function. retina; eye; immunohistochemistry; polymerase chain reaction