Corinne Moundras

Effect of propionate on fatty acid and cholesterol synthesis and on acetate metabolism in isolated rat hepatocytes

In the present study the actual role of propionic acid in the control of fatty acid and cholesterol synthesis was investigated in isolated liver cells from fed rats maintained in the presence of near-physiological concentrations of glucose, glutamine and acetate. Using 3H2O for lipid labelling, propionate appears as an effective inhibitor of fatty acid synthesis and to a lesser extent of cholesterol synthesis, even at the lowest concentration used (0.6 mmol/l). Butyrate is a potent activator of both synthetic pathways, and the activating effect was not counteracted by propionate. Using 1-[14C]acetate, it was observed that propionate at a moderate concentration, or 1 mmol oleate/l, are both very effective inhibitors of 14C incorporation into fatty acid and cholesterol. This incorporation was drastically inhibited when propionate and oleate were present together in the incubation medium. The net utilization of acetate by rat hepatocytes was impaired by propionate, in contrast to oleate. 1-[14C]butyrate was utilized at a high rate for fatty acid synthesis, but to a lesser extent for cholesterol synthesis; both processes were unaffected by propionate. Intracellular citrate concentration was not markedly depressed by propionate, whereas it was strongly elevated by butyrate. In conclusion, propionate may represent an effective inhibitor of lipid synthesis when acetate is a major source of acetyl-CoA, a situation which is encountered with diets rich in readily-fermentable fibres. The present findings also suggest that propionate may be effective at concentrations close to values measured in vivo in the portal vein.

Effectiveness of resistant starch, compared to guar gum, in depressing plasma cholesterol and enhancing fecal steroid excretion

Amylase-resistant starch (RS) represents a substrate that can be administered in substantial amounts in the diet, in contrast to gel-forming polysaccharides, such as guar gum (GG). The aim of this work was thus to compare the effects of GG and RS on cholesterol metabolism in rats adapted to 0.4% cholesterol diets, using dietary GG or RS levels (8 or 20%, respectively) that led to a similar development of fermentations, as assessed by the degree of enlargement of the cecum. The RS diet elicited a marked rise in the cecal pool of short-chain fatty acids, especially acetic and butyric acid, whereas the GG diet favored high-propionic acid fermentations. Both polysaccharides markedly altered the cholesterol excretion, from 50% of ingested cholesterol in controls, up to about 70% in rats adapted to the RS or GG diets. With these diets, the fecal excretion of bile acids was enhanced (67 and 144% with the RS and GG diets, respectively). RS and GG diets were effective in lowering plasma cholesterol (about −40%) and triglycerides (−36%). There was practically no effect of the diets on cholesterol in d>1.040 lipoproteins (high density lipoproteins), whereas RS (and to a larger extent, GG) were very effective to depress cholesterol in d<1.040 lipoproteins (especially in triglyceride-rich lipoproteins). Fermentable polysaccharides counteracted the accumulation of cholesterol in the liver, especially cholesterol esters. In parallel, liver acyl CoA:cholesterol acyltransferase was depressed in rats fed the RS or GG diets, whereas only the GG diet counteracted the downregulation of 3-hydroxy-3-methylglutaryl-CoA by cholesterol. These data suggest that RS may be practically as effective as a gel-forming gum, such as GG, on steroid excretion and on cholesterol metabolism.

Cholesterol-lowering effects of guar gum: Changes in bile acid pools and intestinal reabsorption

Soluble fibers such as guar gum (GG) may exert cholesterol-lowering effects. It is generally accepted that bile acid (BA) reabsorption in portal blood is reduced, thus limiting the capacity of BA to down-regulate liver cholesterol 7α-hydroxylase, the rate-limiting enzyme of BA synthesis. In the present work, rats were adapted to fiber-free (FF) or 5% GG diets (supplemented or not with 0.25% cholesterol), to investigate various aspects of enterohepatic BA cycling. GG in the diet at a level of 5% elicited a significant lowering of plasma cholesterol during the absorptive period, in cholesterol-free (−13%) or 0.25% cholesterol (−20%) diet conditions. In rats adapted to the GG diets, the small intestinal and cecal BA pools and the ileal vein-artery difference for BA were markedly enhanced; reabsorption in the cecal vein was also enhanced in these rats. [14C]Taurocholate absorption, determined in perfused ileal segments, was not significantly different in rats adapted to the FF or GG diet, suggesting that a greater flux of BA in the ileum might support a greater ileal BA reabsorption in rats adapted to the GG diet. In contrast, capacities for [14C]cholate absorption from the cecum at pH 6.5 were higher in rats adapted to the GG diet than to the FF diet. Acidification of the bulk medium in isolated cecum (from pH 7.1 down to pH 6.5 or 5.8) or addition of 100 mM volatile fatty acids was also found to stimulate cecal [14C]cholate absorption. These factors could contribute to accelerated cecal BA absorption in rats fed the GG diet. The effects of GG on steroid fecal excretion thus appear to accompany a greater intestinal BA absorption and portal flux to the liver. These results suggest that some mechanisms invoked to explain cholesterol-lowering effect of fibers should be reconsidered.

Methionine deficiency in rats fed soy protein induces hypercholesterolemia and potentiates lipoprotein susceptibility to peroxidation

A number of studies have provided evidence that plant proteins, especially soy protein, have a cholesterol-lowering effect as compared with casein. However, dietary supply of sulfur amino acids may be deficient when soy protein is present in the diet at a suboptimal level, which could affect lipid metabolism. Accordingly, in rats fed 13% protein diets, soy protein feeding resulted in a cholesterol-increasing effect (+18%), which could be counteracted by methionine supplementation (0.4%). In contrast, soy protein was effective in decreasing plasma triglyceride, as compared with levels in rats fed casein; this triglyceride-lowering effect was entirely abolished by methionine supplementation. The hypercholesterolemic effect of soy protein was characterized by a higher cholesterol content in low-density lipoprotein (LDL) and high-density lipoprotein 1 (HDL1) fractions, together with a marked induction of hepatic hydroxymethyl glutaryl coenzyme A (HMG CoA) reductase activity and to a lesser extent cholesterol 7 alpha-hydroxylase. There was practically no induction of these enzymes, as compared with levels in rats fed casein diets, when the soy protein diet was supplemented with methionine. Very-low-density lipoprotein (VLDL) plus LDL susceptibility to peroxidation was higher in rats fed soy protein than in casein-fed rats, which could reflect in part the lack of sulfur amino acid availability, since methionine supplementation led to a partial recovery of lipoprotein resistance to peroxidation. These findings suggest that amino acid imbalance could be atherogenic by increasing circulating cholesterol and leading to a higher lipoprotein susceptibility to peroxidation.

Effect of cyclodextrin on plasma lipids and cholesterol metabolism in the rat

beta-Cyclodextrin (beta-CD) is a bile acid and sterol sequestrant produced by enzymatic modification of starch; this product has the potential to decrease plasma cholesterol. In contrast to the sequestrants having resin- or saponin-like properties, beta-CD is rapidly broken down by the large intestine microflora. beta-CD effects on cecal fermentations and lipid metabolism were thus investigated in rats adapted to semipurified diets containing 0%, 2.5%, or 5% beta-CD. In rats fed beta-CD diets, there was an enlargement of the cecum together with a dramatic increase in the cecal concentration of propionic acid (even with the 2.5% level, in moderately acidic pH conditions). Propionic acid produced in the cecum was readily absorbed and entirely taken up by the liver, whereas there was no significant acetic acid uptake. Dietary beta-CD was highly effective in enhancing bile acid entry into to the cecum: the cecal bile acids pool was 2.2 and 3.6-fold enlarged in rats fed the 2.5% and 5% beta-CD diets, respectively. The solubility percentage of bile acids decreased to approximately 25% in rats fed the beta-CD diets (v 51% in controls); the cecal concentration of soluble bile acids was thus relatively low in these animals. The fecal excretion of steroids was strongly enhanced by beta-CD, and bile acids excretion was practically proportional to the dietary beta-CD level. There was a net lipid-lowering effect of beta-CD, even at the 2.5% level. The effect was more pronounced on triglycerides than on cholesterol.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of selenium deficiency on hepatic lipid and lipoprotein metabolism in the rat

Since experimental Se deficiency results in a significant increase in plasma cholesterol concentration the present investigation was undertaken to assess further the influence of this deficiency on the expression of proteins involved in hepatic lipid metabolism. Se deficiency was induced by feeding weanling male Wistar rats on a deficient diet for 6 weeks. Hypercholesterolaemia associated with Se deficiency was related to increased 3-hydroxy-3-methylglutaryl-coA (HMG-CoA) reductase (EC 1.1.1.34) activity in liver microsomes as compared with control animals. Hepatic lipoprotein receptor levels (LDL-receptor and HDL-binding proteins, HB1 and HB2) were not significantly affected by Se deficiency, as assessed by immunoblotting. Plasma triacylglycerol concentrations tended to decrease in Se-deficient rats in concert with their reduced post-Triton secretion. There was no significant effect of Se deficiency on the hepatic synthesis of apolipoproteins. These results point to the need for further investigations into the mechanism related to the increased activity of HMG-CoA reductase and the enhanced cholesterogenesis in the liver of Se-deficient rats likely to result from this.

Fermentable carbohydrates exert a more potent cholesterol-lowering effect than cholestyramine

The purpose of this work was to assess the respective role of bile acid excretion and of the end-products of cecal fermentations in the cholesterol-lowering effect of complex carbohydrates. The effects of two different fermentable carbohydrates (guar gum, beta-cyclodextrin), and sequestrant resin (cholestyramine) have been investigated in male Wistar rats. Guar gum and beta-cyclodextrin are broken down in the large bowel, with fermentation rich in propionic acid (37% against 26% for control), whereas cholestyramine did not enhance cecal fermentation. beta-Cyclodextrin and guar gum were less potent than cholestyramine to enhance bile acids and sterol excretion. Nevertheless, fermentable carbohydrates exerted a more potent cholesterol-lowering effect than cholestyramine. beta-Cyclodextrin also depressed triacylglycerol-rich lipoprotein (TGRLP). Fermentable carbohydrates lowered cholesterol of LDL and HDL1 fractions. The induction of hepatic HMG-CoA reductase was practically proportional to rate of fecal steroid excretion. Moreover, with beta-cyclodextrin, hepatic HMG-CoA reductase induction was concomitant to a decrease in fatty acid synthase (FAS) activity. Thus, the cholesterol-lowering effect of fermentable carbohydrates could be related to a depressed lipogenesis, as well as to an accelerated removal of HDL1, in relation to an elevated hepatic demand of cholesterol. In conclusion, fermentable carbohydrates could favour cholesterol elimination and have a general lipid-lowering effect by exerting more complex physiological effects than cholestyramine.

Influence of methionine availability on glutathione synthesis and delivery by the liver

Abstract The aim of this study was to define dietary conditions liable to elevate the circulating and tissue levels of glutathione (GSH). For this purpose, GSH synthesis and availability have been compared after supplementation of a low protein diet (10% casein) with 0.6% methionine or after adaptation to a high protein diet (30% casein), the final dietary methionine levels were similar in both cases. To compare the effects of an acute addition of sulfur amino acids to the diet with long-term adaptative changes on GSH status, rats adapted to the 10% casein diet received as a final meal (i) the diet supplemented with 0.6% methionine or (ii) a 30% casein diet. With a 10% casein diet, the plasma and tissue concentrations of methionine and cysteine seem to constitute limiting factors for GSH synthesis. However, in animals adapted to a high protein diet or to a diet supplemented in methionine, the hepatic GSH content was quite enhanced (respectively 1.9 and 2.6 fold that measured in control diet), in keeping with the marked rise of the hepatic cysteine (respectively 2 and 5 fold that measured in the control diet). Acute administration of a final 30% casein meal or 10% casein meal supplemented in methionine induced greater changes in the hepatic GSH content than observed in long-term experiments. Compared with the liver, the muscle and heart GSH content was poorly affected by changes in sulfur amino acids availability or nutritional state. The liver appears as a net site of GSH release, the magnitude of this release was not entirely proportional to the hepatic pool, particularly during the postabsorptive period. Moreover, during this period the hepatic GSH content was markedly depressed compared with the fed state, whatever the dietary conditions. With diets providing a high availability in sulfur amino acids, the activity of the liver γ-glutamyl cysteine synthetase was markedly depressed, even in acute experiments. In vitro experiments confirmed in vivo results, and showed that until its extracellular concentration reached 0.4 mM, methionine may constitute a limiting factor for GSH synthesis in isolated hepatocytes.

Effect of dietary supplementation with glutamic acid or glutamine on the splanchnic and muscle metabolism of glucogenic amino acids in the rat

Abstract The aim of the present study was to examine the metabolic effects of a high availability of dietary glutamic acid or glutamine. A 15% casein diet was supplemented with 7.2% glutamic acid or glutamine. The present results show that both supplementations produced only a slight modification in the circulating concentrations of glutamate. Glutamic acid supplementation noticeably enhanced (+ 19%) the arterial concentrations of glutamine, but to a lesser extent than glutamine supplementation itself (+ 54%). Large amounts of alanine were released by the digestive tract in rats fed the various diets; this release was enhanced by glutamine (+ 33%) and, more markedly, by glutamic acid (+ 80%) supplementation. In the liver, glutamic acid and glutamine supplementation produced an increase in the catabolism of glycine, serine, and threonine that resulted in a drop of their peripheral concentrations, especially in muscles. The decrease in serine and threonine concentrations could be ascribed to the elevation of the serine(threnine)dehydratase activity (three fold), and the drop in glycine concentration seems to be connected to serine metabolism. It appears that the administration of glutamine (but not of glutamic acid) by the oral route could be effective in increasing its availability in peripheral tissues.

LIPID METABOLISM AND LIPOPROTEIN SUSCEPTIBILITY TO PEROXIDATION ARE AFFECTED BY A PROTEIN-DEFICIENT DIET IN THE RAT

Abstract To investigate the influence of the dietary protein level on plasma cholesterol homeostasis and lipoprotein susceptibility to peroxidation, rats were adapted to semi-purified diets with different levels of casein (8, 16, or 32%). Increasing the dietary protein level had a positive influence on the plasma and liver availability of sulfur amino acids, as well as on taurine and glutathione concentrations. Rats fed the protein-deficient diet (8%) exhibited a slight hypercholesterolemia, together with an induction of liver 3-hydroxy-3-methylglutaryl CoA (HMG CoA) reductase and a low activity of fatty acid synthase, compared with controls (16% casein diet). In contrast, feeding high-protein diets resulted in a cholesterol-lowering effect together with a lowered activity of liver HMG CoA reductase. The susceptibility of isolated lipoprotein (VLDL + LDL) fractions to copper-induced oxidation was also investigated: there was a shortened lag time period for conjugated diene production in rats fed the 8% casein diet. Thus, with a protein-deficient diet, a slight hypercholesterolemia could be aggraved by a high susceptibility of lipoprotein to peroxidation, probably because of a limiting bioavailability of sulfur amino acids in plasma and a depletion of intracellular glutathione and taurine. Copyright