Claudio A. Bernal

Long-term hypertriglyceridemia and glucose intolerance in rats fed chronically an isocaloric sucrose-rich diet

We have previously shown that short-term feeding [20 to 25 day induction period (IP)] normal rats a sucrose-rich diet (SRD) results in an increase of plasma (P), liver (L), and heart (H) triacylglycerol (TG) levels, accompanied by a drop in plasma postheparin total (T-TGL) and hepatic (H-TGL) triglyceride lipases activities, IV glucose intolerance (low Kg) and hyperinsulin responses both in vivo and in vitro, suggesting that a state of insulin resistance had developed. Since normalization of P-TG ensued in the medium term [40 to 55 day adaptation period (AP)] we decided to carry out a longitudinal, long-term (90 to 120 day) follow-up study to observe the dynamic behavior of the above metabolic and hormonal parameters as compared to the appropriate time course control rats were fed the standard chow (STD).(ABSTRACT TRUNCATED AT 250 WORDS)

Involvement of oxidative stress in the impairment in biliary secretory function induced by intraperitoneal administration of aluminum to rats.

We have shown that aluminum (Al) induces cholestasis associated with multiple alterations in hepatocellular transporters involved in bile secretory function, like Mrp2. This work aims to investigate whether these harmful effects are mediated by the oxidative stress caused by the metal. For this purpose, the capability of the antioxidant agent, vitamin E, to counteract these alterations was studied in male Wistar rats. Aluminum hydroxide (or saline in controls) was administered ip (27 mg/kg body weight, three times a week, for 90 d). Vitamin E (600 mg/kg body weight) was coadministered, sc. Al increased lipid peroxidation (+50%) and decreased hepatic glutation levels (-43%) and the activity of glutation peroxidase (-50%) and catalase (-88%). Vitamin E counteracted these effects total or partially. Both plasma and hepatic Al levels reached at the end of the treatment were significantly reduced by vitamin E (-40% and -44%, respectively;p< 0.05). Al increased 4 times the hepatic apoptotic index, and this effect was fully counteracted by vitamin E. Bile flow was decreased in Al-treated rats (-37%) and restored to normality by vitamin E. The antioxidant normalized the hepatic handling of the Mrp2 substrates, rose bengal, and dinitrophenyl-S-glutathione, which was causally associated with restoration of Mrp2 expression. Our data indicate that oxidative stress has a crucial role in cholestasis, apoptotic/necrotic hepatocellular damage, and the impairment in liver transport function induced by Al and that vitamin E counteracts these harmful effects not only by preventing free-radical formation but also by favoring Al disposal.

Dietary di(2-ethylhexyl)phthalate-impaired glucose metabolism in experimental animals

The effects of chronic intake of di(2-ethylhexyl)phthalate (DEHP) on the main intermediate glycolytic metabolites in liver and gastrocnemius muscle were investigated in experimental animals. Male Wistar rats (90 -100 g) were fed for 21 days either with a standard chow or the same diet supplemented with 2% (w/w) of DEHP. The DEHP-fed rats had an altered in vivo glucose tolerance associated with abnormal glucose intermediate metabolite contents in liver and skeletal muscle. In these rats, the hepatic content of glucose-6 -phosphate (G-6 -P), fructose-6 -phosphate, pyruvate, lactate, glucose-1 -phosphate and glycogen decreased. At the same time, the G-6 -P content decreased while the pyruvate and lactate levels increased in skeletal muscle. These data, along with the high plasma glucose concentration and the normal lactate blood levels of this group, could indicate that DEHP-fed rats could present a deficiency in muscle glucose and lactate transport, a reduction of the flux through muscle hexokinase and hepatic glucokinase, and a reduction in glycogen synthesis.

Effects of dietary conjugated linoleic acid at high-fat levels on triacylglycerol regulation in mice.

OBJECTIVE Our aim was to investigate the effects of dietary conjugated linoleic acid (CLA) at high-fat (HF) levels on parameters related to triacylglycerol (TG) regulation and some potential impacts on liver damage. METHODS Growing mice were fed a control diet (7% corn oil), an HF diet containing 20% corn oil, or an HF diet containing 3% CLA (HF + CLA) for 30 d. Tissue and organ weights, plasma and tissue TG levels, and parameters related to their regulation were evaluated. Liver oxidative status was also assessed. RESULTS Dietary CLA showed detrimental and beneficial effects. CLA added to the HF diet caused hepatomegaly (+32%) and exacerbated the hepatic TG accumulation (+168%) observed with the HF diet without inducing liver damage; however, it significantly reduced plasma TG concentrations (-37%) and normalized muscular TG content. An increase in glutathione was associated with total normalization of liver lipid peroxidation. In addition, HF + CLA caused dystrophy of epididymal fat pads, even when the HF diet had increased the adipose tissue mass (30%). The biochemical mechanisms involved in the regulation of lipid levels were related to reduced (-20%) hepatic very low-density lipoprotein-TG secretion and decreased muscle (-35%) and adipose (-49%) tissue contributions to the removal of plasma TG by lipoprotein lipase enzymes. CONCLUSION Examination of CLA at HF levels showed hepatomegaly and exacerbation of lipid accretion as a negative impact; however, some positive aspects such as hypotriglyceridemia and protection against oxidative stress were also induced. Even the fat reduction is nutritionally important for weight control; the biochemical mechanisms whereby CLA mediates the potential effects could produce undesirable metabolic alterations.

The duration of feeding on a sucrose-rich diet determines variable in vitro effects of insulin and fructose on rat liver triglyceride metabolism

The aim of the present study was to investigate under controlled conditions the in vitro metabolic effects of fructose and insulin on the triglyceride formation by the isolated perfused livers obtained from hypertriglyceridemic rats that had been fed a sucrose-rich diet for a long-term (15 week) period as compared with those fed sucrose for a short-term (3 week) period. Our findings indicate a significantly higher increase in triglyceride formation by perfused livers of rats fed the sucrose-rich diet for a long-term period in the presence of oleate as a triglyceride-forming substrate (15 weeks, 6-fold increase; 3 weeks, 2-fold increase). Though the contribution of net triglyceride secretion to this increase in triglyceride formation was about twice as high at both durations of feeding on a sucrose-rich diet, a strikingly elevated liver triglyceride accumulation was recorded for a long-term period (15 week, 10-fold increase; 3 week, 4-fold increase). The addition of fructose in the perfusate further increased the output of triglycerides from livers of animals fed the sucrose diet at both durations of feeding. Despite this finding, long-term sucrose led to even higher hepatic triglyceride storage under the present experimental conditions ( μ mollliver at 15 week; 318.2 ± 17.2 vs. 63.0 ± 10.0 at 3 week; P

Effect of the dietary exposure of rat to di(2-ethyl hexyl) phthalate on their metabolic efficiency

The nutritional impact of di(2-ethyl hexyl) phthalate (DEHP), specifically its energy efficiency and nitrogen utilization, was studied in the experimental rat. Groups of male Wistar rats were fed over 21 days with a standard diet alone or a standard diet supplemented with 2% (w/w) DEHP. Food intake, body weight and nitrogen compounds excretion were measured daily. The composition and energetic content of the carcass were determined in animals of both dietary groups after the feeding period, as well as in a separate group on day 0. The food and energy intakes were similar in both groups, however, the efficiencies of energy and nitrogen use were significantly reduced in the DEHP-fed rat. These alterations were reflected by a reduction of 31% on carcass energy retention and a decrease of 26% on cumulative nitrogen balance, without changes in the body composition. The increase of urinary nitrogen excretion, mainly as urea compound, is the major contributing factor to the lower nitrogen retention. These results indicate that DEHP decreases energy efficiency and nitrogen utilization, leading to a pronounced reduction in body weight gain. In addition, this study provides a possible conceptual framework that could explain the metabolic changes induced by DEHP and related compounds in experimental animals.

DIETARY EFFECTS OF PARTIAL OR TOTAL SUBSTITUTION OF SUCROSE FOR STARCH ON GLUCOSE AND LIPID METABOLISM IN DYSLIPIDEMIC RATS

Rats fed on a sucrose-rich diet (SRD) (63 % w/w) up to 30 weeks develop stable hypertriglyceridemia, impaired glucose tolerance and insulin insensitivity. At present only scarce and non-systematic information is available concerning the possible reversion of these metabolic derangements by nutritional or metabolic interventions. The present study was designed to investigate the effect of the isocaloric substitution of sucrose for starch (during 15 weeks) on lipid and glucose metabolisms in rats in which a well-established hypertriglyceridemia and glucose intolerance were present before the amount of sucrose (63% w/w) was partially (33% w/w) or totally (0% w/w) replaced by an isocaloric amount of starch. Our findings show that: i) when the amount of fructose (sucrose moieties) is partially replaced by starch (from 33 to 18% of the total calories) most of its undesirable effects on lipid metabolism are still present and no significant improvement in glucose regulation is noticed; ii) hypertriglyceridemia and glucose intolerance can be completely reversed, without detectable changes in circulating insulin levels, by shifting completely to starch as the source of carbohydrate in the diet. Moreover, when sucrose was removed from the diet, it took 7 weeks for plasma triglyceride levels to become completely normal while plasma free fatty acids and glucose levels needed twice that time. These findings suggest that manipulation of dietary fructose may play a role in the management of lipid disorders associated with glucose intolerance and insulin resistance.

Effects of isomeric fatty acids on reproductive parameters in mice.

Problem  The aim of this study was to determine if dietary fatty acids (FA) level or isomeric FA type may affect reproductive parameters in mice.

Effect of di(2-ethylhexyl) phthalate (DEHP) on lipolysis and lipoprotein lipase activities in adipose tissue of rats.

The di(2-ethylhexyl) phthalate (DEHP) is an ubiquitous environmental chemical with detrimental health effects. The present work was designed to asses some potential mechanisms by which DEHP causes, among others, a reduced body fat retention. Since this effect could be related to an alteration of adipocyte triacylglycerol (TG) metabolism, we evaluated the effects of dietary DEHP in adipose tissues upon (1) the number and size of fat cells; (2) the basal and stimulated lipolysis and (3) the lipoprotein lipase (LPL) activity. Groups of male Wistar rats were fed for 21 days a control diet alone (control group) or the same control diet supplemented with 2% (w/w) of DEHP (DEHP group). The LPL activity of DEHP-fed rats was increased in lumbar and epididymal adipose tissues. These rats had significantly reduced weight in epididymal and lumbar tissues, together with reduced size of epididymal adipocytes. These alterations do not seem to be associated with higher lipid mobility because neither basal lipolysis nor ‘in vitro’ stimulated lipolysis by noradrenaline (NA) showed to be modified by DEHP. Based on these results, we concluded that the adipose tissue size reduction induced by DEHP intake is not due to changes in lipolysis nor to a decreased LPL activity. More research is needed to achieve a comprehensive understanding of the potential mechanisms by which DEHP causes, among others, a reduced body fat retention.

Effects of Trans-Fatty Acids on Liver Lipid Metabolism in Mice Fed on Diets Showing Different Fatty Acid Composition

Aim: Our aim was to investigate the effects of trans-fatty acids (TFA) on liver lipid metabolism in mice fed on experimental diets rich in either oleic or linoleic acid. Methods: Twenty-two male CF1 mice (22.0 ± 0.1 g) were fed with diets rich in corn oil or olive oil, supplemented or not with TFA (0.75 g TFA/100 g diet), for 4 weeks. Changes in triacylglycerol content, the activity and expression of enzymes involved in lipogenesis and fatty acid oxidation were measured. Results: Supplementation of an olive oil-rich diet with TFA increased liver triacylglycerols, the activity and expression of lipogenic enzymes and sterol regulatory element-binding protein SREBP-1a expression. By contrast, when TFA were added to a corn oil-rich diet, they did not modify these parameters. No significant differences were observed among the experimental groups in the activity and expression of carnitine palmitoyltransferase-Ia, body and liver weights or serum triacylglycerol concentrations. Conclusions: The effect of TFA on liver fat accumulation depends on the dietary fatty acid composition. Steatosis induced by TFA when included in an olive oil diet (but not in a corn oil diet) was associated with an increased lipogenesis but not with a decreased fatty acid oxidation in animals fed on the olive oil diet. This metabolic change is mediated by SREBP-1a but not by SREBP-1c, and seems to be independent of insulin.