1. Investigación

Hepatectomy and nephrectomy in the rat produced an increase in blood glycerol levels which was observed before the fall in blood glucose. The disappearance of total radioactivity from plasma after the i.v. injection of either (U- 14C)-glycerol or (U- 14C)-glucose in these animals was slower than in their sham operated controls. Total radioactivity in plasma was always lower after (U- 14C)-glycerol administration than after (U- 14C)-glucose. The loss of either tracer from plasma in the eviscerated animals was followed by the appearance of 14C-lactate, demonstrating their rapid metabolism. The radioactivity appearing in the water soluble fraction of lumbar fat pads was increased in eviscerated animals while in the lipid fraction it was reduced from both tracers. These results show that adipose tissue is able to metabolize small quantities of glycerol directly in viva. The use of both (1 4C)-glycerol and (14C)- glucose by skeletal and heart muscles was enhanced in hepatectomized rats but the effect on synthesis of labelled glyceride glycerol was greater for (1 4C)-glycerol, suggesting its important role in the esterification of fatty acids in these tissues.

Plasma-triglyceride levels were higher in pregnant than in virgin rats. The glucose infusion did not modify this parameter, probably because ef the changes in LPL activity in other tissues which are known to occur in the opposite direction to those observed in this study for adipose tissue and mammary gland. The present results support the notion that the insulin resistant condition which normally occurs during late gestation is responsible for the decreased LPL acti11ity in adipose tissue, but that the mammary gland remains sensitive to insulin and so maternal hyperinsulinemia would contribute to the induction of LPL activity in this organ prior to parturition.

Triglyceride-rich lipoproteins (chylomicrons and very low density lipoproteins) were labelled "in vivo" by injecting (U-14C)-glycerol and (9-10(n)-3H)-palmitate in female rats. After purification, these lipoproteins contained most of the 3 H in esterified fatty acids and the 14c in glyceride glycerol of neutral lipids. This preparation was incubated "in vitro" in the presence of either isolated adipocytes or epididymal fat pad pieces from male rats. With the incubation, a certain proportion of both 3 H-esterified fatty acids and 14C-glyceride glycerol disappeared from the medium, the effect being greater when the incubations were performed with adipocytes than with fat pad pieces. Much greater radioactivity appeared in the lipids of adipocytes than in those of fat pad pieces at the end of 60 or 120 min incubation, and the incorporation of 3H being relatively greater than that of 14c. With the latter isotope, the label appeared not only in the glyceride glycerol fraction but also in the free and esterified fatty acids. Although it is known that lipoprotein lipase activity is lower in adipocytes than in fat pad pieces, our results indicate that, in the former preparation, the enzyme may be more accessible for the substrate. These data also demonstrate that glycerol released by the hydrolysis of lipoprotein glycerides may be partially incorporated into lipids by adipose tissue.

During early pregnancy, long-chain polyunsaturated fatty acids (LC-PUFA) may accumulate in maternal fat depots and become available for placental transfer during late pregnancy, when the fetal growth rate is maximal and fetal requirements for LC-PUFAs are greatly enhanced. During this late part of gestation, enhanced lipolytic activity in adipose tissue contributes to the development of maternal hyperlipidaemia; there is an increase in plasma triacylglycerol concentrations, with smaller rises in phospholipid and cholesterol concentrations. Besides the increase in plasma very-low-density lipoprotein, there is a proportional enrichment of triacylglycerols in both low-density lipoproteins and high-density lipoproteins. These lipoproteins transport LC-PUFA in the maternal circulation. The presence of lipoprotein receptors in the placenta allows their placental uptake, where they are hydrolysed by lipoprotein lipase, phospholipase A2 and intracellular lipase. The fatty acids that are released can be metabolized and diffuse into the fetal plasma. Although present in smaller proportions, maternal plasma non-esterified fatty acids are also a source of LC-PU FA for the fetus, their placental transfer being facilitated by the presence of a membrane fatty acid-binding protein. There is very little placental transfer of glycerol, whereas the transfer of ketone bodies may become quantitatively important under conditions of maternal hyperketonaemia, such as during fasting, a high-fat diet or diabetes. The demands for cholesterol in the fetus are high, but whereas maternal cholesterol substantially contributes to fetal cholesterol during early pregnancy, fetal cholesterol biosynthesis rather than cholesterol transfer from maternal lipoproteins seems to be the main mechanism for satisfying fetal requirements during late pregnancy.

In vitro effects of pantethine on adipose tissue lipolysis and on both hepatic and intestinal cholesterol and fatty acid synthesis in normolipidemic rats are determined and related to their respective in vivo hypolipidemic effects after acute oral administration. At 3, 5, 7 and 24 h after a single high dose of pantethine to rats, free fatty acids (FFA), cholesterol and triglycerides levels decreased whereas plasma glycerol increased, the effect becoming significant at 7 h. The release of glycerol and FF A by epididymal fat pad pieces from rats was measured in Krebs Ringer bicarbonate-albumin buffer supplemented or not with epinephrine and several concentrations of pantethine (0, 10 - 5 , 10 -4, or 10 - 3 M), and it turned out to be enhanced as pantethine concentration increased. Besides, when glucose was present in the medium, this drug lowered fatty acid re-esterification in a dose-dependent manner, the effect being specially evident in the presence of epinephrine. In vitro synthesis of both cholesterol and fatty acids by slices of liver or intestinal epithelial cells was depressed as the concentration of pantethine increased in the medium. Thus, an inhibition of both cholesterolgenesis and lipogenesis seems to contribute to the hypocholesterolemic and hypotriglyceridemic effects of pantethine. On the other hand, the stimulation of lipolysis and the inhibition of fatty acid re-esterification on adipose tissue caused by pantethine must be counteracted by a high fatty acid oxidation in the liver which would explain the decrease in FF A and the increase in glycerol levels detected in the plasma of the pantethine-treated animals. © 1998 Elsevier Science B.V. All rights reserved.

The effects of 2-deoxy-D-glucose (2DG), oligomycin and theophylline on the in vitro production and metabolism of glycerol and its response to insulin and epinephrine were studied in epididymal fat pads from fed rats. 2-DG failed to affect basal or epinephrine stimulated glycerol production but it decreased the uptake of 1-14C-glycerol by the tissue and its conversion to glyceride-glycerol Oligomycin also failed to affect the basal production of glycerol but it in· hibited the effect of epinephrine on this parameter as well as the uptake and utilization of 1-14-C-glyceroL Theophylline enhanced the production of glycerol by the tissue and this effect was not further augmented by epinephrine. Theophylline also inhibited the uptake and utilization of 1-14C-glycerol; the most pronounced effect of theophylline was observed in the formation of 14C-fatty acids from 1.14c. glycerol in the presence of glucose. Insulin, but not epinephrine, decreased the inhibitory effect of theophylline on glycerol utilization. It is concluded that these compounds af. feet more intensely the ability of adipose tissue to metabolize glycerol than to release it through lipolysis. The pathway for glycerol utilization in adipose tissue appears to be more sensitive to changes in the availability of ATP than the mechanisms responsible for the release of glycerol fom the tissue.

The present study in rats was aimed at determining the specific day of pregnancy on which maternal body fat accumulation starts and which tissues are involved. Most of the body weight increase at day 12 of gestation corresponded to conceptus-free maternal weight which progressively increased until the 19th day of gestation after which maternal weight stabilized and the rate of conceptus weight gain became maximal. Maternal carcass fat content progressively increased until day 18 of gestation, increased very markedly on day 19, stabilized between day 19 and 20 and then decreased on day 21. These changes were the opposite of the course of the specific-gravity values. The fresh weight oflumbar fat-pads and mesenteric adipose tissue reflected the changes in carcass fat content throughout gestation. Periuterine adipose-tissue mass declined on day 12 of gestation to be recuperated later, subcutaneous adipose tissue increased on day 12 to decline progressively thereafter and interscapular brown adipose tissue remained stable until day 20 and increased on day 21. With only a few exceptions, the lipid concentration in all these adipose tissues remained stable throughout gestation. Mammary glands and liver weights increased intensely from day 12 and, whereas the lipid concentration in the former was stable, in the latter it decreased on day 12 and increased on days 18 and 19. These results show that in the rat (a) maternal carcass fat accumulation during gestation is not paralleled by the size of the different fatstoring tissues and (b) mammary-gland fat accumulation also contributes to maternal fat storage.