Plasma lipoproteins and regulation of hepatic metabolism of fatty acids in altered thyroid states.

This article reviews our understanding of effects of thyroid hormone excess and deficiency on hepatic metabolism of FFA, and consequent effects on production, secretion, and metabolism of plasma lipoproteins. In the hyperthyroid state the following alterations are observed. Fatty acid oxidation and ketogenesis are stimulated simultaneously with a paradoxical stimulation of fatty acid synthesis, which may be linked by virtue of a blunted response of mitochondrial carnitine palmitoyltransferase I (CPT-I) to malonyl coenzyme A (CoA). Esterification of fatty acid to triglyceride (TG) is reduced, as is the secretion of the very low density lipoprotein (VLDL) (including VLDL TG, cholesterol, and apoprotein); this may be due, in part, to decreased concentrations of glycerol-3-phosphate (G3P) in the hepatic cell. In the intact animal or patient, however, serum TG concentration is variable, which may reflect increased adipose tissue lipolysis and elevated concentrations of plasma FFA, which would tend to drive VLDL secretion by the liver. Clearance of the VLDL and its metabolic product, the low density lipoprotein (LDL), is increased, resulting in decreased plasma total and LDL cholesterol. Although high density lipoprotein (HDL) cholesterol may also be reduced, the ratio of LDL/HDL cholesterol is further decreased. The regulatory role of the lipoprotein apoproteins is less clear, but hepatic apolipoprotein (apo) B secretion (required for VLDL) is diminished, while apo-AI secretion (required for HDL) is stimulated, perhaps both reflecting rates of synthesis. Plasma concentrations of apo-AI are variable, dependent on relative rates of secretion and clearance. In the hypothyroid, many of these effects are reversed, which results in hyperlipoproteinemias and greater risk for the development of atherosclerotic cardiovascular disease.

Influence of thyroid status on lipid metabolism in the perfused rat liver.

Thyroid disease is often accompanied by changes in the concentrations of serum lipids and lipoproteins. To evaluate the hepatic contribution to the serum abnormalities in thyroid disease, we examined fatty acid metabolism in perfused livers from pair-fed rats made hypothyroid with propylthiouracil (PTU) or made hyperthyroid by treatment with triiodothyronine (T(3)). The animals treated with T(3) became hyperphagic, depending on dose of drug and duration of hyperthyroidism. It was necessary, therefore, for appropriate controls, that food intake of T(3)-treated rats be restricted to quantities consumed by euthyroid rats. Animals treated with PTU for 2 wk became hypophagic, and therefore, food consumption of controls was restricted to that eaten by rats receiving PTU. Dependent on dose of T(3) and duration of treatment, the output of triglyceride and glucose was diminished, whereas output of ketone bodies was increased by livers from hyperthyroid animals. In contrast, livers from PTU-treated animals secreted increased amounts of triglyceride and glucose, whereas ketogenesis was diminished. The best models for study proved to be animals treated with either 10 mug T(3)/100 g body wt per d or 1 mg PTU/100 g body wt per d for 7 d. Under these conditions, all animals consumed the same quantity of food as did the euthyroid rats, but continued to display the metabolic alterations outlined above. The effects of PTU on hepatic metabolism were readily reversible by simultaneous administration of T(3). It is clear from these data that the thyroid status of the rat regulates hepatic triglyceride formation and secretion, and ketogenesis.

Effects of hyperthyroidism on synthesis, secretion and metabolism of the VLDL apoproteins by the perfused rat liver.

Livers from fed male Sprague-Dawley rats, made hyperthyroid by treatment with triiodothyronine (T3), were isolated and perfused in vitro. T3 (9.6 micrograms/day) was administered by osmotic minipump implanted intraperitoneally. Treatment with T3 for either 7 or 28 days reduced hepatic output of very-low-density lipoprotein (VLDL) and net synthesis of total associated apoproteins. After 7 days treatment, incorporation of [4,5-3H]leucine by livers from hyperthyroid rats into VLDL apo E was reduced while incorporation into apo B100, apo B48, and apo C's did not differ from euthyroid controls. The depressed incorporation of radioactivity into total VLDL protein was accounted for almost entirely on the basis of apo E. Incorporation of leucine into the total lipoprotein apo E isolated in the d less than 1.210 was also diminished by the hyperthyroid state, while that into apo B100, apo B48, and apo C in the total perfusate lipoprotein was similar to that of the euthyroid, as was found for the VLDL. Increased amounts of radioactive apo B100 and apo B48, however, were detected in the HDL fraction isolated from the medium perfusing livers from hyperthyroid rats. Hepatic uptake of VLDL protein and lipid was similar in euthyroid and hyperthyroid rats. Reduction of VLDL lipid and protein in the medium perfusing livers from T3-treated rats, therefore reflects hormonal action on synthesis and secretion, rather than uptake. Since the availability of apo B is thought to be required for secretion of VLDL, our observation suggests that synthesis of apo B is not depressed by treatment with T3 and that apoprotein synthesis is not a significant factor in the decreased output of VLDL by the liver, but that, as reported earlier, the lower output is a consequence of decreased synthesis of TG, the result of a diminished supply of hepatic glycero-3-phosphate in the hyperthyroid. The diminished amount of VLDL protein appears to be accounted for by the decreased quantity of apo E associated with a smaller VLDL particle secreted by livers from T3-treated rats.

Hepatic metabolism of [1-14C]oleate in pregnancy.

Livers from normal ad libitum fed nonpregnant and 20 day pregnant rats were perfused in vitro. Uptake and utilization of [1-14C]oleate were measured while the concentrations of free fatty acids in the erythrocyte-free perfusate was maintained at a steady-state level (mean 0.42 +/- 0.03 (S.E.) mM). Pregnancy increased the incorporation by the isolated liver of [1-14C]oleate into perfusate triacylglycerol and cholesteryl esters and into hepatic triacylglycerol, phospholipid, diacylglycerol and cholesteryl esters. The conversion of [1-14C]oleate by the livers into ketone bodies and CO2 was depressed by pregnancy. With pregnancy, the output of the very low density lipoprotein triacylglycerol, cholesterol, cholesteryl esters and phospholipid was increased significantly by the perfused liver. The relative molar composition of the lipid components of the very low density lipoproteins, however, was not altered by gestation, suggesting that the number of particles secreted was increased by pregnancy. The total output of glucose was decreased by livers from pregnant rats. It may be concluded from these data that livers from pregnant rats in late gestation channel fatty acid ([1-14C]oleate) preferentially into products of esterification rather than into products of oxidation.

Lack of effect of somatostatin on the glucagon-induced alterations of hepatic metabolism of [1-14C]oleate.

Livers from normal fed male rats were perfused in a recycling system in vitro. Glucagon was infused in varying quantities to give final concentration in the cell-free perfusate of 4.9 . 10(-10)-4.9 . 10(-7) M after 3 h of perfusion, assuming no degradation of the hormone. Where indicated, cyclic somatostatin was infused simultaneously to give a final concentration of 3.0 . 10(-6) M. In the absence of somatostatin, glucagon at a concentration as low as 4.9 . 10(-10) M increased the release of glucose and increased ketogenesis, but impaired the synthesis and release of perfusate triacylglycerol and very low density lipoprotein lipids. Somatostatin did not affect these actions of glucagon. Somatostatin alone, however, did reduce the output of very low density lipoprotein. It is suggested that the alteration of fatty acid metabolism by somatostatin in vivo results from modulation of pancreatic glucagon secretion, not from interference by somatostatin of the action of glucagon on the liver.

Relationships between fatty acid synthesis and lipid secretion in the isolated perfused rat liver: effects of hyperthyroidism, glucose and oleate.

Various studies on the effects of thyroid status on hepatic fatty acid synthesis have produced conflicting results. Several variables (e.g., plasma free fatty acid and glucose concentrations) are altered simultaneously by thyroid status and can affect fatty acid synthesis. To evaluate the effects of these variables, hepatic fatty acid synthesis (lipogenesis) was studied in isolated perfused livers from normal and triiodothyronine-treated rats. Livers were perfused with media containing either 5.5 or 25 mM glucose without fatty acid, or 5.5 mM glucose and 0.7 mM oleate. Rates of lipogenesis were determined by measurement of incorporation of 3H2O into fatty acids. Lipogenesis in livers from hyperthyroid animals exceeded that of controls, when perfused with 5.5 mM glucose with or without oleate. Perfusion with 25 mM glucose increased lipogenesis in both euthyroid and hyperthyroid groups to the same level, abolishing this difference between them. Perfusion with oleate reduced rates of lipogenesis by livers from euthyroid and hyperthyroid rats to a similar extent, but stimulated secretion of radioactive fatty acid in phospholipid and free fatty acid fractions. Oleate increased ketogenesis by livers from normal and triiodothyronine-treated rats, with higher rates of ketogenesis in the triiodothyronine-treated group. When oleate was omitted, ketogenesis in the presence of 5.5 mM glucose by the hyperthyroid group was similar to that of euthyroid controls, while ketogenesis was decreased in the hyperthyroid group relative to controls when perfused with 25 mM glucose. About 30% of the radioactivity incorporated into the total fatty acid of both groups was recovered in palmitate, with the remainder in longer chain saturated and unsaturated fatty acids. In both euthyroid and hyperthyroid groups, the ratio of triacylglycerol:phospholipid fatty acid radioactivity was not only less than predicted (based on synthetic rates of PL and TG) but also was decreased in perfusions with exogenous oleate compared to perfusions without oleate. In perfusions with oleate, both groups incorporated twice as much radioactivity into phospholipid as into triacylglycerol. The data suggest the following concepts: while hepatic fatty acid synthesis and oxidation are increased simultaneously in the hyperthyroid state, de novo synthesized fatty acids seem to be poorer substrates for oxidation than are exogenous fatty acids, and are preferentially incorporated into phospholipid, while exogenous fatty acids are better substrates for oxidation and esterification to triacylglycerol. The preferential utilization of de novo synthesized fatty acid for phospholipid synthesis may be an important physiologic adaptation insuring a constant source of fatty acid for membrane synthesis.

Effects of high-dose ethinyl estradiol on serum concentrations and hepatic secretion of the very-low-density lipoprotein, triacylglycerol, cholesterol, and apolipoprotein A-I in the rat.

Female and male rats were treated with ethinyl estradiol (5.0 mg/kg daily for 5 days). Control animals were pair fed to compensate for the reduction in food intake induced by the estrogen, or were fed ad libitum. Treatment with ethinyl estradiol reduced total cholesterol and apolipoprotein A-I concentrations in the serum of female and male animals. The concentrations of serum and hepatic triacylglycerol were depressed markedly in animals of both sexes in groups treated with ethinyl estradiol, compared to the control group fed ad libitum. Compared to the pair-fed controls, however, ethinyl estradiol had only a very minor further reduction on serum triacylglycerol concentration. In male and female rats, the synthesis and secretion of triacylglycerol by the liver was, in comparison to the pair-fed controls, stimulated by estrogen, whereas the secretion of unesterified cholesterol was unaffected by any of the treatment regimens. The synthesis and secretion of total cholesteryl esters by livers from male and female rats was increased by treatment with ethinyl estradiol. The hepatic synthesis and secretion of VLDL triacylglycerol and cholesteryl ester was stimulated by ethinyl estradiol in male and female rats, and the VLDL particle was enriched with cholesteryl ester. Treatment with the high-dose estrogen increased the secretion of apolipoprotein A-I by livers from female rats. It is suggested that the depression in the serum concentrations of cholesteryl esters and apolipoprotein A-I is the result of increased rates of hepatic and/or peripheral catabolism of these components and that the hepatic production rates were increased or unaffected in animals administered high doses of ethinyl estradiol. Since the secretion of apolipoprotein A-I by livers from male rats was unaffected by treatment with ethinyl estradiol, the response to estrogen may be sex related.

Effects of several common long chain fatty acids on the properties and lipid composition of the very low density lipoprotein secreted by the perfused rat liver.

1. Livers from normal fed male rats were perfused in vitro with a bloodless medium which contained intially 3% bovine serum albumin and 100 mg% glucose. Albumin alone, or myristate (14 : 0), palmitate (16 : 0), palmitoleate (16 : 1), stearate (18 : 0), oleate (18 : 1), or linoleate (18:2) was infused at a constant rate (496 mumol/4 h), as a complex with albumin, during the experiment. 2. The very low density lipoprotein secreted by the liver after infusion of unsaturated fatty acids (16 : 1, 18 :1, 18 : 2) has a faster rate-zonal mobility in the ultracentrifuge and is, therefore, probably a larger particle with fewer moles of phospholipid and cholesterol relative to triacyglycerol (triacyglycerol/phospholipids/cholesterol = 100/25.1/16.4) than the very low density lipoproteins produced after infusion of saturated (14 : 0, 16 : 0, 18 : 0) fatty acids (triacyglycerol/phospholipids/cholesterol = 100/30.1/19.1). The molar ratio of phosphoipids/cholesterol of the very low density lipoprotein was similar regardless of which fatty acid was infused. The predominant fatty acid of the very low density lipoprotein or hepatic triacyglycerol, in all cases, was the infused acid. 3. We conclude that free fatty acid regulates the quantity and proportions of triacyglycerol, phospholipids, and cholesterol secreted by the liver in the very low density lipoprotein, and therefore, may secondarily influence concentrations of lipids in the very low density lipoprotein and other plasma lipoproteins circulating in vivo.

Enrichment of rat tissue lipids with fatty acids that are prostaglandin precursors.

The effects of supplementation of a complete diet with ethyl arachidonate and with ethyl dihomo-gamma-linolenate (20 : 3Omega6) on the fatty acid composition of plasma and tissue lipid classes were studied in normal rats. 2. These prostaglandin precursors were incorporated in varying degrees into all lipid classes of the tissues that were investigated. The largest elevations were seen in plasma and tissue triacylglycerols. Significant increases were also observed in phospholipids, cholesteryl esters and the free fatty acid fraction. 3. Following the feeding of the ester of 20 : 3Omega6, arachiodonate levels also rose in the lipids of some tissues. In others, such as the renal medulla and platelets, and increase in 20 : 3Omega6 content occurred without a rise in 20 : 4. 4. Platelet aggregation is known to be stimulated by 20 : 4 (via active metabolites), but not by 20 : 3Omega6. The ability to modify 20 : 3Omega6 levels selectively in certain tissues is of interest in light of such pharmacologic differences from 20 : 4.

Hepatic secretion and turnover of serum phosphatidylcholine-cholesterol acyltransferase in male and female rats.

The activity of serum phosphatidylcholine-cholesterol acyltransferase (LCAT), output of the enzyme by the perfused rat liver, and the effect of pretreatment with colchicine on LCAT were studied in male and female rats. It was observed that: 1. Serum LCAT activity in the female exceeded that of the male in fasted animals, whereas in fed animals, LCAT activity was higher in the male than the female. With both sexes, however, serum LCAT activity in fed animals was greater than that in fasted animals. Data are presented which suggest that the observed sex differences were due to concentration and/or composition of the substrate rather than to differences in the serum concentration of the enzyme. 2. The release of LCAT by perfused livers from fasted female rats exceeded that of the male animals. The output of LCAT was inhibited by pretreatment (male) with colchicine, which suggests that hepatic secretion of LCAT is dependent on vesicular transport. 3. The decay of serum LCAT activity in vivo following injection of colchicine was more rapid in fasted female rats than in male animals. These observations lead us to postulate that the turnover rate of LCAT is higher in female rats than in male animals.

Alterations of plasma HDL lipids and apolipoproteins in female rats treated with ethynyl estradiol.

Female Sprague-Dawley rats (160-200 g body weight) were injected subcutaneously daily for 14 days with 1, 5, 15, 50 or 100 micrograms ethynyl estradiol in sesame oil/kg body weight, or with oil alone. Food consumption was restricted to 15 g/day, and animals were fasted overnight before exsanguination. Concentrations of plasma cholesteryl ester decreased with increasing dosage of ethynyl estradiol, as a result of decreases in HDL cholesteryl ester. Concentrations of plasma unesterified cholesterol were not altered by treatment; the tendency for a decrement in the HDL cholesterol was not statistically significant. Plasma and HDL phospholipid also tended to decrease with increasing dose of ethynyl estradiol. Plasma levels of apolipoprotein A-I increased on treatment with 5 micrograms ethynyl estradiol/kg but were diminished at a dose of 50 micrograms ethynyl estradiol/kg. The ultracentrifugally isolated high density lipoproteins were delipidated and the distribution of apolipoproteins was characterized by SDS-polyacrylamide gel electrophoresis and esoelectric focusing. The proportion of apolipoprotein E in HDL was depressed, while apolipoprotein A-I was increased and apolipoprotein C unchanged by treatment with ethynyl estradiol. Clearly, the alterations in plasma HDL lipid levels resulting from treatment with ethynyl estradiol were accompanied by distinct changes in composition of the apolipoproteins of HDL, and a biphasic response dependent on dose of the drug. A possible mechanism for the diminution in the proportion of HDL apolipoprotein E may be the enhanced removal of the apolipoprotein E-rich (HDL1) subfraction of the HDL from the circulation on treatment with ethynyl estradiol.

Cholesterol is required for the secretion of the very-low-density lipoprotein: in vivo studies.

Rats were fed for 1 week with a standard chow diet, a diet supplemented with lovastatin (0.1%), or a diet supplemented with both lovastatin and cholesterol (0.1/0.1%), to study effects of depletion of a putative hepatic metabolic pool of cholesterol on secretion of the very-low-density lipoprotein (VLDL) in the intact animal. Triton WR-1339 (50 mg/100 g body wt.) or the 0.9% NaCl vehicle alone was given intravenously via a sacral vein. Treatment with lovastatin decreased the secretion of all plasma VLDL lipids, the average decrease after 2 h for VLDL triacylglycerol, phospholipid, cholesterol and cholesteryl ester being 45%. When both lovastatin and cholesterol were included in the diet, the secretion of VLDL triacylglycerol and phospholipid was similar to that of control animals, while secretion of VLDL cholesterol and cholesteryl ester was increased. Treatment with lovastatin reduced the hepatic concentration of cholesteryl esters 42% without affecting free cholesterol. In separate experiments, in vivo synthesis of cholesterol was determined 1 h after intraperitoneal administration of 3H2O. Incorporation into hepatic and plasma free cholesterol and cholesteryl esters was greater in the rats fed lovastatin than in control animals, concurrent with decreased VLDL secretion. The metabolism of VLDL was determined in vivo by intravenous administration of 125I-VLDL. The fractional clearance rates of 125I-VLDL from the plasma were similar among the three experimental groups. Synthesis of hepatic triacylglycerol from [1-14C]oleate in vivo was similar in all treatment groups; incorporation into plasma triacylglycerol was reduced with lovastatin treatment and reversed partially by inclusion of 0.1% cholesterol in the lovastatin diet. Plasma concentrations of triacylglycerol followed patterns of incorporation of [1-14C]oleate. Triacylglycerol concentration in the liver increased when cholesterol was included in the diet. In companion experiments, incorporation of [1-14C]oleate into perfusate triacylglycerol in vitro was reduced with perfused livers from lovastatin-treated animals. In these experiments, oxidation of fatty acid into CO2 and perchloric acid-soluble counts was not affected by lovastatin, added either to the diet or to the perfusate in vitro. It appears, therefore, that lovastatin does not affect triacylglycerol synthesis or fatty acid oxidation, which per se might reduce formation and secretion of VLDL. These data, therefore, strengthen the hypothesis that reduced availability of cholesterol in a putative hepatic metabolic pool, required for secretion and transport of triacylglycerol in the VLDL, is a factor contributing to decreased secretion of the VLDL.

Deprivation and repletion of androgen in vivo modifies triacylglycerol synthesis by rat hepatocytes.

Given the same quantity of fatty acid, livers from male rats esterify less fatty acid and secrete less triacylglycerol in very-low-density lipoprotein than do livers from female animals. To elucidate the role of testosterone in maintenance of this male pattern, conversion of [1-14C]oleic acid into triacylglycerol was assessed in vitro by rat hepatocytes (male) following gonadectomy and replacement with testosterone. Following castration, incorporation of fatty acid into triacylglycerol was increased. In contrast, esterification of exogenous fatty acid into phospholipid, cholesteryl esters, and diacylglycerol was unchanged. Treatment with testosterone (75 micrograms/day) reduced incorporation of exogenous fatty acid into triacylglycerol. Higher doses of testosterone (200 or 100 micrograms/day) modified the effect, such that inhibition was observed only at low oleate (0.5 mM) concentrations. At higher substrate concentrations (1.0-2.0 mM) the inhibitory effect was no longer observed. Further, a similar dose-dependent effect of testosterone was observed following in vivo treatment of castrate females with testosterone. These data support the concept of a regulatory role of testosterone in hepatic triacylglycerol synthesis. These findings also demonstrate a biphasic effect of testosterone, an effect that is dependent not only upon the dose of testosterone administered, but also on the concentration of fatty acid to which the hepatocyte is exposed in vitro.

The effects of 17 beta-estradiol and progesterone on the metabolism of free fatty acid by perfused livers from normal female and ovariectomized rats.

To determine whether any interaction occurs between progesterone and 17 beta-estradiol in the regulation of FFA metabolism by the liver, normal female rats were injected sc daily for 14 days with 7.5, 15, 50, or 100 microgram 17 beta-estradiol/kg, 25 mg progesterone/kg, 15 microgram 17 beta-estradiol plus 25 mg progesterone/kg, or vehicle (sesame oil) alone. To determine the effects of these hormones in the absence of endogenous estradiol and progesterone, ovariectomized rats were treated with the steroids. Livers were removed from the variously treated rats and perfused in vitro in a recycling system. An albumin-oleate complex was infused into the perfusate, providing a steady state concentration of 0.3--0.5 nM oleate. The uptake of FFA and output of ketone bodies and glucose by the livers were generally not altered by any of the steroid treatments. Neither these parameters nor triglyceride secretion was altered by ovariectomy. The administration of 15, 50, and 100 microgram estradiol/kg to normal, but not ovariectomized rats, increased triglyceride secretion. Progesterone alone had no effect on the secretion of triglyceride, but did antagonize the estradiol-mediated stimulation of triglyceride secretion in normal rats. The molar ratios of phospholipid to triglyceride and cholesterol to triglyceride of the very low density lipoprotein secreted by livers from normal females treated with estradiol were not altered, suggesting that the livers secreted more very low density lipoprotein particles of the same size. Based on the same criteria, livers from ovariectomized rats secreted smaller very low density lipoprotein particles compared to livers from control animals, an effect which was reversed by the administration of estradiol. We conclude that progesterone antagonizes the stimulation of hepatic triglyceride secretion by estradiol in normal female rats and that ovariectomy prevents this effect of progesterone.

In vivo growth hormone treatment stimulates secretion of very low density lipoprotein by the isolated perfused rat liver.

We have previously demonstrated in hepatocyte suspensions prepared after in vivo GH deprivation [hypophysectomy (hypox)] that rates of esterification of [1-14C]oleic acid into triglyceride (TG) and phospholipid (PL) were diminished, and that these esterification rates were correspondingly restored by repletion with recombinant GH. The current studies were designed to determine if GH exerts a similar effect on the secretion of very low density lipoprotein (VLDL), the primary plasma carrier of TG. We assessed rates of secretion of VLDL lipid and apoprotein by perfused livers prepared from cortisol/T3-replaced hypox female rats in the presence and absence of recombinant human (h) GH infusion. We also determined rates of synthesis and secretion of VLDL TG from infused [1-14C]oleic acid. After hypox, rates of secretion of VLDL lipid (TG, PL, and cholesterol) and apoprotein (total) were significantly decreased. In addition, VLDL secreted under these conditions was depleted of PL, relative to the other lipid components. Secretion of newly synthesized VLDL TG from [1-14C]oleic acid was also decreased; however, neither intracellular accumulation of labeled TG nor absolute tissue levels of TG were significantly changed. Conversely, GH treatment of hypox rats effectively restored rates of secretion of VLDL TG, PL, cholesterol (C) and apoprotein to control levels. These findings support the putative role of GH in regulating VLDL secretion in vivo by demonstrating that alterations in plasma GH are accompanied by changes in VLDL secretion. The findings further suggest that GH may regulate VLDL secretion by altering the amount of PL and/or apoprotein available for formation of the VLDL particle.

Effects of thyroid status on the structure of the very low density lipoprotein secreted by the perfused liver.

To determine whether hyperthyroidism or hypothyroidism affected the structure of the very low density lipoprotein (VLDL) secreted by the perfused rat liver, rats were pretreated with 0.9% NaCl, T3, or propylthiouracil. The livers were perfused with 0-332 mumol bovine serum albumin-oleate/h for 4 h. The structure of the secreted VLDL was determined by compositional analysis and the use of the fluorescence probe molecules diphenylhexatriene, trans-parinaric acid, and cis-parinaric acid. Compositional analysis indicated that the VLDL secreted by livers from rats pretreated with T3 had lower unsaturated to saturated fatty acid ratios than did controls, regardless of the amount of oleate infused. Fluorescence polarization of diphenylhexatriene indicated that the interior core lipids of VLDL secreted by livers from T3-treated rats were more rigid than those secreted by livers of euthyroid animals. Arrhenius plots of polarization and corrected fluorescence of diphenylhexatriene and trans-parinaric acid, but not cis-parinaric acid, revealed that characteristic temperatures were shifted 8 to 10 C higher in VLDL secreted by livers from rats pretreated with T3. Treatment of rats for 7 days with propylthiouracil under these mild conditions did not alter the structure or composition of the VLDL secreted by the perfused liver. In summary, the lipid changes in the VLDL secreted by perfused rat livers from hyperthyroid animals were consistent with these VLDL being more rigid particles than those secreted by livers from euthyroid rats, independent of the infusion rate of the FFA.

Modulation by thyroid status of hepatic low Km phosphodiesterase.

The effects of thyroid status on the activity of hepatic cAMP phosphodiesterase (PDE) were studied in the rat. Male rats were rendered hyperthyroid by treatment with T3 or hypothyroid by treatment with propylthiouracil. The hepatic particulate low Km PDE was solubilized, and its activity was measured at concentrations of 0.12-1.3 microM cAMP. The Km decreased in hypothyroidism and tended to increase in hyperthyroidism with respect to individual controls. The maximal velocity (Vmax) was unaffected by changes in thyroid status. The increases in Km correlated with increasing plasma T3, whereas the Vmax did not. Concentrations of cAMP increased in the livers from hyperthyroid rats and decreased in those from hypothyroid, in comparison with euthyroid rat livers. The effects of thyroid status on various aspects of hepatic lipid metabolism reported from this laboratory may, in part, have resulted from alterations in hepatic cAMP concentrations. These alterations, may have resulted secondarily from changes in the activity of hepatic PDE by changes in the Km for cAMP with little change in the Vmax.

Stimulation of in vitro triglyceride synthesis in the rat hepatocyte by growth hormone treatment in vivo.

Hepatic fatty acid metabolism in the rat is sexually differentiated. Rates of esterification by the liver of fatty acid into triglyceride and other esterification products (phospholipid, diglyceride, cholesteryl esters) are higher in the female than in the male. There is evidence to suggest that GH feminizes other hepatic systems that exhibit sexual dimorphism, including hepatic steroid metabolism, PRL receptors, and estrogen binding. To investigate the role of GH in maintenance of the high rates of fatty acid esterification observed in the female, we assessed rates of [1-14C]oleic acid utilization by hepatocytes prepared from hypophysectomized (hypox) cortisol/T3-replaced female rats with an without continuous in vivo infusion of human (h) GH (5 micrograms/h). In addition, we assessed the effect of in vivo hGH treatment (5 micrograms/h) on [1-14C]oleic acid utilization in the normal male rat. Hypophysectomy was accompanied by a reduction in incorporation of [1-14C]oleic acid into products of esterification (triglyceride, phospholipid, diglyceride) and oxidation (CO2, ketone bodies). Continuous infusion of hGH (5 micrograms/h; 14 days) restored rates of fatty acid esterification in the hypox-cortisol/T3-replaced female rat, with the exception of cholesteryl esters. hGH infusion partially restored rates of fatty acid oxidation in the hypox cortisol/T3-replaced female rat. Treatment of the adult male rat with continuous infusion of hGH (5 micrograms/h; 7 days) resulted in increased rates of incorporation of [1-14C] oleic acid into triglyceride. In contrast, incorporation of oleic acid into phospholipid, diglyceride, and cholesteryl esters was unaltered. These results suggest that GH may be an important regulator of hepatic fatty acid metabolism.

Effects of diuretic and propranolol on plasma lipoprotein lipids.

Ticrynafen (TCNF), a nonthiazide diuretic, has been reported to be nonhyperlipidemic. To define the effects of these drugs on plasma lipoproteins, experiments were performed in hypertensive subjects after placebo therapy, 4 wk after therapy with either hydrochlorothiazide (HCTZ) or TCNF, 3 mo after diuretic with propranolol, and 1 mo after therapy with propranolol alone. Plasma lipoproteins were separated by ultracentrifugation and the lipid fractions isolated by extraction and silicic acid thin-layer chromatography. Plasma low-density lipoprotein (LDL) total cholesterol fell and high-density lipoprotein (HDL) total cholesterol rose in subjects receiving TCNF. TCNF had no effect on plasma low-density lipoprotein (VLDL) triglyceride or phospholipid. There was no significant changes in LDL or HDL total cholesterol in subjects on HCTZ. HCTZ tended to increase plasma VLDL triglyceride and phospholipid. The addition of propranolol to either diuretic had no effect on LDL or HDL total cholesterol but increased VLDL triglyceride, especially in subjects on HCTZ. Propranolol alone had no effect on any of the lipids measured.

Increased arachidonate in lipids after administration to man: effects on prostaglandin biosynthesis.

Ethyl arachidonate was administered orally to 4 healthy male volunteers in a dose of 6 gm daily for a 2 to 3 wk period after 10-day control period. The increased intake of this precursor of the dienoic prostaglandins resulted in significant increases in the relative and absolute amount of arachidonate in plasma triglycerides, phospholipids, and cholesteryl esters. Similar changes in lipid composition were noted in platelets. The excretion of 7alpha-hydroxy-5,11-diketotetranoprostane-1,16-dioic acid, the major urinary metabolite of E prostaglandins in man, was increased by an average of 47% in 3 of the 4 volunteers. Platelet reactivity was assessed by determining the threshold concentration of adenosine diphosphate (ADP) necessary to induce secondary, irreversible aggregation of platelet-rich plasma. This threshold concentration dropped significantly in all volunteers (10% to 60% of control values). It is concluded that the biosynthesis and function of prostaglandins can be augmented in man by oral administration of an esterified precursor fatty acid.