Effect of chlorpromazine on rat arterial lipid synthesis,in vitro

The effect of chlorpromazine, a major tranquilizer, on arterial lipid metabolism was studied in vitro in rat aortas incubated with [¹⁴C] acetate and [¹⁴C] mevalonate as lipid precursors. Chlorpromazine at a level of 0.25 mM in the incubation medium significantly reduced the incorporation of [¹⁴C] acetate into free fatty acids (p<0.01) and total phospholipids (p<0.001) but not triglycerides. Chlorpromazine also altered the pattern of arterial phospholipids synthesized from [¹⁴C] acetate by significantly increasing the relative proportion of phosphatidylinositol plus phosphatidylserine (p<0.02) and reducing the relative proportion of sphingomyelin (p<0.001). [¹⁴C] Acetate incorporation into the combined fractions of steryl esters plus hydrocarbons and sterols plus diglycerides was also significantly reduced (p<0.001) by 0.25 mM chlorpromazine. Studies with [¹⁴C] mevalonate showed that chlorpromazine is also an inhibitor of sterol biosynthesis in arterial tissues as evidenced by 35–40% reductions (p<0.05) in the formation of¹⁴C-labeled squalene and C₂₇ sterols.     

Asymmetric distribution of decanoate in triacylglycerol synthesized in vitro by mammary glands of lactating mice

Slices, prepared from the mammary glands of lactating mice, were incubated with either [1-¹⁴C]acetate, [U-¹⁴C]glucose, or [1-¹⁴C]decanoate. From all 3 substrates, radioactivity in the synthesized lipids was found mainly in triacylglycerols (TG). When acetate or glucose served as substrate, decanoate (C₁₀) accounted for 24% of the fatty acids in TG. Hydrolysis of the TG by pancreatic lipase yielded [¹⁴C] fatty acids which had relatively more C₁₀ (38%) than did either of the other hydrolysis products mono- or diacylglycerol (14–17%). However, when TG produced by slices from C₁₀ were hydrolyzed, the acid was found to be esterified equally at the C-1, C-2 and C-3 of glycerol. Thus, when fatty acids are synthesized de novo and are converted to TG by gland slices, C₁₀ is predominantly located in the C-3 position, a finding in accord with the situation in milk TG, although such preferential incorporation does not occur when the free acid is presented to the tissue slices.     

Inhibition of sterologenesis in brain and liver of fetal and suckling rats from dams fed di-2-ethylhexyl phthalate plasticizer

The effect of di-2-ethylhexyl phthalate (DEHP) on lipid metabolism was studied in liver and brain from fetal rats taken 3 days before parturition from dams receiving dietary DEHP during gestation. In fetuses from rats receiving 0.5% or 1.0% DEHP in a stock diet, the incorporation of¹⁴C-acetate and labeled mevalonate (³H or¹⁴C) into the C₂₇ sterols, C₃₀ sterols, and squalene fractions of brain tissue incubated in vitro was significantly reduced between the confidence limits P<0.05 to P<0.001. When liver from fetuses was incubated with labeled mevalonate, incorporation of label into the C₂₇ sterol and C₃₀ sterol fractions was significantly reduced as well (P<0.02 to P<0.001), whereas incorporation of labeled mevalonate into the squalene fraction was not significantly altered. The incorporation of¹⁴C-acetate into total hepatic lipids of the fetal rats was also studied, and statistically significant reductions in incorporation were observed in the lanosterol fraction (P<0.001), the combined fraction of sterol esters + squalene (P<0.02), and the combined fraction of cholesterol + diglycerides (P<0.01). No significant changes were observed in the incorporation of¹⁴C-acetate into phospholipids, free fatty acids, or triglycerides. In 8-day old suckling rats delivered from dams fed 0.5% DEHP for the last 16 days of gestation and maintained on the same diet during the nursing period, the incorporation of¹⁴C-mevalonate into hepatic C₂₇ sterols, in vitro, was significantly depressed (P<0.05) whereas in corporation into C₃₀ sterols and squalene was similar to control values. In these same suckling rats, body weights were significantly lower in the control group (21.7 vs. 18.8 g, P<0.01), whereas liver weight as a % of body weight was significantly higher (P<0.01) in rats nursing from the DEHP-fed dams. The results indicate that the inhibitory effect of dietary DEHP on lipid metabolism in the mature rat is transmitted across the placental barrier to the developing fetus and that the abnormal pattern of lipid metabolism in rats delivered from DEHP-fed females is only partially restored to normal during the suckling periods.     

A possible regulatory role of squalene epoxidase in chinese hamster ovary cells

Growth of Chinese Hamster Ovary (CHO) cells in the presence of 20% lipid depleted serum (LDS) for only 2 hr results in an increase in the synthesis of [¹⁴C] sterols from [¹⁴C] mevalonate and from [¹⁴C] squalene compared with cells grown under normal growth conditions in the presence of 10% fetal calf serum (FCS). This enhanced sterol synthesis increases with time of exposure of the cells to LDS. However, exposing these cells for time periods up to 42.5 hr to a growth medium containing 20% LDS did not result in enhanced [¹⁴C] sterol synthesis from [¹⁴C] 2,3-oxidosqualene. Incubation of these cells with [¹⁴C] mevalonate resulted in the accumulation of [¹⁴C] squalene regardless of the presence of either LDS or FCS. These results suggest that squalene epoxidase is a regulatory enzyme in the cholesterol biosynthetic pathway in CHO. This research was supported by Grant 1426 from the Council for Tobacco Research-USA, Inc.     

Effect of dietary safflower oil upon lipogenesis in neonatal lamb

The effect of dietary safflower oil upon lipogenesis has been investigated in neonatal lambs. Preliminary experiments with lambs suckled by their mothers showed that there was a 10-fold increase in the rate of incorporation of [¹⁴C] from acetate into fatty acids in adipose tissue slices during the first 10 days post partum. Barely detectable rates of [¹⁴C] acetate incorporation into fatty acids were found in liver slices from lambs during the same period. In lambs given cows' milk from birth until 11 days of age, there was also a 10-fold increase in the rate of lipogenesis in adipose tissue slices. Supplementing the diet of cows' milk with safflower oil (5 ml/lamb/day) resulted in significantly lower rates of lipogenesis in adipose tissue slices from 11 day old lambs. Administration of safflower oil had no effect upon the concentration of unesterified fatty acids, including linoleic acid, in the lamb adipose tissue slices. The data show that lipogenesis in ovine adipose tissue, like that in rodent liver and adipose tissue, is sensitive to dietary polyunsaturated fatty acids, and that, for the neonatal lamb, the effect of polyunsaturated fatty acids upon lipogenesis is not dependent upon an increase in the tissue concentration of polyunsaturated fatty acids.     

Fatty acid biosynthesis in the developing endosperm ofCocos nucifera

Endosperm tissue of developing coconut endosperm incorporated [¹⁴C] acetate and [¹⁴C]-malonate into [¹⁴C]C₈-C₁₈ fatty acids. The distribution of [¹⁴C] label into the various fatty acids mimicked the distribution of endogenous fatty acids at early and middle stages of endosperm development. Although [¹⁴C] C₈-C₁₈ fatty acids were taken up rapidly by endosperm tissue slices, no elongation occurred; [¹⁴C] stearic acid was not desaturated to oleic. Cell free preparations have also been obtained from this tissue that readily incorporated [¹⁴C] malonyl-CoA into a range of [¹⁴C] fatty acids in the presence of ACP and NADH at pH 7.0. Employing this system, a number of experiments were designed to determine the mechanism of chain length termination. In contrast to intact tissue slice experiments, cell-free extracts yielded as principal products palmitic and stearic acid, although up to 20% were shorter chain acids. A number of possible mechanisms for chain length termination were proposed and tested. Supported in part by NSF Grant No. PCM76 01495.     

Action of the new hypolipidemic agent lifibrol (K12.148) on lipid homeostasis in normal rats: Plasma lipids,hepatic sterologenesis,and the fate of injected [14C] acetate

Lifibrol, a new hypocholesterolemic agent with activity in humans, was examined in normal rats for its short-term and long-term effects on lipid homeostasis. Cholesterol (Chol) synthesis inhibition by lifibrol was demonstratedin vitro in liver minces from normal rats by following [1-¹⁴C]acetate ([¹⁴C]Ac) and DL-[2-¹⁴C]mevalonate ([¹⁴C]-MVA) incorporation into [¹⁴C]Chol. When administered at 50 mg/kg/d, lifibrol reduced plasma total Chol and triglycerides (TG) (P<0.001) within 24 h. The Chol reduction was largely a result of reduction of low density and very low density lipoprotein cholesterol (LDL+VLDL-chol) and a smaller decrease in high density lipoprotein cholesterol (HDL-chol). After 10 d, however, a rebound effect emerged, and after 41 d, plasma Chol, LDL+VLDL-chol, and HDL-chol were restored. In contrast, plasma TG remained at reduced levels (P<0.01). The rebound is attributed to counter-regulation of hepatic sterologenesis that was assessed bothex vivo andin vivo. Theex vivo incorporation of [¹⁴C]MVA and [¹⁴C]octanoate into [¹⁴C]Chol and total digitonin-precipitable [¹⁴C]sterols ([¹⁴C]DPS) in liver minces was increased 2-and 6-fold, respectively, in rats treated 6 d at 50 mg/kg. Similarly,in vivo incorporation of intraperitoneally injected [¹⁴C]Ac into hepatic [¹⁴C]DPS (2 h post-injection) was increased 2- to 5-fold at 50 mg/kg, and evidence for increased sterologenesis in nonhepatic tissue was also obtained. The increased hepatic sterologenesis, evident within 48 h, persisted out to 41 d of treatment by which time increases (P<0.002) in hepatic Chol and carcass total sterols were observed. Additionally, incorporation of injected [¹⁴C]Ac into hepatic [¹⁴C]TG was inhibited 60% by lifibrol (P<0.001), and the appearance of [¹⁴C]TG in plasma was reduced. Circulating free [¹⁴C]fatty acids ([¹⁴C]FFA) were also reduced, but hepatic [¹⁴C]FFA synthesis was unaffected, thus suggesting either a lesser release of newly formed FFA from liver or an enhanced removal from plasma.     

Inhibition of cholesterol synthesis by β-benzal butyric acid and derivatives

Cholesterol biosynthesis has been examined using rat liver slices in vitro from 2-¹⁴C-acetate and 2-¹⁴C-mevalonate, in the presence of β-benzal butyric acid (BBA) and its metabolite, α-hydroxy β-benzal butyric acid (HBBA), both of which are postulated to act as potential hypocholesterolemic agents. Procedures have been devised to follow radioactivity incorporation of these precursors into the squalene, lanosterol and cholesterol fractions. The results show that cholesterol synthesis from labeled acetate is noticeably inhibited by BBA final concentrations as small as 10 μM, while the rate of labeling is much less inhibited by HBBA. When acetate is replaced by labeled mevalonate, cholesterol synthesis is hardly inhibited by both BBA and HBBA. The results indicate that BBA probably affects some of the reactions which lead acetate to mevalonate formation. Acetyl-CoA: ligase (E.C.6.2.1.1) and acetyl-CoA acetyl transferase (E.C.2.3.1.) therefore have been examined. Ligase activity is substantially inhibited only by 1 mM concentration of BBA and HBBA, whereas the transferase enzyme is unaffected. BBA probably affects other reactions in the metabolic sequence which converts acetate into mevalonate. Presented in part at the International Symposium on Problems and Prospectives in Medical Therapy of Atherosclerosis, Parma, November, 1966.     

Inhibition of cholesterol synthesis by cyclopropylamine derivatives of squalene in human hepatoblastoma cells in culture

Two squalene derivatives, trisnorsqualene cyclopropylamine and trisnorsqualeneN-methylcyclopropylamine, were synthesized and tested for inhibition of lanosterol and squalene epoxide formation from squalene in rat hepatic microsomes, and for the inhibition of cholesterol syntheses in human cultured hepatoblastoma (HepG2) cells. Trisnorsqualene cyclopropylamine inhibited [³H]-squalene conversion to [³H]squalene epoxide in microsomes (IC₅₀=5.0 μM), indicating that this derivative inhibited squalene mono-oxygenase. Trisnorsqualenen-methylcyclopropylamine inhibited [³H]squalene conversion to [³H]lanosterol (IC₅₀=12.0 μM) and caused [³H]-squalene epoxide to accumulate in microsomes, indicating that this derivative inhibited 2,3-oxidosqualene cyclase. Cholesterol biosynthesis from [¹⁴C]acetate in HepG2 cells was inhibited by both derivatives (IC₅₀=1.0 μM for trisnorsqualene cyclopropylamine; IC₅₀=0.5 μM for trisnorsqualeneN-methylcyclopropylamine). Cells incubated with trisnorsqualene cyclopropylamine accumulated [¹⁴C]squalene, while cells incubated with trisnorsqualeneN-methylcyclopropylamine accumulated [¹⁴C]squalene epoxide and [¹⁴C]squalene diepoxide. The concentration range of inhibitor which caused these intermediates to accumulate coincided with that which inhibited cholesterol synthesis. The results indicate that cyclopropylamine derivatives of squalene are effective inhibitors of cholesterol synthesis, and that substitutions at the nitrogen affect enzyme selectivity and thus the mechanism of action of the compounds.     

Ketone bodies,glucose and glutamine as lipogenic precursors in human diploid fibroblasts

Incorporation of [¹⁴C] from acetoacetate, D(-)- and L(+)-3-hydroxybutyrate, glucose, glutamine, acetate and palmitate in cellular lipids were studied in cultures in human diploid fibroblasts (HDF). The results showed that acetoacetate was 2–10 times more effective as a lipogenic precursor than was either D- or L-3-hydroxybutyrate. Its extent of incorporation into lipids was 2- to 8-fold more than the other precursors examined under conditions when the overall rates of nonsaponifiable and saponi-fiable lipogenesis as measured by³H₂O incorporation were essentially unchanged. Acetoacetate supported both saponifiable and nonsaponifiable lipid syntheses with half-saturation values (K_m app.) of 185 μM and 30 μM, respectively. Glucose stimulated acetoacetate incorporation into lipids whereas, conversely, acetoacetate inhibited [¹⁴C] glucose incorporation into lipids. The presence of low density lipoproteins (LDL) cholesterol (@40 μg cholesterol/mL) inhibited the incorporation of [¹⁴C] from acetoacetate 56% into nonsaponifiable lipids; the inhibition was consistently higher (75%) when [¹⁴C] glucose or glutamine were the precrusors. The loss of 3-hydroxy-3-methyl-glutaryl CoA (HMG CoA) reductase activity upon addition of LDL-cholesterol was greater than the suppression of [¹⁴C] incorporation from acetoacetate or glucose into nonsaponifiable lipids. In the presence of glucose, [¹⁴C] acetoacetate was incorporated into 3-βOH sterols (digitonin precipitable). 7.7±1.1 times more effectively than was [¹⁴C] glucose. The results suggest that HDF would be a suitable model to investigate the effects of various precrusors of HMG CoA on the rate of cholesterol biosynthesis.     

Differential trypsin effect upon (1-14C) acetate incorporation into choline and ethanolamine glycerophosphatides of rat brain and liver

Preincubation of rat brain and liver slices in a medium (5% glucose, 5% fructose, 1% albumin, 1% trypsin, 10 mM phosphate buffer pH 6.0) used to pretreat brain tissue for the separation of cell types was found to uncouple the incorporation of (1-¹⁴C) acetate into ethanolamine phosphoglycerides from that of the choline phosphoglycerides. Incorporation into ethanolamine phosphoglycerides was stimulated in both brain (330%) and liver (780%) slices, while the incorporation of (1-¹⁴C) acetate into choline phosphoglycerides was reduced for both brain (71%) and liver (63%) slices, compared to control values from nonpreincubated material. With (1-¹⁴C) linolenic acid as a precursor, no significant differences were found in incorporation into ethanolamine phosphoglycerides and choline phosphoglycerides.     

Effect of tibric acid on hepatic cholesterol synthesis in rats

Male albino rats were administered various oral doses of tibric acid daily for 1 week. Serum cholesterol and triglyceride levels were reduced, but total liver content of cholesterol, phospholipids, and triglycerides was increased. Tibric acid treatment suppressed the incorporation of both [¹⁴C] acetate and [³H] mevalonate into cholesterol by liver homogenates.     

Mechanisms Underlying Decreased Hepatic Triacylglycerol and Cholesterol by Dietary Bitter Melon Extract in the Rat

In these studies, we focused on finding the mechanism(s) underlying the bitter melon (Momordica charantia L.) methanol fraction (MF)-dependent reduction in the concentration of hepatic triacylglycerol (TAG) and cholesterol in the rat. Rats were fed diets containing low (5 %) fat for 2 weeks (experiment 1), or low (5 %) and high (15 %) fat for a longer period of 8 weeks (experiment 2). MF was supplemented at 1 % level in both experiments. After feeding, rats were sacrificed, and their livers were prepared as slices and hepatocytes, followed by incubation with [1(2)-¹⁴C] acetate or [1-¹⁴C] oleic acid (18:1 n-6). Under these conditions, we found that rats fed diets containing MF, as compared to those without MF, showed: (1) no adverse effects on food intake and growth, (2) a decreased hepatic TAG and total cholesterol, irrespective of the difference in dietary fat level or feeding period, and (3) a decreased incorporation of [1(2)-¹⁴C] acetate and [1-¹⁴C] oleic acid into TAG of liver slices and hepatocytes. MF-supplemented rats also showed no altered incorporation of labeled acetate into cholesterol and cholesterol ester, an increased fecal excretion of neutral steroids, but not of acidic steroids, and an enhanced mRNA abundance of carnitine palmitoylacyltransferase I, which is the rate-limiting enzyme for fatty acid oxidation. These results suggest that dietary MF decreases hepatic TAG synthesis while enhancing fatty acid oxidation, thereby reducing the concentration of hepatic TAG. The liver cholesterol-lowering effect of MF, however, is probably mediated through an increased fecal excretion of neutral steroids, without an effect on cholesterogenesis.     

Sterol biosynthesis in the oyster,Crassostrea virginica

Sterol biosynthesis in the oyster,Crassostrea virginica, was examined by injection of [2-¹⁴C]acetate and [2,3-³H] lanosterol. The oyster incorporated [2-¹⁴C] acetate into squalene 4,4′-dimethylsterols, 4-monomethylsterols, cholesterol, desmosterol, isofucosterol, and 24-methylenecholesterol, and also converted [2-³H] lanosterol to cholesterol. Therefore, the oyster was concluded to synthesize cholesterol, desmosterol, isofucosterol, and 24-methylenecholesterol from acetate via squalene, probably using the lanosterol pathway.     

Biosynthesis of (Z,Z)-6,9-heptacosadiene in the American cockroach

The biosynthesis of (Z,Z)-6,9-heptacosadiene, the major cuticular hydrocarbon component of the American cockroach, was examined by radiotracer and¹³C-nuclear magnetic resonance (NMR) techniques. Sodium [1-¹⁴C] acetate was incorporated about equally into the saturated and diunsaturated hydrocarbons, whereas [1-¹⁴C] linoleate preferentially labeled the C₂₇ alkadiene and [9,10-³H] oleate labeled the C₂₇ alkadiene almost exclusively.¹³C-NMR demonstrated that [2-¹³C] acetate labeled carbons 25 and 27 but not carbon 3 of the C₂₇ alkadiene. In addition, ozonolysis of the diene labeled from [1-¹⁴C] acetate followed by radio-gas liquid chromatography showed that carbons 1–6 were not labeled, whereas the fragment containing carbons 10–27 was labeled. The data presented in this paper indicate that linoleate from the diet or synthesized de novo is elongated by the addition of acetate units and is then decarboxylated.     

4,8-Dimethyldecanal,the Aggregation Pheromone of <Emphasis Type="Italic">Tribolium castaneum</Emphasis>, is Biosynthesized Through the Fatty Acid Pathway

4,8-Dimethyldecanal (4,8-DMD) is the aggregation pheromone produced by male red flour beetles (RFB), Tribolium castaneum. To elucidate the biosynthetic origin of 4,8-DMD, the following studies were performed: (1)effects of juvenile hormone (JH) III, and pathway inhibitors mevastatin, an inhibitor of the mevalonate pathway, and 2-octynoic acid, an inhibitor of the fatty acid pathway, were tested to determine whether 4,8-DMD is derived from the fatty acid pathway or the mevalonate pathway; (2) incorporation of ¹³C-labeled acetate, propionate, and mevalonolactone into 4,8-DMD was measured to directly determine the biosynthetic origin of 4,8-DMD; and (3) incorporation of deuterium-labeled precursors, including 2-methylbutanoate (C5D), 4-methylhexanoate (C7D), 2,6-dimethyloctanoate (C10D), and 4,8-dimethyldecanoate (C12D) was tested to determine whether 4,8-DMD is biosynthesized in the sequence AcPrAcPrAc (Ac; acetate, Pr; propionate). JH III was topically applied to males at various doses. Inhibitors and isotopically labeled substrates were administered orally by feeding the beetles flour treated with the substrates of interest, after which volatiles were collected from both sexes of RFBs. The amount of 4,8-DMD produced was significantly reduced with increasing doses of JH III. Also, 2-octynoic acid inhibited the production of 4,8-DMD, but mevastatin did not. Exposure of RFBs to [1-¹³C]acetate and [1-¹³C]propionate, but not [2-¹³C]mevalonolactone, resulted in incorporation of the labeled compounds into 4,8-DMD. RFBs fed flour treated with deuterium-labeled C5D, C10D, and C12D, but not C7D, incorporated these compounds into 4,8-DMD. The findings that the production of 4,8-DMD was inhibited by 2-octynoic acid but unaffected by mevastatin, combined with the high incorporation of [1-¹³C]acetate and [1-¹³C]propionate into 4,8-DMD and the incorporation of deuterated precursors, unambiguously demonstrated that 4,8-DMD is of fatty acid rather than terpene biosynthetic origin, and that the biosynthesis of 4,8-DMD proceeds in the sequence Ac-Pr-Ac-Pr-Ac.     

Desaturation of endogenous and exogenous palmitate in lung tissue in vitro

Lung slices from rats fed a fat-free diet supplemented with safflower oil (control) or tripalmitoyl-glycerol (essential fatty acid [EFA]-deficient) were incubated with [¹⁴C] acetate, [¹⁴C] palmitate, or [¹⁴C] stearate. Of the¹⁴C recovered in phospholipids after incubation with [¹⁴C] acetate, more than 87% was in 16-carbon fatty acids. Desaturation, as assayed by the percentage of radioactivity in monoenoates in phospholipid fatty acids, was generally double in EFA-deficient slices compared to control slices, regardless of substrate. Desaturation was significantly greater in slices incubated with acetate or octanoate compared to palmitate, indicating that endogenously synthesized palmitate was desaturated more actively than that derived from an exogenous source. Presented in part to the American Physiological Society, Toronto, Canada, October 1980, and published in abstract form inPhysiologist (1980) 23, 135.     

Stearic acid unlike shorter-chain saturated fatty acids is poorly utilized for triacylglycerol synthesis and β-oxidation in cultured rat hepatocytes

Utilization of stearate as compared to various saturated fatty acids for cholesterol and lipid synthesis and β-oxidation was determined in primary culture of rat hepatocytes. At 0.5 mmol/L in the medium, stearate (18:0) adequately solubilized by albumin was less inhibitory to cholesterol synthesis from [2-¹⁴C] acetate than myristate (14:0) and palmitate (16:0) (68% vs. 91 and 88% inhibition, respectively). The rate of incorporation into cholesterol from [1-¹⁴C] stearate (3.0±0.6 nmol/mg protein/4 h) was 37-, 1.8-, and 7.8-fold of that from myristate, palmitate, and oleate, respectively. Conversely, the rate of [1-¹⁴C] stearate incorporation into total glycerolipids was 88–90% lower than that of labeled palmitate, myristate, and oleate. The rate of [1-¹⁴C] stearate incorporation into triacylglycerol (3.6±0.4 nmol/mg protein/4 h) was 6–8% of that from myristate, palmitate, oleate, and linoleate. The rate of stearate incorporation into phospholipids was the lowest among tested fatty acids, whereas the rate of mono- and diacylglycerol synthesis was the highest with stearate treatment. The rate of β-oxidation as measured by CO₂ and acid soluble metabolite production was also the lowest with [1-¹⁴C] stearate treatment at 22.7 nmol/mg protein/4 h, which was 35–40% of those from other [1-¹⁴C] labeled fatty acids. A greater proportion of stearate than other fatty acids taken up by the hepatocytes remained free and was not metabolized. Clearly, stearate as compared to shorter-chain saturated fatty acids was less efficiently oxidized and esterified to triacylglycerol in cultured rat hepatocytes.     

Inhibition of hepatic sterol and squalene biosynthesis in rats fed di-2-ethylhexyl phthalate

Di-2-ethylhexyl phthalate (DEHP), a commonly used plasticizer, was found to be an inhibitor of the biosynthesis of hepatic nonsaponifiable lipids in the rat. The addition of DEHP at levels of 0.5% or 1.0% to a stock diet of rats resulted in a decreased conversion of acetate-1-¹⁴C and mevalonate-5-³H into squalene, C₃₀ sterols, and C₂₇ sterols by liver minces or slices, in vitro. In studies conducted with 0.5% DEHP feeding from 2 to 11 days, the degree of inhibition was found to increase with the duration of DEHP feeding; the inhibition of³H-mevalonate conversion to squalene and sterols developed more slowly, being reduced to ca. 70% of control values in 11 days, whereas¹⁴C-acetate conversion was reduced to ca. 35% of control values during the same period.³H-mevalonate conversion to sterols and squalene was, however, found to be suppressable to the same extent as¹⁴C-acetate conversion when diets containing 1.0% DEHP were fed for 18 days. The inhibitory effect of dietary DEHP on sterol and squalene biosynthesis from¹⁴C-acetate and³H-mevalonate by rat liver preparations is unlikely to be accounted for by the negative feedback of cholesterol secondary to hepatic sterol accumulation since, in these studies, hepatic total lipid and hepatic total sterol levels were simialr in control and DEHP-fed rats.     

Stimulation of hepatic lipogenesis by eicosa-5,8,11,14-tetraynoic acid in mice fed a high linoleate diet

Liver slices, from mice fasted for one day and then refed for three days either a 15% corn oil diet or a 15% corn oil diet containing eicosa-5,8,11,14-tetraynoic acid (TYA), were incubated with [1-¹⁴C] acetate or [³H]H₂O to determine lipogenic capacity. Dietary TYA produced a twofold stimulation in fatty acid and cholesterol synthesis. TYA also caused an increase in the relative proportion of linoleate (C_18∶2) and a decrease in that of arachidonate (C_20∶4) in liver. Thus, (a) despite high levels of C_18∶2, hepatic lipogenesis can be increased, and (b) even short term feeding of TYA can alter the hepatic fatty acid composition presumably by inhibition of arachidonate synthesis from linoleate.