Hepatic Cholesterol Accumulation Research Articles

Effects of tibolone and conventional hormone replacement therapies on arterial and hepatic cholesterol accumulation and on circulating endothelin-1, vascular cell adhesion molecule-1, and E-selectin in surgically menopausal monkeys.

Menopause and aging are associated with a marked increase in the incidence of coronary heart disease as well as reductions in circulating estrogen, progestogen, and androgen levels. The synthetic compound tibolone and its metabolites have estrogenic, progestogenic, and androgenic characteristics. In the present study, we compared the effects of tibolone, estrogen replacement therapy, and estrogen plus progestogen replacement therapy on arterial and hepatic lipid accumulation and on circulating soluble adhesion molecule and endothelin-1 concentrations in surgically postmenopausal cynomolgus monkeys. Animals were fed an atherogenic diet for 2 years while receiving either no hormone treatment (control, n = 31) or the following treatments at doses designed to mimic the human dose on a daily caloric intake basis: tibolone at 2.5 mg/day (HiTib, n = 31), tibolone at 0.625 mg/day (LoTib, n = 29), conjugated equine estrogens (CEE) alone at 0.625 mg/day (CEE, n = 29), or CEE plus continuous medroxyprogesterone acetate (MPA) at 2.5 mg MPA/day (CEE + MPA, n = 30). Relative to the control group, iliac artery total cholesterol content was not different in the HiTib, LoTib, and CEE + MPA groups but was significantly lower in the group receiving CEE only (P < 0.05). In contrast, hepatic free cholesterol content was reduced in all treatment groups [HiTib (P < 0.01), LoTib (P < 0.05), CEE (P < 0.01), and CEE + MPA (P < 0.05)], whereas hepatic total and esterified cholesterol content were reduced in the HiTib, CEE, and CEE + MPA groups (all P < 0.05). HiTib and CEE groups had lower hepatic triglyceride levels per milligram of protein (P < 0.05). Iliac arterial cholesterol content was highly correlated with hepatic cholesterol content and with previously published histomorphometrically determined coronary artery atherosclerosis, supporting the use of the iliac artery as a surrogate for the coronary artery in the monkey. Circulating levels of soluble vascular cell adhesion molecule-1 were significantly reduced in the HiTib (P < 0.02) and CEE (P < 0.05) groups, whereas soluble E-selectin was reduced in the CEE group only (P < 0.01). Plasma endothelin-1 was significantly reduced in the LoTib (P < 0.05), CEE (P < 0.01), and CEE + MPA (P < 0.01) groups. These results suggest that while tibolone caused marked depression of high-density lipoprotein cholesterol and a resultant twofold increase in the total plasma cholesterol/high-density lipoprotein cholesterol ratio, those effects did not result in exacerbation of iliac artery atherosclerosis, perhaps because of beneficial effects on vascular biology or hepatic metabolism.

The identification of intestinal scavenger receptor class B, type I (SR-BI) by expression cloning and its role in cholesterol absorption

The molecular mechanisms of cholesterol absorption in the intestine are poorly understood. With the goal of defining candidate genes involved in these processes a fluorescence-activated cell sorter-based, retroviral-mediated expression cloning strategy has been devised. SCH354909, a fluorescent derivative of ezetimibe, a compound which blocks intestinal cholesterol absorption but whose mechanism of action is unknown, was synthesized and shown to block intestinal cholesterol absorption in rats. Pools of cDNAs prepared from rat intestinal cells enriched in enterocytes were introduced into BW5147 cells and screened for SCH354909 binding. Several independent clones were isolated and all found to encode the scavenger receptor class B, type I (SR-BI), a protein suggested by others to play a role in cholesterol absorption. SCH354909 bound to Chinese hamster ovary (CHO) cells expressing SR-BI in specific and saturable fashion and with high affinity ( K d∼18 nM). Overexpression of SR-BI in CHO cells resulted in increased cholesterol uptake that was blocked by micromolar concentrations of ezetimibe. Analysis of rat intestinal sections by in situ hybridization demonstrated that SR-BI expression was restricted to enterocytes. Cholesterol absorption was determined in SR-B1 knockout mice using both an acute, 2-h, assay and a more chronic fecal dual isotope ratio method. The level of intestinal cholesterol uptake and absorption was similar to that seen in wild-type mice. When assayed in the SR-B1 knockout mice, the dose of ezetimibe required to inhibit hepatic cholesterol accumulation induced by a cholesterol-containing ‘western’ diet was similar to wild-type mice. Thus, the binding of ezetimibe to cells expressing SR-B1 and the functional blockade of SR-B1-mediated cholesterol absorption in vitro suggest that SR-B1 plays a role in intestinal cholesterol metabolism and the inhibitory activity of ezetimibe. In contrast studies with SR-B1 knockout mice suggest that SR-B1 is not essential for intestinal cholesterol absorption or the activity of ezetimibe.

Ezetimibe selectively inhibits intestinal cholesterol absorption in rodents in the presence and absence of exocrine pancreatic function.

1. Ezetimibe potently inhibits the transport of cholesterol across the intestinal wall, thereby reducing plasma cholesterol in preclinical animal models of hypercholesterolemia. The effect of ezetimibe on known absorptive processes was determined in the present studies. 2. Experiments were conducted in the hamster and/or rat to determine whether ezetimibe would affect the absorption of molecules other than free cholesterol, namely cholesteryl ester, triglyceride, ethinylestradiol, progesterone, vitamins A and D, and taurocholic acid. In addition, to determine whether exocrine pancreatic function is involved in the mechanism of action of ezetimibe, a biliary anastomosis model, which eliminates exocrine pancreatic function from the intestine while maintaining bile flow, was established in the rat. 3. Ezetimibe reduced plasma cholesterol and hepatic cholesterol accumulation in cholesterol-fed hamsters with an ED(50) of 0.04 mg kg(-1). Utilizing cholesteryl esters labelled on either the cholesterol or the fatty acid moiety, we demonstrated that ezetimibe did not affect cholesteryl ester hydrolysis and the absorption of fatty acid thus generated in both hamsters and rats. The free cholesterol from this hydrolysis, however, was not absorbed (92 - 96% inhibition) in the presence of ezetimibe. Eliminating pancreatic function in rats abolished hydrolysis of cholesteryl esters, but did not affect the ability of ezetimibe to block absorption of free cholesterol (-94%). Ezetimibe did not affect the absorption of triglyceride, ethinylestradiol, progesterone, vitamins A and D, and taurocholic acid in rats. 4. Ezetimibe is a potent inhibitor of intestinal free cholesterol absorption that does not require exocrine pancreatic function for activity. Ezetimibe does not affect the absorption of triglyceride as a pancreatic lipase inhibitor (Orlistat) would, nor does it affect the absorption of vitamin A, D or taurocholate, as a bile acid sequestrant (cholestyramine) would.

The cholesterolaemic effects of dietary fats in cholesteryl ester transfer protein transgenic mice.

In order to investigate the role of cholesteryl ester transfer protein (CETP) in the cholesterolaemic response to dietary fats, we analysed plasma lipid profiles of CETP-transgenic and control C57BL/6 mice fed standard chow (AIN-93G; AIN), a low-fat diet, and diets high in butter (saturated fatty acids; SFA), high-oleic acid safflower oil (monounsaturated fatty acids; MUFA), and safflower oil (polyunsaturated fatty acids; PUFA) for 5 weeks. Each group contained four or five mice. There were significant diet and dietxgenotype effects on plasma total cholesterol (TC; and respectively), liver TC ( and respectively), and esterified cholesterol (EC; and respectively); diet effects on plasma triacylglycerol liver free cholesterol and body weight a genotype effect on body-weight gain and a dietxgenotype effect on energy intake In transgenic mice the SFA diet caused significantly higher plasma TC than the PUFA diet In control mice MUFA and PUFA diets, but not the SFA diet, caused significantly higher plasma TC than the low-fat and AIN diets Transgenic mice fed PUFA had lower plasma TC while transgenic mice fed MUFA had lower LDL+VLDL-cholesterol than controls in the same dietary groups. Transgenic mice fed MUFA and PUFA diets also had significantly higher liver TC and respectively) and EC and respectively) than controls fed the same diets. In the present study we showed that: (1) CETP transgenic mice had a cholesterolaemic response to dietary fats similar to that in human subjects; (2) CETP transgenic mice fed PUFA showed significantly lower plasma TC, while those fed MUFA had lower LDL+VLDL-cholesterol than controls; (3) hepatic accumulation of cholesterol, possibly resulting from the combination of the enhanced cholesteryl ester transfer to apolipoprotein B-containing lipoproteins and increased hepatic uptake of cholesterol, may contribute to the cholesterol-lowering effect of MUFA and PUFA in CETP-transgenic mice; (4) CETP may play a role in appetite and/or energy regulation.

Effect of δ 22-5β-taurocholenic acid on hepatic cholesterol and fatty acid in gold thioglucose obese mice fed low- or high-fat diets

We previously demonstrated that hyperglycemic-obese (obob) mice fed a 1% corn oil diet accumulated 10 times as much hepatic cholesterol as did their non-obese (+/?) littermates fed this diet because of difficulty in removal of cholesterol from the liver rather than from increased synthesis. Furthermore, feeding the bile acid analog Δ 22-5β-taurocholenic acid completely prevented the accumulation of hepatic cholesterol in obob mice fed the 1% corn oil diet. The hypothesis to be tested in the current study is that these aspects of cholesterol metabolism in the obob mouse do not occur in the hyperinsulinemic and insulin-resistant gold thioglucose obese mouse. Gold thioglucose obese (gtgo) and non-obese (ngtgo) mice were fed diets containing either 1% corn oil or 40% lard each with or without added taurocholenic acid for 6 weeks and then given a 250 mg meal of [U- 14C]-glucose with incorporation of label into hepatic cholesterol and fatty acid measured 2 hours later. Consistent with earlier results in the obob model, incorporation of labeled glucose was significantly increased in obese compared with non-obese mice fed 1% corn oil and significantly reduced either by feeding 40% lard or by adding taurocholenic acid to the diet. In addition, taurocholenic acid greatly increased incorporation of labeled glucose into hepatic cholesterol in obese or non-obese mice fed either diet. In contrast to obob mice, the percentage of fat in the liver of gtgo mice was increased only 50% compared with ngtgo mice. The comparable increase in obob mice was 480%. Hepatic cholesterol did not increase significantly in the liver of gtgo mice fed 1% corn oil when compared with the ngtgo controls. The comparable increase in obob mice fed 1% corn oil was 350%. Also in marked contrast to obob mice, feeding taurocholenic acid increased hepatic cholesterol compared with non-obese controls fed either diet. The results are discussed in the light of the presence of circulating leptin in gtgo but not in obob mice.

Effect of dietary taurine on endogenous hypercholesterolemia in rats fed on phenobarbital-containing diets.

The effect of dietary taurine on endogenous hypercholesterolemia induced by a phenobarbital-containing diet was investigated. Supplemented taurine did not affect the concentrations of serum cholesterol, but further potentiated the accumulation of hepatic cholesterol in the hypercholesterolemic state induced by phenobarbital. It is suggested that taurine might amplify the hepatic cholesterogenesis in phenobarbital-induced hypercholesterolemia.

Hepatic microsomal triglyceride transfer protein messenger RNA concentrations are increased by dietary cholesterol in hamsters

In hamsters fed high fat diets enriched in trimyristin, tripalmitin or tristearin, increased dietary cholesterol content was associated with increased plasma concentrations of very low density lipoprotein (VLDL) cholesterol and triacylglycerol ( p < 0.0001 and p = 0.0017, respectively). Hepatic microsomal triglyceride transfer protein (MTP) mRNA concentration also increased ( p < 0.0001), independent of the nature of dietary fat, and was significantly correlated with the plasma VLDL lipid concentrations ( p = 0.0002 and p = 0.0106 for cholesterol and triacylglycerol, respectively) and hepatic cholesterol concentrations. Increased expression of the MTP gene may be part of a coordinated response to hepatic cholesterol accumulation leading to increased VLDL lipid secretion.

Hypocholesterolemic Potential of Oat Bran Treated with an Endo‐β‐D‐glucanase from Bacillus subfilis

ABSTRACTOat bran was treated with endo‐β‐D‐glucanases to hydrolyze β(1 ± 3) (1 ± 4)‐glucans. Hydrolysis of β‐glucans reduced the in vitro viscosity of a suspension of oat bran. Untreated oat bran and hydrolyzed oat bran were incorporated into hypercholesterolemic purified diets and fed to rats for 4 wk; a diet containing cellulose was used as a control. Hepatic cholesterol accumulation was affected by dietary treatment and was less in the oat bran group than in the enzyme‐treated oat bran or cellulose groups. Thus, functional properties of (1±3) (1±4)‐β‐D‐glucan, such as viscosity, appear to be related to its hypocholesterolemic potential.

N-[2-[ N′-pentyl-(6,6-dimethyl-2,4-heptadiynyl) amino]ethyl]-(2-methyl-1-naphthylthio)acetamide (FY-087) : A new acyl coenzyme A:Cholesterol acyltransferase (ACAT) inhibitor of diet-induced atherosclerosis formation in mice

FY-087 ( N-[2-[ N′-pentyl-(6,6-dimethyl-2,4-heptadiynyl)amino]ethyl]-(2-methyl-1-naphthyl-thio)acetamide) was found to be a competitive inhibitor of human microsomal acyl coenzyme A:cholesterol acyltransferase (ACAT) with an ic 50 value of 0.11 μM. Under our assay conditions, other ACAT inhibitors tested, specifically YM-750, E-5324, and melinamide, all of which are now in phase I clinical trials or in clinical use in Japan, inhibited this enzyme with ic 50 values of 0.18, 0.14, and 3.2 μM, respectively. FY-087 also inhibited ACAT in acetyl-low density lipoprotein loaded human macrophages (THP-1 cells) with an ic 50 of 0.17 μM. Following the oral administration of FY-087 (30 mg/ kg) to rats, the plasma concentration of FY-087 reached 0.42 μg/mL after 2 hr. This concentration of FY-087 was enough to inhibit blood vessel ACAT activity. Cholesterol-lowering and anti-atherogenic effects of FY-087 were examined using C57BL/6J mice fed an atherogenic diet. In this mouse model, treatment with FY-087 (28 mg/kg) inhibited the increase in plasma cholesterol levels by about 20% and decreased the hepatic accumulation of free and esterified cholesterol by 61 and 67%, respectively. FY-087 also significantly inhibited the atherogenic diet-induced increase in the fatty-streak lesion area of the proximal aorta by 57% in C57BL/6J mice. These results indicate that FY-087 is not only a therapeutically bioavailable ACAT inhibitor that lowers plasma cholesterol levels, but also an effective anti-atherogenic agent in mice fed an atherogenic diet.

Identification of novel differentially expressed hepatic genes in cholesterol-fed rabbits by a non-targeted gene approach.

Several key genes involved in cholesterol metabolism are known to be directly regulated by cholesterol. The possible indirect effect, however, of increased levels of cellular cholesterol on gene expression and its possible role in cholesterol metabolism and atherosclerosis has not been thoroughly explored. In order to determine the overall effect of cholesterol on gene expression, we isolated differentially expressed genes from a PCR-based subtraction library prepared from the liver of chow-fed and cholesterol-fed rabbits. A total of nine upregulated and four down-regulated cDNA fragments were isolated. As determined by Northern blot analysis, the expression of the isolated cDNAs began to change as early as the first week on the cholesterol-rich diet or as late as 4 weeks, which corresponded with hepatic cholesterol accumulation. Three of the cDNAs were identified by DNA sequence homology, whereas the remaining cDNAs had no significant homology match. CYP1A1, a cytochrome P450 isoenzyme, was found to be down-regulated in hepatocytes by cholesterol feeding. Osteopontin and Mac-2, which are produced by macrophages, were found to be up-regulated in Kupffer cells by cholesterol feeding. Overall these results demonstrate the usefulness of the subtraction library approach for identifying new candidate genes for exploring the pathogenesis of atherosclerosis.

Increased messenger RNA levels for low-density lipoprotein receptor and 3-hydroxy-3-methylglutaryl coenzyme a reductase in rat liver after long-term ethanol ingestion

Because long-term alcohol intake leads to severe alterations of cholesterol metabolism resulting in both elevated serum cholesterol levels and increased hepatic concentrations of cholesterol esters, we investigated the effect of long-term ethanol consumption on the hepatic messenger RNA (mRNA) content of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and low-density lipoprotein receptor, two major regulatory factors in cholesterol metabolism, and of apoprotein E. Twenty-four male Sprague-Dawley rats were pair-fed nutritionally adequate liquid diets containing 36% of total calories as either ethanol or isocaloric carbohydrates for 3 wk. In addition, the lipid content of the diets was varied, resulting in 35%, 17.5%, and 8.8% of total calories corresponding to a daily intake of cholesterol of between 1.2 and 6.3 mg/kg body wt. Although increasing dietary cholesterol intake resulted in a significant decrease of hepatic mRNA for low-density lipoprotein receptor and HMG-CoA reductase (p < 0.05), long-term ethanol consumption led to a significant increase of the mRNA for both proteins (p < 0.01), and this increase was predominantly obvious in animals fed a low-cholesterol diet. In contrast, mRNA content of apoprotein E was found to be significantly lower in livers from rats fed ethanol for a prolonged period of time as compared with controls (p < 0.01), and this effect was found to be still present, although less pronounced, after low cholesterol intake.(ABSTRACT TRUNCATED AT 250 WORDS)

Dietary Fat Type and Cholesterol Quantity Interact to Affect Cholesterol Metabolism in Guinea Pigs

Interactions of dietary fat saturation with dietary cholesterol on cholesterol homeostasis in guinea pigs were studied by feeding 15% (wt/wt) fat diets containing lard, olive oil or corn oil, with 0.00, 0.08, 0.17 or 0.33% added cholesterol. Plasma total and LDL cholesterol concentrations significantly increased with increasing dietary cholesterol, with pronounced increments occurring at the pharmacologic (0.33%) level. An interaction between fat type and dietary cholesterol was seen for HDL cholesterol concentrations. Saturated fat and the pharmacologic level of dietary cholesterol increased plasma HDL concentrations, whereas polyunsaturated fat minimized the dietary cholesterol-mediated increase. Interactions were also observed for hepatic cholesterol: dietary cholesterol increased both free and esterified hepatic cholesterol concentrations in all groups fed all the dietary fats, and fat type influenced the extent of hepatic cholesterol accumulation. Lard-fed animals accumulated the least hepatic cholesterol over the range of dietary cholesterol intakes. Dietary cholesterol suppressed hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity, with maximal suppression at all levels of cholesterol intake. Dietary cholesterol had a greater effect on plasma and hepatic cholesterol concentrations and hepatic HMG-CoA reductase activity than did fat type. The data indicated limited interactions of fat type and cholesterol quantity in altering mechanisms regulating plasma cholesterol homeostasis.

Lactose Protects against Estrogen-Induced Pigment Gallstones in Hamsters Fed Nutritionally Adequate Purified Diets

To evaluate the impact of dietary factors on gallstone induction in hamsters, male Syrian hamsters were fed for 2-8 wk purified diets that varied in type and amount of simple sugar (glucose vs. lactose, 17.5-72%), fat (2-5%), fiber (0-15%) and estrogen (0 or 300 micrograms/kg diet). Plasma and liver cholesterol and plasma triglycerides were measured, daily weight gain was determined, cecal weights were obtained, and gallbladder bile was scored by light microscopy and analyzed chemically for its lithogenicity and gallstone incidence. Lactose reduced plasma lipids, especially triglycerides, and hepatic cholesterol accumulation, and maintained a lower biliary cholesterol concentration. When fed at 30% or more, lactose reduced weight gain, increased cecal volume 2- to 4-fold and prevented gallstone formation. Diarrhea and death from 'wet tail' was associated with gallstones and was frequent in hamsters fed glucose without fiber, but its incidence was essentially eliminated by rice flour plus fiber or lactose. Under these experimental conditions of time and diet, estrogen supplementation was required for the formation of gallstones. These appeared to be pigment stones containing a minimal amount of cholesterol. In summary, pigment gallstones were induced in less than 8 wk in hamsters fed estrogen-supplemented purified diets. Lactose feeding improved lipid metabolism and reduced gallstone formation, apparently through its impact on large bowel metabolism.

Reduced cholesterol 7α-hydroxylase activity in hypercholesterolemic hamsters. Preventive effect of a fruit-enriched diet

The influence of a fruit-enriched diet on cholesterol 7α-hydroxylase activity was determined in two types of hypercholesterolemic hamsters, hamsters which develop hypercholesterolemia with aging (FEC hamsters) and hamsters with hypercholesterolemia artificially induced by cholesterol feeding. Cholesterol 7α-hydroxylase activity was reduced in hypercholesterolemic hamsters, the decline reached 50% in the animals fed 0.1% cholesterol diet. Addition of fruits (apples) to the standard diet normalized the activity of cholesterol 7α-hydroxylase in FEC hamsters but in the animals receiving the cholesterol-enriched diet, a wide interindividual variation was observed in response to consumption of fruits. The hepatic accumulation of cholesterol was prevented in half of the animals but persisted at a variable degree in the others. The activity of cholesterol 7α-hydroxylase was negatively correlated with the levels of free and esterified cholesterol in the liver. These findings suggested that augmented levels of cholesterol in the liver can inhibit the conversion of cholesterol into 7α-hydroxycholesterol. The fruit-enriched diet can stimulate the 7α-hydroxylation of cholesterol probably by facilitating the supply of substrate for bile acid synthesis.

Mechanism of the hepatic lipase induced accumulation of high-density lipoprotein cholesterol by cells in culture.

Hepatic lipase can enhance the delivery of high-density lipoprotein (HDL) cholesterol to cells by a process which does not involve apoprotein catabolism. The incorporation of HDL-free (unesterified) cholesterol, phospholipid, and cholesteryl ester by cells has been compared to establish the mechanism of this delivery process. Human HDL was reconstituted with 3H-free cholesterol and [14C]sphingomyelin, treated with hepatic lipase in the presence of albumin to remove the products of lipolysis, reisolated, and then incubated with cultured rat hepatoma cells. Relative to control HDL, modification of HDL with hepatic lipase stimulated both the amount of HDL-free cholesterol taken up by the cell and the esterification of HDL-free cholesterol but did not affect the delivery of sphingomyelin. Experiments utilizing HDL reconstituted with 14C-free cholesterol and [3H]cholesteryl oleoyl ether suggest that hepatic lipase enhances the incorporation of HDL-esterified cholesterol. However, the amount of free cholesterol delivered as a result of treatment with hepatic lipase was 4-fold that of esterified cholesterol. On the basis of HDL composition, the cellular incorporation of free cholesterol was about 10 times that which would occur by the uptake and degradation of intact particles. The preferential incorporation of HDL-free cholesterol did not require the presence of lysophosphatidylcholine. To correlate the events observed at the cellular level with alterations in lipoprotein structure, high-resolution, proton-decoupled 13C nuclear magnetic resonance spectroscopy (90.55 MHz) was performed on HDL3 in which the cholesterol molecules were replaced with [4-13C]cholesterol by particle reconstitution.(ABSTRACT TRUNCATED AT 250 WORDS)

Hepatic Lipid Accumulation and Intracellular Cholesterol Crystal Formation in Hypercholesterolemic Diabetic Rats

Hypercholesterolemia, induced by feeding rats a diet of cholesterol and cholic acid, can be markedly accentuated by diabetes, resulting in serum cholesterol levels greater than 1000 mg/dl and extensive accumulate of cholesterol in the liver. To examine the cellular events of hepatic cholesterol accumulation in cholesterol-fed diabetic rats, we used a fixative containing digitonin and water-soluble embedment to preserve native lipid structures.Sprague Dawley male rats were made diabetic by intravenous injection of streptozotocin (45 mg/kg body weight) and fed a powdered diet containing 2% cholesterol, 10% lard and 1% cholic acid (CLC) for 3 wk. Non-diabetic rats were also fed CLC diet. Cholesterol, glucose and insulin levels were monitored for both groups. Livers were fixed by perfusion with 3% glutaraldehyde (30°C) and 0.2% digitonin in cacodylate buffer (1 hr), then by 1% OsO4 in buffer for 2 hr. Staining was in 2% uranyl acetate and embedding in water soluble urea glutaraldehyde. Thin sections were stained with lead citrate and viewed in a Zeiss EM-9S.

Comparison of hydrophobic surfactant and cholestyramine on lipid and sterol balance in the rat

The effects of dietary supplement with hydrophobic surfactant, Pluronic L-81, on sterol and lipid metabolism in rats consuming a high-fat high-cholesterol diet was determined. Results were compared both to rats on the high-fat high-cholesterol diet alone and to other rats on a similar diet supplemented with cholestyramine. All treatment programs were well tolerated. Pluronic L-81 therapy produced reductions of plasma and hepatic cholesterol levels compared to rats on the diet alone. Additionally, plasma triglycerides were reduced. These changes were associated with relatively high fecal outputs of neutral steroids but acidic steroid excretion was less than observed for rats on diet alone. No malabsorption of neutral lipid was observed for detergent-treated rats. Cholestyramine was also effective in limiting hepatic cholesterol accumulation. Fecal losses of both neutral and acidic steroids were greater in this group compared to rats on diet alone. This was associated with a marked increase of hepatic 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase activity in the cholestyramine-treated group. Both types of treatment limit cholesterol accumulation in rats on a high cholesterol diet. Pluronic L-81 treatment, however, does not significantly increase endogenous cholesterol synthesis while it is greatly increased in response to cholestyramine as reflected by hepatic activities of HMG-CoA reductase.

The influence of cholesterol and fat in maternal diet of rats on the development of hepatic cholesterol metabolism in the offspring

The influence of a high cholesterol-high fat (HC-HF) diet on the hepatic synthesis and catabolism of cholesterol in the offspring was investigated at various stages of growth and development in the rat. Pregnant rats were fed HC-HF or the control diet throughout the period of pregnancy and newborn pups were suckled by mothers fed either the control or experimental diet throughout the period of lactation. The animals were killed at birth, weaning or at maturity for the determination of serum cholesterol, hepatic HMG-CoA reductase, cholesterol 7α-hydroxylase, and microsomal cholesterol accumulation. Feeding HC-HF diet to pregnant or lactating animals caused a 34% inhibition of HMG-CoA reductase in newborn pups and an inhibition of over 75% in suckling and mature animals. Hepatic 7α-hydroxylase in the newborn from mothers on the experimental diet rose progressively when compared to those from the control group which remained unaffected. Microsomal cholesterol in newborn pups was not influenced by maternal diet. Weanlings nursed by mothers fed the HC-HF diet showed a 66% reduction in HMG-CoA reductase activity and 150% increase in cholesterol 7α-hydroxylase when compared to those nursed by animals on the control diet. The results are compatible with the thesis that exposure to high dietary cholesterol and fat in gestation has a significant influence on the development of enzymes regulating cholesterol metabolism at weaning.

Feed-back control of cholesterol synthesis in partially hepatectomized rats

Partial hepatectomy caused a marked stimulation of cholesterol and fatty acid syntheses without affecting serum total cholesterol, total phospholipid and triacylglycerol concentrations of rats so far examined 48 h after the operation. Serum free cholesterol level, however, was increased by the treatment and the ratio of lysophosphatidylcholine to phosphatidylcholine was concomitantly decreased, suggesting the impairment of serum lecithin: cholesterol acyltransferase activity. The lipid content in the liver, especially triacylglycerol and ester cholesterol, was increased markedly by the operation. Feeding of a high cholesterol diet which elevated serum cholesterol and phospholipid levels to the partially hepatectomized rats, accelerated the accumulation of hepatic triacylglycerol and ester cholesterol by the partial hepatectomy. The weight of the regenerating liver was not influenced by cholesterol feeding, which suggested that cholesterol feeding did not inhibit the regeneration mechanism of the liver. The increase of cholesterol synthesis after partial hepatectomy was inhibited by cholesterol feeding. Therefore, it is conceivable that the negative feed-back control of cholesterol synthesis is induced by cholesterol feeding under the stimulated cell divisions of the liver after partial hepatectomy. It is suggested from the present data that a large amount of the cholesterol which is necessary for cell growth can be taken up from serum, when serum cholesterol concentration is high.

The influence of exogenous cholesterol on the fatty acid composition of liver lipids in the rats given linoleate and gamma-linolenate.

AbstractThree groups of rats were given cholesterol and cholic acid for four weeks, and then fed a diet with 20% lard (group 1), a diet with 4% of the lard substituted by methyl linoleate (group 2), and a diet with 2% substituted by methyl linoleate and another 2% by methyl γ‐linolenate (group 3) during the same period. Two control groups (4 and 5) received the same diets as did groups 2 and 3, respectively, but without cholesterol and cholic acid. The hepatic cholesterol accumulation was significantly less in group 3 than in groups 1 and 2, and distinct differences in the degree of plasma cholesterol elevation were found among the cholesterol fed groups (1&gt;2&gt;3). Supplement of γ‐linolenate resulted in a reduction of the ratio of arachidonate to homo‐γ‐linolenate in both the cholesterol‐fed group and the control group, but the ratio was much lower in the former. Incorporation of 1‐14C‐linoleate and 1‐14C‐γ‐linolenate into the liver lipids of groups 2 and 4, and groups 3 and 5 was observed respectively at 1 and 3 hr after the intravenous injection. The specific activity of arachidonate from both of the labeled acids in phospholipid and triglyceride was apparently lower in groups 2 and 3 than in groups 4 and 5, respectively. The distribution rate of radioactivities in tetraenoic acids was also low in groups 2 and 3 among the fatty acids of phospholipid and triglyceride. The results indicated impairment of conversion of homo‐γ‐linolenate into arachidonate in the cholesterol‐fed rats.