Inhibition Of Lipoprotein Lipase Activity Research Articles

Recombinant human tumor necrosis factor does not inhibit lipoprotein lipase in primary cultures of isolated human adipocytes

Previous studies have demonstrated that cachectin/tumor necrosis factor (TNF) inhibits lipoprotein lipase (LPL) activity in cultures of 3T3-L1 cells. To determine whether TNF also inhibits LPL in human adipocytes, primary cultures of isolated human adipocytes were exposed to a spectrum of concentrations of recombinant human TNF. TNF concentrations up to 1000 pM had no effect on either LPL activity or LPL immunoreactive mass in the human adipocytes. Specific binding of 125I-labeled TNF was demonstrated in human adipocytes, and a TNF concentration of 100 pM competed for approximately 50% of the 125I-labeled TNF binding sites. In contrast, the same TNF in the same concentrations progressively inhibited LPL activity and immunoreactive mass in 3T3-L1 cells. Thus, human adipocytes respond to TNF in a different manner than 3T3-L1 cells. TNF may not cause the cachexia of cancer or chronic infection by directly inhibiting LPL in adipose tissue.

Effects of tumor necrosis factor (TNF) on lipolytic activities of rat heart.

Tumor Necrosis Factor (TNF) inhibits lipoprotein lipase activity in cultured myocytes and in the Langendorff rat heart after 3 h perfusion with TNF of glucocorticoid-pretreated rats. TNF acutely stimulates glyc(ogen)olysis and concomitantly endogenous lipolysis. The latter was significantly increased only when rats had been pretreated with glucocorticoid or fed a trierucate-rich diet. Under these conditions, contractile activity of the Langendorff hearts was acutely increased by TNF. The mechanism of the acute increase of contractile function and the accompanying increased glycolytic and lipolytic activities, by TNF, may be explained by increased cytosolic Ca2+ and cAMP levels.

Inhibition of lipoprotein lipase by plasma from children with the steroid responsive nephrotic syndrome.

The study was designed to investigate the hyperlipidaemia associated with the steroid responsive nephrotic syndrome in children and in particular to examine the mechanism for the delayed clearance of the circulating triglyceride-rich lipoproteins. The possibility that plasma from patients with steroid responsive nephrotic syndrome may contain an inhibitor of lipoprotein lipase activity was studied by examining the effect of the addition of plasma from patients, on normal postheparin lipoprotein lipase activity. Plasma from children with steroid responsive nephrotic syndrome significantly inhibited lipoprotein lipase activity (p less than 0.001), whereas that from patients with familial hypercholesterolaemia and normal children had no significant effect. The inhibition of lipoprotein lipase activity by plasma from patients with steroid responsive nephrotic syndrome correlated significantly with their increased plasma cholesterol and reduced plasma albumin concentrations (p less than 0.001 and less than 0.02, respectively), but there was no significant correlation with plasma triglyceride concentrations. Thus, the degree of inhibition probably reflected the severity of the condition at the time of study. Neither the cholesterol, albumin nor triglyceride concentrations appeared to directly influence the lipoprotein lipase activity of postheparin plasma.

Triacylglycerol kinetics in endotoxic rats with suppressed lipoprotein lipase activity.

Hypertriglyceridemia observed in animals after bacterial endotoxin administration and some forms of sepsis can result from increased hepatic triacylglycerol (TG) output or decreased TG clearance by extrahepatic tissues. To differentiate between these two possibilities, TG and free fatty acid (FFA) kinetics were determined in control and endotoxin-injected (0.1-0.5 mg/100 g) rats 18 h after treatment. Plasma TG and FFA kinetics were assessed by a constant intravenous infusion with [9,10-3H]palmitate-labeled very low-density lipoprotein and [1-14C]palmitate bound to albumin, respectively. In addition, lipoprotein lipase (LPL) activity was determined in heart, skeletal muscle, and adipose tissue as well as in postheparin plasma of functionally hepatectomized, adrenalectomized, and gonadectomized rats. Plasma FFA acid concentrations were slightly increased in endotoxin-treated rats but their turnover did not differ from control. Endotoxin-treated rats had a threefold increase in plasma TG concentrations and decreased heart, skeletal muscle, and post-heparin plasma LPL activity. Plasma TG turnover was decreased, indicating that hypertriglyceridemia was not due to an increased TG output by the liver. Instead, the endotoxin-induced increase in plasma TG concentration was a consequence of the 80% reduction in TG metabolic clearance rate. Thus, suppression of LPL activity in endotoxic animals impairs TG clearance resulting in hypertriglyceridemia. Furthermore, endotoxin administration reduced the delivery of TG-FFA to extrahepatic tissues because hepatic synthesis and secretion of TG from plasma FFA was decreased and LPL activity was suppressed.

Classification of obese patients and complications related to the distribution of surplus fat

The relation between obesity and noninsulin-dependent diabetes mellitus is established. The weak association between obesity and cardiovascular disease or stroke might be attributable to a risk present only in a subgroup of obesity patients. Recent prospective studies have shown such a group to be characterized by abdominal localization of adipose tissue, reviving old empiric observations of such links. The sex-linked adipose tissue distribution is probably dependent on a balance between glucocorticoids and sex steroid hormones. The former are active mainly on intraabdominal adipose tissues through the high density of a specific receptor expressing lipoprotein lipase activity. This effect is counteracted by female sex steroid hormones, mainly progesterone, which promote fat deposition in the gluteal-femoral regions, utilized mainly during pregnancy and lactation. Testosterone stimulates lipid mobilization through transcriptional expression of beta-adrenergic receptors via a specific androgen receptor and also inhibits lipoprotein lipase activity. Intraabdominal adipose tissues, drained by the portal vein, have a very sensitive lipolytic system in men, based on an increased beta-adrenoceptor activity. This is probably a testosterone effect via the mechanisms mentioned. With testosterone deficiency, these mechanisms are less active, permitting accumulation of fat that can be reversed by testosterone substitution. Abdominal distribution of fat in men thus is probably a sign of relative testosterone deficiency.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibody against human lipoprotein lipase

A polyclonal antibody against human lipoprotein lipase (LPL) was prepared. LPL from post-heparin plasma was first purified by heparin Sepharose 4B affinity chromatography. Protein impurities co-eluted with LPL were then eliminated by electrophoresis in the presence of ampholytes. Antithrombin III was identified in this fraction of protein impurities by immunodiffusion against a human antithrombin antiserum, while no antithrombin III could be detected in the purified LPL fraction. Immunodiffusion revealed a single line of precipitation between this antibody and human post-heparin plasma LPL. When pre-incubated with a constant activity of highly purified post-heparin plasma LPL (2.7 mU/75 μl), an equal volume of the anti-LPL antiserum, either pure or diluted to 1/32 caused complete inhibition of the enzyme activity. Half maximal inhibition was observed at a dilution of approximately 1/200. By using a secondary antibody, it was shown that antiserum inhibited LPL activity by means of its immunoglobulins. This antibody was able to inhibit LPL from human adipose tissue, indicating that human LPL released from endothelial cell membranes has common antigenic determinants with adipose tissue

Inhibition of purified bovine milk lipoprotein lipase by propranolol and other β-adrenergic blockers in vitro

Numerous clinical studies have shown that propranolol administration causes hypertriglyceridemia and a decrease in high-density lipoprotein in man. Although these findings have been attributed to diminution of triacylglycerol-rich lipoprotein catabolism by lipoprotein lipase, biochemical studies of the effects of propranolol on lipoprotein lipase activity in vitro have not been previously reported. We purified lipoprotein lipase from raw bovine skimmed milk and examined the effect of propranolol using as substrate phospholipid-stabilized, triolein emulsions containing purified human apolipoprotein C-II. These studies demonstrate that propranolol inhibits lipoprotein lipase activity. The inhibition was found to be noncompetitive with a K i for propranolol of 0.55 mM. In addition, propranolol was shown to bind to phospholipid-stabilized triolein emulsions reaching local concentrations at the particle surface many times higher than its bulk concentration. Metoprolol, timolol and practolol, which are less hydrophobic than propranolol, were less inhibitory. Atenolol was the weakest inhibitor of purified bovine lipoprotein lipase in vitro.

慢性腎不全患者血清中に存在するLPL阻害物質の分析

In uremics, hypertriglyceridemia is common. The cause is mainly due to an impairement of the removal of triglycerides (TG) by lipoprotein lipase (LPL) reaction.Thus, the present study was undertaken to clarify the inhibitors from uremic serum for LPL activity, in vitro.Serum from both uremics (hemodialysis patients: HD) and normal controls (C) was obtained from venous blood and isolated by ultracentrifugation at d. 1.21g/ml to separate into lipoproteins (Lp) and Lp deficient serum (LPDS).The LPDS was extensively dialyzed against 0.15M NaCl, 1mM EDTA, pH 7.4 and concentrated by lyophilization. The concentrated LPDS was in part further fractionated into subfractions by Sephacryl S-200 column chromatography.LPL was purified from postheparin plasma of 50ml using Heparin-Sepharose Affinity Chromatography.LPL assay system was consist of 1.5mM triolein, 0.02% Triton X-100, 0.1M Tris-HCl, pH 8.0, apolipoprotein (apo) C-II of 2.5μg, LPL of 10μl and inhibitors, and the total volume was 80μl. The LPL activity was determined by the method of Hernell et al.LPL activity was depressed by increasing amounts of serum added in the assay system in both C and HD, especially in HD.On the other hand, only extremely concentrated LPDS, about 10-fold, inhibited LPL activity and the inhibition was significantly higher in HD than in C.However, subfractions of LPDS such as 7S and 4S showed a similar inhibitory effect on LPL activity between C and HD, even though the highly concentrated samples of 7S fractions were used.The results in the present study suggest that there are at least 2 inhibitors in serum in both C and HD, and one is dialyzable and another is not dialyzable.

Regulation of lipoprotein lipase synthesis and 3T3-L1 adipocyte metabolism by recombinant interleukin 1

When fully differentiated 3T3-L1 adipocytes were exposed to purified, recombinant murine interleukin 1 (rIL-1), a dose-dependent suppression of lipoprotein lipase activity was observed. The loss of activity reached a maximum of 60–70% of control and appeared to be due to an effect on the synthesis of the enzyme as judged by a suppression of the ability to incorporate [ 35S]methionine into immunoprecipitable lipoprotein lipase. There was no general effect on protein synthesis as determined by radiolabel incorporation into acid precipitable protein; however, after a 17 h exposure of the 3T3-L1 cells to recombinant interleukin 1, the synthesis of two proteins (molecular weights, 19 400 and 165 000 daltons) was enhanced several-fold. When the effect of Il-1 on the major metabolic pathways of the adipocyte was investigated, lipolysis as measured by glycerol release from the cells was markedly enhanced after a 17 h incubation with the hormone, while no effect was observed on de novo fatty acid synthesis. These effects on the metabolism of the adipocytes occur at concentration on a basis of molecules per cell, similar (only a 3-fold difference) to those required for stimulation of [ 3H]thymidine incorporation into mouse thymocyte DNA, suggesting that IL-1 may be a physiologically significant effector of adipocyte metabolism.

Lipoprotein lipase-suppressing mediator in serum of endotoxin-treated rats.

The conditions under which lipoprotein lipase-suppressing mediator is present in serum of endotoxin-treated rats was determined in this study. The suppression of lipoprotein lipase activity in 3T3-L1 cells was used as a bioassay for mediator in serum. Endotoxin (0.1-10 micrograms/ml) and serum from control rats did not suppress lipoprotein lipase activity. Maximum suppression of cell lipoprotein lipase activity (70%) by serum from endotoxic rats required a cell exposure time of 5 h. At the highest dose of endotoxin used (1 mg/100 g), significant suppression was achieved when cells were incubated with 0.5% serum from endotoxic rats (P less than 0.05). Serum obtained 2-3 h after endotoxin injection possessed the maximal ability to suppress lipase activity, but suppressing activity was not present in serum collected 8 h after endotoxin. Rats rendered tolerant to endotoxin by 5 daily injections (0.1 mg/100 g) did not contain detectable levels of mediator in serum after endotoxin injection. The results demonstrate that the presence of lipoprotein lipase activity-suppressing mediator is transitory after in vivo exposure of naive rats to endotoxin, but does not appear in serum of endotoxin tolerant rats.

Effects of exercise training on regional adipose tissue metabolism in pregnant rats.

To investigate the effects of intense exercise training on maternal adiposity and fetal development, normal lean Zucker (Fa/Fa) rats were divided into three groups: control nonpregnant (CNP), control pregnant (CP), and exercise-trained pregnant (TRP). Trained rats swam three continuous hours per day, 6 days/wk, throughout pregnancy. Food efficiency increased with pregnancy, the highest values being observed during the last trimester of the gestation period. On day 21, pup number, weight, and body composition were similar for CP and TRP. By day 21 of gestation, in both TRP and CP rats, inguinal adipose tissue weight, cell size, and lipoprotein lipase (LPL) activity increased; although TRP values were smaller (P less than 0.05). In CP rats, parametrial and retroperitoneal adipose tissue weights, cell size, and LPL activities increased throughout pregnancy, whereas TRP values were similar to CNP (P less than 0.05). Parametrial fat cell lipolysis showed, however, no effect of training. These results indicate that the extensive energy expenditure occurring during endurance training selectively reduced the dams' adiposity in a regionally specific fashion without any apparent alteration in pup development. The increased substrate utilization required by training during pregnancy did not, however, modify fat cell lipolysis but rather prevented lipid filling through suppression of LPL activity of some depots.

Comparison of apolipoprotein C-II-deficient triacylglycerol-rich lipoproteins and trioleoylglycerol/phosphatidylcholine-stabilized particles as substrates for lipoprotein lipase

The effect of apolipoproteins C-II and C-III on the lipoprotein lipase-catalyzed hydrolysis of apolipoprotein C-II-deficient triacylglycerol-rich lipoproteins and particles of trioleoylglycerol stabilized with a phosphati-dylcholine monolayer was investigated. For both triacylglycerol-rich lipoproteins and artificial lipid particles, maximal lipoprotein lipase activity occurred at a constant apolipoprotein C-II/phospholipid mol ratio of 2.0 Δ 10 −4 and was independent of particle size, indicating that the amount of apolipoprotein C-II bound to the surface of the substrate is important for enzyme activation. The effect of apolipoprotein C-II on lipoprotein lipase activity with apolipoprotein C-II-deficient lipoproteins as substrate was to decrease the apparent Michaelis constant ( K app m ) from 7.1 to 1.0 mM with minor changes on the apparent maximal velocity ( V max ) (22.2 mmol free fatty acid released/h per mg enzyme). In contrast, apolipoprotein C-II increased the apparent V max from 2.4 to 20.0 mmol free fatty acid/h per mg enzyme for the lipoprotein lipase-catalyzed hydrolysis of trioleoylglycerol/phospholipid particles with little change in K app m (1.0 mM). Addition of apolipoprotein C-II-deficient triacylglycerol-rich lipoproteins or high-density lipoproteins to trioleoylglycerol/phospholipid particles in the presence of apolipoprotein C-II inhibited lipoprotein lipase activity. Lipoprotein lipase activity was also inhibited by the addition of a large excess of lipid-free apolipoprotein C-III to the artificial particles. The decrease in lipoprotein lipase activity correlated with the amount of bound apolipoprotein C-II. We suggest that the reported discrepancies on the effect of apolipoproteins C-II and C-III on lipoprotein lipase catalysis is related to differences in substrates and to the amount of added apolipoproteins.

Heparin binding to lipoprotein lipase and low density lipoproteins

Heparin was fractionated on an affinity column of bovine milk lipoprotein lipase (LpL) immobilized to Affi-Gel-15. The bound heparin, designated high-reactive heparin (HRH), enhanced LpL activity, presumably by stabilizing the enzyme against denaturation. The unbound heparin fraction had no observable effect on the initial rate of enzyme activity. However, at longer times of incubation there was inhibition of LpL activity. LpL-specific HRH also showed a high, Ca 2+-dependent precipitating activity towards human plasma low density lipoproteins (LDL). Since LpL and LDL both bind to heparin-like molecules at the surface of the arterial wall, we suggest that their similar heparin-binding specificity may have physiological consequences as it relates to the development of atherosclerosis. Heparin binding Lipoprotein lipase LDL Apolipoprotein Lipolysis

Diethylaminoethoxyhexestrol causes hypertriglyceridemia in guinea pigs

Treatment of rats, monkeys and man with diethylaminoethoxyhexestrol causes phospholipid storage in liver and other tissues. However, this drug has not been reported to alter plasma lipoprotein levels. When guinea pigs were treated with diethylaminoethoxyhexestrol, the fasting plasma triacylglycerol levels increased dramatically, from 43 to 1281 mg/dl, after only five doses of 12.5 mg/kg. Diethylaminoethoxyhexestrol-treated guinea pigs had reduced postheparin lipoprotein lipase activity. In addition, in vitro assays showed that this agent inhibited guinea pig postheparin lipoprotein lipase. It is hypothesized that diethylaminoethoxyhexestrol causes hypertriglyceridemia in guinea pigs because these animals are known to have low levels of serum activator for lipoprotein lipase and may be unusually susceptible to agents that inhibit lipoprotein lipase activity. The ability to produce hypertriglyceridemia in guinea pigs provides an animal model in which the metabolic consequences of hypertriglyceridemia can be studied.

Post-heparin hepatic and lipoprotein lipase activities in nephrotic syndrome.

Lipid metabolism was studied in 18 patients with nephrotic syndrome due to various glomerulonephritides. Nephrotic patients had hypercholesterolemia with or without hypertriglyceridemia. The mean serum high-density lipoprotein cholesterol concentration was not significantly reduced in nephrotic patients. Hepatic lipase and lipoprotein lipase activities were measured selectively in post-heparin plasma from all 18 patients using a substrate-specific method. The mean lipoprotein lipase activity in nephrotic patients was markedly reduced while the mean hepatic lipase activity was not significantly different from that of controls. Lipoprotein lipase activities correlated inversely with serum triglyceride concentrations, but positively with in vivo post-heparin fractional clearance rates of Intralipid and with serum high-density lipoprotein concentrations. Nephrotic serum inhibited lipoprotein lipase activity in normal subjects. The percentage inhibition, however, did not correlate with the degree of hypertriglyceridemia. A relatively weak inverse correlation was shown to exist between plasma albumin concentration and hepatic lipase activities.

Why does experimental insulin deficiency lead to a decrease in removal of very low density-triglyceride from plasma?

We have previously suggested that mechanisms other than reduced lipoprotein lipase (LPL) activity might contribute to the defect in plasma removal of very low density lipoprotein (VLDL)-triglyceride (TG) observed in insulin-deficient rats. To further evaluate this phenomenon, removal rates of TG in nonfractionated plasma, as well as in isolated lipoprotein fractions obtained from insulin-deficient and control rats, were compared in a new, sensitive in vivo bioassay system (estradiol-treated male rats with a consistently low endogenous VLDL-TG pool). Removal of TG in nonfractionated plasma from insulin-deficient rats was slower than that of control rats: 3.0 +/- 0.3 vs 1.6 +/- 0.2 min (P less than 0.001). No difference was found in removal rate of isolated VLDL-TG (2.5 +/- 0.3 vs 2.6 +/- 0.4 min), or in removal rates of TG carried in other lipoprotein fractions. We next determined the effect of injection into normal rats of aliquots of dialyzed lipoprotein-free (D greater than 1.215) plasma from insulin-deficient and control rats on the removal rate of normal VLDL-TG, and found that lipoprotein-free plasma from insulin-deficient rats significantly (P less than 0.01) prolonged removal of normal VLDL-TG (4.3 +/- 0.4 to 6.8 +/- 0.7 min). This same fraction did not interfere with the in vitro hydrolysis of normal VLDL-TG by post-heparin LPL. Thus, a factor in the D greater than 1.215 plasma fraction of insulin-deficient rats is present which interferes with the rate of removal of TG from plasma, unrelated to inhibition of LPL activity.

Characterization of human chylomicrons

Apolipoprotein and lipid composition of differently sized chylomicrons from healthy volunteers was determined. During their intravasal catabolism the chylomicrons lose triacylglycerol and apolipoproteins. Decreasing particle size results in a loss of apolipoprotein C and apolipoprotein E peptides and an increase in apolipoproteins B and A-I, which constitutes more than 20% of the moiety of small chylomicrons. The C peptides do not seem to behave as a functional entity. Apolipoprotein C-III, the inhibitor of lipolytic activities, is catabolized independently of the other C peptides. Albumin constitutes about 15–25% of the protein moiety of all chylomicrons. The different chylomicron fractions were incubated with lipolytic activities of lipoprotein lipase and hepatic triacylglycerol lipase. At lower substrate concentrations the reactions were of first-order. Large chylomicrons were the favored substrate for both enzymes with Michaelis Menten constant K m = 1.1 mM for hepatic triacylglycerol lipase and 0.48 mM for lipoprotein lipase. After incubation with hepatic triacylglycerol lipase or lipoprotein lipase the shape of chylomicrons differs from that of control particles as demonstrated by electron microscopy. C peptides were completely dissociated and found in the infranatant. In the enzyme assay with triolein gum arabic substrate several apolipoproteins showed an influence on the activities of hepatic triacylglycerol lipase and lipoprotein lipase. Apolipoprotein C-III peptides were the most effective inhibitors of both enzymes. Also, apolipoprotein A-II, A-I and apolipoprotein C-I inhibited lipoprotein lipase activity, whereas only apolipoprotein A-II was able to decrease hepatic triacylglycerol activity.

Lipoprotein lipase suppression in 3T3-L1 cells by an endotoxin-induced mediator from exudate cells.

Conditioned medium from cultures of mouse peritoneal exudate cells incubated wih endotoxin contains a mediator that markedly suppresses (greater than 90%) lipoprotein lipase (triacylglycero-protein acylhydrolase, EC 3.1.1.34) activity in differentiating 3T3-L1 mouse preadipocytes. The effect is dependent upon the amount of mediator and is evident as early as 30 min after the addition of the mediator-containing medium to 3T3-L1 cell cultures. Neither endotoxin nor conditioned medium from cultures of exudate cells not exposed to endotoxin shows the presence of the mediator. Lysates of the exudate cells are also unable to suppress the lipase activity. Increasing the amount of insulin does not reverse this suppression, even at 1000 times the concentration used for standard experiments. The lipoprotein lipase suppression mediator present in the conditioned medium of endotoxin-treated exudate cells is heat labile and has an apparent molecular weight of at least 12,000. The mediator does not inhibit lipoprotein lipase activity directly nor does it affect the half-life of enzyme activity released in the medium. The present study demonstrates that endotoxin promotes the release of a mediator from exudate cells that suppresses the activity of lipoprotein lipase in 3T3-L1 preadipocytes.

低HDL血症患者のHDL代謝における肝性トリグリセリドリパーゼの役割

It has been indicated that lipoprotein lipase (LPL) and hepatic triglyceride lipase (H-TGL) play a role not only in the metabolism of triglyceride rich lipoproteins but also in HDL metabolism. The purpose of the present study was to investigate the role of LPL and H-TGL in the metabolism of HDL-subfractions, HDL2 and HDL3 in the subjects with hypo-high density lipoproteinemia. Clinical diagnosis of the 29 patients with hypohigh density lipoproteinemia studied in the present study were postapoplexy in the steady state, coronary heart disease and hypertension etc. Furthermore, seventeen hypo-high density lipoproteinemia with diabetes mellitus, 19 subjects with normal levels of HDL-cholesterol (C) and 8 diabetics with normal levels of HDL-C were studied. Hypo-high density lipoproteinemia was defined by (HDL2+HDL3)-cholesterol less than 40mg/100ml. Plasma lipoproteins were fractionated by sequential ultracentrifugation at densities 1.063 and 1.110g/ml to obtain VLDL+LDL(d<1.063g/ml), HDL2(1.063<d<1.110g/ml) and HDL3 (d>1.110g/ml). Concentrations of cholesterol, triglyceride and phospholipid in plasma and each lipoprotein fraction were measured. Activities of LPL and H-TGL in postheparin plasma obtained 10min after injection of heparin, 10U/Kg body weight, were assayed utilizing the substrate containing [14C] triolein. Separate assay of LPL and H-TGL activities were performed by inhibition of LPL activity by 1M NaCl. Cholesterol concentrations in HDL2 and HDL3 from hypohigh density lipoproteinemia were significantly lower than those from the patients with normal levels of HDL-C. Decrease of HDL2-cholesterol was more remarkable than that of HDL3-cholesterol. Therefore, HDL2-Ch/HDL3-Ch ratio was significantly lower in hypo-high density lipoproteinemia than in the patients with normal levels of HDL-Ch. As far as phospholipid is concerned, the same tendency was observed as cholesterol. There was no significant difference in triglyceride concentration in HDL2 and HDL3 between hypo- and normo-high density lipoproteinemia. LPL activity was significantly lower in hypo-high density lipoproteinemia without diabetes mellitus than in normo-high density lipoproteinemia. There were no significant differences in H-TGL activities among 4 groups. In hypo-high density lipoproteinemia, significantly negative correlations were observed between H-TGL activities and concentrations of cholesterol and phospholipid in HDL2. Significantly positive correlations were observed between LPL activities and concentrations of triglyceride and phospholipid in HDL2. However, there were no significant correlations among LPL activity, H-TGL activity and lipids concentrations in HDL3. These correlations were much more obvious in diabetics with hypo-high density lipoproteinemia. The data presented indicate the important role of LPL and H-TGL in the metabolism of HDL2 and HDL3 in the subjects with hypo-high density lipoproteinemia.

The in vitro inhibition of bovine milk lipoprotein lipase by a glycoprotein preparation from human atherosclerotic intima

Intimal tissue from human atherosclerotic aortae was collected by the Dermatome procedure. The tissue was extracted with 5 mM Tris · HCl buffer containing 0.3 M NaCl and 1 mM EDTA, pH 7.4. The ammonium sulfate precipitate between 0.4–0.8 saturation obtained from the extract was fractionated on a DEAE-cellulose column and the effluent was monitored for lipoprotein lipase inhibition employing purified bovine milk enzyme. The substrate used was an emulsion of purified olive oil and tritiated triolein. Human serum was the source of activator of the substrate. A peak of inhibitory activity was eluted between 0.15–0.17 M NaCl. The major component in the purified material had properties similar to a glycoprotein (lipolipin) which has previously been purified from porcine aorta and shown to inhibit lipoprotein lipase activity. The partially purified human inhibitor decreased both the basal and the serum-stimulated activity of milk lipoprotein lipase. The inhibition was non-competitive with respect to serum. However, high levels of triglyceride substrate appeared to relieve the inhibitory effect. It is postulated that the inhibitor may be involved in an interaction with the emulsified lipid denying the enzyme access to its substrate.