Metabolism of 125I-labeled lipoproteins by the isolated rat lung.

The capacity of the isolated perfused rat lung to metabolize the protein moieties of serum lipoproteins was assessed using homologous (rat) and heterologous (human) plasma lipoproteins. The protein and lipid moieties of the plasma lipoproteins were labeled in vivo with Na[125I]. In selected cases the lipoprotein peptides were labeled in vivo with 14C- or 3H-labeled amino acids. Uptake of lipoprotein label during perfusion was monitored by measure of losses in perfusate label and by rises in pulmonary tissue labeling as shown by radioassay and by light and electron microscope radioautography. Lipoprotein degradation was assessed by fractionation of perfusate and lung tissue radioactive material into trichloroacetic acid (TCA)-isoluble, TCA-soluble, and ether-ethanol-soluble fractions. When heparin was included in the perfusion medium, there was selective degradation of the protein portion of very low density lipoprotein (VLDL) in the perfusate and concomitant uptake of radioactive label by the lungs. Low density lipoprotein (LDL)) was neither taken up nor catabolized by the isolated rat lung in the absence or presence of heparin. By light and electron microscopy, the label was localized over the interalveolar septa, predominantly the capillary endothelium. Disappearance of TCA-insoluble radioactivity from the perfusate was associated with the generation of both TCA-soluble iodide and noniodide radioactivity. Greater than 50% of the radioactive label taken up by the lungs was found in the delipidated TCA-insoluble fraction. This study provides in vitro evidence for pulmonary catabolism of VLDL apolipoproteins and uptake of peptide catabolic products of VLDL by the lung.

Hepatic secretion of lipoproteins in the rat and the effect of experimental nephrosis.

Livers from normal and nephrotic rats were perfused by the nonrecirculating technique. Nephrosis was studied on the 7th d after the injection of puromycin animonucleoside. Amino acid-labeled lipoproteins (d < 1.21) were isolated from the perfusion medium by agarose column chromatography or by sequential density ultracentrifugation. In both groups of animals, in addition to very low density lipoproteins and nascent high density lipoproteins, column chromatography revealed the presence of a peak of 2-3 x 10(6) daltons. This peak contained lipoproteins of densities corresponding to <1.006, 1.006 < d < 1.02, and 1.02 < d < 1.06, which indicated that rat liver secretes a heterogeneous mixture of triglyceride-rich lipoproteins. The amount of these lipoprotein density classes was measured and their lipid and apoprotein composition and their apoprotein specific activity were determined. In both groups of rats there was a progressive rise in phospholipid and decrease in triglyceride content as the isolation density increased from 1.006 and 1.06. The lipoproteins from the nephrotics had higher amounts of cholesterol. The livers from the nephrotic rats secreted two to three times as much lipoprotein as controls in all density classes in the first 20 min, but during the next 40 min only the 1.02 < d < 1.06 and nascent high density lipoproteins remained at this high level compared to controls. A larger total liver pool of apolipoproteins in nephrotic livers was inferred from their lower specific activities during the first 20 min. The apoprotein composition of liver perfusate lipoproteins from nephrotics differed from controls. There was a 40% decrease in the amount of low molecular weight apoproteins in all density classes, with corresponding increases in apo B and apo E in the triglyceride-rich fractions. The apo A-1 content of nascent HDL was increased from 16% in controls to 52% in nephrotics, with corresponding decreases in apo C and apo E. When these results were combined with specific activity measurements of the individual apoproteins and the net secretion rate of total protein in each lipoprotein class, it was possible to estimate the total amount of each apoprotein secreted and the total incorporation of labeled amino acids into each. The incorporation of label gave results similar to those obtained by direct measurement of the amounts of apoproteins. Apo E secretion was increased by a factor of 1.8, apo B by 2.8, and apo A-1 by 8.4, whereas the secretion of apo C was not significantly altered. We explain these results by postulating that the primary stimulus to hepatic plasma protein synthesis in response to proteinuria is general and that subsequent negative feedback regulation affects individual apolipoprotein synthesis rates. A corollary of this hypothesis is that the biosynthesis and secretion of an apoprotein may be regulated independently of the lipoprotein density class in which it is found.

Catabolism of very low density lipoproteins in experimental nephrosis.

The effects of experimental nephrosis in rats, produced by puromycin aminonucleoside, include an elevation of plasma levels of all lipoprotein density classes and the appearance of high density lipoprotein (HDL) rich in apoprotein (apo) A-I and deficient in apo A-IV and apo E. The hyperlipoproteinemia is associated with an increase in hepatic synthesis of lipoproteins. The possible role of decreased very low density lipoprotein (VLDL were obtained from nonfasting animals by ultracentrifugation at d 1.006 and included chylomicrons) catabolism and its relationship to the apolipoprotein composition of nephrotic high density lipoproteins (1.063 less than d less than 1.210, or 1.072 less than d less than 1.210 [HDL]) was explored. When 125I-VLDL was injected, the faster plasma clearance of lower molecular weight apolipoprotein B (apo BL) compared with that of higher molecular weight apo BH which is seen in normal rats was not observed in nephrotic rats. Less labeled phospholipid, apo C, and apo E were transferred from VLDL to higher lipoprotein density classes. Heparin-releasable plasma lipoprotein lipase and hepatic lipase activities were decreased by 50% in nephrotic rats compared with pair-fed controls. Perfusion of livers with medium that contained heparin released 50% less lipase activity in nephrotic rats than in controls. When heparin was injected intravenously, significant decreases in plasma levels of triglycerides and significant increases in levels of free fatty acids were observed in both groups of animals. In the nephrotic rats, 86% of the free fatty acids were in the lipoprotein fractions, as compared with 16% in the controls. Heparin treatment did not restore to normal the decreased apo BL clearance in nephrotic rats but it produced an increased amount of apo A-IV and apo E in the plasma HDL. In vitro addition of partially pure lipoprotein lipase to whole serum from nephrotic rats significantly increased the content of apo E in HDL. We conclude that the abnormal apoprotein composition of HDL in experimental nephrosis is the result of altered entry of apolipoproteins from triglyceride-rich lipoproteins, probably because of decreased lipolysis.

Macrophages from nephrotic rats regulate apolipoprotein E biosynthesis and cholesterol content independently.

The effects of the nephrotic syndrome in rats on the cholesterol content and the biosynthesis of apolipoprotein E (apoE) by resident peritoneal macrophages have been investigated. Since the nephrotic syndrome has been associated with an increased risk of coronary atherosclerosis, we hypothesized that macrophages from nephrotic rats would accumulate cholesterol and undergo transformation into foam cells, with a concomitant increase in apoE biosynthesis. The nephrotic syndrome was induced in rats with puromycin aminonucleoside. Peritoneal macrophages exposed in vivo for 7-21 d to ascites fluid derived from plasma containing sixfold elevations of lipoproteins did not accumulate unesterified or esterified cholesterol. Nevertheless, immunoprecipitation assays after incubation of the isolated cells with [35S]methionine, or immunoblot analysis of the incubation medium demonstrated a 2.6-fold increase in apoE secretion compared with normal macrophages. This increase was accompanied by 5- to 10-fold increases in cellular apoE messenger RNA as determined by quantitative solution hybridization assay. Peritoneal macrophages cultured from nephrotic rats during the period of hypercholesterolemia also showed distinct and highly reproducible morphologic changes. The dissociation between apoE biosynthesis and macrophage cholesterol content provides new insight into the response of peritoneal macrophages in vivo to endogenous hyperlipemia.

The full induction of human apoprotein A-I gene expression by the experimental nephrotic syndrome in transgenic mice depends on cis-acting elements in the proximal 256 base-pair promoter region and the trans-acting factor early growth response factor 1.

To identify molecular factors regulating apo A-I production in vivo, we induced in transgenic mice the experimental nephrotic syndrome, which results in elevated levels of HDL cholesterol (HDL-C), plasma apo A-I, and hepatic apo A-I mRNA. Human (h) apo A-I transgenic mice with different length 5' flanking sequences (5.5 or 0.256 kb, the core promoter for hepatic-specific basal expression) were injected with nephrotoxic (NTS) or control serum. With nephrosis, there were comparable (greater than twofold) increases in both lines of HDL-C, h-apo A-I, and hepatic h-apo A-I mRNA, suggesting that cis-acting elements regulating induced apo A-I gene expression were within its core promoter. Hepatic nuclear extracts from control and nephrotic mice footprinted the core promoter similarly, implying that the same elements regulated basal and induced expression. Hepatic mRNA levels for hepatocyte nuclear factor (HNF) 4 and early growth response factor (EGR) 1, trans-acting factors that bind to the core promoter, were measured: HNF4 mRNA was not affected, but that of EGR-1 was elevated approximately fivefold in the nephrotic group. EGR-1 knockout (EGR1-KO) mice or mice expressing EGR-1 were injected with either NTS or control serum. Levels of HDL-C, apo A-I, and hepatic apo A-I mRNA were lowest in nonnephrotic EGR1-KO mice and highest in nephrotic mice expressing EGR-1. Although in EGR1-KO mice HDL-C, apo A-I, and apo A-I mRNA levels also increased after NTS injection, they were approximately half of those in the nephrotic EGR-1-expressing mice. We conclude that in this model, basal and induced apo A-I gene expression in vivo are regulated by the trans-acting factor EGR-1 and require the same cis-acting elements in the core promoter.

Ascites fluid lipoproteins in experimental nephrotic syndrome.

Lipoprotein content and composition were studied in ascites fluid of puromycin aminonucleoside-nephrotic rats. All of the lipoprotein density classes were found in ascites fluid. Protein levels compared to plasma were: very low density lipoprotein (VLDL, d less than 1.006), 1.2%; intermediate density lipoprotein (IDL, 1.006 less than d less than 1.02), 2.6%; low density lipoprotein (LDL, 1.02 less than d less than 1.063), 1.0%; and high density lipoprotein (HDL, 1.063 less than d less than 1.21), 1.1%. The predominant protein in ascites fluid was albumin, present at 1.9% of the plasma level. Radioiodinated VLDL and HDL injected intravenously into nephrotic rats appeared in lipoprotein fractions of the ascites fluid. VLDL and IDL triacylglycerol content and particle diameter were low compared with plasma particles, suggesting peritoneal triacylglycerol lipase activity; such lipase activity could account for the increased proportion of LDL in the ascites fluid. Ascites fluid LDL and HDL phospholipid and free cholesterol were high and cholesteryl ester was low. Ascites lipoproteins contained the same apolipoproteins as plasma, but in different proportions. Ascites VLDL had higher apolipoprotein B and lower apolipoprotein E, while LDL and HDL had higher apolipoprotein E. Ascites HDL could be separated by heparin-Sepharose affinity column chromatography into a retained and a non-retained fraction, while nearly all nephrotic plasma HDL was non-retained. These data suggest that modification of ascites fluid lipoproteins occurs prior to their entry into the lymph and return to the blood, perhaps mediated by peritoneal macrophages.

Catabolism of the apolipoproteins of HDL in control and nephrotic rats.

125I-labeled high density lipoprotein (HDL) from control rats, or from rats made nephrotic by puromycin aminonucleoside, was injected into control or nephrotic rats. At 5 and 20 h, the amount and distribution of label remaining in apolipoproteins HDL, A-I, E, A-IV and the C apolipoproteins was measured after ultracentrifugal isolation of HDL and SDS-polyacrylamide gel electrophoretic separation of each apolipoprotein. There were no significant differences in the removal rates of apolipoprotein HDL or of the individual apolipoproteins when the removal of HDL of controls was compared to HDL of nephrotics. HDL from nephrotic rats contains less than 10% of either the apolipoprotein A-IV or apolipoprotein E content of control HDL, indicating that neither apolipoprotein A-IV nor apolipoprotein E play a significant role in determining the catabolic fate of rat HDL. In severely nephrotic animals the apoliprotein C content of HDL was reduced to 50% of control values and the apolipoprotein A-I content of HDL rose to 87% of the total apolipoprotein. The individual apolipoproteins of HDL from either nephrotics or controls were catabolized at the same rates irrespective of the degrees of nephrosis or altered HDL apolipoprotein composition. The apparent fractional catabolic rates for apolipoprotein HDL and for each of the apolipoproteins, determined after 20 h, did not differ from one another, and all were reduced by half in the nephrotic rats compared to the normal controls. These results support the concept that HDL is catabolized as a particle mediated by apolipoprotein A-I recognition, and they reinforce earlier work indicating that increased synthesis is the dominant factor responsible for increased plasma HDL concentrations in experimental nephrosis.

Comparative effects of dietary fish oil and carbohydrate on plasma lipids and hepatic activities of phosphatidate phosphohydrolase, diacylglycerol acyltransferase and neutral lipase activities in the rat.

In rats fed a fish oil-enriched diet, plasma triacylglycerols were lowered 51%. At the same time there was a mean 45% reduction in Mg2+-dependent phosphatidate phosphohydrolase activity in liver microsomes and a mean 20% decrease in microsomal triacylglycerol (neutral) and diacylglycerol hydrolase activities, but not of diacylglycerol acyltransferase. These observations support the hypothesis that decreases in the activities of phosphatidate phosphohydrolase and of both lipases are involved in the expression of the inhibitory effects of fish oil feeding on hepatic lipoprotein triacylglycerol secretion. Conversely, the feeding of a sucrose-enriched diet resulted in a mean 39% rise in plasma triacylglycerols, a 19% increase in triacylglycerol hydrolase and a mean 45% increase in Mg2+-dependent microsomal phosphohydrolase activity. The effects of the two nutritional interventions on phosphatidate phosphohydrolase activity confirm a key function for this enzyme in triacylglycerol formation.

Characterization of a bile salt-dependent cholesteryl ester hydrolase activity secreted from HepG2 cells.

HepG2 cells and medium were assayed for cholesteryl ester hydrolase (CEH) activity in the presence and absence of sodium cholate. Although bile salt-dependent CEH activity was measured in the medium at 6 to 96 h (up to 4500 pmol/h per mg cell protein), there was very little activity detected in the corresponding cell homogenates (less than 70 pmol/h per mg cell protein). Activity in the medium was expressed only in the presence of trihydroxy bile salts and was maximal at 40 mM cholate and pH 7.5. Incubation of HepG2 cells with brefeldin A resulted in an 80 to 90% inhibition of secretion of the bile salt-dependent CEH activity, while only inhibiting total protein secretion by 42%. Bile salt-dependent CEH activity could also be detected in rat liver perfusates. Although there was measurable activity in all of 14 livers analyzed (47 +/- 10 and 53 +/- 17 nmol/h per g liver per h perfusion during two 5-min collections after 15 and 30 min of perfusion, respectively), it did not correlate with the activity found in corresponding liver homogenates, as only four livers had detectable bile salt-dependent CEH activity. These results provide evidence for the secretion of a bile salt-dependent CEH activity, from both a hepatic cell line and the intact liver, that has similar properties to the enzyme previously isolated from rat liver homogenates and rat pancreas.

Proteolysis of very low density lipoprotein in perfused lung.

Perfusion of homologous 125I-labeled rat very low density lipoprotein through isolated rat lungs in the presence of heparin resulted in apoprotein proteolysis. At least the apoprotein C was degraded into two peptides smaller than 7500 daltons as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The lung uptake of radioactivity was small and due mainly to the presence of the larger of the two peptides. The lung protease was not active against an 125-I-labeled albumin substrate and was not released into the medium by heparin.

Sex differences in African-Americans regarding sensitivity to insulin's glucoregulatory and antilipolytic actions.

OBJECTIVE: The purpose of this study was to determine if there are sex differences in African-Americans regarding the effect of obesity on sensitivity to insulin as a glucoregulatory and antilipolytic hormone. RESEARCH DESIGN AND METHODS: Data from study participants, 127 nondiabetic African-Americans (mean age 32 +/- 4 years), included anthropometric measurements, an oral glucose tolerance test (OGTT), a 2-h euglycemic-hyperinsulinemic clamp, and a fasting triglyceride level. Sensitivity to insulin as a glucoregulatory hormone was determined by M/FFM, where M is the mean glucose infusion rate during the second hour of the clamp and FFM is fat-free mass. Sensitivity to insulin's antilipolytic action was assessed during the OGTT by the percent suppression of free fatty acid (FFA) concentrations between 0 and 120 min. The higher the suppression of FFAs, the greater the sensitivity to insulin's antilipolytic action. RESULTS: The participants were classified by BMI into three groups: nonobese (31 men, 24 women), obese (17 men, 14 women), and severely obese (12 men, 29 women). The women had higher percentages of body fat (P < 0.001), and the men had greater FFM (P < 0.001). The M/FFM values for men versus women in each BMI group were nonobese, 8.8 +/- 2.8 vs. 10.8 +/- 4.4; obese, 7.2 +/- 3.4 vs. 8.5 +/- 3.4; and severely obese, 4.7 +/- 2.1 vs. 6.1 +/- 2.2. The difference between the BMI groups was significant (P < 0.001), as was the difference between men and women (P < 0.01). In addition, there was a significant sex difference in percent suppression of FFAS (P < 0.001). The men and women had similar fasting insulin and FFA concentrations; however, in the men only, the percent suppression of FFA declined with increasing obesity (nonobese, 83 +/- 15%; obese, 73 +/- 18%; and severely obese, 69 +/- 19%; P = 0.02). The women in all three BMI groups had lower FFA levels of 86-88%. CONCLUSIONS: Obese African-American men and women are resistant to insulin as a glucoregulatory hormone, but only obese men are resistant to insulin's antilipolytic action; obese African-American women are sensitive to insulin's antilipolytic action. The combined presence of sensitivity to insulin's antilipolytic action with resistance to insulin's glucoregulatory action in obese African-American women may contribute to their high prevalence of obesity and type 2 diabetes.

High density lipoprotein binding by rat Fu5AH hepatoma cells is not related to cholesterol content.

The binding of 125I-HDL obtained from nephrotic rats (HDLne) containing only apo A-I and apo C, to rat hepatoma cells (Fu5AH) grown to confluency was studied under conditions which increased the free cholesterol or the cholesteryl ester content. The high affinity binding (Kd = 5 nM) measured at 4 degrees C was unchanged. This transformed cell line also exhibited greater specificity for rat HDL compared to human HDL than has been reported for other types of cultured cells. When the cells were allowed to internalize and degrade HDLne at 37 degrees C, the acid-soluble products were derived almost entirely from the breakdown of apo A-I. Competition experiments with an LDL fraction from nephrotic rat plasma (LDLne) which contained 20% of apo A-I indicated that it was as effective as other rat HDL preparations in competing for the binding of HDLne at 4 degrees C, based on its content of apo A-I. Control experiments indicated that labeled apo A-I in HDLne exchanged less than 1% when incubated with a 50-fold excess of unlabeled LDLne for 2 h at 4 degrees C. These results point to a critical role of cell type in HDL binding. They support the view that apo A-I is a ligand. The up-regulation of high affinity HDL binding by cholesterol which has been reported with cultured human fibroblasts and Hep G2 cells does not occur in the Fu5AH rat hepatoma cell line.

Hepatic lipoprotein biosynthesis.

Methods for the study of hepatic lipoprotein synthesis and secretion have been described, and the advantages of each system discussed. Attention has been focused on intact cell systems. The isolated perfused liver constitutes a standard of comparison for all others, even though it, too, has limitations. Table IV below gives our assessment of some of the advantages and disadvantages of the techniques for studying hepatic lipoprotein biosynthesis.

Lipoproteins in experimental nephrosis: plasma levels and composition.

Experimental nephrosis was induced in rats by administration of puromycin aminonucleoside and the levels of plasma lipoproteins were examined 7 days later and compared to controls. As determined by density ultracentrifugation, VLDL, IDL, LDL, and HDL protein levels were increased by 8, 4, 5, and 5 times, respectively. These increases were accompanied by changes in lipid and apoprotein composition. The VLDL, IDL, and LDL fractions contained less triglyceride and more phospholipid and cholesterol, while HDL lipid composition was not altered. The apoprotein composition of VLDL and IDL were not measurably altered, but LDL contained less apoE. HDL had a markedly abnormal composition characterized by an almost complete absence of apoA-IV and apoE, increased apoA-1, and decreased apoC. While increased hepatic synthesis can account for much of the observed hyperlipoproteinemia in nephrosis, the changes in lipoprotein composition suggest impaired catabolism as a contributory factor.

Inhibition of apolipoprotein secretion and phosphatidate phosphohydrolase activity by eicosapentaenoic and docosahexaenoic acids in the perfused rat liver.

Infusion of albumin-bound eicosapentaenoic acid (EPA) docosahexaenoic acid (DHA), or oleic acid (OA) in perfused rat livers was carried out for two hours at a rate that maintained the perfusate concentration at 1 mmol/L. When compared with fatty acid-poor albumin alone, triacylglycerol (TAG) output was not significantly increased with DHA or EPA, whereas OA infusion resulted in a twofold increase. Incorporation of labeled leucine into VLDL apo B-100, apo B-48, apo E, and apo Cs was decreased by 50% by DHA or EPA compared with OA. The total phosphatidate phosphohydrolase activity was decreased by 35% with DHA or EPA compared to oleic acid or albumin alone. In no case was there a significant change in the distribution of activity between the microsomal and cytosolic factions. Fatty acid infusion did not significantly change the liver TAG content. Total liver lipids, microsomal lipids, and lipids of secreted VLDL were enriched with the infused fatty acids. The degree of enrichment for secreted TAG averaged 24% for OA and 36% for DHA or EPA. The effects of DHA and EPA on PPH activity and on apo B secretion in feeding experiments with marine oils rich in these acids may relate to changes in the fatty acid composition of liver membranes.

Isolation of nascent high-density lipoprotein from rat liver perfusates by immunoaffinity chromatography: effects of oleic acid infusion.

Immunoaffinity chromatography on a column of rabbit IgG anti-rat apolipoprotein (apo) A-I covalently bonded to agarose was used to isolate nascent high-density lipoprotein (nHDL) from recirculated perfusates of rat livers. After passage through the affinity column, the bound material was eluted with sodium thiocyanate and analyzed for apolipoproteins and lipids. The protein content was 52% and the lipid composition was 37% triglyceride, 40% phospholipid, and 23% cholesterol. Apolipoproteins E and A-I each comprised approximately one third of the total, and very little apo B was detectable as judged by SDS-PAGE analysis. The affinity-isolated particles were therefore similar in composition to the major apo A-I:apo E-rich subfraction of nHDL isolated by ultracentrifugation in earlier work. It is concluded that the apo E in this class of nHDL (containing both apo E and apo A-I) is present in the secreted particle and is not a consequence of a loss of apo E from very-low-density lipoprotein (VLDL) during ultracentrifugation. The high triglyceride content in the virtual absence of apo B confirms and extends previous analyses and reinforces the conclusion that nHDL particles are enriched in triglyceride compared to plasma HDL. The inclusion of 4% albumin in the perfusion medium did not significantly change the total triglyceride output of 115 micrograms/g liver/h, but it decreased the triglyceride output isolated by anti-apo A-I affinity chromatography from 3.2 to 0.48 micrograms/g liver/h. The addition of oleic acid complexed to albumin increased the total triglyceride output by 70% and that associated with the immunoaffinity column increased from 0.48 to 2.7 micrograms/g liver/h.(ABSTRACT TRUNCATED AT 250 WORDS)

The relationship in African-Americans of sex differences in insulin-mediated suppression of nonesterified fatty acids to sex differences in fasting triglyceride levels.

Insulin is a potent antilipolytic hormone that promotes the deposition of fat and decreases the release of nonesterified fatty acids (NEFA) from adipose tissue. The purpose of this study was to investigate in African-Americans (AAs) sex differences in insulin-mediated suppression of plasma NEFA and fasting triglyceride (TG) levels. Ninety AAs, 44 men and 46 women with a mean age of 34 +/- 8 years were classified by body mass index (BMI) into three groups: non-obese (22 men and 18 women), obese (12 men and 10 women), and severely obese (10 men and 18 women). In each BMI group, women versus men had greater percent body fat (non-obese, 30 +/- 6 v 18 +/- 6, P < .001; obese, 36 +/- 3 v 26 +/- 2, P < .001; and severely obese, 39 +/- 4 v 29 +/- 4, P < .001). An oral glucose tolerance test (OGTT) was performed with fasting TG levels and plasma insulin and NEFA concentrations obtained at 0, 30, 60, and 120 minutes. In women, insulin-mediated NEFA suppression was similar in each of the three BMI groups (non-obese, 85% +/- 14%; obese, 88% +/- 11%; and severely obese, 87% +/- 10%; P = .8). In men, the percent suppression of NEFA declined with increasing obesity (non-obese, 83% +/- 14%; obese, 71% +/- 21%; and severely obese, 68% +/- 16%; P = .04). Changes in NEFA suppression were reflected in the fasting TG levels. TG levels in women were similar in each BMI group (non-obese, 71 +/- 39 mg/dL; obese; 69 +/- 21; severely obese, 79 +/- 30; P = .7). In contrast, fasting TG levels for men were higher in the higher BMI groups. Plasma TG levels in men were 87 +/- 41 mg/dL for obese, 113 +/- 65 for obese, and 169 +/- 81 for severely obese (P = .001). These data demonstrate sex differences in insulin-mediated NEFA metabolism. In AA women, the maintenance of sensitivity to insulin-mediated suppression of NEFA regardless of the degree of obesity may contribute to the normal plasma TG levels. For AA men, the resistance to insulin-mediated suppression of NEFA in the higher BMI categories may allow more NEFA to be released from adipose tissue into the circulation and available to the liver for synthesis into TG-containing lipoproteins.

Gender differences in insulin-induced free fatty acid suppression: studies in an African American population.

This study of African Americans (AA) was designed to investigate gender differences in insulin-induced free fatty acid (FFA) suppression. Sixty AA (34 women, mean age 34 +/- 7.6 years, and 26 men, mean age 30 +/- 2.9 years) participated. All subjects had an oral glucose tolerance test (OGTT). Nineteen women and 18 men also underwent a euglycemic hyperinsulinemic clamp (IC) study. Plasma insulin and FFA concentrations were obtained during both tests at 0, 60, and 120 min. While there was no gender difference in body mass index (P = 0.21), women had greater percent body fat (P < 0.001) calculated by the Siri formula. There was no gender difference in fasting FFA levels, but during the OGTT, women compared to men had significantly greater FFA suppression. Both nonobese and obese women suppressed FFA concentration by 88%, and nonobese and obese men suppressed FFA concentration by 80 and 66% respectively. This gender difference in FFA suppression was significant (P = 0.001) and independent of obesity and insulin concentration. During the IC studies, there were no gender or obesity differences in FFA suppression, with women and men suppressing FFA levels by 87-89% (P = 0.7). Fasting insulin concentrations were higher in obese vs. nonobese (P = 0.03), but fasting FFA concentrations were not different (P = 0.15). For nonobese and obese females, fasting FFA levels were 0.55 +/- 0.24 and 0.44 +/- 0.26 mEq/L, respectively, and for nonobese and obese males, 0.45 +/- 0.2 and 0.35 +/- 0.18 mEq/L, respectively. In women, development of obesity may be enhanced by greater sensitivity to insulin-induced FFA suppression as measured during an OGTT. To detect gender differences in FFA metabolism, the OGTT is superior to the IC. The lack of elevation in fasting FFA levels in obese AA women and men has not been reported in other racial groups and may indicate a greater adipocyte sensitivity to insulin in AA.