Net flux of nonesterified fatty acids, cholesterol, triacylglycerol, and glycerol across the portal-drained viscera and liver of pregnant ewes

The objective of this study was to determine the pattern of nutrient flux across portal-drained viscera (PDV) and liver in ewes with varying numbers of fetuses. Catheters were placed in the hepatic portal vein, a branch of the hepatic vein, a mesenteric vein, and the abdominal aorta of each ewe. Plasma flow and net cholesterol, nonesterified fatty acids (NEFA), and glycerol release across the PDV and liver were determined prior to exposure to rams. Ewes were subsequently mated. Two ewes were not pregnant, six ewes gave birth to singles, and 11 ewes gave birth to twins. Additional measurements were taken 103, 82, 61, 39, 19, and 6 d before parturition. There was a net PDV uptake of nonesterified cholesterol in the nonpregnant ewes and a net release in the ewes with singles and twins. Net nonesterified cholesterol hepatic release did not differ with days from parturition (P = .77). There was a net hepatic release of nonesterified cholesterol in the ewes with twins and a net hepatic uptake in the ewes with singles and in nonpregnant ewes (P = .03). There was a net PDV release of NEFA; however, it did not differ with litter size (P = .59) or days from parturition (P = .63). Hepatic NEFA uptake increased with litter size (P = .03) and increased as gestation progressed (P = .006). There was an interaction (P = .04) between litter size and days from parturition for net PDV glycerol release. Net PDV glycerol release in the nonpregnant ewes decreased over time, but release in pregnant ewes tended to increase over time. Hepatic glycerol uptake increased with litter size and increased as gestation progressed. There was a net PDV uptake of triacylglycerol, but it did not differ with litter size (P = .11) or with days from parturition (P = .06). There was a net hepatic release of triacylglycerol, but it did not differ with litter size (P = .59) or with days from parturition (P = .67). Liver utilization of glycerol and NEFA as substrates for metabolism increases as pregnancy progresses. In the nonpregnant ewe, the combined contribution of glycerol and NEFA carbon accounted for 10% of the carbon taken up by the liver, and in ewes pregnant with twins, the combined contribution accounted for 42% of the carbon uptake 19 d before parturition. In conclusion, these data demonstrate NEFA are an important metabolite when determining carbon balance across the liver and their relative contribution to carbon balance increases as pregnancy progresses.     

Hepatic triacylglycerol synthesis during a period of fatty liver development in sheep

Fatty liver was induced in six sheep by fasting and treatment with phloridzin and epinephrine. Treatment was associated with a rise in the concentration in serum and hepatic uptake of nonesterified fatty acid (NEFA) compared with pretreatment (P less than .01). At 24 h after the start of the treatment, concentrations of serum lipoprotein in each density class were not different from baseline, but all were elevated (P less than .01) by 120 h of treatment. Hepatic triacylglycerol (TG) concentration increased (P less than .01) 17-fold within the first 48 h of treatment, with no additional increase during the remaining 72 h. The activity of phosphatidate phosphohydrolase (PAP) in hepatic microsomes increased (P less than .01) fourfold over baseline by 48 h of treatment, then declined slightly by 120 h. The activities of diacylglycerol acyltransferase (DGAT) and glycerolphosphate acyltransferase (GPAT) in hepatic microsomes increased during treatment but appeared to follow a slightly different pattern from that of PAP. Activity of GPAT was not above baseline at 48 h, but was at 120 h (P less than .05); DGAT activity was increased (P less than .05) twofold at 48 h, with an apparent continued increase (P less than .01) to threefold over baseline by 120 h of treatment. Fatty liver appeared to develop during a period of rapid hepatic uptake of NEFA without a corresponding increase in serum lipoprotein concentrations. The activities of PAP, GPAT and DGAT, putative regulators of TG synthesis rate, all increased in liver microsomes during a period of high hepatic NEFA uptake, but that of PAP appeared to coincide most closely with the development of fatty liver.     

Regulation of hepatic triacylglycerol synthesis and secretion

Triacylglycerols are the most concentrated storage form of energy for the mammalian organism. These lipids are synthesized and secreted by the liver and serve as a fuel for other tissues. This paper presents a brief review of the regulation of hepatic triacylglycerol synthesis and secretion. Particular attention will be given to the dissociation of the synthesis of triacylglycerols from that of the metabolically closely related nitrogenous phospholipids. Recent evidence is presented which suggests that triacylglycerol synthesis (and secretion) is regulated, at least partially, at the diacylglycerol branchpoint.     

Regulation of hepatic triacylglycerol synthesis and secretion

Triacylglycerols are the most concentrated storage form of energy for the mammalian organism. These lipids are synthesized and secreted by the liver and serve as a fuel for other tissues. This paper presents a brief review of the regulation of hepatic triacylglycerol synthesis and secretion. Particular attention will be given to the dissociation of the synthesis of triacylglycerols from that of the metabolically closely related nitrogenous phospholipids. Recent evidence is presented which suggests that triacylglycerol synthesis (and secretion) is regulated, at least partially, at the diacylglycerol branchpoint.     

Effects of nonesterified fatty acids and beta-hydroxybutyrate on functions of mononuclear cells obtained from ewes

OBJECTIVE: To assess the effects of nonesterified fatty acids (NEFA) and beta-hydroxybutyrate (BHBA) on functions of mononuclear cells obtained from ewes. ANIMALS: 6 Sardinian ewes. PROCEDURE: Mononuclear cells were cultured with concentrations of NEFA (0, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, or 2,000 micromol/L) and BHBA (0, 0.45, 0.9, 1.8, or 3.6 mmol/L). Concentrations of NEFA and BHBA were intended to mimic those of ketotic or healthy ewes, and NEFA and BHBA were tested alone and in combination. Synthesis of DNA was stimulated by use of concanavalin A (Con A) or pokeweed-mitogen (PWM). Secretion of IgM was stimulated by use of PWM. RESULTS: Synthesis of DNA stimulated by Con A and PWM was significantly inhibited by high concentrations of NEFA (> or = 250 micromol/L) or by a combination of high concentrations of NEFA (> or = 250 micromol/L) and all concentrations of BHBA (> or = 0.45 mmol/L). In contrast, DNA synthesis was not inhibited by low concentrations of NEFA (< or = 125 micromol/L) or by a combination of low concentrations of NEFA (< or = 125 micromol/L) and the lowest concentration of BHBA (0.45 mmol/L). Secretion of IgM was significantly inhibited by all concentrations of NEFA and by all combinations of NEFA and BHBA concentrations. When used alone, none of the concentrations of BHBA inhibited DNA synthesis or IgM secretion. CONCLUSIONS AND CLINICAL RELEVANCE: Reduced immunoresponsiveness during ketosis is likely to be associated with an increase in plasma concentration of NEFA and not with an increase in plasma concentration of BH BA.     

Paradoxical increases of circulating nonesterified fatty acids in somatotropin treated cattle undergoing mild disturbances

Effects of various doses of bovine somatotropin (bST) on plasma concentrations of nonesterified fatty acids (NEFA) were studied in a 14-d Latin square with six Holstein heifers. Animals were given daily injections of excipient or bST at 12:00 p.m. and fed twice daily at 7:00 a.m. and 7:00 p.m. On Day 14, plasma NEFA remained low through the day except around the 7:00 p.m. feeding when they were substantially elevated. The elevation was significantly greater in bST-treated animals and corresponded to the excitement of the animals in anticipation of the evening feeding. To further investigate this phenomenon, a second experiment was conducted in which nine growing Holstein steers were fed hourly and received either daily intramuscular (i.m.) injection of excipient or bST (120 mg/kg BW) for 15 d in a crossover design. Daily profiles of NEFA were obtained under undisturbed conditions or concurrently with intensive handling. Although no elevations could be detected in any case in control animals, bST caused a substantial rise in NEFA concentration only when animals were subjected to intensive handling. This suggested that NEFA peaks noted in bST-treated heifers in the first experiment resulted from increased ability of adipose tissue to respond to adrenergic stimulation associated with the anticipation of feeding. Consistent with this hypothesis, plasma NEFA concentrations in bST-treated steers were increased to a greater extent during a challenge involving i.v. injection of epinephrine. This amplification of adipose tissue response by bST must be considered when conducting intensive studies. Even the minimal excitement associated with blood sampling can confound the results regarding lipid mobilization, and this may have contributed to the notion that ST is a lipolytic hormone.     

金属硫蛋白对应激猪血液和肌肉中非酯化脂肪酸及肌乳酸含量的影响

采用驱赶法使乳猪产生应激,探讨金属硫蛋白对应激猪血清与肌肉中非酯化脂肪酸及肌乳酸含量的影响.结果表明:在应激状态下,金属硫蛋白可使血清和运动初期肌肉中的非酯化脂肪酸含量升高,肌乳酸的生成降低,因而可增强有氧代谢,改善机体对运动负荷的压力,提高动物的抗应激能力,因此金属硫蛋白对乳猪具有一定的应激保护作用.     

Nutritional anestrus in beef cows: concentrations of glucose and nonesterified fatty acids in plasma and insulin in serum

Twenty-two nonlactating Hereford cows exhibiting normal estrous cycles were fed either maintenance (M) or restricted (R) diets until most of the R cows became anestrus; R cows then were fed 160% of the M diet until estrous cycles resumed. Concentrations of progesterone, glucose, insulin and nonesterified fatty acids (NEFA) were determined in weekly blood samples. Blood also was collected frequently, before and after i.v. infusion of 300 ml of a 40% glucose solution, to evaluate responses in blood concentrations of glucose and insulin when cows were exhibiting normal estrous cycles, when R cows were initiating anestrus, during anestrus, and at the reinitiation of estrous cycles. Losses in BW and body condition score in R cows were associated with reduced (P less than .01) concentrations of glucose and insulin and greater (P less than .01) concentrations of NEFA in blood plasma compared with those of M cows. During normal estrous cycles, disappearance of infused glucose from plasma and concentrations of insulin in serum were similar for R and M cows. Glucose disappearance from plasma was retarded and serum concentrations of insulin remained increased for a longer time after glucose infusion in R at the start of anestrus compared with M cows (P less than .01). Similarly, during anestrus, the rate of glucose disappearance was slower for R cows (P less than .01). During refeeding of R cows, disappearance of infused glucose was similar for R and M cows. In conclusion, reduced concentrations of glucose and insulin and increased concentrations of NEFA in blood were associated with nutritional anestrus and the glucoregulatory effects of insulin were compromised during nutritional anestrus.     

Effect of different exogenous fatty acids on the cytosolic triacylglycerol content in bovine mammary cells

The objective of this study was to determine how cytosolic triacylglycerols (TAG) are stored in mammary cells and whether this depends on the individual chemical configuration of fatty acids (FA). This objective was accomplished by addition of different FA to a FA-free medium used to culture mammary alveolar cells-large T antigen cells (MAC-T). Treatments consisted of adding FA (palmitate, stearate, oleate, linoleate, rumenic acid [CLA], elaidate and vaccinate) solutions to the medium at 100, 200, 300 and 400 mmol/L concentrations for a 24-h incubation period. At the end of each incubation period, cytosolic TAG, DNA and protein contents were measured. Palmitate, vaccenate, linoleate and CLA increased (P < 0.05) cytosolic TAG (μg/mg protein). Palmitate and CLA increased (P < 0.05) cytosolic TAG adjusted for DNA content. Overall, effects on cytosolic TAG accumulation depended on individual FA structure (chain length, degree of saturation, and number and orientation of FA double bonds). In addition, the long-chain FA used in this study did not have a detrimental effect on MAC-T cells as indicated by cytosolic protein and DNA contents reflecting their biological role in lipid accumulation.     

Dietary (n-3) fatty acids from menhaden fish oil alter plasma fatty acids and leukotriene B synthesis in healthy horses

The study objective was to determine the effect of feeding corn oil or fish oil to horses on plasma fatty acid profiles and leukotriene B (LTB) synthesis by stimulated peripheral blood neutrophils. Two groups of horses (n = 5) were randomly assigned to diets supplemented with either 3.0% (by weight) corn oil or fish oil for a period of 14 weeks. The ratio of (n-6) to (n-3) fatty acids in oil supplements was 68.1:1 for corn oil and 0.12:1 for fish oil. Production of LTB4 and LTB, by peripheral blood neutrophils stimulated with calcium ionophore A23187 and plasma cholesterol, triacylglycerol, and alpha-tocopherol concentrations were measured. At 12 weeks, horses fed fish oil had increased plasma concentrations of eicosapentaenoic acid (27-fold; 8.5 versus 0.3 g/100 g fatty acids; P < .0001), docosahexaenoic acid (34-fold; 5.1 versus 0.1 g/100 g fatty acids; P < .0001), and arachidonic acid (8.3-fold; 4.1 versus 0.5 g/100 g fatty acids; P < .0001) compared with horses fed corn oil. Neutrophils from horses fed fish oil produced 78-fold (P = .01) more LTB5 and 9.5-fold (P = .003) more LTB4 compared with predietary levels, and 17.6-fold (P = .01) and 3.3-fold (P = .02), respectively, more than horses fed corn oil, and the ratio of LTB5 to LTB4 concentrations was 4.0-fold (P = .002) higher in horses fed fish oil. This study suggests that dietary polyunsaturated fatty acids modulate the leukotriene inflammatory response of horses. If the ratio of LTB5 to LTB4 concentrations is important in determining how inflammatory processes are mediated, then fish oil supplementation may have value in treatment of equine inflammatory diseases.     

Detection of nonesterified (free) fatty acids in bovine serum: comparative evaluation of two methods.

A method comparison study for the determination and quantitation of nonesterified fatty acids (NEFAs) in serum, using the commercial "NEFA C" enzymatic test kit, was performed using the spectrophotometric method recommended by the manufacturer and a modified procedure optimized for the use of a microplate reader, a 96-well microtiter plate, and small sample volumes (10 microl). Linearity, sensitivity, and precision using the test kit were determined for each method of detection. The assay was linear from 0 to 1.97 mEq/liter for both procedures, and the limits of detection were determined to be 0.22 (+/- 0.074) and 0.05 (+/- 0.002) mEq/liter for the spectrophotometer and microplate reader, respectively. Pairs of measurements for bovine serum samples were compared and evaluated by a mean difference plot method and not regression analysis, a method that has been shown to be inappropriate for method comparison studies. The difference plot was used to evaluate the systematic bias between the 2 methods. Random error is reported on the basis of SD differences, and "limits of agreement" are used to describe the maximum differences likely to occur between the 2 methods. Results suggest that the microplate reader method can be used reliably in place of the recommended spectrophotometric method. The microplate reader method is preferred because of its high throughput capabilities, simultaneous analysis of all the standards and samples, use of small sample and reagent volumes, and reduction in labor requirements and costs.     

Effect of withholding feed on concentration and composition of plasma very low density lipoprotein and serum nonesterified fatty acids in horses

OBJECTIVE: To measure and compare the concentration and composition of very low-density lipoprotein (VLDL) in plasma and selected lipids in serum of horses fed mixed grass hay ad libitum or denied feed for 36 hours. ANIMALS: 4 healthy adult mares. PROCEDURE: Mares were either fed mixed grass hay ad libitum or denied feed for 36 hours beginning at 8:00 AM. Blood samples were collected every 2 hours during the study period and analyzed for nonesterified fatty acid (NEFA), triglyceride (TG), VLDL, and glucose concentrations and composition of VLDL. RESULTS: Withholding feed significantly increased mean serum concentrations of NEFA. By 36 hours, a 16-fold increase in mean serum NEFA concentration and 2-fold increase in mean plasma VLDL concentration, compared with baseline values, were detected. Mean plasma TG concentrations significantly increased with time in feed-deprived horses. Significantly lower overall mean plasma glucose concentrations were detected in feed-deprived horses. Mean percentage of protein in VLDL was significantly lower in feed-deprived horses. Plasma VLDL concentrations varied widely among horses in response to withholding feed. Plasma TG and VLDL concentrations remained unaltered in 2 horses. CONCLUSIONS AND CLINICAL RELEVANCE: Withholding feed significantly increases blood lipid concentrations in horses, but individual horses respond differently. Serum NEFA concentrations were increased in all 4 horses denied feed, indicating mobilization of tissue triglyceride stores. Variation in plasma VLDL concentration in response to withholding feed suggests that its metabolism is strongly influenced by other, as yet undetermined, factors in horses. Differences in the plasma VLDL concentrations among horses in response to withholding feed may be used as an indication of susceptibility to the hyperlipemic syndrome of Equidae.     

Spectrum of nonesterified fatty acids in blood and muscle tissue of swine. 3. Effects of target-oriented deflection of hormonal control circuits

Gas chromatography was used to study both the relative composition and level of nonesterified fatty acids in the blood of pigs, following target-oriented deflection of two hormonal systems, hypothalamus-pituitary gland-thyroid and hypothalamus-pituitary gland-adrenal cortex. The findings originally expected were not made. Percentual C-18 was increased, following long-time application of tri-iodothyronine/thyroxine, while c-12 was percentually lowered, as compared to the perchlorate group. Deflection of the hypothalamus-pituitary gland-adrenocortical system caused slight alteration in both the composition and overall concentration of blood-borne non-esterified fatty acids.     

Links between de novo fatty acid synthesis and leptin secretion in bovine adipocytes

Leptin secretion by adipose tissue is involved in many physiological control systems, including those that determine growth, development, body composition, milk production, and reproductive function. In the adipocyte of monogastric animals, malonyl CoA (coenzyme A) seems to link the flux of energy substrates to the control of leptin production. In this study, we tested this for ruminants by examining the effect of cerulenin, an inhibitor of de novo fatty acid synthesis at the step from malonyl CoA to palmitate, on leptin production by cultured bovine adipocytes derived from intermuscular fat. Purified preadipocytes were obtained by the ceiling culture method, and adipogenic media were used to induce their differentiation into adipocytes. We found that leptin concentrations increased significantly with time in culture, and with increases in glucose concentration. Addition of 2-deoxy-D-glucose to the medium, a competitive inhibitor of glucose transport and metabolism, suppressed leptin secretion. In media with high glucose concentrations, cerulenin enhanced leptin secretion. We conclude that, as in monogastrics, malonyl CoA may play a key role in the control of leptin secretion in ruminants.