Effects of ovine growth hormone and other anterior pituitary hormones on lipolysis of rat and ovine adipose tissue in vitro

Ovine growth hormone ( oGH ) was tested for its effects on lipolysis of rat and ovine adipose tissue in vitro. Ovine growth hormone at 1, 5 and 25 micrograms/ml stimulated lipolysis (P less than .05) of chopped rat adipose tissue and isolated rat adipocytes incubated in the presence of 100 mU/ml adenosine deaminase and .2 micrograms/ml dexamethasone, but had no effect on lipolysis of chopped ovine adipose tissue or isolated ovine adipocytes. Isoproterenol, a beta-adrenergic agonist, stimulated lipolysis (P less than .05) of both rat and ovine adipose tissue. Contaminants of the oGH preparation used were examined for lipolytic effects. Thyroid-stimulating hormone (TSH), luteinizing hormone (LH) and adrenocorticotropic hormone (ACTH) content in oGH were measured by radioimmunoassay. When quantities of these hormones contaminating 5 and 25 micrograms oGH were tested for lipolysis in rat adipose tissue, the TSH contamination could account for some (30%) of the lipolysis observed with oGH , while the other hormones had no effect. Also, preincubation of oGH with anti-GH, but not with anti-TSH or anti-LH, removed the principle in oGH responsible for the lipolytic effect on rat adipose tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of in vitro ronnel on metabolic activity in subcutaneous adipose tissue and skeletal muscle from steers

Ronnel [0,0-dimethyl 0-(2,4,5-trichlorophenyl) phosphorothioate] is an organophosphate pesticide with growth-promoting properties. Experiments were conducted to determine effects of ronnel on oxidation of and fatty acid synthesis from acetate and glucose as indices of metabolic activity in subcutaneous adipose tissue and skeletal muscle from 6-, 12- and 18-mo-old steers. Ronnel depressed metabolic activity in adipose tissue from 6- and 12-mo-old steers without concomitantly decreasing metabolic activity in skeletal muscle. Production of CO2 and fatty acids from acetate and glucose in tissues from 18-mo-old steers was influenced less by ronnel than in tissues from younger steers. Interactions of ronnel with thyroxine or growth hormone on acetate oxidation and conversion to fatty acids in adipose tissue also were investigated. Thyroxine increased acetate oxidation and decreased fatty acid synthesis. Ronnel interfered with the metabolic effects of thyroxine. Growth hormone, with or without ronnel, did not affect metabolic activity of adipose tissue. Ronnel seemingly alters the partitioning of acetate and glucose between major metabolic processes in adipose tissue and skeletal muscle.     

Effects of isoprothiolane and phytosterol on lipogenesis and lipolysis in adipocytes from rats of dietary fat necrosis

To study effects of isoprothiolane and phytosterol on dietary fat necrosis, 3 groups of rats were fed hardened-tallow (HT) diet. Two groups of rats received either isoprothiolane (50 mg/kg) or phytosterol (20 mg/kg) orally once a day consecutively for 10 weeks. One group of rats received standard diet (CE-2) as a control. Fat necrotic lesions were observed in epididymal and perirenal adipose tissues from all rats in the 3 groups fed HT diet. Rats with fat necrosis were characterized by visceral type obesity and saturation in fatty acid composition of triglyceride in adipose tissue. The highest glucose conversion to total lipids was seen in adipocytes from the rats given phytosterol. There was no lipolytic response to epinephrine stimulation (1-100 microM) in adipocytes from the rats given only HT diet, while similar response of adipocytes from the 2 groups treated with either drug to those from the rats fed standard diet was observed. The levels of total saturated fatty acids of phospholipid in adipose tissue from the rats given either drug were lower than that of the rats given only HT diet. These data suggest that either drug alters fatty acid composition of phospholipid in fat cell membrane and enhances lipolysis of the cells.     

Comparison of plasma free-fatty-acid and blood-glycerol concentrations with measurement of lipolysis in porcine adipose tissue in vitro

Rates of adipose tissue lipid metabolism in vitro are often measured to evaluate the function in vivo of metabolic pathways and thus appraise the accretion or loss of depot fat. This study directly addressed the comparison of degradative metabolism in vitro and in vivo. The concentrations of plasma free-fatty-acids and blood-glycerol as putative representatives of lipolysis in vivo and the lipolytic rate in adipose tissue in vitro obtained at the time of blood sampling were both measured in the same pig. Concentrations of plasma free-fatty-acids and blood-glycerol were increased or decreased by infusion of the norepinephrine analog, isoproterenol or by infusion of the adrenergic antagonist, propranolol, respectively. Although lipolytic rates and sensitivity to isoproterenol in vitro changed during some acute hormonal manipulations of the pig, the modulation in vitro was usually small relative to the large changes observed in plasma free-fatty-acid and blood-glycerol concentrations. Some of the subtle changes in vitro may reflect biological responses to hormone infusion, e.g., desensitization of the response to adrenergic agonists, but the magnitude of rate changes in vitro negates prediction of the rates in vivo from rates in vitro. Extrapolation of lipolytic rates in vitro and several adipose tissue anabolic rates obtained from the literature indicate the improbability for prediction of rates and degree of fat accretion in pigs from metabolic rates in vitro.     

Size and lipolytic capacity of bovine adipocytes from subcutaneous and internal adipose tissue

Adipose tissue samples were taken from five dairy cows in early lactation. Various external and internal sites were chosen, and after isolation of the adipocytes, their size and lipolytic capacity were measured. It was found that cells from all the sites showed high lipolytic activity, although there was a four-fold difference between cells from perirenal fat and those from omental fat, which showed the highest activity. It was not possible to detect any relationship between diameter and lipolytic capacity of the adipocytes.     

Effects of glucagon and insulin on lipolysis and ketogenesis in sheep.

The hepatic and portal productions of acetoacetate and beta-hydroxybutyrate and lipolysis were studied in normal and insulin-controlled alloxan-diabetic sheep. Since hyperinsulinemia is associated with glucagon administration, the latter group of sheep were used to maintain constant plasma insulin levels. After control values were obtained glucagon was infused intraportally at 90 mug/hr for two hours. The ketone body production by portal drained viscera was not significantly affected by glucagon. In alloxanized sheep, glucagon significantly (P less than 0.01) increased net hepatic production of acetoacetate (from -0.54 +/- 0.08 to 0.46 +/- 0.07 g/hr). Lipolysis also increased. However, in the normal sheep, hyperinsulinemia prevented any stimulatory effect of glucagon on hepatic ketogenesis and lipolysis. Therefore, while glucagon appears capable of stimulating ketogenesis andlipolysis, these effects are readily suppressed by insulin.     

Effects of adrenergic agonists and insulin on porcine adipose tissue lipid metabolism in vitro

Because this laboratory has been able to demonstrate only a small and somewhat inconsistent stimulation of glucose metabolism by insulin in porcine adipose tissue in vitro, the tissue was preincubated with insulin to attempt to enhance the hormone effect. Preincubation with or without insulin did not increase insulin stimulation. Furthermore, insulin did not stimulate triacylglycerol biosynthesis. Adrenergic hormones stimulated lipolysis in porcine adipose tissue in vitro. Several analogs of norepinephrine incubated with porcine adipose tissue in vitro did not inhibit glucose incorporation into CO2 or total lipids, in contrast to inhibition observed in adipose tissue from other species. Isoproterenol inhibited glycerol-3-phosphate incorporation into lipids; the maximal inhibition was 50% for the initial stages of the pathway. Palmitate incorporation into lipids also was inhibited 50% by isoproterenol but this may have been an artifact. Preincubation of adipose tissue, with no exogenous hormone, might decrease the concentration of endogenous adrenergic hormones and thus make the tissue more responsive to exogenous adrenergic hormones. Preincubation of porcine adipose tissue did not consistently lower the basal lipolytic rate but enhanced the stimulated lipolytic rate; the mechanism is not known. These experiments provide no evidence that preincubation is beneficial to measurement of lipolysis or glucose metabolism in porcine adipose tissue in vitro.     

GH对猪脂肪组织和Leptin分泌的调节

一般认为生长激素 (Growthhormone,GH)对动物的生长的调节是通过其受体 (GH -R)介导产生胰岛素样生长因子 - 1 (Insulinlikegrowthfactor -1 ,IGF - 1 ) ,再以内分泌方式作用于靶器官。而GH对脂肪的作用似乎是直接的 ,与IGF - 1无关 ,因为GH在体内试验的结果均可在体外试验中重现 ,而经IGF - 1处理的猪胴体组成没有重现GH处理的效应 (Etherton和Bauman ,1 998;Kindt等 ,1 998)。然而有试验表明 ,GH处理使生长猪皮下脂肪组织IGF - 1基因表达显著增高 (Coleman等 ,1 994;Brameld等 ,1 996) ,去垂体大鼠经GH处理其白色脂肪组织I…     

Porcine leptin alters insulin inhibition of lipolysis in porcine adipocytes in vitro

The present study determined whether porcine leptin can alter the lipolytic rate in porcine adipocytes produced in vitro. The stromal-vascular cell fraction of neonatal subcutaneous adipose tissue was isolated by collagenase digestion, filtration, and subsequent centrifugation. These stromal-vascular cells were seeded on 25-cm2 tissue culture flasks and proliferated to confluency in 10% fetal bovine serum in DMEM/F12 (50:50). Cultures were differentiated using 2% pig serum + 10 mM isobutyl methylxanthine + 1 microM dexamethasone for 48 h. This medium was replaced with 5% pig serum + 1 microM insulin to promote lipid filling of adipocytes for 7 d. Adipocyte-containing cultures were incubated overnight in serum-free medium and then used for experiments. Acute experiments assessed lipolysis in cultures exposed to porcine leptin (0 to 1,000 ng/mL medium) for 2 h. Chronic experiments used cultures incubated with 100 ng porcine leptin/mL of medium for 72 h prior to lipolysis measurements. Direct effects of leptin were examined by incubating cultures in DMEM/F12, 25 mM HEPES, 3% bovine serum albumin, 20 mU of adenosine deaminase/mL of medium in the presence of 0 to 1,000 ng of porcine leptin/mL of medium. Indirect effects of leptin were examined using the same incubation medium but also supplemented with 1 microM isoproterenol +/- 10 nM insulin in the presence of 0 to 1,000 ng of porcine leptin/mL of medium. Media glycerol concentration was measured at the end of 2-h incubations. Acute leptin exposure induced up to a 76% increase in lipolysis (P < 0.05) but had no effect on insulin's inhibition of lipolysis. Chronic exposure to leptin produced up to a 56% increase in lipolysis (P < 0.05) and reduced insulin's inhibition ofisoproterenol-stimulated lipolysis by up to 31% (P < 0.05). These data demonstrate leptin functions to promote the partitioning of energy away from lipid accretion within porcine adipose tissue by promoting lipolysis directly and indirectly by reducing insulin-mediated inhibition of lipolysis.     

Pharmacokinetics of ceftiofur crystalline free acid after single subcutaneous administration in lactating and nonlactating domestic goats (Capra aegagrus hircus)

Doré, E., Angelos, J. A., Rowe, J. D., Carlson, J. L., Wetzlich, S. E., Kieu, H. T., Tell, L. A. Pharmacokinetics of ceftiofur crystalline free acid after single subcutaneous administration in lactating and nonlactating domestic goats (Capra aegagrus hircus). J. vet. Pharmacol. Therap. 34 , 25–30. Six nonlactating and six lactating adult female goats received a single subcutaneous injection of ceftiofur crystalline free acid (CCFA) at a dosage of 6.6 mg/kg. Blood samples were collected from the jugular vein before and at multiple time points after CCFA administration. Milk samples were collected twice daily. Concentrations of ceftiofur and desfuroylceftiofur‐related metabolites were measured using high‐performance liquid chromatography. Data were analyzed using compartmental and noncompartmental approaches. The pharmacokinetics of CCFA in the domestic goat was best described by a one compartment model. Mean (±SD) pharmacokinetic parameters were as follows for the nonlactating goats: area under the concentration time curve_0–∞ (159 h·μg/mL ± 19), maximum observed serum concentration (2.3 μg/mL ± 1.1), time of maximal observed serum concentration (26.7 h ± 16.5) and terminal elimination half life (36.9 h; harmonic). For the lactating goats, the pharmacokinetic parameters were as follows: area under the concentration time curve_0–∞ (156 h·μg/mL ± 14), maximum observed serum concentration (1.5 μg/mL ± 0.4), time of maximal observed serum concentration (46 h ± 15.9) and terminal elimination half life (37.3 h; harmonic). Ceftiofur and desfuroylceftiofur‐related metabolites were only detectable in one milk sample at 36 h following treatment. There were no significant differences in the pharmacokinetic parameter between the nonlactating and lactating goats.     

重组瘦蛋白对大鼠内分泌代谢的影响

利用重组瘦蛋白进行动物饲养试验研究。结果表明:重组瘦蛋白对大鼠内分泌代谢指标有一定影响。腹腔注射6μg/g体重,连续注射瘦蛋白7d后,大鼠血清胰岛素水平明显下降,与对照组相比,降低了15.44%(P<0.05),但对大鼠血清葡萄糖水平则没有显著影响;禁食可明显抑制大鼠生长激素的分泌,同时禁食大鼠的血清游离T4、T3水平与正常进食的对照组相比分别下降了17.39%和18.88%(P<0.05),与禁食大鼠相比,注射瘦蛋白大鼠的T4水平升高了17.11%(P<0.01),接近正常对照组水平,但给予瘦蛋白的大鼠的T3水平与对照组相比仅升高了8.25%(P<0.05),表明给予重组瘦蛋白可对禁食引起的血清生长激素和游离T4、T3水平的降低起到纠正作用。     

重组瘦蛋白对猪蛋白质代谢的影响

利用重组瘦蛋白进行动物饲养试验研究。结果表明:重组瘦蛋白对猪蛋白质代谢指标有不同程度的影响。腹腔注射0.5 mg/kg和1.0 mg/kg体重重组瘦蛋白后,血清总蛋白(TP)和球蛋白(GPs)的含量高于对照组,其中0~3周差异显著(P<0.05),而血清白蛋白(ALB)的含量只在注射后第3周有明显提高;血清天门冬氨酸转氨酶(AST)有不同程度的提高,在注射后1~3周和5~7周内明显高于对照组(P<0.05);血清碱性磷酸酶(ALP)和丙氨酸转氨酶(ALT)只有在注射0.5 mg/kg体重重组瘦蛋白后0~3周明显提高(P<0.05);注射0.1 mg/kg体重重组瘦蛋白对猪蛋白质代谢指标无明显影响(P>0.05)。     

Insulin responsiveness to glucose and tissue responsiveness to insulin in lactating, pregnant, and nonpregnant, nonlactating beef cows

Insulin responsiveness to glucose and tissue responsiveness to insulin, using the hyperglycemic clamp and the hyperinsulinemic euglycemic clamp techniques, were measured in lactating, late pregnant, and nonpregnant, nonlactating (NPNL) beef cows. The glucose infusion rate (GIR) in the hyperglycemic clamp technique was higher (P less than .05). in lactating cows than in NPNL cows. The plateau in plasma insulin concentration (insulin responsiveness) was higher (P less than .05) in lactating cows than in late pregnant and NPNL cows. Pregnant cows tended to have higher GIR and lower plateau in plasma insulin concentration than NPNL cows. In the hyperinsulinemic euglycemic clamp technique, GIR (tissue responsiveness to insulin) was higher (P less than .05) in lactating cows than in late pregnant cows; values for NPNL cows were intermediate. We conclude that insulin responsiveness to glucose and tissue responsiveness to insulin were enhanced during lactation but tended to be decreased during late pregnancy in beef cows.     

Pharmacokinetics of erythromycin in nonlactating and lactating goats after intravenous and intramuscular administration

The objectives of this work were to compare the pharmacokinetics of erythromycin administered by the intramuscular (i.m.) and intravenous (i.v.) routes between nonlactating and lactating goats and to determine the passage of the drug from blood into milk. Six nonpregnant, nonlactating and six lactating goats received erythromycin by the i.m. (15 mg/kg) and the i.v. (10 mg/kg) routes of administration. Milk and blood samples were collected at predetermined times. Erythromycin concentrations were determined by microbiological assay. Results are reported as mean +/- SD. Comparison of the pharmacokinetic profiles between nonlactating and lactating animals after i.v. administration indicated that significant differences were found in the mean body clearance (8.38 +/- 1.45 vs. 3.77 +/- 0.83 mL/kg x h respectively), mean residence time (0.96 +/- 0.20 vs. 3.18 +/- 1.32 h respectively), area under curve from 0 to 12 h (AUC(0-12)) (1.22 +/- 0.22 vs. 2.76 +/- 0.58 microg x h/mL respectively) and elimination half-life (1.41 +/- 1.20 vs. 3.32 +/- 1.34 h); however, only AUC(0-12) showed significant differences after the i.m. administration. Passage of erythromycin in milk was high (peak milk concentration/peak serum concentration, 2.06 +/- 0.36 and AUC(0-12milk)/AUC(0-12serum),6.9 +/- 1.05 and 2.37 +/- 0.61 after i.v. and i.m. administrations respectively). We, therefore, conclude that lactation affects erythromycin pharmacokinetics in goats.     

Evaluation of prostaglandin E2 as a regulator of lipolysis in bovine adipose tissue

Effects of exogenous prostaglandin E2 (PGE2) on rates of lipolysis in sections of subcutaneous adipose tissue biopsied from fed and fasted Holstein steers were determined. The interaction of PGE2 with several exogenous effectors of lipolysis and of the adenylate cyclase-cAMP system also was measured. Epinephrine increased basal (nonstimulated) lipolysis approximately one-fold. Prostaglandin E2 had no effect on either basal or epinephrine-stimulated lipolysis. Dibutyryl cAMP increased rate of lipolysis .4-fold, whereas theophylline increased lipolysis more than one-fold. Theophylline had an additive effect on epinephrine-stimulated lipolysis. Dibutyryl cAMP increased theophylline-stimulated lipolysis but not epinephrine-stimulated lipolysis. Prostaglandin E2 had no effect on epinephrine-, dibutyryl cAMP- or theophylline-stimulated lipolysis. Fasting decreased basal lipolysis by 40%. Furthermore, lipolysis in tissue incubated with PGE2, epinephrine or PGE2 plus epinephrine decreased from 30 to 50% upon fasting. As also shown with tissue from fed steers, PGE2 did not alter basal or epinephrine-stimulated lipolysis in tissue from fasted steers. Influences of exogenous effectors on lipolysis in adipose tissue from fed and fasted steers indicate that PGE2 does not control the adenylate cyclase-cAMP system that regulates lipolysis in bovine adipose tissue.