Potent Lipolytic Activity of Lactoferrin in Mature Adipocytes

Momilactone A, a major rice diterpene phytoalexin, could be synthesized by dehydrogenation at the 3-position of 3β-hydroxy-9β-pimara-7,15-dien-19,6β-olide in rice leaves. The presence of 3β-hydroxy-9β-pimara-7,15-dien-19,6β-olide in UV-irradiated rice leaves was confirmed by comparing the mass spectra and retention times after a GC/MS analysis of the natural and synthetic compounds. The soluble protein fraction from UV-irradiated rice leaves showed dehydrogenase activity to convert 3β-hydroxy-9β-pimara-7,15-dien-19,6β-olide into momilactone A. The enzyme required NAD⁺ or NADP⁺ as a hydrogen acceptor. The optimum pH for the reaction was 8. The K _m value to 3β-hydroxy-9β-pimara-7,15-dien-19,6β-olide was 36 μM when NAD⁺ was supplied as a cofactor at a concentration of 1 mM. 3β-Hydroxy-9β-pimara-7,15-dien-19,6β-olide and its dehydrogenase activity were induced in a time-dependent manner by UV irradiation.     

Resistin Gene Expression in Human Adipocytes Is Not Related to Insulin Resistance

Objectives: Obesity is an important risk factor for the development of insulin resistance and type 2 diabetes. Recently, a newly described circulating hormone resistin, which is expressed primarily in adipocytes, has been shown to antagonize insulin action in mice. Resistin, therefore, has been suggested to play a role in the pathogenesis of insulin resistance. Research Methods and Procedures: We studied the expression of the resistin gene in primary cultured human adipocytes and preadipocytes. We also examined resistin gene expression in subcutaneous abdominal adipocytes in women (n = 24) over a wide range of body weight and insulin sensitivity. Results: Whereas resistin gene expression was barely detectable in mature adipocytes, it was highly expressed in preadipocytes. Adipogenic differentiation of preadipocytes was associated with a time-dependent down-regulation of resistin gene expression. There was no relationship between body weight, insulin sensitivity, or other metabolic parameters and adipocyte resistin gene expression in the clinical study. Discussion: Together these findings do not support an important role of adipose-tissue resistin gene expression in human insulin resistance.     

Gender Differences in Lipoprotein Lipase Activity after Acute Exercise

Objective: To determine whether gender differences exist in lipoprotein lipase (LPL) activity in response to exercise and/or insulin. Exercise and insulin are known modulators of LPL activity in men, but this is less clear in women. LPL activity may predict propensity for obesity; therefore, understanding its modulators is of considerable importance. Research Methods and Procedures: Gender differences in skeletal muscle and adipose tissue LPL activity were determined after a single bout of exercise followed by a hyperinsulinemic/euglycemic clamp and compared with an identical rest day in healthy lean men (n = 10) and women (n = 10). Muscle and adipose tissue biopsies were obtained pre‐ (post‐exercise vs. rest) and post‐clamp. Results: Basal levels of muscle and adipose tissue LPL activity were not different between men and women. There was, however, a significant gender by day interaction for muscle LPL activity (p = 0.023) and adipose tissue LPL activity (p = 0.013). In muscle, this was because of a significant increase in LPL activity on the exercise vs. rest day in men (p < 0.001) but not women. Adipose tissue LPL activity also increased significantly in men on the exercise day relative to rest day (p = 0.04) but decreased in women (p = 0.10). The hyperinulinemic/euglycemic clamp had no independent effect on tissue LPL activity, in either gender, after rest or exercise. Discussion: In the 3 to 4 hours after exercise, muscle and adipose tissue LPL activity increased significantly in men, whereas LPL activity remained unchanged in women.     

Comparison of the Lipolytic Effects of Norepinephrine and BRL 37344 in Rat Brown and White Adipocytes

The lipolytic effects of norepinephrine (a non-selective β-agonist) and BRL 37344 (a selective β₃-agonist) were compared in isolated rat brown and white adipocytes. Norepinephrine and BRL 37344 maximally stimulated lipolysis in brown and white adipocytes, approximately 10 times above basal values. However, adipocyte sensitivity for BRL 37344 was greater than that for norepinephrine, particularly in brown adipocytes [the EC₅₀ values (nM) for BRL 37344 and norepinephrine were 5 ± 1 and 103 ± 31 in brown adipocytes (P <0.01) versus 56 ± 9 and 124 ± 17 in white adipocytes (P <0.05), respectively]. On the other hand, the lipolytic effects of norepinephrine were totally blocked by 20–40 times superior concentrations of propranolol or bupranolol in brown as well as in white adipocytes. In contrast, the lipolytic effects of BRL 37344 were fully inhibited by concentrations of propranolol or bupranolol that were 200–1000 superior to the β₃ agonist concentration. The results demonstrate that: (1) the (β₃-agonist BRL 37344 is as effective as norepinephrine for maximally stimulating lipolysis in rat brown and white adipocytes, (2) both adipocyte types are more sensitive to the lipolytic effects of BRL 37344 than to those of norepinephrine, (3) although bupranolol is a better antagonist than propranolol on BRL 37344-stimulated lipolysis, it cannot be considered as a specific β₃-antagonist, (4) brown adipocytes are 10 times more sensitive than white adipocytes to the lipolytic effects of BRL 37344, suggesting an important role of β₃-receptors in brown adipose tissue.     

Adipocyte Differentiation‐related Protein in Human Skeletal Muscle: Relationship to Insulin Sensitivity

Objective: To determine whether adipocyte differentiation‐related protein (ADRP), a lipid droplet—associated protein that binds to and sequesters intracellular fatty acids, is 1) expressed in human skeletal muscle and 2) differentially regulated in human skeletal muscle obtained from obese non‐diabetic (OND) and obese diabetic (OD) subjects. Research Methods and Procedures: Ten OND subjects and 15 OD subjects underwent a weight loss or pharmacological intervention program to improve insulin sensitivity. Anthropometric data, hemoglobin A_1C, fasting glucose, lipids, and glucose disposal rate were determined at baseline and at completion of studies. Biopsies of the vastus lateralis muscle (SkM) were obtained in the fasting state from OND and OD subjects. Protein expression was determined by Western blotting. Results: ADRP was highly expressed in SkM from OND (4.4 ± 1.54 AU/10 μg, protein, n = 10) and OD (5.02 ± 1.33 AU/10 μg, n = 12) subjects. OND subjects undergoing weight loss had decreased triglyceride levels and improved insulin action. SkM ADRP content increased with weight loss from 5.14 ± 2.15 AU/10 μg to 9.92 ± 1.57 AU/10 μg (p < 0.025). OD subjects were treated with either troglitazone or metformin, together with glyburide, for 3 to 4 months. Both treatments attained similar levels of glycemic control. OD subjects with lower baseline ADRP content (2.85 ± 1.07 AU/10 μg, n = 6) displayed up‐regulation of ADRP expression (to 9.27 ± 2.76 AU/10 μg, p < 0.025). Discussion: ADRP is the predominant lipid droplet—associated protein in SkM, and low ADRP expression is up‐regulated in circumstances of improved glucose tolerance. Up‐regulation of ADRP may act to sequester fatty acids as triglycerides in discrete lipid droplets that could protect muscle from the detrimental effects of fatty acids on insulin action and glucose tolerance.     

Ethnic differences in body composition and other markers of cardiovascular disease risk: study in matched Haitian and White subjects from Quebec

Objectives: People of African descent may be at greater risk of metabolic syndrome (MS) compared with whites. We examined the associations among MS markers, body composition, and resting metabolic rate (RMR) in black Haitians and in white subjects living in Quebec, Canada. Research Methods and Procedures: Forty randomly selected Haitians were matched with 40 white subjects for age, sex, and BMI. Glycemic status and insulin resistance were assessed based on a 3‐hour glucose tolerance test. Blood lipids, blood pressure, abdominal fat (computed tomography), and waist circumference (WC) were measured. RMR was estimated by indirect calorimetry. Results: Triglycerides were significantly correlated with blood pressure only in Haitians and with the area under the curve for insulin only in whites. Haitians had significantly (p < 0.05) lower triglycerides and higher high‐density lipoprotein‐cholesterol concentrations but higher blood pressure than whites at any given WC value. General linear models showed that Haitians had less visceral adipose tissue than whites for the same WC. RMR was lower among Haitians for any given value of BMI or WC than in whites. Also, WC was more strongly associated with glucose area under the curve and to log‐homeostasis model assessment in white than in Haitian subjects. Discussion: The MS may be ethnospecific in its features and etiology. The standard anthropometric indices of obesity may not be as effective in populations of African descent compared with whites, unless appropriate cut‐off values are defined.     

Insulin stimulates turnover of phosphatidylcholine in rat adipocytes

The present study investigated the effect of insulin on phosphatidylcholine turnover in rat adipocytes labelled to equilibrium with [¹⁴C]-choline. Insulin induced a rapid turnover of this major phospholipid that was maximal by 1 min and transient in nature. Following a 1 min stimulation of the cells with insulin at a maximally effective concentration (7 nM), a 4–6% decrease in the percentage of total cellular choline associated with this phospholipid was observed. This reflected a significant transient increase in the percentage of total cellular choline associated with phosphorylcholine, which together with diacylglycerol are the phospholipase C cleavage products of phosphatidylcholine. These effects were observed over a physiological range of insulin concentrations. No effect of insulin on any other choline phospholipid or metabolite (sphingomyelin, lysophophatidylcholine, glycerophosphocholine or choline) was seen. These results suggest that insulin stimulates a phospholipase C-mediated turnover of phosphatidylcholine in rat adipocytes. The rapid nature of this turnover suggests a potential role in signal transduction.     

Racial Differences in Insulin Secretion and Sensitivity in Prepubertal Children: Role of Physical Fitness and Physical Activity

Objective: To investigate in prepubertal children whether physical fitness and/or physical activity are: 1) associated with insulin secretion and sensitivity and 2) account for racial differences in insulin secretion and sensitivity. Research Methods and Procedures: Subjects included 34 African American and 34 white nondiabetic children aged 5 to 11 years. Data were divided into two sets according to the availability of VO_2max and physical activity data. Body composition was measured by dual‐energy X‐ray absorptiometry. Subcutaneous abdominal adipose tissue and intra‐abdominal adipose tissue were examined by computed tomography. Insulin sensitivity (S_I) and acute insulin response (AIR) were determined by a frequently sampled intravenous glucose tolerance test. An all‐out, progressive treadmill exercise test was used for measuring VO_2max. Physical activity data were collected by questionnaire. Results: African American children had lower S_I and higher AIR than white children, after adjusting for total body fat mass. African Americans reported higher levels of physical activity (hours/wk) than whites, but had a lower VO_2max. In multiple linear regression analysis, hours/wk of activity and hours/wk of vigorous activity, but not moderate activity, were independently related to S_I and AIR after adjusting for race, total body fat mass or fat distribution, and total lean tissue mass. VO_2max was not related to AIR, and was inversely related to S_I, after adjusting for body composition. Race remained significantly associated with both S_I and AIR, even after adjusting for body composition, fat distribution, and hours/wk of activity or hours/wk of vigorous activity. Discussion: In summary, overall physical activity and, especially, vigorous activity were associated with insulin secretion and sensitivity. However, neither physical activity nor VO_2max explained the racial difference in insulin secretion (higher in African Americans) and sensitivity (lower in African Americans). Thus, racial (African American to white) differences in aspects of insulin action seem to be due to factors other than body composition, fat distribution, cardiovascular fitness, and amount of physical activity.     

胰岛素对3T3-L1脂肪细胞中极低密度脂蛋白受体基因表达的影响

体外培养3T3-L1细胞分化模型,研究不同浓度胰岛素及慢性胰岛素刺激对3T3-L1脂肪细胞中极低密度脂蛋白受体（VLDLR）基因表达的影响.在不同浓度胰岛素及胰岛素慢性刺激的干预下,用半定量RT-PCR检测细胞VLDLR mRNA水平的变化.微量化GOD-PAP法检测培养基中残存的葡萄糖.在细胞诱导分化过程中,胰岛素浓度的增高促进VLDLR的表达;胰岛素慢性刺激下,VLDLR表达因浓度差异呈现不同变化.研究结果表明,胰岛素的浓度及慢性刺激对3T3-L1脂肪细胞的成熟和VLDLR基因的表达有显著作用,而胰岛素抵抗明显减低成熟脂肪细胞VLDLR的表达.     

葡萄糖与胰岛素对3T3-F442A脂肪细胞中Leptin表达的调节

为了解葡萄糖与胰岛素对３Ｔ３Ｆ４４２Ａ脂肪细胞中Ｌｅｐｔｉｎ表达的调节,应用ＲＴＰＣＲ方法以ｂｅｔａａｃｔｉｎ为内对照对不同葡萄糖和／或胰岛素浓度培养条件下３Ｔ３Ｆ４４２Ａ脂肪细胞中ＬｅｐｔｉｎｍＲＮＡ表达水平进行相对定量分析。结果表明葡萄糖与胰岛素对３Ｔ３Ｆ４４２Ａ脂肪细胞中Ｌｅｐｔｉｎ表达有促进作用,过高浓度的葡萄糖抑制Ｌｅｐｔｉｎ的表达及胰岛素对Ｌｅｐｔｉｎ表达的促进。葡萄糖和胰岛素对Ｌｅｐｔｉｎ表达的促进作用无协同效应,且这种促进作用表现出饱和性特点。葡萄糖浓度的变化对脂肪细胞中Ｌｅｐｔｉｎ的表达与调控具有十分重要的影响。     

Effect of Changes in Fat Distribution on the Rates of Change of Insulin Response in Children

Objective: To develop mixed models for examining longitudinal associations between rates of change in visceral, subcutaneous abdominal, and total body fat with rates of change in fasting insulin (FI) and insulin sensitivity (SI) over 3 years in children. Research Methods and Procedures: Seventy-seven children (mean age, 8.3 years at baseline) from Birmingham, Alabama, with three or more annual measures of FI and SI were included. Abdominal fat was measured by computed tomography, and total body fat and lean tissue mass were measured by DXA. Mixed models examined the longitudinal associations between the baseline level/rate of change of different fat compartments and the rate of change in FI or SI. Results: An annual increase of ∼5% in FI was associated with 1 cm²/yr of visceral fat gain per year (p < 0.05), independent of subcutaneous abdominal fat. A 1-cm² difference in initial subcutaneous abdominal fat was associated with an ∼0.2% increase per year in FI (p < 0.02), independent of visceral fat. None of the rates of change in any of the fat measures was associated with the rate of change of SI. Discussion: The rate of change in visceral fat was positively associated with the rate of change in FI, independent of increasing subcutaneous abdominal fat; however, subcutaneous abdominal fat may be more predictive of the rate of change of FI than visceral or total fat. Therefore, growth-related increases in abdominal fat, particularly subcutaneous abdominal fat, may contribute to accelerating increases in FI, but have no effect on SI.     

Normal Insulin Sensitivity in Lean Offspring of Obese Parents

Objective: Offspring of diabetic or hypertensive patients are insulin resistant at a prediabetic/prehypertensive stage. We tested the hypothesis that insulin action may be impaired in the offspring of obese nondiabetic parents. Research Methods and Procedures: Twenty‐one lean offspring of nonobese subjects [(OL) 22 ± 3 years of age] were matched to 23 lean offspring of obese subjects (OOb) by gender distribution, age, BMI, and waist circumference. Anthropometry, oral glucose tolerance, in vivo insulin sensitivity [by a euglycemic insulin clamp (6 pmol/min per kilogram_FFM; where FFM represents fat‐free mass)], and thermogenesis (by indirect calorimetry) were measured in each subject. The study subjects were from a population of 267 nuclear families (one offspring and both his/her parents) in which there was statistically significant (χ² = 30.2, p = 0.001) concordance of BMI between parents and offspring. Results: In comparing OOb with OL, no statistically significant difference or trend toward a difference was detected in fasting plasma glucose and insulin concentrations, glucose and insulin responses to oral glucose, insulin sensitivity [metabolism value = 45 ± 12 (OOb) vs. 47 ± 17 μmol/min per kilogram_FFM (OL)], insulin‐induced inhibition of protein and lipid oxidation, stimulation of glucose oxidation and nonoxidative glucose disposal, respiratory quotient, resting energy expenditure, and glucose‐induced thermogenesis. Discussion: The metabolic similarity between lean offspring of obese parents and those of nonobese parents suggests that insulin resistance and its correlates are not co‐inherited with the predisposition to develop obesity.     

Polybrominated diphenyl ethers as endocrine disruptors of adipocyte metabolism

Objective: Obesity is thought to result from poor diet and insufficient exercise. An additional factor may be endocrine‐disrupting environmental chemicals that contaminate the air, water, and food supply. We tested the hypothesis that a class of lipid‐soluble flame retardant chemicals known to accumulate in adipose tissue, polybrominated diphenyl ethers (PBDEs), disrupts insulin and isoproterenol sensitivity of isolated rat adipocytes. Research Methods and Procedures: Six‐week‐old Sprague‐Dawley rats were gavaged daily with 14 mg/kg body weight (BW) pentabrominated diphenyl ether (penta‐BDE) in corn oil (n = 24) or corn oil alone (n = 24). At 2 and 4 weeks of treatment, epididymal fat pad adipocytes were isolated, and isoproterenol‐stimulated lipolysis, insulin‐stimulated glucose oxidation, and adipocyte size were measured. Results: There was no alteration in adipocyte metabolism after 2 weeks of in vivo penta‐BDE treatment, but after 4 weeks of treatment, adipocytes averaged a 30% increase in isoproterenol‐stimulated lipolysis and a 59% decrease in insulin‐stimulated glucose oxidation, compared with control. There were no differences in average rat BW and adipocyte size between treated and control rats, but plasma total thyroxine level in 2‐ and 4‐week treated rats was 30% of control. Discussion: Daily exposure of rats to 14 mg/kg BW penta‐BDE for 4 weeks has no effect on animal or adipocyte size but significantly alters insulin and isoproterenol‐stimulated metabolism of isolated adipocytes. These alterations, hallmark features of metabolic obesity, suggest the need for further research on the contribution of lipid‐soluble, endocrine‐disrupting environmental chemicals to the obesity epidemic.     

A Mouse Homolog of a Human TP53 Germline Mutation Reveals a Lipolytic Activity of p53

1. Download : Download high-res image (96KB)
2. Download : Download full-size image

     

Lipolytic response of adipocytes to epinephrine in sedentary and exercise-trained subjects: sex-related differences

Adipose tissue lipolytic activity is increased in endurance-trained subjects, but little is known about the mechanisms of this increase. To understand more fully the mechanisms involved and to discover whether sex-related differences exist, biopsies of fat were performed in the periumbilical region of 20 sedentary subjects (10 women (W) and 10 men (M)) and 20 trained subjects (10 W, 10 M); the in vitro response to epinephrine of the collagenase-isolated fat cells was studied. Glycerol release, chosen as an adipocyte lipolysis indicator, was measured by bioluminescence. Dose-response curves with epinephrine (alpha 2 and beta agonist), with isoproterenol (beta agonist) and epinephrine + propranolol and adenosine deaminase, were studied. Epinephrine-induced lipolysis was enhanced in trained subjects and this was due to an increased efficiency of the beta-adrenergic pathway. However, differences were found between the two sexes. In trained men, the lipolysis increase resulted from the enhancement of the beta-adrenergic pathway efficiency without any significant decrease in the alpha 2-adrenergic pathway efficiency. In trained women, the lipolysis increase was not only due to the enhancement of the beta-adrenergic pathway efficiency (which was greater than in trained men), but also to a significant decrease in the alpha 2-adrenergic pathway efficiency. Despite the decrease, the alpha 2-adrenergic pathway remained more efficient in trained women than in trained men, as was the case in sedentary subjects. It is concluded that endurance training led to better lipid mobilization and that this effect seemed greater in women than in men.     

Insulin Sensitivity as a Predictor of Weight Regain

A recent study found that increases in insulin sensitivity following weight loss and stabilization were strongly related to subsequent weight regain. The present paper analyzed this relationship in two behavioral weight-loss programs. In the first study, 125 nondiabetic subjects were followed over 30 months; weight losses averaged 10 kg at six months, and subjects had regained 8 kg of their weight loss by their 30-month follow-up. Neither fasting insulin levels at six months nor changes in fasting insulin from zero to six months were related to subsequent weight regain. Similarly, insulin levels measured two hours after a 75 g glucose load were unrelated to subsequent weight regain. The second study followed 33 individuals with Type II diabetes, treated with behavior modification, and either a low calorie diet or a very low calorie diet. Weight losses averaged 18 kg at six months, and subjects had regained 10 kg by their 24-month follow-up. The Bergman minimal model was used to assess insulin sensitivity at 6-month intervals. Initial analyses suggested that changes in insulin sensitivity from zero to six months were related to subsequent weight regain, but this effect was strongly influenced by an outlier. After removing this individual, there were no significant relationships between the changes in insulin sensitivity that accompanied weight loss and future weight regain. Likewise, insulin sensitivity at 12 months did not predict weight regain from 12 to 24 months. These data do not support the hypothesis that increases in insulin sensitivity with weight loss are associated with subsequent weight regain.     

Insulin resistance: Review of the underlying molecular mechanisms

Most human cells utilize glucose as the primary substrate, cellular uptake requiring insulin. Insulin signaling is therefore critical for these tissues. However, decrease in insulin sensitivity due to the disruption of various molecular pathways causes insulin resistance (IR). IR underpins many metabolic disorders such as type 2 diabetes and metabolic syndrome, impairments in insulin signaling disrupting entry of glucose into the adipocytes, and skeletal muscle cells. Although the exact underlying cause of IR has not been fully elucidated, a number of major mechanisms, including oxidative stress, inflammation, insulin receptor mutations, endoplasmic reticulum stress, and mitochondrial dysfunction have been suggested. In this review, we consider the role these cellular mechanisms play in the development of IR.     

组蛋白H3甲基转移酶Ezh2与小鼠脂肪细胞分化关系研究

目的:探索组蛋白H3K27me3甲基转移酶Ezh2对小鼠白色、棕色和米色脂肪细胞分化的影响。方法:构建诱导型Ezh2全身敲除小鼠(Ezh2~(flox/flox) CAGcre)并于6周龄时腹腔注射他莫昔芬诱导敲除,以同窝、同性别、相同基因型假诱导(腹腔注射玉米油)小鼠作为对照。诱导完成后在光镜下观察脂肪细胞形态,采用Western Blot法检测脂肪组织中H3K27me3、Ezh2和Ucp1的蛋白表达量。采用Realtime PCR法检测不同部位脂肪组织的脂肪分化相关基因(Pparγ、Adipoq和Fabp4)、棕色脂肪标志基因(Ucp1、Cidea和Prdm16)和米色脂肪标志基因(CD137、Tmem26和Tbx1)的表达。检测敲除组小鼠的冷耐受能力,并予以高脂饮食诱导肥胖,观察小鼠体重增长情况、诱导结束后小鼠的糖耐量和胰岛素敏感性指标。结果:Ezh2敲除小鼠Ezh2和H3K27me3的蛋白含量降低,背部棕色脂肪细胞脂滴明显小于对照组,Ucp1的基因和蛋白表达明显高于对照组(P0.05);敲除组小鼠白色脂肪细胞分化较差,米色脂肪分化增加,米色脂肪的Ucp1和Tbx1基因表达增加(P0.05)。敲除小鼠可以更好地耐受冷刺激,并抵抗高脂饮食诱导的肥胖和胰岛素抵抗。结论:Ezh2在体内促进白色脂肪细胞的分化,抑制棕色和米色脂肪细胞分化。