p38 MAPK regulates calcium signal-mediated lipid accumulation through changing VDR expression in primary preadipocytes of mice

In the present study we have examined whether p38 mitogen activated protein kinase (p38 MAPK) signal pathway interacts with calcium signal on lipid accumulation in primary preadipocytes of mice. The primary preadipocytes were treated with p38 MAPK inhibitor SB203580, blockers and excitomotors of calcium channel for 24 h, respectively. Intracellular triglyceride (TG) content was measured by triglyceride kit and lipid accumulation was determined by Oil Red O staining. Meanwhile, the mRNA expressions of peroxisome proliferators-activated receptor gamma (PPARγ) gene, fatty acid synthetase (FAS) gene, lipoprotein lipase (LPL) gene, vitamin D receptor (VDR) gene and extracellular Ca²⁺-sensing receptor (CaSR) gene were analyzed with real-time PCR. The protein content and phosphorylation of VDR and p38 were tested with Western Blotting. The data showed that intracellular TG content and the mRNA expression levels of PPARγ, FAS, LPL in N group and L group as well as FAS, LPL in C group were increased significantly (P < 0.01) compared to the control. On the contrary, intracellular TG content and the mRNA expression levels of PPARγ, FAS in B group as well as intracellular TG content and PPARγ, FAS, LPL in SB group and B+SB group were decreased significantly (P < 0.01). VDR mRNA expression and protein content were decreased in B, C, and SB added groups (P < 0.01). In addition, p38 phosphorylation levels increased in N and L groups (P < 0.01) and decreased in SB added groups (P < 0.01). These findings suggest that p38 MAPK pathway through regulating VDR mRNA expression participates in mediation of calcium signal and affects calcium signal regulating lipid accumulation in mice preadipocytes through changing PPARγ, FAS and LPL mRNA expression. In addition, calcium signal have a feedback effect in phosphorylation of p38.     

The contrasting roles of PPARδ and PPARγ in regulating the metabolic switch between oxidation and storage of fats in white adipose tissue

Background  

The nuclear receptors peroxisome proliferator-activated receptor γ (PPARγ) and peroxisome proliferator-activated receptor δ (PPARδ) play central roles in regulating metabolism in adipose tissue, as well as being targets for the treatment of insulin resistance. While the role of PPARγ in regulating insulin sensitivity has been well defined, research into PPARδ has been limited until recently due to a scarcity of selective PPARδ agonists.     

The Adipose Tissue Phenotype of Hormone‐Sensitive Lipase Deficiency in Mice

Objective: To directly ascertain the physiological roles in adipocytes of hormone‐sensitive lipase (HSL; E.C. 3.1.1.3), a multifunctional hydrolase that can mediate triacylglycerol cleavage in adipocytes. Research Methods and Procedures: We performed constitutive gene targeting of the mouse HSL gene (Lipe), subsequently studied the adipose tissue phenotype clinically and histologically, and measured lipolysis in isolated adipocytes. Results: Homozygous HSL^?/? mice have no detectable HSL peptide or cholesteryl esterase activity in adipose tissue, and heterozygous mice have intermediate levels with respect to wild‐type and deficient littermates. HSL‐deficient mice have normal body weight but reduced abdominal fat mass compared with normal littermates. Histologically, both white and brown adipose tissues in HSL^?/? mice show marked heterogeneity in cell size, with markedly enlarged adipocytes juxtaposed to cells of normal morphology. In isolated HSL^?/? adipocytes, lipolysis is not significantly increased by β₃‐adrenergic stimulation, but under basal conditions in the absence of added catecholamines, the lipolytic rate of isolated HSL^?/? adipocytes is at least as high as that of cells from normal controls. Cold tolerance during a 48‐hour period at 4 °C was similar in HSL^?/? mice and controls. Overnight fasting was well‐tolerated clinically by HSL^?/? mice, but after fasting, liver triglyceride content was significantly lower in HSL^?/? mice compared with wild‐type controls. Conclusions: In isolated fat cells, the lipolytic rate after β‐adrenergic stimulation is mainly dependent on HSL. However, the observation of a normal rate of lipolysis in unstimulated HSL^?/? adipocytes suggests that HSL‐independent lipolytic pathway(s) exist in fat. Physiologically, HSL deficiency in mice has a modest effect under normal fed conditions and is compatible with normal maintenance of core body temperature during cold stress. However, the lipolytic response to overnight fasting is subnormal.     

Obesity Is Accompanied by Disturbances in Peripheral Glucocorticoid Metabolism and Changes in FA Recycling

The glucocorticoid activating enzyme 11β‐hydroxysteroid dehydrogenase type 1 (11βHSD1) is of major interest in obesity‐related morbidity. Alterations in tissue‐specific cortisol levels may influence lipogenetic and gluco/glyceroneogenetic pathways in fat and liver. We analyzed the expression and activity of 11βHSD1 as well as the expression of phosphoenolpyruvate carboxykinase (PEPCK), sterol regulatory element binding protein (SREBP), and fatty acid synthase (FAS) in adipose and liver and investigated putative associations between 11βHSD1 and energy metabolism genes. A total of 33 obese women (mean BMI 44.6) undergoing gastric bypass surgery were enrolled. Subcutaneous adipose tissue (SAT), omental fat (omental adipose tissue (OmAT)), and liver biopsies were collected during the surgery. 11βHSD1 gene expression was higher in SAT vs. OmAT (P = 0.013), whereas the activity was higher in OmAT (P = 0.009). The SAT 11βHSD1 correlated with waist circumference (P = 0.045) and was an independent predictor for the OmAT area in a linear regression model. Energy metabolism genes had AT depot–specific expression; higher leptin and SREBP in SAT than OmAT, but higher PEPCK in OmAT than SAT. The expression of 11βHSD1 correlated with PEPCK in both AT depots (P = 0.05 for SAT and P = 0.0001 for OmAT). Hepatic 11βHSD1 activity correlated negatively with abdominal adipose area (P = 0.002) and expression positively with PEPCK (P = 0.003). In human obesity, glucocorticoid regeneration in the SAT is associated with central fat accumulation indicating that the importance of this specific fat depot is underestimated. Central fat accumulation is negatively associated with hepatic 11βHSD1 activity. A disturbance in peripheral glucocorticoid metabolism is associated with changes in genes involved in fatty acid (FA) recycling in adipose tissue (AT).     

Cloning,structural characterization and expression analysis of a novel lipid storage droplet protein-1 (LSD-1) gene in Chinese honeybee (<Emphasis Type="Italic">Apis cerana cerana</Emphasis>)

Lipid storage droplet 1 (LSD-1), a PAT family protein located around lipid droplets in insects, is intimately linked to lipid droplets formation and lipid metabolism. Conjugated linoleic acid (CLA) and rosiglitazone (Rosi) have previously been shown to modulate the expression of several PAT family proteins through peroxisome proliferator-activated receptor-γ (PPARγ). In the present study, we isolated and characterized a novel LSD-1 gene, referred to AccLSD-1, from Chinese honeybee (Apis cerana cerana). Sequence analysis indicated that the central region of LSD-1 protein had significant sequence similarity and a typical LSD-1 gene was composed of 8 exons and 7 introns. Interestingly, the first intron of AccLSD-1 including several PPARγ-response elements (PPREs) was located in 5′ UTR. Analysis of 5′-flanking region of AccLSD-1 revealed a number of putative cis-acting elements, including three PPREs. Quantitative real-time PCR showed that AccLSD-1 expressed ubiquitously from feeding larva to adult, and its expression level was highest at brown-eyed pupae (Pb) stage. The effect of CLA, Rosi and combination on AccLSD-1 expressions indicated 1% CLA and 0.5 mg/ml Rosi were considered as the suitable diets for rearing adult workers in laboratory, and AccLSD-1 was down-regulated by CLA whereas up-regulated by Rosi. Furthermore, the combination of CLA and Rosi remarkly rescued the suppression of AccLSD-1 expression by CLA alone. These results suggest that AccLSD-1 is associated with A. cerana cerana development, especially during pupal metamorphosis, and can be regulated by CLA or Rosi possibly via activating PPARγ.     

Depot-Specific Changes in Fat Metabolism with Aging in a Type 2 Diabetic Animal Model

Visceral fat accretion is a hallmark of aging and is associated with aging-induced metabolic dysfunction. PPARγ agonist was reported to improve insulin sensitivity by redistributing fat from visceral fat to subcutaneous fat. The purpose of this study was to investigate the underlying mechanisms by which aging affects adipose tissue remodeling in a type 2 diabetic animal model and through which PPARγ activation modulates aging-related fat tissue distribution. At the ages of 21, 31 and 43 weeks, OLETF rats as an animal model of type 2 diabetes were evaluated for aging-related effects on adipose tissue metabolism in subcutaneous and visceral fat depots. During aging, the ratio of visceral fat weight to subcutaneous fat weight (V/S ratio) increased. Aging significantly increased the mRNA expression of genes involved in lipogenesis such as lipoprotein lipase, fatty acid binding protein aP2, lipin 1, and diacylglycerol acyltransferase 1, which were more prominent in visceral fat than subcutaneous fat. The mRNA expression of adipose triglyceride lipase, which is involved in basal lipolysis and fatty acid recycling, was also increased, more in visceral fat compared to subcutaneous fat during aging. The mRNA levels of the genes associated with lipid oxidation were increased, whereas the mRNA levels of genes associated with energy expenditure showed no significant change during aging. PPARγ agonist treatment in OLETF rats resulted in fat redistribution with a decreasing V/S ratio and improved glucose intolerance. The genes involved in lipogenesis decreased in visceral fat of the PPARγ agonist-treated rats. During aging, fat distribution was changed by stimulating lipid uptake and esterification in visceral fat rather than subcutaneous fat, and by altering the lipid oxidation.     

De novo lipogenesis and lipolysis activities in normal (Dw) and dwarf (dw) White Leghorn laying hens

1. In vitro activities of glucose oxidation, de novo lipogenesis and lipolysis were compared in normal (Dw) and dwarf (dw) laying hens. 2. Dwarfism reduced the hepatic glucose oxidation while de novo lipogenesis was not altered. As liver weight was depressed, total liver lipogenesis capacity was probably reduced by dwarfism. 3. As compared to normal hens, de novo lipogenesis and basal or stimulated lipolysis were lower in dwarf adipose tissue while its lipid content was enhanced in dwarfs. 4. Results suggest that in laying hens dwarfism reduces the adipose tissue lipid mobilization but probably also the liver de novo lipogenesis.     

Effect of the feeding system on the fatty acid composition,expression of the Δ<Superscript>9</Superscript>-desaturase,Peroxisome Proliferator-Activated Receptor Alpha,Gamma, and Sterol Regulatory Element Binding Protein 1 genes in the semitendinous muscle of light lambs of the Rasa Aragonesa breed

Background  

Conjugated linoleic acids (CLAs) are receiving increasing attention because of their beneficial effects on human health, with milk and meat products derived from ruminants as important sources of CLA in the human diet. SCD gene is responsible for some of the variation in CLA concentration in adipose tissues, and PPARγ, PPARα and SREBP1 genes are regulator of SCD gene. The aim of this work was to evaluate the effect of the feeding system on fatty acid composition, CLA content and relative gene expression of Δ⁹-desaturase (SCD), Peroxisome Proliferator-Activated Receptor Gamma (PPARγ), Peroxisome Proliferator-Activated Receptor Alpha, (PPARα) and Sterol Regulatory Element Binding Protein (SREBP1) in Rasa Aragonesa light lambs in semitendinous muscle. Forty-four single-born male lambs were used to evaluate the effect of the feeding system, varying on an intensity gradient according to the use of concentrates: 1. grazing alfalfa, 2. grazing alfalfa with a supplement for lambs, 3. indoor lambs with grazing ewes and 4. drylot.     

The role of SOCS2 in recombinant human growth hormone (rhGH) regulating lipid metabolism in high-fat-diet-induced obesity mice

In addition to regulate body growth and development process, growth hormone (GH) also involved in lipid metabolism, decreasing fat mass and improving lipolysis. To normal mice, GH could reduce their fat content, but events turned uncertain coming to the pattern of feeding high-fat-diet. In order to investigate the role of GH in adipogenesis of mice with high-fat-diet, the high-fat-diet feeding mice were randomly assigned into three groups and treated with recombinant human growth hormone (rhGH) and the somatostatin analogue octreotide respectively. Results demonstrated that both rhGH and octreotide could reduce the body weight but the trends diminished in the end. HDL-C level was increased in octreotide treated groups but the activity of lipase was increased significantly in both two groups. RhGH remarkable increased the expression of SOCS2, FAS (P < 0.01) and SREBP-1c (P < 0.05), decreased the expression of SOCS1, SOCS3 (P < 0.05) and HSL (P < 0.01) in subcutaneous fat mass. In visceral fat tissue, all genes were increased except SOCS2 (P < 0.01), at the same time the visceral fat mass was decreased. The protein phosphorylation of JAK2 and STAT5 which were treated with octreotide were increased in subcutaneous fat, visceral fat and liver (P < 0.01) and were increased significant in visceral fat by rhGH treated (P < 0.01). In liver, only JAK2 protein phosphorylation was raised (P < 0.01). In conclusion, rhGH and octreotide could decrease the whole body mass before 6 days; the trend was weaken in later period with high-fat-diet. RhGH could increase the subcutaneous fat mass and reduce the visceral fat mass, and SOCS2 might be involved in regulation of the mechanism through JAK2/STAT5 signaling pathway.     

The Farnesoid X Receptor Regulates Adipocyte Differentiation and Function by Promoting Peroxisome Proliferator-activated Receptor-γ and Interfering with the Wnt/β-Catenin Pathways

The bile acid receptor farnesoid X receptor (FXR) is expressed in adipose tissue, but its function remains poorly defined. Peroxisome proliferator-activated receptor-γ (PPARγ) is a master regulator of adipocyte differentiation and function. The aim of this study was to analyze the role of FXR in adipocyte function and to assess whether it modulates PPARγ action. Therefore, we tested the responsiveness of FXR-deficient mice (FXR^−/−) and cells to the PPARγ activator rosiglitazone. Our results show that genetically obese FXR^−/−/ob/ob mice displayed a resistance to rosiglitazone treatment. In vitro, rosiglitazone treatment did not induce normal adipocyte differentiation and lipid droplet formation in FXR^−/− mouse embryonic fibroblasts (MEFs) and preadipocytes. Moreover, FXR^−/− MEFs displayed both an increased lipolysis and a decreased de novo lipogenesis, resulting in reduced intracellular triglyceride content, even upon PPARγ activation. Retroviral-mediated FXR re-expression in FXR^−/− MEFs restored the induction of adipogenic marker genes during rosiglitazone-forced adipocyte differentiation. The expression of Wnt/β-catenin pathway and target genes was increased in FXR^−/− adipose tissue and MEFs. Moreover, the expression of several endogenous inhibitors of this pathway was decreased early during the adipocyte differentiation of FXR^−/− MEFs. These findings demonstrate that FXR regulates adipocyte differentiation and function by regulating two counteracting pathways of adipocyte differentiation, the PPARγ and Wnt/β-catenin pathways.     

Endocrine regulation of the fasting response by PPARalpha-mediated induction of fibroblast growth factor 21

Peroxisome proliferator-activated receptor α (PPARα) regulates the utilization of fat as an energy source during starvation and is the molecular target for the fibrate dyslipidemia drugs. Here, we identify the endocrine hormone fibroblast growth factor 21 (FGF21) as a mediator of the pleiotropic actions of PPARα. FGF21 is induced directly by PPARα in liver in response to fasting and PPARα agonists. FGF21 in turn stimulates lipolysis in white adipose tissue and ketogenesis in liver. FGF21 also reduces physical activity and promotes torpor, a short-term hibernation-like state of regulated hypothermia that conserves energy. These findings demonstrate an unexpected role for the PPARα-FGF21 endocrine signaling pathway in regulating diverse metabolic and behavioral aspects of the adaptive response to starvation.     

Molecular Cloning, Genomic Organization, and Expression of a Testicular Isoform of Hormone-Sensitive Lipase

By catalyzing the rate-limiting step in adipose tissue lipolysis, hormone-sensitive lipase (HSL) is an important regulator of energy homeostasis. The role and importance of HSL in tissues other than adipose are poorly understood. We report here the cloning and expression of a testicular isoform, designated HSL_tes. Due to an addition of amino acids at the NH₂-termini, rat and human HSL_tesconsist of 1068 and 1076 amino acids, respectively, compared to the 768 and 775 amino acids, respectively, of the adipocyte isoform (HSL_adi). A novel exon of 1.2 kb, encoding the human testis-specific amino acids, was isolated and mapped to the HSL gene, 16 kb upstream of the exons encoding HSL_adi. The transcribed mRNA of 3.9 kb was specifically expressed in testis. No significant similarity with other known proteins was found for the testis-specific sequence. The amino acid composition differs from the HSL_adisequence, with a notable hydrophilic character and a high content of prolines and glutamines. COS cells, transfected by the 3.9-kb human testis cDNA, expressed a protein of the expected molecular mass (M_r≈ 120,000) that exhibited catalytic activity similar to that of HSL_adi. Immunocytochemistry localized HSL to elongating spermatids and spermatozoa; HSL was not detected in interstitial cells.