The specific expression of TGH and HSL genes in different tissues of Bamei pig was investigated by RT-PCR and Western blot in this study. The result of RT-PCR showed that the expression of HSL could be detected in all these seven tissues examined, and which was higher expressed in fat, lower in heart, liver, lung, spleen and kidney. Expression of TGH gene could also be detected in seven tissues, and higher in liver and fat, lower in heart and kidney and lowest in spleen and lung. The result of Western blot showed that, HSL gene was highest expressed in epiploica fat and subcutaneous fat, higher in other tissues, but couldn' t be detected in kidney. Expression of TGH was detected in epiploica fat, subcutaneous fat, liver, lung and spleen, and highest in fat and liver, but it hadn't be found in heart and kidney. These results suggested that both HSL and TGH could be regulated by post-transcriptional, and their function was involved in different tissues.
Animals ; Gene Expression Regulation ; Lipase ; genetics ; metabolism ; Male ; Sterol Esterase ; genetics ; metabolism ; Swine ; Tissue Distribution

Estrogen-related receptor a (ERRalpha) is a key regulator for energy metabolism and adipogenesis. However, its role in lipolysis is unknown. To study the function of ERRalpha in lipolysis, primary cultured differentiated porcine adipocytes were treated by a specific inverse agonist XCT790 or infected with adenoviral vector expressed ERRalpha for 48 h, in the absence and/or presence of specific protein kinase A (PKA) inhibitor or extracellular signal-related kinase (ERK) inhibitor. Then, we measured the triglyceride (TG) content and the glycerol release into the culture media to analysis the effect of ERRalpha on lipolysis; Further, we analyzed the expression of PPARgamma, perilipin A, p-perilipin A, HSL and ATGL with Western blotting. Here, we found that ERRalpha significantly increased adipocytes differentiation, TG accumulation and glycerol release. Separately or simultaneously block the PKA and ERK pathway do not significantly altered the effect of ERRalpha on glycerol release. ERRalpha significantly up-regulated the proteins expression of PPARgamma, perilipin A, HSL and ATGL, while the p-perilipin A protein level was not significantly changed. These findings imply that ERRalpha could increase lipolysis via up-regulating HSL and ATGL, thereby to supply more FFA as substrate for a larger turnover of cellular triglyceride pool during adipocytes differentiation.
Adipocytes ; cytology ; metabolism ; Animals ; Animals, Newborn ; Cells, Cultured ; Glycerol ; analysis ; Lipase ; metabolism ; Lipolysis ; physiology ; Receptors, Estrogen ; metabolism ; physiology ; Sterol Esterase ; metabolism ; Swine ; Triglycerides ; analysis

The purpose of this study was to find out effects of treatment of ginsenoside Re, Rc and EGCG on mRNA expressions of leptin, hormone sensitive lipase (HSL) and resistin in 3T3-L1 adipocytes. The concentrations of EGCG were treated with 0.01 x 10(-7), 0.1 x 10(-7), 1 x 10(-7) and 1 x 10(-6) M or 100 microgram/ml ginsenoside Re, Rc in culture cell for 13 days. mRNA expression of leptin wasn't expressed in preadipocyte but according to differentiation of adipocyte, the that of mRNA expression was decreased at gensenosids or EGCG treated cells compared with non treated adipocyte. Expression of HSL mRNA was increased in G-Re, G-Rc and EGCG treated cells compared with non treated cells. The resistin level was significantly decreased in adipocytes treated with G-Re, G-Rc and EGCG. These pattern was similar to leptin expression.These results support that treatment of gensenosides or EGCG in 3T3-L1 adipocyte resulted to affect of leptin and resistin as well as HSL mRNA levels, accordingly, levels of leptin and HSL will be acted by signalling body fat stores to the hypothalamus which in turn regulates food intake and energy expenditure to maintain body weight homeostasis. And also regulation of resistin mRNA will prevent to diabetics attacked with obesity. In conclusion, we suggest that consumption of ginseng saponine or EGCG might prevent human diabetics or/and obesity.
3T3-L1 Cells ; Adipocytes* ; Adipose Tissue ; Body Weight ; Catechin* ; Eating ; Energy Metabolism ; Homeostasis ; Humans ; Hypothalamus ; Leptin* ; Obesity ; Panax* ; Resistin* ; RNA, Messenger* ; Saponins ; Sterol Esterase* ; Tea*

OBJECTIVETo explore the effects of zinc-alpha2-glycoprotein (ZAG) on body weight and body fat in high-fat-diet (HFD)-induced obesity in mice and the possible mechanism.

METHODSThirty-six male mice were fed with standard food (SF) (n = 9) and HFD (n = 27), respectively. Five weeks later, 9 mice fed with HFD were subjected to ZAG expression plasmid DNA transfection by liposome transfection method, and another 9 mice to negative control plasmid transfection. Two weeks later, serum ZAG level in the mice was assayed by Western blot, and the effects of ZAG over-expression on body weight, body fat, serum biochemical indexes, and adipose tissue of obese mice were evaluated. The mRNA expressions of fatty acid synthase (FAS) and hormone-sensitive lipase (HSL) in liver tissue were determined by reverse transcription-polymerase chain reaction.

RESULTSSerum ZAG level significantly lowered in simple HFD-fed mice in comparison to SF-fed mice (0.51 +/- 0.10 AU vs. 0.75 +/- 0.07 AU, P < 0.01). Further statistical analysis demonstrated that ZAG level was negatively correlated with body weight (r = -0.56, P < 0.001), epididymal fat mass (r = -0.67, P < 0.001), percentage of epididymal fat (r = -0.65, P < 0.001), and increased weight (r = -0.57, P < 0.001) in simple SF- and HFD-fed mice. ZAG over-expression in obese mice reduced body weight and the percentage of epididymal fat. Furthermore, FAS mRNA expression decreased (P < 0.01) and HSL mRNA expression increased (P < 0.001) in the liver in ZAG over-expressing mice.

CONCLUSIONSZAG is closely related to obesity. Serum ZAG level is inversely correlated with body weight and percentage of body fat. The action of ZAG is associated with reduced FAS expression and increased HSL expression in the liver of obese mice.

Adipose Tissue ; metabolism ; Animals ; Fatty Acid Synthases ; genetics ; physiology ; Liver ; enzymology ; Male ; Mice ; Mice, Obese ; Seminal Plasma Proteins ; blood ; physiology ; Sterol Esterase ; genetics ; physiology ; Weight Loss

OBJECTIVEIn order to explore the expression of hormone-sensitive lipase (HSL) mRNA in white adipose tissue of diet-induced obesity-resistant (DIO-R) rats.

METHODSFifty male Sprague-dawley (SD) rats were randomly divided into control group and high-fat group and they were fed with basic diet and high-fat diet respectively for 13 weeks. DIO-R and DIO rats were selected according to their body weight and the quantity of energy intake, then observing changing of the body fat content, the level of serum growth hormone was determined by radioimmunoassay and the level of the HSL mRNA in white adipose tissue was determined by RT-PCR.

RESULTSThe body fat content of DIO-R rats were lower than those of DIO rats (P < 0.05). The level of serum growth hormone of DIO-R rats were higher than those of DIO rats (P < 0.05). High fat diets increased the level of the HSL mRNA in DIO-R rats.

CONCLUSIONIncreasing expression of HSL in white adipose tissue may play a part of role in resisting diet-induced obesity of rats.

Adipose Tissue ; drug effects ; enzymology ; metabolism ; Animals ; Body Weight ; drug effects ; Dietary Fats ; administration & dosage ; Male ; Obesity ; enzymology ; etiology ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sterol Esterase ; biosynthesis ; genetics

Higher levels of body fat are associated with an increased risk for development numerous adverse health conditions. FTY720 is an immune modulator and a synthetic analogue of sphingosine 1-phosphate (S1P), activated S1P receptors and is effective in experimental models of transplantation and autoimmunity. Whereas immune modulation by FTY720 has been extensively studied, other actions of FTY720 are not well understood. Here we describe a novel role of FTY720 in the prevention of obesity, involving the regulation of adipogenesis and lipolysis in vivo and in vitro. Male C57B/6J mice were fed a standard diet or a high fat diet (HFD) without or with FTY720 (0.04 mg/kg, twice a week) for 6 weeks. The HFD induced an accumulation of large adipocytes, down-regulation of phosphorylated AMP-activated protein kinase alpha (p-AMPKalpha) and Akt (p-Akt); down-regulation of hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL) and perilipin mRNA as well as up-regulation of phosphorylated HSL (p-HSL, Ser563) and glycogen synthase kinase 3 alpha/beta (p-GSK3alpha/beta). All these effects were blunted by FTY720 treatment, which inhibited adipogenesis and promoted lipolysis. Also, FTY720 significantly decreased lipid accumulation in maturing preadipocytes. FTY720 down-regulated the transcriptional levels of the PPARgamma, C/EBPalpha and adiponectin, which are markers of adipogenic differentiation. FTY720 significantly increased the release of glycerol and the expression of the HSL, ATGL and perilipin, which are regulators of lipolysis. These results show that FTY720 prevented obesity by modulating adipogenesis and lipolysis, and suggest that FTY720 is used for the treatment of obesity.
3T3-L1 Cells ; AMP-Activated Protein Kinases/metabolism ; Adipocytes/*drug effects/physiology ; Adipogenesis/drug effects ; Animals ; Anti-Obesity Agents/*pharmacology/therapeutic use ; Antigens, Differentiation/genetics/metabolism ; Carrier Proteins/genetics/metabolism ; Cell Size ; Diet, High-Fat/adverse effects ; Disease Models, Animal ; Enzyme Activation ; Gene Expression Regulation, Enzymologic/drug effects ; Glycogen Synthase Kinase 3/genetics/metabolism ; Lipase/genetics/metabolism ; Lipolysis/drug effects ; Male ; Mice ; Mice, Inbred C57BL ; Obesity/etiology/metabolism/*prevention & control ; Phosphoproteins/genetics/metabolism ; Phosphorylation ; Propylene Glycols/*pharmacology/therapeutic use ; Protein Processing, Post-Translational ; Proto-Oncogene Proteins c-akt/metabolism ; Sphingosine/*analogs & derivatives/pharmacology/therapeutic use ; Sterol Esterase/metabolism