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1.
Electron. j. biotechnol ; 18(3): 215-220, May 2015. ilus, graf, tab
Article in English | LILACS | ID: lil-750650

ABSTRACT

Background The objective of this study was to compare the level differences of mRNA transcription and protein expression of PPARγ, FAS and HSL in different parts of the carcass in different tail-type sheep. Six Tan sheep and six Shaanbei fine-wool sheep aged 9 months were slaughtered and samples were collected from the tail adipose, subcutaneous adipose, and longissimus dorsi muscle. The levels of mRNA transcription and protein expression of the target genes in these tissues were determined by real-time quantitative PCR and western blot analyses. Results The results showed that PPARγ, FAS, and HSL were expressed with spatial differences in tail adipose, subcutaneous adipose and longissimus dorsi muscle of Tan sheep and Shaanbei fine-wool sheep. Differences were also observed between the two breeds. The mRNA transcription levels of these genes were somewhat consistent with their protein expression levels. Conclusion The present results indicated that PPARγ, FAS and HSL are correlated with fat deposition, especially for the regulating of adipose deposition in intramuscular fat, and that the mRNA expression patterns are similar to the protein expression patterns. The mechanism requires clarification in further studies.


Subject(s)
Animals , Sheep , Sterol Esterase/genetics , PPAR gamma/genetics , Fatty Acid Synthases/genetics , Tail , Transcription, Genetic , RNA, Messenger , Blotting, Western , Sterol Esterase/metabolism , PPAR gamma/metabolism , Fatty Acid Synthases/metabolism , Real-Time Polymerase Chain Reaction
2.
Electron. j. biotechnol ; 18(2): 122-127, Mar. 2015. ilus, graf, tab
Article in English | LILACS | ID: lil-745580

ABSTRACT

Background The objective of this study was to investigate proliferator-activated receptor (PPARγ), fatty acid synthase (FAS) and hormone-sensitive lipase (HSL) mRNA and protein expression in fat tails of Tan sheep. Rams from different developmental stages (aged 3, 6, 9, 12, 15 and 18 months) were selected, and their tail measurements including length (L), width (W) and girth (G) were recorded. The mRNA and protein expressions of PPARγ, FAS and HSL were examined by quantitative real-time polymerase chain reaction (PCR) and Western blot. Results The tail measurements increased with age. We observed no significant differences (P > 0.05) of PPARγ mRNA expression between ages 9 and 15 months, and between 12 and 15 months; FAS mRNA expression levels at each developmental stage were observed significantly in Tan sheep (P < 0.05); HSL mRNA expression with no significant differences were only observed between 6 and 15 months (P > 0.05). Significant differences (P < 0.05) of PPARγ, FAS and HSL protein expressions at each developmental stage were observed in Tan sheep. Conclusion We observed that the mRNA expression patterns of PPARγ and FAS decreased first before they increased again and then this process repeated. Conversely, the mRNA expression patterns of HSL increased first before they decreased and then this process repeated. The protein expression patterns of PPARγ and FAS decreased first before they increased again and then this process repeated. Conversely, the protein expression pattern of HSL increased first before it decreased again and then increased again.


Subject(s)
Animals , Sheep/growth & development , Sheep/genetics , Proteins/metabolism , Sterol Esterase/metabolism , PPAR gamma/metabolism , Fatty Acid Synthases/metabolism , Transcription Factors , RNA, Messenger , Blotting, Western , Sterol Esterase/genetics , PPAR gamma/genetics , Fatty Acid Synthases/genetics , Real-Time Polymerase Chain Reaction
3.
Electron. j. biotechnol ; 17(6): 251-261, Nov. 2014. ilus, graf, tab
Article in English | LILACS | ID: lil-730255

ABSTRACT

Background Fatty acid synthase (FAS) is a key enzyme of de novo lipogenesis (DNL), which has been cloned from several species: Gallus gallus, Mus musculus, Homo sapiens, but not from Anas platyrhynchos. The current study was conducted to obtain the full-length coding sequence of Peking duck FAS and investigate its expression during adipocyte differentiation. Results We have isolated a 7654 bp fragment from Peking duck adipocytes that corresponds to the FAS gene. The cloned fragment contains an open reading frame of 7545 bp, encodes a 2515 amino acid protein, and displays high nucleotide and amino acid homology to avian FAS orthologs. Twelve hour treatment of oleic acid significantly up-regulated the expression of FAS in duck preadipocytes (P < 0.05). However, 1000 µM treatment of oleic acid exhibited lipotoxic effect on cell viability (P < 0.05). In addition, during the first 24 h of duck adipocyte differentiation FAS was induced; however, after 24 h its expression level declined (P < 0.05). Conclusion We have successfully cloned and characterized Peking duck FAS. FAS was induced during adipocyte differentiation and by oleic acid treatment. These findings suggest that Peking duck FAS plays a similar role to mammalian FAS during adipocyte differentiation.


Subject(s)
Animals , Adipose Tissue/metabolism , Ducks , Fatty Acid Synthases/genetics , Fatty Acid Synthases/metabolism , Phylogeny , RNA/analysis , Gene Expression , Cell Differentiation , Cell Survival , Cloning, Molecular , Sequence Analysis , DNA, Complementary/chemical synthesis , Oleic Acid , Computational Biology , Lipogenesis
4.
Article in English | IMSEAR | ID: sea-135460

ABSTRACT

Background & objectives: Conjugated linoleic acid (CLA) reduces fat deposition in the body, but the mechanism of action is not clear. The present study was conducted to investigate the effects of CLA on body fat metabolism. Since milk fat is the best natural source of dietary CLA, intervention of non-fat milk constituents on CLA treatment was also investigated. Methods: Diets containing CLA (1%) with or without skim milk powder (SMP) was fed to male Swiss albino mice for 60 days. Adipose depots weight, faecal fat and the activities of selected enzymes of lipid metabolism were determined. Results: The mice on CLA and CLA+SMP diets gained weight similar to those on control diet, despite higher feed intake in the former two groups. Total fat pad mass was significantly (P<0.05) less in CLA group than in control group, and inclusion of SMP in the diet enhanced the fat reducing effect of CLA. Adiposity index was also less on CLA and CLA+SMP diets than on control diet, and CLA+SMP was more efficacious in reducing adiposity index. The weight of liver and spleen was increased by CLA, and this effect was eliminated by inclusion of SMP in the diet. The fatty acid synthase (FAS) activity in liver and retroperitoneal adipose tissue decreased substantially on CLA and CLA+SMP diets compared to that on control diet. Interpretation & conclusions: Our preliminary data show that dietary CLA reduces body fat mass by decreasing fatty acid biosynthesis, and the effect is enhanced by inclusion of SMP in the diet.


Subject(s)
Adipose Tissue/metabolism , Animals , Body Weight , Drug Synergism , Fatty Acid Synthases/metabolism , Fatty Acids/metabolism , Female , Linoleic Acids, Conjugated/pharmacology , Lipids/chemistry , Liver/metabolism , Male , Mice , Milk/metabolism , Models, Biological , Models, Statistical , Spleen/metabolism
5.
Genet. mol. res. (Online) ; 7(2): 284-294, 2008. tab
Article in English | LILACS | ID: lil-641011

ABSTRACT

The objective of the present study was to determine the effects of trans-10, cis-12 conjugated linoleic acid (CLA) in adipose tissue explant cultures of growing pigs on the following responses: lipogenesis (measured as rate of 14C-labeled glucose incorporation over a subsequent 2-h incubation in the presence or absence of insulin), lipolysis (release of non-esterified fatty acid over a 2-h incubation in the presence or absence of isoproterenol), activities of lipogenic enzymes, and mRNA abundance of fatty acid synthase (FAS). Adipose tissue explants from nine growing pigs (78 ± 3 kg) were cultured in 199 medium with insulin, dexamethasone and antibiotics for 4, 12, 24, and 48 h. The treatments were 1) control: 100 μM polyvinyl alcohol (PVA); 2) pGH: 100 ng/mL porcine growth hormone (pGH) plus 100 μM PVA; 3) CLA200: 200 μM trans-10, cis-12 CLA; 4) CLA50: 50 μM trans-10, cis-12 CLA, and 5) LA: 200 μM linoleic acid. Fatty acids were added along with PVA (2:1), respectively, for 24 h. Explants were collected after each culture period and assayed for lipogenesis. Transcripts of FAS mRNA were quantified by real-time RT-PCR after 24 and 48 h. Lipolysis and activities of FAS, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and NADP-malate dehydrogenase were determined after 48 h. As expected, glucose incorporation was decreased (P < 0.05) in response to pGH treatment (positive control). LA had no effect on any parameter evaluated. Treatment with trans-10, cis-12 CLA decreased FAS activity (P < 0.05), but NADPH-generating enzymes were unaffected by treatments. Consistent with reduction in FAS activity, both lipid synthesis and FAS mRNA abundance were reduced with chronic CLA treatment, pGH increased baseline and stimulated lipolysis (P < 0.05) after 48 h of culture, while CLA treatment had no effect on non-esterified fatty acid release. Results of this study showed that trans-10, cis-12 CLA alters lipogenesis but has no effect on lipolysis...


Subject(s)
Animals , Male , Linoleic Acids, Conjugated/pharmacology , Lipid Metabolism , Gene Expression Regulation, Developmental , Swine/genetics , Adipose Tissue , Fatty Acids/metabolism , Fatty Acid Synthases/genetics , Fatty Acid Synthases/metabolism , Lipogenesis , Lipolysis , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger/genetics , RNA, Messenger/metabolism , Tissue Culture Techniques , Adipose Tissue/growth & development , Adipose Tissue/metabolism
6.
Article in English | WPRIM | ID: wpr-171364

ABSTRACT

This study was conducted to examine the effects of dietary grape extracts on preneoplastic foci formation in rat hepatocarcinogenesis, and related hepatic enzymes. Male Sprague-Dawley rats were fed basal diet or grape diet containing 15% concentrated grape extracts (68 bricks). The grape diet groups were divided into whole-period grape diet group (DEN-GW; grape diet group fed throughout experimental period) and postinitiation grape diet group (DEN-GP; grape diet group fed from post initiation stage) according to the starting time point of the grape diet. Hepatocarcinogenesis was induced by diethylnitrosamine (DEN; 200 mg/kg bw) and 2/3 partial hepatectomy (DEN-B; DEN-treated basal diet group, DEN-GW, and DEN-GP groups), while the control group treated with saline and sham operation (Control group). The formation of placental glutathione (GSH) S-transferase positive (GST-P(+)) foci in DEN-GW group was moderately but significantly suppressed, however, not in DEN- GP group. Thiobarbituric acid reactive substances content of DEN-GW group was significantly lower than that of DEN-B group. The activity of fatty acid synthase (FAS) in the grape diet groups was decreased about 1/2 of the DEN-B group. The content of GSH and GSH peroxidase activity were increased by carcinogen treatment, but not modulated by grape diet. The activities of GSH S-transferase, p-nitrophenol hydroxylase, and catalase were not affected by diet or treatment. Conclusively, the grape diet-induced reduction of FAS activity that was expressed highly in neoplastic tissues, might be one of the contributing mechanisms of hepatic cancer prevention.


Subject(s)
Administration, Oral , Animals , Body Weight/drug effects , Catalase/metabolism , Cell Transformation, Neoplastic/drug effects , Diet , Dietary Supplements , Fatty Acid Synthases/metabolism , Glutathione/metabolism , Glutathione Peroxidase/metabolism , Liver/drug effects , Liver Neoplasms/diet therapy , Male , Organ Size/drug effects , Plant Extracts , Rats , Rats, Sprague-Dawley , Vitis/chemistry
7.
Indian J Biochem Biophys ; 1999 Apr; 36(2): 63-8
Article in English | IMSEAR | ID: sea-26717

ABSTRACT

Pigeon liver fatty acid synthetase (FAS) was inactivated irreversibly by stoichiometric concentration of o-phthalaldehyde exhibiting a bimolecular kinetic process. FAS-o-phthalaldehyde adduct gave a characteristic absorption maxima at 337 nm. Moreover this derivative showed fluorescence emission maxima at 412 nm when excited at 337 nm. These results were consistent with isoindole ring formation in which the -SH group of cysteine and epsilon-NH2 group of lysine participate in the reaction. The inactivation is caused by the reaction of the phosphopantetheine -SH group since it is protected by either acetyl- or malonyl-CoA. The enzyme incubated with iodoacetamide followed by o-phthalaldehyde showed no change in fluorescence intensity but decrease in intensity was found in the treatment of 2,4,6-trinitrobenzenesulphonic acid (TNBS), a lysine specific reagent with the enzyme prior to o-phthalaldehyde addition. As o-phthalaldehyde did not inhibit enoyl-CoA reductase activity, so nonessential lysine is involved in the o-phthalaldehyde reaction. Double inhibition experiments showed that 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), a thiol specific reagent, binds to the same cysteine which is also involved in the o-phthalaldehyde reaction. Stoichiometric results indicated that 2 moles of o-phthalaldehyde were incorporated per mole of enzyme molecule upon complete inactivation.


Subject(s)
Animals , Columbidae , Fatty Acid Synthases/metabolism , Liver/metabolism , o-Phthalaldehyde/metabolism
8.
Indian J Biochem Biophys ; 1990 Feb; 27(1): 43-7
Article in English | IMSEAR | ID: sea-28614

ABSTRACT

Levels of fatty acid binding proteins (FABPs), lipids as well as activities of fatty acid synthesizing enzymes such as fatty acid synthase and ATP-citrate lyase increase with gestation showing maximum at term in human fetal lung. However, the activity of ATP-citrate lyase showed the same trend up to 30 weeks of gestation before declining slightly at term. These results indicate the importance of supply and/or synthesis of fatty acids when lung surfactant synthesis begins; thereby showing a correlation between the FABPs, lipid pattern and the activities of fatty acid synthesizing enzymes during prenatal lung development.


Subject(s)
ATP Citrate (pro-S)-Lyase/metabolism , Carrier Proteins/metabolism , Embryonic and Fetal Development/physiology , Fatty Acid Synthases/metabolism , Fatty Acid-Binding Proteins , Fatty Acids/metabolism , Humans , Lung/enzymology , Neoplasm Proteins , Tumor Suppressor Proteins
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