Molecular Differences Caused by Differentiation of 3T3-L1 Preadipocytes in the Presence of either Dehydroepiandrosterone (DHEA) or 7-Oxo-DHEA

Gómez, F. Enrique; Miyazaki, Masaru; Kim, Young‐Cheul; Marwah, Padma; Lardy, Henry A.; Ntambi, James M.; Fox, Brian G.

doi:10.1021/bi012177r

Cited by 32 publications

(23 citation statements)

References 58 publications

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“…Quite the opposite, palmitoleic acid (16:1 n-7) was reduced in s.c. and brown AT (and in serum, with marginal significance). These results are in agreement with those obtained by Imai et al (2001), who observed that DHEA stimulates oleate synthesis, resulting in increased 18:1 n-9 contents in liver, and with Gómez et al (2002), who also reported reduced 16:1 n-7 and elevated 18:1 n-9 in 3T3-L1 adipocytes treated with DHEA.…”

Section: Discussionsupporting

confidence: 93%

“…By contrast, our experiment spanned a 13-week period that allowed us to observe a significant effect of DHEA on AT composition. Similarly to us, other authors have reported differences in FAs in muscle (Abadie et al 2001), liver (Imai et al 2001), and 3T3-L1 adipocytes (Gómez et al 2002) due to DHEA administration.…”

Section: Discussionsupporting

confidence: 84%

“…Despite this fact, little is known about the effects of DHEA on FA composition in different tissues (Abadie et al 2001, Imai et al 2001, Gómez et al 2002, and no studies to our knowledge have shown significant changes in AT fatty acid profile due to DHEA action.…”

Section: Introductionmentioning

confidence: 99%

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Dehydroepiandrosterone modifies rat fatty acid composition of serum and different adipose tissue depots and lowers serum insulin levels

Heredia¹,

Larqué²,

Zamora³

et al. 2009

Journal of Endocrinology

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Dehydroepiandrosterone (DHEA) is reported to exert beneficial effects, such as protection from cardiovascular risk and lowering serum insulin levels. Adipose tissue (AT) is a target for DHEA actions, and the hormone can also affect hepatic fatty acid (FA) metabolism. FAs are involved in the development of insulin resistance; thus, there might be a relationship between DHEA, FA, and insulin. However, few data are available regarding DHEA and FA composition, especially concerning AT. Seventeen-month old female Sprague-Dawley rats (nZ11; controls: nZ10) were treated with DHEA (0 . 5% w/w in the diet) for 13 weeks, after which serum, periovarian, mesenteric, s.c., and brown AT were analyzed for FA composition. DHEA treatment resulted in significant changes in FA profiles in serum and adipose depots, like reduced 16:1n-7 (s.c. and brown AT; P!0 . 01), elevated n-9 monounsaturated FA (serum and s.c. AT; P!0 . 05), diminished n-6 polyunsaturated FA (PUFA; general; P!0 . 05) and increased n-3 PUFA (brown AT; P!0 . 01), along with lower n-6/n-3 ratios (s.c. and brown AT; P!0 . 05, P!0 . 01 respectively). DHEA modified estimates of desaturase activities, decreasing stearoyl-CoAdesaturase markers in s.c., and brown AT (P!0 . 05) and increasing those of delta-6-desaturase in serum and AT (P!0 . 05). In addition, DHEA-treated rats showed lower serum insulin levels (P!0 . 05). We have demonstrated for the first time that DHEA induces significant modifications in AT fatty acid composition in vivo, mainly concerning unsaturated FAs, and changes occurred in a tissue-dependent manner. We propose that these changes may be related to the capacity of DHEA to lower serum insulin levels.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 84%

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Dehydroepiandrosterone modifies rat fatty acid composition of serum and different adipose tissue depots and lowers serum insulin levels

Heredia¹,

Larqué²,

Zamora³

et al. 2009

Journal of Endocrinology

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“…Protein Extraction and Western Blot-Proteins from 3T3-L1 cells were extracted as described previously (31). Briefly, the cell monolayer was washed with ice-cold PBS and subsequently scraped into 1 ml of ice-cold lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EDTA, 0.25% sodium deoxycholate, 1% Nonidet P-40, 0.1% SDS, 1 mM phenylmethylmethanesulfonyl fluoride, 2 g/ml aprotinin, and 1 g/ml leupeptin).…”

Section: Methodsmentioning

confidence: 99%

Small Interfering RNA Knockdown of Calcium-independent Phospholipases A2 β or γ Inhibits the Hormone-induced Differentiation of 3T3-L1 Preadipocytes

Mancuso

Bickel

et al. 2004

Journal of Biological Chemistry

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Alterations in lipid secondary messenger generation and lipid metabolic flux are essential in promoting the differentiation of adipocytes. To determine whether specific subtypes of intracellular phospholipases A 2 (PLA 2 s) facilitate hormone-induced differentiation of 3T3-L1 cells into adipocytes, we examined alterations in the mRNA level, protein mass, and activity of three previously characterized mammalian intracellular PLA 2 s. Hormone-induced differentiation of 3T3-L1 cells resulted in 7.3 ؎ 0.5-and 7.4 ؎ 1.4-fold increases of mRNA encoding the calcium-independent phospholipases, iPLA 2 ␤ and iPLA 2 ␥, respectively. In contrast, the temporally coordinated loss of at least 90% of cPLA 2 ␣ mRNA was manifest. Western analysis demonstrated the near absence of both iPLA 2 ␤ and iPLA 2 ␥ protein mass in resting 3T3-L1 cells that increased dramatically during differentiation. In vitro measurement of PLA 2 activities demonstrated an increase in both iPLA 2 ␤ and iPLA 2 ␥ activities that were discriminated using the chiral mechanism based inhibitors ( Recently, there has been a dramatic increase in the incidence of obesity in industrialized and newly developed countries (1). Obesity results from abnormal increases in white adipose tissue (WAT) 1 mass leading to alterations in whole organism energy storage and utilization (2-4). Increased adipose tissue mass can result from either an increase in individual adipocyte cell size (hypertrophy) or from an increase in total adipocyte number (hyperplasia). Alterations in whole organism lipid homeostasis leading to increased adipocyte tissue mass are highly correlated with the metabolic syndrome that is accompanied by its lethal sequelae of diabetes, hypertension, and atherosclerosis (2-5). During the last decade, substantial progress has been made in understanding the biochemical events leading to adipocyte differentiation utilizing the hormone-induced 3T3-L1 cell model of adipocyte differentiation (6 -9). Central to this understanding has been the detailed characterization of temporally coordinated changes in the expression of specific genes that collectively define the adipocyte phenotype. Differentiation of adipocytes is accomplished by the programmed activation of transcriptional regulatory proteins that modulate the expression of mRNA and proteins that effectively reprogram 3T3-L1 cell lipid metabolism to that of a mature adipocyte. Such alterations include increased de novo fatty acid synthesis, accumulation of perilipin-coated triglyceride droplets, and the generation of lipid secondary messengers including eicosanoids and lysophosphatic acid that serve as potent and specific regulators of coordinated differentiation programs (3, 7, 10 -14).Phospholipases A 2 (PLA 2 s) catalyze the hydrolysis of the sn-2 fatty acid substituents from glycerophospholipid substrates to yield free fatty acid (e.g. arachidonic acid) and lysophospholipid (15-17). Mammalian phospholipases A 2 have been categorized into several classes based on their requirement for calcium ion in in vitro ...

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“…The assay was carried out according to a published procedure [31]. Aliquots of excised muscles were weighed (ca.…”

Section: Methodsmentioning

confidence: 99%

Glucose oversupply increases Δ9-desaturase expression and its metabolites in rat skeletal muscle

et al. 2003

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Aim/hypothesis. Previous studies have shown that prolonged glucose infusion causes insulin resistance and triglyceride accumulation in rat skeletal muscle. In this study, we investigated a possible relationship between insulin resistance and the composition of different accumulated lipid fractions in rat skeletal muscle. Methods. Continuous glucose infusion was carried out in rats for 7 days. Lipids were extracted from skeletal muscle, separated by thin layer chromatography and fatty acid composition of phospholipids, triglycerides, diglycerides, free fatty acids and cholesterol esters fractions was analysed by gas chromatography. ∆9-Desaturase mRNA was measured by real time polymerase chain reaction. The enzyme activity was measured in the microsomal fractions. Results. Prolonged glucose infusion (5 days) increased the relative content of palmitoleic acid (16:1 N7) several-fold (2.3-to 5.8-fold) in four out of five lipid fractions and enhanced oleic acid (18:1 N9) two-fold in three lipid fractions suggesting increased ∆9-desaturase activity while the content of several polyunsaturated fatty acids was reduced. In parallel, ∆9-Desaturase mRNA contents and enzyme activities in skeletal muscle were increased 10-fold, 75-fold, 2.6-fold and 7.7-fold after 2 and 5 days of glucose infusion, respectively. Conclusion/interpretation. Our results suggest that long-term glucose oversupply induces a rapid increase in ∆9-desaturase expression and enzyme activity in skeletal muscle which leads to fast and specific changes in fatty acid metabolism possibly contributing to the insulin resistance in this animal model. [Diabetologia (2003) 46:203-212]

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Molecular Differences Caused by Differentiation of 3T3-L1 Preadipocytes in the Presence of either Dehydroepiandrosterone (DHEA) or 7-Oxo-DHEA

Cited by 32 publications

References 58 publications

Dehydroepiandrosterone modifies rat fatty acid composition of serum and different adipose tissue depots and lowers serum insulin levels

Dehydroepiandrosterone modifies rat fatty acid composition of serum and different adipose tissue depots and lowers serum insulin levels

Small Interfering RNA Knockdown of Calcium-independent Phospholipases A2 β or γ Inhibits the Hormone-induced Differentiation of 3T3-L1 Preadipocytes

Glucose oversupply increases Δ9-desaturase expression and its metabolites in rat skeletal muscle

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