FA (fatty acid) recycling in adipose tissue appears to be an important pathway for regulating FA release into the blood during fasting. Re-esterification requires G3P (glycerol 3-phosphate), which cannot be synthesized from glucose because glycolysis is much reduced under such circumstances. In addition, G3P can scarcely originate from glycerol since glycerol kinase has a very low activity in white adipose tissue. It was shown about 35 years ago that a metabolic pathway named glyceroneogenesis, which allows G3P synthesis from non-carbohydrate precursors like pyruvate, lactate or amino acids, is activated during fasting. The major enzyme in this pathway was shown to be PEPCK-C [cytosolic phosphoenolpyruvate carboxykinase (GTP); EC 4.1.1.32]. The present review analyses the mechanisms by which a series of hormones and nutrients affect PEPCK-C gene transcription and glyceroneogenesis and describes evidence for dysregulation of this pathway in type 2 diabetes.

Polyunsaturated fatty acids (PUFAs) and 3‐thia fatty acids are hypolipidemic and decrease insulin resistance in Type II diabetic animals. To exert such an action, these FAs could decrease adipose tissue lipolysis or increase esterification. Glyceroneogenesis is an important metabolic pathway in adipocytes for re‐esterification of FAs originating from lipolysis and in hepatocytes for triacylglycerol synthesis during fasting. Cytosolic phosphoenolpyruvate carboxykinase (PEPCK) plays a key role in this pathway. Here we show that the PUFA docosahexaenoic acid (DHA) stimulates PEPCK mRNA in glucose‐deprived adipose tissue explants from fed rats and in 3T3‐F442A differentiated adipocytes. This effect is maximum at 3 h, stable up to at least 11 h of treatment, and affects the transcription of the gene. PEPCK mRNA half‐life is not affected. Among a series of adipocyte transcripts, only the adipocyte lipid binding protein mRNA is also increased by DHA, although later than the PEPCK mRNA and at a much lower extent. DHA has no effect on PEPCK gene expression in the H4IIE hepatoma cells in which this gene is responsive to other inducers like cAMP. This lack of effect is not due to a failure of DHA to act in H4IIE cells since it induces the carnitine palmitoyltransferase 1 (CPT‐1) mRNA. Therefore, the DHA effect appears to be cell‐selective. Results of experiments using either tetradecylthio acetic acid and α‐bromopalmitate, two nonmetabolized Fas, or a series of inhibitors of FA metabolism show that the FA effect on PEPCK mRNA is not due to a product of its metabolism. Hence, polyunsaturated and nonmetabolized FAs stimulate adipose PEPCK, therefore potentially enhancing glyceroneogenesis and reducing FA output. This mechanism could participate in the hypolipidemic action of PUFAs. J. Cell. Biochem. 85: 651–661, 2002. © 2002 Wiley‐Liss, Inc.

Phosphoenolpyruvate carboxykinase (PEPCK) is the key enzyme in glyceroneogenesis, an important metabolic pathway that functions to restrain the release of non-esterified fatty acids (NEFAs) from adipocytes. The antidiabetic drugs known as thiazolidinediones (TZDs) are thought to achieve some of their benefits by lowering elevated plasma NEFAs. Moreover, peroxisome proliferator activated receptor γ (PPARγ) mediates the antidiabetic effects of TZDs, though many TZD responses appear to occur via PPARγ-independent pathways. PPARγ is required for adipocyte PEPCK expression, hence PEPCK could be a major target gene for the antidiabetic actions of TZDs. Here we used tissue culture and transfection assays to confirm that the TZD, rosiglitazone, stimulates PEPCK gene transcription specifically in adipocytes. We made the novel observation that this effect was by far the most rapid and robust among several other genes expressed in adipocytes. Adipocytes were transfected with a PEPCK/chloramphenicol acetyltransferase chimeric gene, in which either of the two previously discovered PPARγ/retinoid X receptor alpha response elements, PCK2 and gAF1/PCK1, had been inactivated by mutagenesis. We demonstrate that PCK2 alone is a bona fide thiazolidinedione response element. We show also that the regulation of PEPCK by PPARs is cell-specific and isotype-specific since rosiglitazone induces PEPCK gene expression selectively in adipocytes, and PPARα- and PPARβ-specific activators are inefficient. Hence, TZDs could lower plasma NEFAs via PPARγ and PEPCK by enhancing adipocyte glyceroneogenesis.

A heterodimer of peroxisome proliferator-activated receptor γ (PPARγ) and retinoid X receptor (RXR) is required for adipocyte differentiation. The gene encoding cytosolic phosphoenolpyruvate carboxykinase (PEPCK) is a PPARγ/RXR target gene in adipose tissue. Of the two PPARγ response elements, gAF1/PCK1 and PCK2, only PCK2 is required for PEPCK expression and responsiveness to the PPARγ agonist, rosiglitazone, in adipose tissue even though both elements bind PPARγ/RXR in vitro. In contrast, gAF1/PCK1 is essential for glucocorticoid inhibition of PPARγ-induced PEPCK gene expression in adipocytes. We report that chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) is the predominant nuclear receptor bound to gAF1/PCK1 in preadipocytes. COUP-TFII declines during adipogenesis in reciprocal fashion to PPARγ. In transiently transfected fibroblasts COUP-TFII acts at gAF1/PCK1 to inhibit PPARγ/RXR activation via PCK2. In contrast COUP-TFs are transcriptional activators of PEPCK in hepatocytes. PPARγ/RXR occupies gAF1/PCK1 in adipocytes, and mutation of gAF1/PCK1 enhances PEPCK promoter transactivation by PPARγ/RXR in fibroblasts, suggesting that this element is also a negative PPARγ response element. These results indicate that gAF1/PCK1 is a pleiotropic element through which COUP-TFII inhibits premature PEPCK expression, and perhaps adipogenesis in general, and PPARγ/RXR uses this same element in adipocytes to participate in PEPCK modulation by glucocorticoids.

Fatty acids (FAs) are energy-rich molecules, which play important metabolic roles. They are also an integral part of cells as membrane components, which can influence fluidity and receptor or channel function. Over the past 10 years, it has become evident that FAs can also act as signaling molecules involved in regulating gene expression. For the most part, these target genes encode proteins with roles in FA transport or metabolism. The corresponding change in the amount of specific proteins is an adaptative process that the cells develop in response to variations in FA concentration in the vicinity of the target tissue. Although interesting progress has been made recently, the mechanism(s) by which FAs modulate gene transcription still remains largely unresolved. The purpose of the present review is to address this important issue.

Hormone-sensitive lipase (HSL) catalyses the rate-limiting step in adipocyte lipolysis. Short-term hormonal regulation of HSL activity is well characterized, whereas little is known about the control of HSL gene expression. We have measured HSL mRNA content of 3T3-F442A and BFC-1 adipocytes in response to the cAMP analogue 8-(4-chlorophenylthio)-cAMP (8-CPT-cAMP) and to the phorbol ester phorbol 12-myristate 13-acetate (PMA) by Northern blot, using a specific mouse cDNA fragment. Treatment of the cells for 12 or 6 h with, respectively, 0.5 mM 8-CPT-cAMP or 1 microM PMA produced a maximal decrease of about 60% in HSL mRNA. These effects were unaffected by the protein-synthesis inhibitor anisomycin, suggesting that cAMP and PMA actions were direct. The reduction in HSL mRNA was accompanied by a reduction in HSL total activity. The intracellular routes that cAMP and PMA follow for inducing such an effect seemed clearly independent. (i) After desensitization of the protein kinase C regulation pathway by a 24 h treatment of the cells with 1 microM PMA, PMA action was abolished whereas cAMP was still fully active. (ii) Treatment with saturating concentrations of both agents produced an additive effect. (iii) The synthetic glucocorticoid dexamethasone had no proper effect on HSL gene expression but potentiated cAMP action without affecting PMA action. cAMP inhibitory action on HSL is unexpected. Indeed, the second messenger of catecholamines is the main activator of HSL by phosphorylation. We envision that a long-term cAMP treatment of adipocytes induces a counter-regulatory process that reduces HSL content and, ultimately, limits fatty acid depletion from stored triacylglycerols.