Are resistant to obesity induced either genetically or by a high

Are resistant to obesity induced either genetically or by a high fat diet [18]. The nuclear receptor pregnane X receptor (PXR; NR1I2), originally isolated as a xenobiotic receptor, is highly expressed in the liver, and plays a major role in drug metabolism and elimination through its regulation of the expression of cytochrome P450 enzymes [19]. Several recent studies suggested that PXR is also involved in hepatic lipid homeostasis. Activation of PXR perturbs lipid homeostasis in mice by decreasing b-oxidation, increasing free acid uptake and lipogenesis, which results in hepatic steatosis in mice [20,21,22,23]. Activation of PXR also decreases the expression of carnitine palmitoyltransferase 1A (CPT1A), which controls the entry of activated long-chain fatty acids into the mitochondria, and mitochondrial 3-hydroxy-3methyl-glutarate-CoA synthase 2 (Hmgcs2), the rate-limiting enzyme of ketogenesis. PXR regulates CPT1A and HmgcsSCD1 Contributes to the Lipogenic CAL 120 Effect by PXRexpression through its crosstalk with the insulin-responsive forkhead factor A2 (FoxA2) [21]. Another study showed that in VPhPXR transgenic mice, the expression of several genes involved in fatty acid b-oxidation, such as PPARa and thiolase, was suppressed [23]. The fatty acid translocase CD36 was established as a direct target gene of PXR. PXR binds to a DR3 type PXRE in the CD36 gene MedChemExpress Oltipraz promoter and induces the expression of CD36, increasing the fatty acid uptake in liver [23]. In human hepatocytes (HHPC), PXR 1315463 activation by rifampicin, a well-known hPXR agonist, stimulates de novo lipogenesis through the activation of S14, a small acidic protein that plays an important role in the induction of lipogenic enzymes [20]. Stearoyl-CoA desaturase-1 (SCD1) is the rate-limiting enzyme that converts palmitoyl- and stearoyl-coenzyme A to palmitoleoyland oleoyl-coenzyme A, respectively [24]. The monounsaturated products of SCD1 are preferred substrates for the synthesis of triglycerides, cholesterol esters, and phospholipids [24]. The expression of SCD1 is regulated by a number of dietary, physiological and hormonal factors including insulin, fructose, glucose, cholesterol and polyunsaturated fatty acids [25]. Activation of several nuclear receptors, such as LXRs [26], TR [27] and PPARa [28], can induce SCD1 gene expression. SCD1 has been reported as a direct target gene of LXRa and SREBP-1c [29]. In mouse models, activation of PXR induced SCD1 gene expression in the liver. However, whether SCD1 is up-regulated upon PXR activation in human hepatocytes and whether the human SCD1 is a direct PXR target gene are still unknown. In this study, we showed that activation of PXR in the human hepatoma HepG2 cells induced the expression of SCD1. We also showed that SCD1 is a direct PXR target gene.Materials and Methods ReagentsRifampicin, Oil Red O, Isopropanol, 49,6-diamidino-2-phenylindole (DAPI), TO901317, penicillin and streptomycin were purchased from Sigma (St. Louis, MO). Dimethyl sulfoxide (DMSO) was purchased from Merck (Darmstadt, Germany). Trizol, Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco-BRL (Grand Island, NY). BCA-100 protein quantitative analysis kit was from Pierce (Rockford, IL). Rabbit polyclonal anti-PXR (H-160, sc25381), rabbit polyclonal anti-SCD1 (H-300, sc-30081), rabbit anti-b-actin (sc-1618), goat anti-rabbit IgG-HRP (ZB-2308) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Protein molecular weight.Are resistant to obesity induced either genetically or by a high fat diet [18]. The nuclear receptor pregnane X receptor (PXR; NR1I2), originally isolated as a xenobiotic receptor, is highly expressed in the liver, and plays a major role in drug metabolism and elimination through its regulation of the expression of cytochrome P450 enzymes [19]. Several recent studies suggested that PXR is also involved in hepatic lipid homeostasis. Activation of PXR perturbs lipid homeostasis in mice by decreasing b-oxidation, increasing free acid uptake and lipogenesis, which results in hepatic steatosis in mice [20,21,22,23]. Activation of PXR also decreases the expression of carnitine palmitoyltransferase 1A (CPT1A), which controls the entry of activated long-chain fatty acids into the mitochondria, and mitochondrial 3-hydroxy-3methyl-glutarate-CoA synthase 2 (Hmgcs2), the rate-limiting enzyme of ketogenesis. PXR regulates CPT1A and HmgcsSCD1 Contributes to the Lipogenic Effect by PXRexpression through its crosstalk with the insulin-responsive forkhead factor A2 (FoxA2) [21]. Another study showed that in VPhPXR transgenic mice, the expression of several genes involved in fatty acid b-oxidation, such as PPARa and thiolase, was suppressed [23]. The fatty acid translocase CD36 was established as a direct target gene of PXR. PXR binds to a DR3 type PXRE in the CD36 gene promoter and induces the expression of CD36, increasing the fatty acid uptake in liver [23]. In human hepatocytes (HHPC), PXR 1315463 activation by rifampicin, a well-known hPXR agonist, stimulates de novo lipogenesis through the activation of S14, a small acidic protein that plays an important role in the induction of lipogenic enzymes [20]. Stearoyl-CoA desaturase-1 (SCD1) is the rate-limiting enzyme that converts palmitoyl- and stearoyl-coenzyme A to palmitoleoyland oleoyl-coenzyme A, respectively [24]. The monounsaturated products of SCD1 are preferred substrates for the synthesis of triglycerides, cholesterol esters, and phospholipids [24]. The expression of SCD1 is regulated by a number of dietary, physiological and hormonal factors including insulin, fructose, glucose, cholesterol and polyunsaturated fatty acids [25]. Activation of several nuclear receptors, such as LXRs [26], TR [27] and PPARa [28], can induce SCD1 gene expression. SCD1 has been reported as a direct target gene of LXRa and SREBP-1c [29]. In mouse models, activation of PXR induced SCD1 gene expression in the liver. However, whether SCD1 is up-regulated upon PXR activation in human hepatocytes and whether the human SCD1 is a direct PXR target gene are still unknown. In this study, we showed that activation of PXR in the human hepatoma HepG2 cells induced the expression of SCD1. We also showed that SCD1 is a direct PXR target gene.Materials and Methods ReagentsRifampicin, Oil Red O, Isopropanol, 49,6-diamidino-2-phenylindole (DAPI), TO901317, penicillin and streptomycin were purchased from Sigma (St. Louis, MO). Dimethyl sulfoxide (DMSO) was purchased from Merck (Darmstadt, Germany). Trizol, Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco-BRL (Grand Island, NY). BCA-100 protein quantitative analysis kit was from Pierce (Rockford, IL). Rabbit polyclonal anti-PXR (H-160, sc25381), rabbit polyclonal anti-SCD1 (H-300, sc-30081), rabbit anti-b-actin (sc-1618), goat anti-rabbit IgG-HRP (ZB-2308) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Protein molecular weight.