Effect of maternal triglycerides and free fatty acids on placental LPL in cultured primary trophoblast cells and in a case of maternal LPL deficiency

Magnusson-Olsson, Anne Liese; Lager, Susanne; Jacobsson, Bo; Jansson, Thomas; Powell, Theresa L.

doi:10.1152/ajpendo.00571.2006

Cited by 41 publications

(30 citation statements)

References 58 publications

(69 reference statements)

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“…Sixty-six hours after plating, to allow for syncytialization, 400 µM oleic acid (dissolved in 10% BSA; SigmaAldrich, St. Louis, MO) was added to the cell culture media. Similar fatty acid concentrations have previously been used by us ( 31 ) and others ( 34 ). Although high for a single fatty acid, the concentration is within the physiological range ( ‫ف‬ 350-600 µM) of total NEFA concentration in maternal circulation during late pregnancy ( 30 ).…”

Section: Measurement Of Amino Acid Uptakesupporting

confidence: 71%

“…Placental villous cytotrophoblasts were isolated as previously described ( 31,32 ) by DNase/trypsin digestion and purifi ed by separation on a Percoll gradient (n = 19 placentas). This protocol has been widely used and validated, and results in the isolation of cells that are expressing cytokeratin 7, a syncytial marker, that do not express vimentin, and that undergo biochemical differentiation as measured by hCG release ( 21,27,32,33 ).…”

Section: Isolation and Maintenance Of Trophoblast Cellsmentioning

confidence: 99%

“…Western blots were performed on NuPAGE precast 4-12% gels (Invitrogen, Carlsbad, CA) or SDS-PAGE as previously described ( 31,32 ). After electrophoresis and transfer, the membranes were blocked in 5% (phosphorylated STAT3 was blocked in 2.5%) nonfat milk in TBS with 0.1% tween (TBS-T) for 1 h. ( Fig.…”

Section: Western Blottingmentioning

confidence: 99%

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Oleic acid stimulates system A amino acid transport in primary human trophoblast cells mediated by toll-like receptor 4

Lager

Gaccioli

Ramirez

et al. 2013

Journal of Lipid Research

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Section: Measurement Of Amino Acid Uptakesupporting

confidence: 71%

Section: Isolation and Maintenance Of Trophoblast Cellsmentioning

confidence: 99%

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Oleic acid stimulates system A amino acid transport in primary human trophoblast cells mediated by toll-like receptor 4

Lager

Gaccioli

Ramirez

et al. 2013

Journal of Lipid Research

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“…Cytotrophoblast cells were isolated from human placentas and cultured as previously described (38,45). Cells were plated in either plastic 6-well plates at a density of ϳ1.…”

Section: Trophoblast Cell Culturementioning

confidence: 99%

Regulation of placental amino acid transporter activity by mammalian target of rapamycin

Roos¹,

Kanai²,

Prasad³

et al. 2009

American Journal of Physiology-Cell Physiology

130

135

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“…The supernatant (0.2 ml) was mixed with 40 l of 10 mM ferrous ammonium sulfate and 20 l of 2.5 M potassium thiocyanate in a 96-well plate, and absorbance was measured at 491 nm (24). To expose ␤TC3 cells to a high-fat condition, cells were treated for 48 h with an intralipos fat emulsion (100 mg/dl; Fresenius Kabi, Homburg, Germany), which consists of 10% soybean oil and 1.2% egg yolk (25).…”

Section: Methodsmentioning

confidence: 99%

Mechanism for antioxidative effects of thiazolidinediones in pancreatic β-cells

Chung

Kim

Lee

et al. 2011

American Journal of Physiology-Endocrinology and Metabolism

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dinediones (TZDs) are synthetic ligands of peroxisome proliferatoractivated receptor-␥ (PPAR␥), a member of the nuclear receptor superfamily. TZDs are known to increase insulin sensitivity and also to have an antioxidative effect. In this study, we tested whether TZDs protect pancreatic ␤-cells from oxidative stress, and we investigated the mechanism involved in this process. To generate oxidative stress in pancreatic ␤-cells (INS-1 and ␤TC3) or isolated islets, glucose oxidase was added to the media. The extracellular and intracellular reactive oxygen species (ROS) were measured to directly determine the antioxidant effect of TZDs. The phosphorylation of JNK/MAPK after oxidative stress was detected by Western blot analysis, and glucose-stimulated insulin secretion and cell viability were also measured. TZDs significantly reduced the ROS levels that were increased by glucose oxidase, and they effectively prevented ␤-cell dysfunction. The antioxidative effect of TZDs was abolished in the presence of a PPAR␥ antagonist, GW9662. Real-time PCR was used to investigate the expression levels of antioxidant genes. The expression of catalase, an antioxidant enzyme, was increased by TZDs in pancreatic ␤-cells, and the knockdown of catalase significantly inhibited the antioxidant effect of TZDs. These results suggest that TZDs effectively protect pancreatic ␤-cells from oxidative stress, and this effect is dependent largely on PPAR␥. In addition, the expression of catalase is increased by TZDs, and catalase, at least in part, mediates the antioxidant effect of TZDs in pancreatic ␤-cells.peroxisome proliferator-activated receptor-␥; reactive oxygen species; catalase OXIDATIVE STRESS HAS BEEN IMPLICATED in the pathogenesis of type 2 diabetes and its vascular complications. Oxidative stress plays an important role in the development of ␤-cell dysfunction and insulin resistance, two major pathophysiological abnormalities of type 2 diabetes. It is well known that excessive reactive oxygen species (ROS) cause ␤-cell dysfunction, such as defects in insulin synthesis and secretion, and apoptosis (9, 23, 32). Pancreatic ␤-cells are very susceptible to oxidative stress because the expression levels of antioxidant enzymes, including superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx), are relatively low in ␤-cells (12, 39). There are several reports showing that the overexpression of antioxidant enzymes protects pancreatic ␤-cells from oxidative stress-induced dysfunction (1). Exposure to a high level of glucose and/or free fatty acid (FFA) results in the dysfunction of isolated islet or ␤-cells, such as a decrease in insulin release and apoptosis (33,36). An antioxidant N-acetyl-cysteine (NAC) protects ␤-cells from the high-glucose-or high-fatinduced impairments (37). These results suggest that ROS are involved in the glucose/FFA-induced ␤-cell dysfunction.Peroxisome proliferator-activated receptor-␥ (PPAR␥), a member of the nuclear receptor superfamily, is expressed predominantly in adipose tissue and plays an im...

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Effect of maternal triglycerides and free fatty acids on placental LPL in cultured primary trophoblast cells and in a case of maternal LPL deficiency

Cited by 41 publications

References 58 publications

Oleic acid stimulates system A amino acid transport in primary human trophoblast cells mediated by toll-like receptor 4

Oleic acid stimulates system A amino acid transport in primary human trophoblast cells mediated by toll-like receptor 4

Regulation of placental amino acid transporter activity by mammalian target of rapamycin

Mechanism for antioxidative effects of thiazolidinediones in pancreatic β-cells

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