29 research outputs found
Incorporation of [<sup>14</sup>C]-acetate and [<sup>14</sup>C]-oleate in the biosynthetic pathway of triglycerides.
No full text<p>Incorporation of [<sup>14</sup>C]-acetate and [<sup>14</sup>C]-oleate in the biosynthetic pathway of triglycerides.</p
Effect of camphene on the incorporation of [<sup>14</sup>C]-acetate into <i>de novo</i> synthesized fatty acids.
No full text<p>On day 7, HepG2 cells were incubated for 24 h with camphene (25, 50 and 100 μM) in medium containing 10% (v/v) human LPDS. Four hours prior to the end of the incubation period, the cells were pulse-labeled with [<sup>14</sup>C]-acetic acid, sodium salt. The incorporation of acetate into fatty acids was determined as described. Results were normalized to the amount of cellular protein and expressed in percent of the respective control incubations. Results are from three independent experiments in triplicates and expressed as the mean ± SD. Significantly different compared to control: <i>p</i><0.05 (*); <i>p</i> <0.001 (***) and ns (non significant) vs control by Student-Newmann-Keuls test.</p
Effect of camphene, mevinolin and atorvastatin on the incorporation of [<sup>14</sup>C]-acetate into <i>de novo</i> synthesized triglycerides.
No full text<p>On day 7, HepG2 cells were incubated for 24 h with each compound in medium containing 10% (v/v) human LPDS. The cells received camphene, mevinolin and atorvastatin at the concentrations indicated. Four hours prior to the end of the incubation period, the cells were pulse-labeled with [<sup>14</sup>C]-acetic acid, sodium salt. The incorporation of acetate into triglycerides was determined as described in Materials and Methods section. Results were normalized to the amount of cellular protein and expressed in percent of the respective control incubations. Results are from three independent experiments in triplicates and expressed as the mean ± SD. Significantly different compared to control: <i>p</i><0.05 (*); <i>p</i><0.01 (**); <i>p</i><0.001 (***); <i>p></i>0.05 and ns (non significant) vs control by Student-Newmann-Keuls test.</p
Promoter Elements and Factors Involved In Hepatic Transcription of the Human Apoa-i Gene Positive and Negative Regulators Bind To Overlapping Sites
No full textDNase I footprinting analysis of the proximal apoA-I promoter sequences with rat liver nuclear extracts identified four protected regions: A, -22 to +17; B, -128 to -77; C, -175 to -148; and D, -220 to -190. Region D (-220 to -190) binds at least two distinct activities, designated AID1 and AID2, respectively, which can be separated by ion exchange chromatography. Region C (-175 to -148) forms five DNA protein complexes. Three of the complexes (2, 4, and 5) originate from the binding of more than one heat-stable nuclear factor, and two (1 and 3), from the binding of two heat-labile factors. The heat-stable factors bind in the -175 to -148 region and can be distinguished from C/EBP, which recognizes the same region, with DNA binding gel electrophoretic assays. Both factors 1 and 3 bind in the -168 to -148 apoA-I region. Despite the lack of a CCAAT motif in this region, the binding of factor 1 is competed out by oligonucleotides containing the binding sites of NFY and NFY*. Mutagenesis of the promoter region showed that mutations in the -171 to -166 and -158 to -153 regions diminished the binding of the heat-stable factors and reduced hepatic transcription to 14 and 8% of control, respectively. In contrast, a mutation in the -164 to -159 region abolished the binding of factor 1 and was associated with a 4.6-fold increase in hepatic transcription. These findings suggest that the heat-stable factors act as positive regulators, whereas factor 1 acts as a negative regulator in apoA-I gene transcription
Effect of camphene on lipid profile and metabolism in HepG2 cells-Comparison with well known hypolipidemic compounds.
No full text<p>On day 7, the cells were incubated with the compounds for 24 h in DMEM containing 10% (v/v) human LPDS. HepG2 cells were treated with camphene (lanes: 2–4), mevinolin (lane: 5), atorvastatin (lane: 7), the OSC inhibitor, U18666A (lane: 6) and the ACAT inhibitor, F1394 (lane: 8). Control-non treated cells migrate in places 1 and 9. Camphene was used at concentration of 25, 50 and 100 μM lanes 2, 3 and 4 respectively. Four hours prior to the end of the incubation period, the cells were pulse-labeled with [<sup>14</sup>C]-acetic acid, sodium salt. This panel shows the autoradiograph of synthesized intracellular lipids separated by TLC after labelling of cells with [<sup>14</sup>C]-acetate. The positions of the different lipids have been determined using non radioactive standards. The arrows indicate the positions of migration of cholesterol, fatty acids, triglycerides, cholesterol esters, MOS and DOS.</p
Effect of camphene, mevinolin and atorvastatin on the incorporation of acetate into <i>de novo</i> synthesized cholesterol.
No full text<p>On day 7, the cells were incubated with the compounds for 24 h in DMEM containing 10% (v/v) human LPDS. Camphene, mevinolin and atorvastatin were received at the concentrations indicated. Four hours prior to the end of the incubation period, the cells were pulse-labeled with [<sup>14</sup>C]-acetic acid, sodium salt. The incorporation of acetate into cholesterol was determined as described. Results were normalized to the amount of cellular protein and expressed in percent of the respective control incubations. Results are from three independent experiments in triplicates and expressed as the mean ± SD. Significantly different compared to control: <i>p<</i>0.05 (*); <i>p<</i>0.01(**); <i>p<</i>0.001 (***) vs control by Student-Newmann-Keuls test. 25 μM vs 50 μM <i>p</i><0.05; 25 μM vs 100 μM <i>p</i><0.01; 50 μM vs 100 μM <i>p</i>>0.05.</p
rev protein of human immunodeficiency virus type 1 affects the stability and transport of the viral mRNA.
No full textEffect of camphene on the esterification of fatty acids into triglycerides.
No full text<p>On day 7, HepG2 cells were incubated for 24 h with each compound in medium containing 10% (v/v) human LPDS. The cells received camphene (25, 50 and 100 μM), mevinolin (37 μM) or atorvastatin (10 μM). Four hours prior to the end of the incubation period, the cells were pulse-labeled with [<sup>14</sup>C]-oleic acid, sodium salt. The incorporation of oleate into triglycerides was determined as described. Results were normalized to the amount of cellular protein and expressed in percent of the respective control incubations. Results are from three independent experiments in triplicates and expressed as the mean ± SD. Results not statistically significant vs control, ns (non significant) by Student-Newmann-Keuls test.</p
Effect of camphene on MTP expression.
No full text<p>On day 7, HepG2 cells were incubated with indicated concentrations of camphene for 24 h in DMEM containing 10% (v/v) human LPDS. <b>Panel A.</b> Total cell protein was extracted and 50 μg of protein were separated on a 12% polyacrylamide gel and analyzed by Western immunoblotting using goat anti-MTP as described in Materials and Methods section. Mouse β-actin was used to control for equal loading and normalization. Relative intensity of the bands was quantified by Phosphorimager. Values express the MTP/actin ratio and are calculated by comparison to control samples the value of which is defined as 1. <b>Panel B.</b> A representative Western blot is shown. <b>Panel C.</b> Total RNA was extracted and subjected to Real Time PCR. The results represent the relative MTP mRNA normalized to endogenous YWHAZ mRNA. The same results were observed after normalization to the housekeeping gene actin (data not shown). The abundance of mRNA in the control-untreated cells was set at 1. Data represent means ± SD of three independent experiments. <i>p<</i>0.001 (***) and <i>p<</i>0.01 (**) vs control cells by the Student-Newmann-Keuls test.</p
Effect of camphene and mevinolin on the protein expression of SREBP-1. A and B panels.
No full text<p><b>Effects of the compounds on the mature and the precursor form of SREBP-1 respectively.</b> On day 7, HepG2 cells were incubated for 24 h with camphene (37 μM) and mevinolin (37 μM) in DMEM containing 10% LPDS. 25 μg/ml ALLN were added for the last 5 h before harvesting, in order to stabilize the short-lived nuclear forms. Total cell protein was extracted and 50 μg of protein were separated on a 8% polyacrylamide gel and analyzed by Western immunoblotting using rabbit anti-SREBP-1 as described. Mouse β-actin was used to control for equal loading and normalization. Relative intensity of the bands was quantified by Phosphorimager. Values express the SREBP-1/actin ratio and are calculated by comparison to control samples the value of which is defined as 1.Values are means ± SD of three independent experiments in triplicates. <i>p<</i>0.05 (*); <i>p<</i>0.001 (***); <i>p></i>0.05 and ns (non significant) vs control by the Student-Newmann-Keuls test. <b>Panel C</b>. A representative Western blot is shown. The upper band is SREBP-1 precursor (125 kDa) and the lower band is mature SREBP-1 (68 kDa).</p
