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Cyclic fluctuations of 3-hydroxy-3-methylglutaryl-CoA reductase in aortic smooth muscle cell cultures

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Lipids

Abstract

The cyclic fluctuations of HMG-CoA reductase activity and mRNA are reportedly related to feeding the cells in culture or to variations in food consumption by the animals over a 24-h cycle. In this work, we demonstrate cyclic increments in HMG-CoA reductase activity in smooth muscle cells (SMC) not associated with the culture feeding. Since reductase activity also shows a marked rise preceding the S phase, one of the major goals of the present work was to evaluate this dual role of reductase activity and mRNA fluctuations related to the cell cycle and to food intake in the SMC-C/SMC-Ch cultures derived from control-fed (SMC-C) and cholesterol-fed (SMC-Ch) chicks. The period and amplitude oscillations in HMG-CoA reductase activity varied depending on culture conditions: lipoprotein-deficient serum vs. FBS, young vs. senescent cells, or confluent vs. nonconfluent cultures. The HMG-CoA reductase mRNA concentration showed a marked rise after feeding not correlated to the fluctuation activity, suggesting posttranscriptional modulation. Reductase activity and mRNA were down-regulated in SMC-Ch. Since the nutritional culture conditions were the same in both cell lines, these findings indicate that consumption of a high-cholesterol diet by the animals prior to the establishment of the SMC cultures induced changes in the HMG-CoA reductase gene expression in aortic SMC.

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Abbreviations

dNTP:

deoxy nucleotide 5′-triphosphate

dThd:

double thymidine

LPDS:

lipoprotein-deficient serum

PI:

propidium iodide

RT-PCR:

reverse transcription-polymerase chain reaction

SMC:

smooth muscle cell

SMC-C:

smooth muscle cell-control fed chicks

SMC-Ch:

smooth muscle cell-cholesterol fed chicks

SRE:

sterol regulatory element

SREBP:

sterol regulatory element-binding protein

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Faculty of Sciences, University of Granada, C/Fuentenueva s/n, 18071, Granada, Spain

    Ma José Alejandre, Sonia Perales, Ángel Carazo, Rogelio Palomino-Morales & Ana Linares

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  1. Ma José Alejandre
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  2. Sonia Perales
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  3. Ángel Carazo
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  4. Rogelio Palomino-Morales
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  5. Ana Linares
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Correspondence to Ana Linares.

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The first two authors listed contributed equally to this work.

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Alejandre, M.J., Perales, S., Carazo, Á. et al. Cyclic fluctuations of 3-hydroxy-3-methylglutaryl-CoA reductase in aortic smooth muscle cell cultures. Lipids 41, 1089–1099 (2006). https://doi.org/10.1007/s11745-006-5058-x

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