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Sterol-mediated regulation of hormone-sensitive lipase in 3T3-L1 adipocytes

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Lipids

Abstract

We previously reported that intracellular free cholesterol at physiological concentrations regulates the activity of neutral cholesterol esterase (N-CEase) in macrophages. The objective of the present study is to investigate whether the regulation of N-CEase by cholesterol is generally observed in other types of cells such as adipocytes with high activity of hormone-sensitive lipase (HSL), the same gene product as N-CEase. 3T3-L1 adipocytes were cultured with and without cholesterol (1–30 μg/mL) or 25-hydroxycholesterol (0.1–10 μg/mL), and changes in the N-CEase activity, expression of HSL mRNA, and protein were examined. Incubation (24 h) of cells with cholesterol did not change N-CEase activity, but incubation with 25-hydroxycholesterol decreased the activity in a concentration-dependent manner by 24 (24 h) and 54% (36 h). Quantitative reverse transcription-PCR indicated that 25-hydroxycholesterol (10 μg/mL) did not influence expression of HSL mRNA. However, Western blot analysis showed that this sterol reduced HSL protein by 72 (24 h) and by 93% (36 h), respectively. It was concluded that sterol-mediated regulation of HSL/N-CEase occurs not only in macrophages but also in adipocytes, and regulation appears to occur not at a transcriptional level but by a post-transcriptional process. Sterol-mediated proteolysis may be involved in the loss of HSL protein.

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Abbreviations

ACAT:

acyl-CoA: cholesterol acyltransferase

CE:

cholesteryl ester

CS:

calf serum

ECL:

enhanced chemiluminescence

HSL:

hormone-sensitive lipase

LPDS:

lipoportein-deficient serum

N-CEase:

neutral cholesterol esterase

RT:

reverse transcription

SCAP:

SREBP cleavage-activating protein

SRE:

sterol regulatory element

SREBP:

sterol regulatory element-binding protein

SSD:

sterol-sensing domain

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Authors and Affiliations

  1. School of Pharmaceutical Sciences, University of Shizuoka, 422, Shizuoka, Japan

    Shinji Miura, Hiromi Nagura, Fusae Sawamura & Isao Tomita

  2. Graduate School of Health Sciences, University of Shizuoka, 422, Shizuoka, Japan

    Eiji Kawai, Norihiro Mochizuki, Masahiko Ikeda & Takako Tomita

  3. Department of Veterans Affairs Medical Center, 94304, Palo Alto, California

    Fredric B. Kraemer

  4. Division of Endocrinology, Gerontology, and Metabolism, Department of Medicine, Stanford University School of Medicine, 94305, Stanford, California

    Fredric B. Kraemer

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  1. Shinji Miura
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  2. Hiromi Nagura
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  3. Fusae Sawamura
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  4. Isao Tomita
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Correspondence to Shinji Miura.

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Miura, S., Nagura, H., Sawamura, F. et al. Sterol-mediated regulation of hormone-sensitive lipase in 3T3-L1 adipocytes. Lipids 38, 743–750 (2003). https://doi.org/10.1007/s11745-003-1122-9

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  • DOI: https://doi.org/10.1007/s11745-003-1122-9

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