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

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 microg/mL) or 25-hydroxycholesterol (0.1-10 microg/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 microg/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.

Hormone-sensitive lipase - new roles for an old enzyme

“…Sterol esters appear to down-regulate the expression of HSL in mouse macrophages [124], a phenomenon recently observed also in 3T3-L1 adipocytes [125], while chronic exposure to elevated concentrations of glucose and insulin and/or leptin cause decreased activity of HSL, but increased activity of ACAT, perhaps contributing to increased foam cell formation in hyperinsulinaemia, hyperleptinaemia and hyperglycaemia [112]. Acute treatment of mouse macrophage cell lines with leptin and insulin also influences the activity of nCEH, as measured in anti-HSL immunoprecipitates.…”