17β-estradiol and anti-estrogen ICI:Compound 182,780 regulate expression of lipoprotein lipase and hormone-sensitive lipase in isolated subcutaneous abdominal adipocytes

doi:10.1053/meta.2003.50088

Metabolism

Volume 52, Issue 4, April 2003, Pages 383-388

https://doi.org/10.1053/meta.2003.50088 Get rights and content

Abstract

We sought to investigate the influence of 17β-estradiol (E₂) on key enzymes of lipogenesis and lipolysis in subcutaneous (SC) abdominal adipocytes isolated from women. In addition, we wished to determine the influence of an anti-estrogen, ICI:compound 182,780 (anti-E), known to act via the estrogen receptor (ER), alone and in combination with E₂. Adipose tissue was obtained from 17 women undergoing elective surgery, with a mean age of 47 years (range, 34 to 62), mean weight of 65.4 kg (range, 58.1 to 75.0), and mean body mass index (BMI) of 25 kg/m² (range, 22 to 27). Isolated adipocytes were treated with varying doses of E₂, anti-E, or E₂ in combination with anti-E 10⁻⁸ mol/L for 48 hours. Following treatment, proteins were extracted and the effects on lipogenesis and lipolysis were assessed, using Western blotting to determine the relative expression of the key enzymes of these processes, lipoprotein lipase (LPL; 56 kd), and hormone-sensitive lipase (HSL; 84 kd), respectively. Glycerol release into the medium was also measured as an index of lipolytic activity. The protein expression studies demonstrated that E₂ altered expression of LPL relative to control, with the highest dose significantly reducing LPL expression and the lower doses significantly increasing LPL expression (mean protein expression relative to control ± SE): E₂ 10⁻¹² mol/L, 1.79 ± 0.16 (P <.001); E₂ 10⁻⁷ mol/L, 0.56 ± 0.08 (P <.05). In contrast, HSL expression was increased relative to control at the higher doses of E₂ but was not significantly altered relative to control at the lower doses: E₂ 10⁻¹² mol/L, 1.02 ± 0.14 (P >.05); E₂ 10⁻⁷ mol/L, 1.55 ± 0.17 (P <.01). Anti-E 10⁻⁸ mol/L alone reduced LPL protein expression relative to control (P <.05) and increased HSL protein expression relative to control (P >.05). In combination with E₂ 10⁻⁷ mol/L, anti-E 10⁻⁸ mol/L did not abrogate the inhibitory effect on LPL expression relative to control (P <.05). Furthermore, E₂ 10⁻⁷ mol/Lin combination with anti-E 10⁻⁸ mol/L, displayed a stimulatory effect on HSL expression relative to control (P <.01). Glycerol release studies following the higher doses of E₂, and also following E₂ 10⁻⁷ mol/L in combination with anti-E 10⁻⁸ mol/L, provided support for the HSL protein expression studies. We conclude that the highest concentration of E₂ (10⁻⁷ mol/L) significantly reduced LPL expression relative to control, while the lower concentrations significantly increased LPL expression relative to control. The highest concentration of E₂ also significantly increased both HSL expression and glycerol release relative to control. The effects of anti-E suggest that the in vitro effects of E₂ on lipogenesis and lipolysis occur, at least in part, through a receptor-mediated pathway. In addition, as recently observed in other tissues, ICI:compound 182,780 does not appear to behave as a pure anti-estrogen in isolated human adipocytes. Copyright 2003 Elsevier, Inc. All rights reserved.

Section snippets

Subjects

SC abdominal adipose tissue was obtained from 17 women undergoing elective abdominal surgery, with the majority of cases being performed for cosmetic reasons. The mean age for the group was 47 years (range, 34 to 62), mean weight was 65.4 kg (range, 58.1 to 75.0), and mean body mass index (BMI) was 25 kg/m² (range, 22 to 27). Subjects using endocrine therapy such as corticosteroids, thyroxine, and either the oral contraceptive pill or HRT were excluded, as were those on lipid-lowering

Effect of E₂

Increasing concentrations of E₂ altered LPL protein expression relative to control, with the highest dose (E₂10⁻⁷ mol/L) significantly reducing LPL protein expression and the lower doses increasing LPL protein expression; mean protein expression relative to control ± SE: E₂ 10⁻¹² mol/L, 1.79 ± 0.16 (P <.001); E₂ 10⁻⁷ mol/L, 0.56 ± 0.08 (P <.05) (Fig 1A).

. Mean relative protein expression (± SE) of E₂ treatments from 10⁻¹² mol/L (M) to 10⁻⁷ mol/L compared to control for (A) LPL (n = 5) and (B) HSL

Discussion

Human epidemiological and prospective clinical studies suggest estrogen deficiency is associated with the development of central obesity, which may be limited or reversed by HRT.6, 7, 29 Our findings provide the first in vitro evidence that E₂ has a regulatory effect on key enzymes of lipogenesis and lipolysis in SC abdominal adipose tissue from women. These results suggest that E₂ may regulate adipose tissue mass by influencing the net amount of adipose tissue present in the adipocyte. Taken

Acknowledgements

We thank all the surgeons and theatre staff at the University Hospitals Trust and the Priory Hospital, Edgbaston, Birmingham who aided in the studies. We especially thank Dr Allan Green of Bassett Healthcare Research Institute, Cooperstown, NY, for providing the HSL antibody.

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^*: Address reprint requests to Dr S.L. Palin, Diabetes Centre (Ward 34), Birmingham Heartlands Hospital, Bordesley Green East, Birmingham B9 5SS, UK.

^**: 0026-0495/03/5204-0027$30.00/0

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17β-estradiol and anti-estrogen ICI:Compound 182,780 regulate expression of lipoprotein lipase and hormone-sensitive lipase in isolated subcutaneous abdominal adipocytes*,**

Abstract

Section snippets

Subjects

Effect of E2

Discussion

Acknowledgements

Am J Clin Nutr

Metabolism

Metabolism

Metabolism

J Steroid Biochem

Am J Obstet Gynecol

Fertil Steril

Biochem Biophys Res Commun

Obesity as a medical problem

Nature

Relationship of body fat distribution to metabolic complications of obesity

J Clin Endocrinol Metab

Abdominal adipose tissue distribution, obesity and risk of cardiovascular disease and death: 13 year follow up of participants in the study of men born in 1913

BMJ

Menopause and risk of cardiovascular disease—The Framingham Study

Ann Intern Med

Body weight, body fat distribution, and hormonal replacement therapy in early postmenopausal women

J Clin Endocrinol Metab

Regulation of adipose cell number in man

Clin Sci

Diet-induced adipocyte number increase in adult rats: A new model of obesity

Am J Physiol

Lipoprotein lipase—A multifunctional enzyme relevant to common metabolic diseases

N Engl J Med

Adipocyte hormone-sensitive lipase: A major regulator of lipid metabolism

Proc Nutr Soc

Regional adiposity and morbidity

Physiol Rev

Promotion of human adipocyte precursor replication by 17β-estradiol in culture

J Clin Invest

Opposite effects of androgens and estrogens on adipogenesis in rat preadipocytes: Evidence for sex and site-related specificities and possible involvement of insulin-like growth factor 1 receptor and peroxisome proliferator-activated receptor γ2*

Endocrinol

Effect of E₂