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. 2017 Feb;61(2):1600574.
doi: 10.1002/mnfr.201600574. Epub 2016 Dec 6.

Metabolic Programming of a Beige Adipocyte Phenotype by Genistein

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Free PMC article

Metabolic Programming of a Beige Adipocyte Phenotype by Genistein

Sadat A Aziz et al. Mol Nutr Food Res. .
Free PMC article

Abstract

Scope: Promoting the development of brown or beige adipose tissue may protect against obesity and related metabolic features, and potentially underlies protective effects of genistein in mice.

Methods and results: We observed that application of genistein to 3T3-L1 adipocytes changed the lipid distribution from large droplets to a multilocular distribution, reduced mRNAs indicative of white adipocytes (ACC, Fasn, Fabp4, HSL, chemerin, and resistin) and increased mRNAs that are a characteristic feature of brown/beige adipocytes (CD-137 and UCP1). Transcripts with a role in adipocyte differentiation (Cebpβ, Pgc1α, Sirt1) peaked at different times after application of genistein. These responses were not affected by the estrogen receptor (ER) antagonist fulvestrant, revealing that this action of genistein is not through the classical ER pathway. The Sirt1 inhibitor Ex-527 curtailed the genistein-mediated increase in UCP1 and Cebpβ mRNA, revealing a role for Sirt1 in mediating the effect. Baseline oxygen consumption and the proportional contribution of proton leak to maximal respiratory capacity was greater for cells exposed to genistein, demonstrating greater mitochondrial uncoupling.

Conclusions: We conclude that genistein acts directly on adipocytes or on adipocyte progenitor cells to programme the cells metabolically to adopt features of beige adipocytes. Thus, this natural dietary agent may protect against obesity and related metabolic disease.

Keywords: Brown adipose tissue; Estrogen receptor; Genistein; SIRT1; White adipose tissue.

Figures

Figure 1
Figure 1
3T3‐L1 cells on day 12 after induction of differentiation to adipocytes in the presence of genistein at the concentrations indicated stained using oil red O to reveal the distribution of intracellular lipid. Scale bar 100 μM.
Figure 2
Figure 2
The effect of genistein (10, 50, or 100 μM) in 3T3‐L1 cells on mRNAs corresponding to genes expressed at high levels in white adipocytes (Panel A), genes that play a role in stimulating the development of brown/beige adipocytes (Panel B), or genes expressed preferentially in brown/beige compared with white adipocytes (Panel C). Genistein was applied on induction of differentiation (d 0) and measurements were made at d 12. Data are expressed relative to the reference genes Topo1 and NONO and normalised to control. Values are mean ± SEM for n = 3–15 (derived from 3 replicates (wells) from 1 to 5 independent experiments). Statistical analysis was by one‐way ANOVA followed by Dunnett's test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. The inset panel in panel B shows a representative western blot of protein extracted from cells at day 10 of treatment with 100 μM genistein and from control cells probed with antibodies against Sirt1 and α‐tubulin. Panel D provides an overview of the pattern of responses of these different categories of genes in the form of a heat map, where red shading indicates an increased quantity of mRNA, yellow shading indicates no difference from control and green shading indicates a reduced quantity of mRNA.
Figure 3
Figure 3
The effect of genistein on the proportional contribution of proton leak to basal and maximal respiratory capacity in 3T3‐L1 cells. The oxygen consumption rate by cells was measured on day 12 after induction of differentiation under control conditions or in the presence of 100 μM genistein. Data are from n = 10 wells for each condition. Statistical analysis was by Student's t‐test. **p ≤ 0.01.
Figure 4
Figure 4
The effect in 3T3‐L1 cells of the estrogen receptor antagonist fulvestrant on responses to genistein of mRNAs corresponding to genes expressed at high levels in white adipocytes (Panel A) or genes that play a role in stimulating the development of brown/beige adipocytes or that are expressed preferentially in brown/beige compared with white adipocytes (Panel B). Genistein (100 μM), with or without fulvestrant (0.1 μM), as indicated, was applied on induction of differentiation (day 0) and measurements were made on day 12. Data are expressed relative to the reference genes Topo1 and NONO and normalised to control. Values are mean ± SEM for n = 3. Statistical analysis was by one‐way ANOVA followed by Bonferroni’ multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.
Figure 5
Figure 5
The effect in 3T3‐L1 cells of the Sirt1 inhibitor Ex‐527 on responses to genistein of mRNAs corresponding to genes expressed at high levels in white adipocytes (Panel A) or genes that play a role in stimulating the development of brown/beige adipocytes or that are expressed preferentially in brown/beige compared with white adipocytes (Panel B). Genistein (100 μM), with or without Ex‐527 (10 μM), as indicated, was applied on induction of differentiation (day 0) and measurements were made on day 12. Data are expressed relative to the reference genes Topo1 and NONO and normalised to control. Values are mean ± SEM for n = 3. Statistical analysis was by one‐way ANOVA followed by Bonferroni’ multiple comparisons test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.

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