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. 2014 Oct 15;19(10):16656-71.
doi: 10.3390/molecules191016656.

Anti-obesity Effects of Hispidin and Alpinia Zerumbet Bioactives in 3T3-L1 Adipocytes

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Anti-obesity Effects of Hispidin and Alpinia Zerumbet Bioactives in 3T3-L1 Adipocytes

Pham Thi Be Tu et al. Molecules. .
Free PMC article

Abstract

Obesity and its related disorders have become leading metabolic diseases. In the present study, we used 3T3-L1 adipocytes to investigate the anti-obesity activity of hispidin and two related compounds that were isolated from Alpinia zerumbet (alpinia) rhizomes. The results showed that hispidin, dihydro-5,6-dehydrokawain (DDK), and 5,6-dehydrokawain (DK) have promising anti-obesity properties. In particular, all three compounds significantly increased intracellular cyclic adenosine monophosphate (cAMP) concentrations by 81.2% ± 0.06%, 67.0% ± 1.62%, and 56.9% ± 0.19%, respectively. Hispidin also stimulated glycerol release by 276.4% ± 0.8% and inhibited lipid accumulation by 47.8% ± 0.16%. Hispidin and DDK decreased intracellular triglyceride content by 79.5% ± 1.37% and 70.2% ± 1.4%, respectively, and all three compounds inhibited glycerol-3-phosphate dehydrogenase (GPDH) and pancreatic lipase, with hispidin and DDK being the most potent inhibitors. Finally, none of the three compounds reduced 3T3-L1 adipocyte viability. These results highlight the potential for developing hispidin and its derivatives as anti-obesity compounds.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Conversion of 5,6-dehydrokawain (DK) to hispidin.
Figure 2
Figure 2
The analytical high performance liquid chromatography (HPLC) chromatograms of A. zerumbet rhizomes (A) DK, (B) dihydro-5,6-dehydrokawain (DDK). HPLC column: ODS-100Z column (5 µm, 150 × 46 mm i.d.). Flow rate: 0.8 mL/min. Injection volume: 5 µL. Monitored wavelength: 280 nm.
Figure 3
Figure 3
The effect of hispidin, DK and DDK on cell viability in 3T3-L1 adipocytes. Differentiated 3T3-L1 cells were treated with various concentrations of isolated compounds and incubated for 72 h at 37 °C in a humidified incubator containing 5% CO2. Data are expressed as the percent growth rate of cells cultured in the presence of samples compared with untreated control cells (mean ± SE; n = 3). Letters with different superscripts indicate samples that are significantly different (p < 0.05) than the control.
Figure 4
Figure 4
The effect of hispidin, DK and DDK on glycerol release in 3T3-L1 adipocytes. Differentiated 3T3-L1 cells were treated with various concentrations of isolated compounds and incubated for 72 h at 37 °C in a humidified incubator containing 5% CO2. Cell supernatants were collected and free glycerol release was assayed. Reported values are the means ± SE (n = 3). Letters with different superscripts indicate samples that are significantly different (p < 0.05) compared with the control.
Figure 5
Figure 5
The effect of hispidin, DK and DDK on intracellular cAMP concentrations in 3T3-L1 adipocytes. Differentiated 3T3-L1 cells were treated with various concentrations of isolated compounds and incubated for 72 h at 37 °C in a humidified incubator containing 5% CO2. Cell supernatants were collected for cAMP assay. Reported values are the means ± SE (n = 3). Letters with different superscripts indicate samples that are significantly different (p < 0.05) than the control.
Figure 6
Figure 6
The effect of hispidin, DK and DDK on lipid accumulation in 3T3-L1 adipocytes. (A) Effects of hispidin, DK and DDK on lipid droplet formation in 3T3-L1 adipocytes assessed by staining with Oil Red O dye and visualization light microscopy. (B) Relative lipid content of each sample determined by quantitative analysis of Oil Red O content. Reported values are the means ± SE (n = 3). Letters with different superscripts indicate samples that are significantly different (p < 0.05) than the control.
Figure 7
Figure 7
The effect of hispidin, DK and DDK on the inhibition of intracellular triglyceride content in 3T3-L1 adipocytes. Cells were treated with 100 and 250 µg/mL tested compounds for 72 h at 37 °C in a humidified incubator containing 5% CO2. Reported values are the mean ± SE (n = 3). Letters with different superscripts indicate samples that are significantly different (p < 0.05) than the control.
Figure 8
Figure 8
The effect of the hispidin, DK and DDK on the inhibition of GPDH activity in 3T3-L1 adipocytes. Cells were treated with 100 and 250 µg/mL tested compounds for 72 h and incubated at 72 h at 37 °C in a humidified incubator containing 5% CO2. Reported values are the mean ± SE (n = 3). Letters with different superscripts indicate samples that are significantly different (p < 0.05) than the control.
Figure 9
Figure 9
The effect of hispidin, DK and DDK on pancreatic lipase activity. Reported values are the mean ± SE (n = 3). Letters with different superscripts indicate samples that are significantly different (p < 0.05) than the control.

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