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Effects of Thymoquinone on Adipocyte Differentiation in Human Adipose-Derived Stem Cells

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Abstract

Inhibition of adipocyte differentiation would be a key strategy to control obesity. Human adipose tissue-derived stem cells (ADSCs) are a promising tool for adipocyte differentiation research. Thymoquinone (TQ) as a potent antioxidant molecule may inhibit adipocyte differentiation. Herein, we aim to investigate the inhibitory effect of TQ on lipid differentiation in ADSCs. Quantification of cell surface markers was used by Flow-Cytometry and the effect of TQ on cell viability was assessed using the AlamarBlue test. ADSCs were subjected to induction of differentiation in the presence of non-cytotoxic concentrations of TQ (6.25, 12.5 and 25 μg/mL). Lipid accumulation was assessed using the Oil-Red O staining technique. Moreover, the expression of PPARγ (Peroxisome proliferator-activated receptor-γ) and FAS (Fatty Acid Synthetase) proteins was evaluated using Western blotting. Flow-cytometry demonstrated the expression of CD44, CD90, and CD73 as mesenchymal stem cell markers on the cell surface. At concentrations ≤100 μg/mL of TQ, no significant difference in cell viability was observed compared to the control. Lipid accumulation in ADSCs significantly decreased at 25 μg/mL (P < 0.001) and 12.5 μg/mL (P < 0.01) of TQ. The findings of the qualitative examination of Lipid Droplets also confirmed these results. Western-blot showed that TQ at 12.5 (p < 0.05) and 25 μg/mL (p < 0.01) reduced FAS/β-actin ratio compared to the positive group. TQ also decreased the expression of PPARγ at 6.25 μg/mL but not at higher concentrations. In conclusion, TQ may reduce differentiation of fat stem cells into fat cells through inhibition of the expression of PPARγ and FAS proteins and might be a potential anti-obesity compound.

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Abbreviations

PPARγ:

Peroxisome proliferator receptor gamma

GAPDH:

Glyceraldehyde phosphate dehydrogenase

FAS:

Fatty Acid Synthetase

TQ:

Thymoquinone

ADSCs:

Adipose tissue-derived stem cells

WAT:

White adipose tissue

BAT:

Brown adipose tissue

SVF:

Stromal vascular fraction

PBS:

Phosphate buffered saline

DMEM:

Dulbecco’s Modified Eagle’s Medium

DMC:

Differentiation medium cocktail

ECL:

Enhanced Chemiluminescent

LDL:

Low-density lipoprotein

HIF1α:

Hypoxia-inducible factor 1α

GSIS:

Stimulated insulin secretion

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Acknowledgements

The authors would like to thank the Mashhad University of Medical Sciences research council for the financial support of the work (Grant No: 950779).

Funding

This study was funded by Research Affairs of Mashhad University of Medical Sciences, Mashhad, Iran (Grant No: 950779).

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Authors and Affiliations

  1. Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Monireh Shahbodi & Zahra Tayarani-Najaran

  2. Department of Traditional Pharmacy, School of Pharmacy, Mashhad University of Medical Sciences, Azadi Square, Pardis University Campus, P.O. Box: 9188617871, Mashhad, Iran

    Seyed Ahmad Emami & Behjat Javadi

  3. Targeted Drug Delivery Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

    Zahra Tayarani-Najaran

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  1. Monireh Shahbodi
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Z.T.N., S.A.E., B.J., and M.S. The first draft of the manuscript was written by M.S. B.J. and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Behjat Javadi or Zahra Tayarani-Najaran.

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Shahbodi, M., Emami, S.A., Javadi, B. et al. Effects of Thymoquinone on Adipocyte Differentiation in Human Adipose-Derived Stem Cells. Cell Biochem Biophys 80, 771–779 (2022). https://doi.org/10.1007/s12013-022-01095-z

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