In vitro targeted screening and molecular docking of stilbene, quinones, and flavonoid on 3T3-L1 pre-adipocytes for anti-adipogenic actions

Abstract

In metabolic disorders like obesity, NAFLD and T2DM, adipocytes are dysfunctional. Hence, pharmacological interventions have importance in preventing differentiation of adipocytes and stimulating lipid uptake. We, therefore, investigated the effects of arbutin (ARB), purpurin (PUR), quercetin (QR), and pterostilbene (PTS) on adipocyte differentiation and lipid uptake using 3T3-L1 adipocytes. Further, in silico docking studies were achieved to investigate interactions of ARB, PUR, QR, and PTS with beta-ketoacyl reductase (KR) and thioesterase (TE) domains of fatty acid synthase (FAS) enzyme. Mature 3T3-L1 adipocytes were used to investigate the anti-adipogenic effect of selected pharmacological agents by Oil Red O staining and in vitro fatty acid uptake analysis. Molecular docking studies were performed to predict the binding interactions of selected compounds with KR and TE domains of FAS enzyme. All these agents significantly decrease the adipocyte differentiation and showed the stimulatory effect on fatty acid uptake in 3T3-L1 adipocytes. However, PTS and PUR proved to be anti-adipogenic, whereas ARB and QR showed significant effect on fatty acid uptake, compared to others. Similarly, all the compounds displayed significant binding interactions with KR and TE domains of FAS enzyme, supporting the results of in vitro studies.

Graphical abstract

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Abbreviations

ACC:

Acetyl-CoA carboxylase

ARB:

Arbutin

DH:

β-Hydroxyacyl-ACP dehydratase

DMEM:

Dulbecco’s modified Eagle’s medium

ER:

Enoyl-ACP reductase

FAS-I:

Fatty acid synthase

FFA:

Free fatty acids

HBA:

Hydrogen bond acceptors

HBD:

Hydrogen bond donors

KR:

β-Ketoacyl-ACP reductase

KS:

β-Ketoacyl-ACP synthase

log P:

Partition coefficient

log Sw :

Water solubility

MAT:

Malonyl/acetyl-CoA-ACP transferase

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

MW:

Molecular weight

NASH:

Non-alcoholic steatohepatitis

NEFA:

Non-esterified fatty acids

NIDDM:

Non-insulin-dependent diabetes mellitus

PTS:

Pterostilbene

PUR:

Purpurin

QR:

Quercetin

TE:

Thioesterase

tPSA:

Topological polar surface area

VLDL:

Very low-density lipoprotein

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Acknowledgments

The authors are thankful to DST-SERB, New Delhi, for providing financial assistance for the accomplished research work. We also acknowledge the Department of Pharmaceutical Sciences & Drug Research and SIC Punjabi University, Patiala, for extending necessary research facilities for this work. We are privileged to extend our thanks to ILBS, New Delhi, for providing continuous lab support for conducting experiments. We also extend our thanks to the management members of Chitkara University, Punjab, for their continuous support. We also show our gratitude to Sami Labs Limited, Bangalore, India for providing the gift sample for experimental research work.

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OB and SA: conducted experiments and wrote the manuscript. NTP and GK: conceived and designed research. ASG: conducted molecular dynamics studies. PK: analyzed data and designed research. All authors read and approved the manuscript. It is also declared that authors did not use a paper mill and all experimental work was performed in-house as per respective authors’ experimental laboratories. All authors reviewed the manuscript.

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Correspondence to Pawan Krishan.

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Chemical compounds studied in this article

Purpurin (PubChem CID: 6683)

Arbutin (PubChem CID: 440936)

Quercetin (PubChem CID: 5280343)

Dexamethasone (PubChem CID: 5743)

Insulin (PubChem CID: 70678557)

3-Isobutyl-1-methylxanthine (PubChem CID: 3758)

Pterostilbene (PubChem CID: 5281727)

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Bedi, O., Aggarwal, S., Trehanpati, N. et al. In vitro targeted screening and molecular docking of stilbene, quinones, and flavonoid on 3T3-L1 pre-adipocytes for anti-adipogenic actions. Naunyn-Schmiedeberg's Arch Pharmacol (2020). https://doi.org/10.1007/s00210-020-01919-w

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Keywords

  • 3T3-L1 adipocytes
  • Arbutin (ARB)
  • Purpurin (PUR)
  • Quercetin (QR)
  • Pterostilbene (PTS)