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


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

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13
Fig. 14



Acetyl-CoA carboxylase




β-Hydroxyacyl-ACP dehydratase


Dulbecco’s modified Eagle’s medium


Enoyl-ACP reductase


Fatty acid synthase


Free fatty acids


Hydrogen bond acceptors


Hydrogen bond donors


β-Ketoacyl-ACP reductase


β-Ketoacyl-ACP synthase

log P:

Partition coefficient

log Sw :

Water solubility


Malonyl/acetyl-CoA-ACP transferase


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


Molecular weight


Non-alcoholic steatohepatitis


Non-esterified fatty acids


Non-insulin-dependent diabetes mellitus










Topological polar surface area


Very low-density lipoprotein


  1. Ahmadi A, Khalili M, Mashaee F, Nahri-Niknafs B (2017) The effects of solvent polarity on hypoglycemic and hypolipidemic activities of Vaccinium arctostaphylos L. Unripe Fruits Pharma Chem J 50:746–752

    CAS  Article  Google Scholar 

  2. Bedi O, Aggarwal S, Trehanpati N, Ramakrishna G, Krishan P (2019) Molecular and pathological events involved in the pathogenesis of diabetes associated non-alcoholic fatty liver disease. J Clin Exp Hepatol 9:607–618

    Article  Google Scholar 

  3. Benet LZ, Hosey CM, Ursu O, Oprea TI (2016) BDDCS, the rule of 5 and drugability. Adv Drug Deliv Rev 101:89–98

    CAS  Article  Google Scholar 

  4. Bergman RN, Ader M (2000) Free fatty acids and pathogenesis of type 2 diabetes mellitus. Trends Endocrinol Metab 11:351–356

    CAS  Article  Google Scholar 

  5. Berman HM (2000) The protein data bank. Nucleic Acids Res 28:235–242

    CAS  Article  Google Scholar 

  6. Bhakkiyalakshmi E, Sireesh D, Sakthivadivel M, Sivasubramanian S, Gunasekaran P, Ramkumar KM (2016) Anti-hyperlipidemic and anti-peroxidative role of pterostilbene via Nrf2 signaling in experimental diabetes. Eur J Pharmacol 777:9–16

    CAS  Article  Google Scholar 

  7. Bhatt Harikrashna B, Smith RJ (2015) Fatty liver disease in diabetes mellitus. Hepatobiliary Surg Nutr 4:101–108

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Bugianesi E, Gastaldelli A, Vanni E, Gambino R, Cassader M, Baldi S, Ponti V, Pagano G, Ferrannini E, Rizzetto M (2005) Insulin resistance in non-diabetic patients with non-alcoholic fatty liver disease: sites and mechanisms. Diabetologia 48:634–642

    CAS  Article  Google Scholar 

  9. Burnett AR, Thomson RH (1968) Naturally occurring quinones. Part XV. Biogenesis of the anthraquinones in Rubia tinctorum L. (Madder). J Chem Soc 2437–2441

  10. Choi KC, Lee SY, Yoo HJ, Ryu OH, Lee KW, Kim SM, Baik SH, Choi KM (2007) Effect of PPAR-delta agonist on the expression of visfatin, adiponectin, andresistin in rat adipose tissue and 3T3-L1 adipocytes. Biochem Bioph Res Co 357:62–67

  11. Daina A, Michielin O, Swiss ZV (2017) ADME: a free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules. Sci Rep 7:42717

    Article  Google Scholar 

  12. Gupta RK, Krishnamurti M, Parthasarathi J (1980) Purpurin, a new flavanone from Tephrosia purpurea Seeds. Phytochemistry 19:1264

    CAS  Article  Google Scholar 

  13. Hajri T, Han XX, Bonen A, Abumrad NA (2002) Defective fatty acid uptake modulates insulin responsiveness and metabolic responses to diet in CD36-null mice. J Clin Invest 109:1381–1389

    CAS  Article  Google Scholar 

  14. Hsu CL, Lin YJ, Ho CT, Yen GC (2012) Inhibitory effects of garcinol and pterostilbene on cell proliferation and adipogenesis in 3T3-L1 cells. Food Funct 3:49–57

    CAS  Article  Google Scholar 

  15. Khadir F, Pouramir M, Joorsaraee SG, Feizi F, Sorkhi H, Yousefi F (2015) The effect of arbutin on lipid peroxidation and antioxidant capacity in the serum of cyclosporine-treated rats. Caspian J Intern Med 6:196

    PubMed  PubMed Central  Google Scholar 

  16. Kianbakht S, Abasi B, Hashem Dabaghian F (2014) Improved lipid profile in hyperlipidemic patients taking Vaccinium arctostaphylos fruit hydroalcoholic extract: a randomized double-blind placebo-controlled clinical trial. Phytother Res 28:432–436

    CAS  Article  Google Scholar 

  17. Kohjima M, Enjoji M, Higuchi N, Kato M, Kotoh K, Yoshimoto T, Fujino T, Yada M, Yada R, Harada N, Takayanagi R (2007) Re-evaluation of fatty acid metabolism-related gene expression in nonalcoholic fatty liver disease. Int J Mol Med 20:351–358

    CAS  PubMed  Google Scholar 

  18. La Fontaine S, Firth SD, Lockhart PJ, Brooks H, Parton RG, Camakaris J, Mercer JF (1998) Functional analysis and intracellular localization of the human menkes protein (MNK) stably expressed from a cDNA construct in Chinese hamster ovary cells (CHO-K1). Hum Mol Genet 7:1293–1300

    Article  Google Scholar 

  19. Lehninger AL, Nelson DL, Cox MM (2005) Principles of biochemistry.

  20. Lillioja S, Mott DM, Spraul M, Ferraro R, Foley JE, Ravussin E, Knowler WC, Bennett PH, Bogardus C (1993) Insulin resistance as precursor of non-insulin dependent diabetes mellitus. Prospective studies of Pima Indians. N Engl J Med 329:1988–1992

    CAS  Article  Google Scholar 

  21. Lipinski CA (2000) Drug-like properties and the causes of poor solubility and poor permeability. J Pharmacol Toxicol Methods 44:235–249

    CAS  Article  Google Scholar 

  22. Lipinski CA, Lombardo F, Dominy BW, Feeney PJ (2001) Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev 46:3–26

    CAS  Article  Google Scholar 

  23. McGarry JD (1992) What if Minkowski had been ageusic? An alternative angle on diabetes. Science (Wash DC) 258:766–770

    CAS  Article  Google Scholar 

  24. Miteva MA, Guyon F, Tuffïry P (2010) Frog 2: efficient 3D conformation ensemble generator for small compounds. Nucleic Acids Res 38:W622–W627

    CAS  Article  Google Scholar 

  25. Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell DS, Olson AJ (2009) Autodock4 and AutoDockTools4: automated docking with selective receptor flexibility. J Comput Chem 16:2785–2791

    Article  Google Scholar 

  26. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol 65:55–63

    CAS  Google Scholar 

  27. Petkou D, Diamantidis G, Vasilakakis M (2002) Arbutin oxidation by pear (Pyrus communis L.) peroxidases. Plant Sci 162:115–119

    CAS  Article  Google Scholar 

  28. Porras D, Nistal E, Martínez-Flórez S, Pisonero-Vaquero S, Olcoz JL, Jover R, González-Gallego J, García-Mediavilla MV, Sánchez-Campos S (2017) Protective effect of quercetin on high-fat diet-induced non-alcoholic fatty liver disease in mice is mediated by modulating intestinal microbiota imbalance and related gut-liver axis activation. Free Radic Biol Med 102:188–202

    CAS  Article  Google Scholar 

  29. Randle PJ, Garland PB, Hales CN, Newsholme EA (1963) The glucose fatty-acid cycle: its role in insulin sensitivity and the metabolic disturbances of diabetes mellitus. Lancet 281:785–789

    Article  Google Scholar 

  30. Randle PJ, Garland PB, Newsholmet EA, Hales CN (1965) The glucose fatty acid cycle in obesity and maturity onset diabetes mellitus. Ann N Y Acad Sci 131:324–333

    CAS  Article  Google Scholar 

  31. Rose PW, Bi C, Bluhm WF, Christie CH, Dimitropoulos D, Dutta S, Green RK, Goodsell DS, Prlić A, Quesada M, Quinn GB (2013) The RCSB protein data bank: new resources for research and education. Nucleic Acids Res 41:75–182

    Article  Google Scholar 

  32. Satheesh MA, Pari L (2008) Effect of pterostilbene on lipids and lipid profiles in streptozotocin–nicotinamide induced type 2 diabetes mellitus. J Appl Biomed 6:31–37

    CAS  Article  Google Scholar 

  33. Shahaboddin ME, Pouramir M, Moghadamnia AA, Parsian H, Lakzaei M, Mir H (2011) Pyrus biossieriana Buhse leaf extract: an antioxidant, antihyperglycaemic and antihyperlipidemic agent. Food Chem 126:1730–1733

    CAS  Article  Google Scholar 

  34. Takahashi E, Marczylo TH, Watanabe T, Nagai S, Hayatsu H, Negishi T (2001) Preventive effects of anthraquinone food pigments on the DNA damage induced by carcinogens in Drosophila. Mutat Res-Fund Mol M 480:139–145

    Article  Google Scholar 

  35. Takahashi E, Fujita KI, Kamataki T, Arimoto-Kobayashi S, Okamoto K, Negishi T (2002) Inhibition of human cytochrome P450 1B1, 1A1 and 1A2 by antigenotoxic compounds, purpurin and alizarin. Mutat Res-Fund Mol M 508:147–156

    CAS  Article  Google Scholar 

  36. Thakur A, Hamrapurkar P (2009) Quantitative densitometric HPTLC analysis of purpurin in the parts of Rubia cordifolia and in pharmaceutical dosage forms. JPC-J Planar Chromat 22:109–113 

  37. Trott O, Olson AJ (2010) Auto Dock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization and multithreading. J Comput Chem 31:455–461

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Williamson RM, Price JF, Glancy S, Perry E, Nee LD, Hayes PC, Frier BM, Van Look LA, Johnston GI, Reynolds RM, Strachan MW (2011) Prevalence of and risk factors for hepatic steatosis and nonalcoholic fatty liver disease in people with type 2 diabetes: the Edinburgh type 2 diabetes study. Diabetes Care 34:1139–1144

    Article  Google Scholar 

  39. Zengin G, Degirmenci NS, Alpsoy L, Aktumsek A (2016a) Evaluation of antioxidant, enzyme inhibition, and cytotoxic activity of three anthraquinones (alizarin, purpurin, and quinizarin). Hum Exp Toxicol 35:544–553

    CAS  Article  Google Scholar 

  40. Zengin G, Locatelli M, Ceylan R, Aktumsek A (2016b) Anthraquinone profile, antioxidant and enzyme inhibitory effect of root extracts of eight Asphodeline taxa from Turkey: can Asphodeline roots be considered as a new source of natural compounds. J Enzyme Inhib Med Chem 31:754–759

    CAS  Article  Google Scholar 

Download references


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.

Author information




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.

Corresponding author

Correspondence to Pawan Krishan.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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)

Electronic supplementary material


(PDF 80 kb)


(PDF 67 kb)


(PDF 67 kb)


(PDF 69 kb)


(PDF 61 kb)


(PDF 68 kb)


(PDF 68 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

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).

Download citation


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