In vitro and In silico Evaluation of the Oestrogenic Activities of Some Edible Vegetables in Nigeria
Article Sidebar
Main Article Content
Abstract
Breast cancer is the most common malignancy in women worldwide with an estimated 2.3 million new cases and 1,380,000 deaths. The steroid hormone; 17-β estradiol plays a significant role in the growth and development of the breast epithelium, and is implicated in the aetiology and progression to breast cancer. The phytoestrogens are plant-derived compounds that mimic endogenous oestrogens, however, reports on their beneficial and adverse health effects remain inconclusive. Therefore, this study aimed to evaluate the oestrogenic activities of some edible vegetables using in vitro and in silico assays. The leaves of Gnetum africanum (GA), Lasianthera Africana (LA), and Vernonia amygdalina (VA) were extracted by cold maceration with ethanol. The Yeast Estrogen Screen (YES) which employed Recombinant Saccharomyces cerevisiae W303 expressing oestrogen receptors (ERs; α and β) was used to assess the oestrogenic potency of the extracts. The in silico investigations involved molecular docking of selected phytochemicals on ERs and prediction of ADMET properties using SwissADME webserver. The YES revealed the following Estradiol Equivalents (EEQs); ERα: LA: 154.05μg/g; VA: 130.20μg/g; GA: 109.40μg/g, and ERβ: LA: 9.26μg/g; VA: 6.22μg/g, GA: 5.73μg/g. Molecular docking studies showed negative binding affinities ranging from -3.3 to -9.0 Kcal/mol, while the SwissADME predictions revealed four compounds with good ADMET properties, interacting with ERα binding pocket similarly to 17-β estradiol. Knowledge of the oestrogenic potencies of edible vegetables is vital as this would help promote safer consumption habits and inform preventive strategies against ER-positive breast cancer.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
1. Bray F, Laversanne M, Sung H, Ferlay J, Siegel RL, Soerjomataram I, Jemal A. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2024; 74(3):229-263.
2. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021; 71(3):209-249.
3. Olayide A, Isiaka A, Ganiyu R, Samuel O, Halimat A, Olalekan O, Julius O, Anya R. Breast Cancer Treatment and Outcomes in Nigeria: A Systematic Review and Meta-analysis. Asian Pac J Cancer Care. 2023; 8 (3):591-598
4. Clusan L, Ferrière F, Flouriot G, Pakdel F. A Basic Review on Estrogen Receptor Signaling Pathways in Breast Cancer. Int J Mol Sci. 2023; 24(7):6834.
5. Liang Y, Zhang H, Song X, Yang Q. Metastatic heterogeneity of breast cancer: Molecular mechanism and potential therapeutic targets. Semin Cancer Biol. 2020; 60:14-27.
6. Starek-Świechowicz B, Budziszewska B, Starek A. Endogenous estrogens-breast cancer and chemoprevention. Pharmacol Rep. 2021; 73(6):1497-1512.
7. Patra S, Gorai S, Pal S, Ghosh K, Pradhan S, Chakrabarti S. A review on phytoestrogens: Current status and future direction. Phytother Res. 2023; 37(7):3097-3120.
8. Rietjens IMCM, Louisse J, Beekmann K. The potential health effects of dietary phytoestrogens. Br J Pharmacol. 2017; 174(11):1263-1280.
9. Bennetau-Pelissero C. Risks and benefits of phytoestrogens: where are we now? Curr Opin Clin Nutr Metab Care. 2016; 19(6):477-483.
10. Morgan HE, Dillaway D, Edwards TM. Estrogenicity of soybeans (Glycine max) varies by plant organ and developmental stage. Endocr Disruptors. 2014; 2(1): 28490-28498
11. Domínguez-López I, Yago-Aragón M, Salas-Huetos A, Tresserra-Rimbau A, Hurtado-Barroso S. Effects of Dietary Phytoestrogens on Hormones throughout a Human Lifespan: A Review. Nutrients. 2020; 12(8):2456.
12. Canivenc-Lavier MC, Bennetau-Pelissero C. Phytoestrogens and Health Effects. Nutrients. 2023; 15(2):317.
13. Rizzo G, Feraco A, Storz MA, Lombardo M. The role of soy and soy isoflavones on women's fertility and related outcomes: an update. J Nutr Sci. 2022; 11: e17.
14. Udoh FV, Eyong UE, Udoh PE, Ebong PE. Dynamic Activity of the Ethanolic and Water Extracts of the Leaf of Gnetum africanum Repeated Treatment on Uterine Muscle Morphology of Rats. Reprod Contracept. 2011; 22(3): 169-175.
15. Agradi E, Vegeto E, Sozzi A, Fico G, Regondi S, Tome F. Traditional healthy Mediterranean diet: estrogenic activity of plants used as food and flavoring agents. Phytother Res. 2006; 20(8): 670-675
16. Wagner M, Oehlmann J. Endocrine disruptors in bottled mineral water: estrogenic activity in the E-Screen. J Steroid Biochem Mol Biol. 2011; 127(1-2):128-135.
17. Akanbi AI, Izevbigie EV, Sherif AO, Dlamini NH, Fadele LO, Oyawaluja BO. Molecular Docking, ADME and SAR Analysis of 383 Phytochemicals in the Quest for Lead Antidiabetic Inhibitors Targeting α-Amylase and α-Glucosidase Enzymes. Trop J Drug. 2025; 2 (1): 6-13
18. Shodehinde SA, Oyeleye SI, Olasehinde TA, Adebayo AA, Oboh G, Boligon AA. Lasianthera Africana leaves inhibits α-amylase α-glucosidase, angiotensin-I converting enzyme activities and Fe2+-induced oxidative damage in pancreas and kidney homogenates. Orient Pharm Exp Med. 2017; 17(1): 41–49.
19. Akpaso MI, Atangwho IJ, Akpantah A, Fischer VA, Igiri AO, Ebong PE. Effect of Combined Leaf Extracts of Vernonia amygdalina (Bitter Leaf) and Gongronema latifolium (Utazi) on the Pancreatic β-Cells of Streptozotocin-Induced Diabetic Rats. Br J Med Med Res. 2011; 1(1), 24–34.
20. Farombi EO, Owoeye O. Antioxidative and chemopreventive properties of Vernonia amygdalina and Garcinia biflavonoid. Int J Environ Res Public Health. 2011; 8(6): 2533–2555.
21. Lawal BAS, Odoala FK. Drug-Phytochemical Interaction: In vitro Investigation of the Effects of Aframomum melegueta Seed Extract on Acetaminophen and Amlodipine Absorption. Trop J Phytochem Pharm Sci. 2024; 3(6): 278-283.
22. Lawal BAS, Odoala FK. Interference in drug assay by phytochemicals: An experience with colorimetric assay of amlodipine in physiological fluids. Trop J Pharm Res. 2023; 22(11):2327-2332.
23. Odoala FK, Lawal BAS. Extract from the Seeds of Aframomum melegueta Alters Acetaminophen Oral Bioavailability in Sprague Dawley Rats. Trop J Nat Prod Res. 2023; 7(8): 3819-3822.
24. Edwards TM, Morgan HE, Balasca C, Chalasani NK, Yam L, Roark AM. Detecting Estrogenic Ligands in Personal Care Products using a Yeast Estrogen Screen Optimized for the Undergraduate Teaching Laboratory. J Vis Exp 2018; (131): 55754.
25. Miller III CA, Tan X, Wilson M, Bhattacharyya S, Ludwig S. Single plasmids expressing human steroid hormone receptors and a reporter gene for use in yeast signaling assays. Plasmid. 2010; 63(2): 73–78.
26. Edet UU, Ehiabhi OS, Ogunwande IA, Walker TM, Schmidt JM, Setzer WN, Ogunbinu AO, Ekundayo O. Analyses of the Volatile Constituents and Antimicrobial Activities of Gongronema Latifolium (Benth.) and Gnetum africanum L. Jeobp. 2013; 8(3): 324 – 329.
27. Ezekwe AS, Ugwuezumba PC, Nwankpa P, Egwurugwu JN, Ekweogu CN, Emengaha FC, Akukwu D. Qualitative Phytochemical Screening, GCMS Studies and In-Vitro Anti-Oxidative Properties of Aqeuous Leaf Extract of Gnetum africanum. J Drug Deliv Ther. 2020; 10(1): 11 – 17.
28. Ekpo DE, Joshua PE, Ogidigo JO, Nwodo OFC. High-resolution UPLC-PDA-QTOF-ESI-MS/MS analysis of the flavonoid-rich fraction of Lasianthera africana leaves and in vivo evaluation of its renal and cardiac function effects. Heliyon. 2020; 6(7): e04154.
29. Obia C, Onyegeme-Okerenta BM, Monago-Igorodje CC. Evaluation of Bioactive Compounds and Antioxidant Activities of Dichloromethane Leaf Extracts of Ficus richocarpa and Lasianthera africana. S Afr J Bot. 2023; 22: 181-194.
30. Luo X, Jiang Y, Fronczek FR, Lin C, Izevbigie EB, Lee KS. Isolation and structure determination of a sesquiterpene lactone (vernodalinol) from Vernonia amygdalina extracts. Pharm Biol. 2011; 49(5):464-470.
31. Alara OR, Abdurahman NH, Ukaegbu CI. Kabbashi NA. Extraction and characterization of bioactive compounds in Vernonia amygdalina leaf ethanolic extract comparing Soxhlet and microwave-assisted extraction techniques. J Taibah Univ Sci. 2019; 13(1): 414–422.
32. Ijoma IK, Okafor CE, Ajiwe VIE. Computational Studies of 5- methoxypsolaren as Potential Deoxyhemoglobin S Polymerization Inhibitor. Trop J Nat Prod Res. 2024; 8(10): 8835 – 8841 https://doi.org/10.26538/tjnpr/v8i10.28
33. Sotoca AM, Ratman D, van der Saag P, Ström A, Gustafsson JA, Vervoort J, Rietjens IM, Murk AJ. Phytoestrogen-mediated inhibition of proliferation of the human T47D breast cancer cells depend on the ERalpha/ERbeta ratio. J Steroid Biochem Mol Biol. 2008; 112(4-5):171-178.
34. Buller CJ, Zang XP, Howard EW, Pento JT. Measurement of beta-galactosidase tissue levels in a tumor cell xenograft model. Methods Find Exp Clin Pharmacol. 2003; 25(9):713-716.
35. Uchil PD, Nagarajan A, Kumar P. Assay for β-Galactosidase in Extracts of Mammalian Cells. Cold Spring Harb Protoc. 2017(10):pdb.prot095778.
36. Anstead GM, Carlson KE, Katzenellenbogen JA. The estradiol pharmacophore: ligand structure-estrogen receptor binding affinity relationships and a model for the receptor binding site. Steroids. 1997; 62 (3):268-303.
37. Brzozowski AM, Pike AC, Dauter Z, Hubbard RE, Bonn T, Engström O, Ohman L, Greene GL, Gustafsson JA, Carlquist M. Molecular basis of agonism and antagonism in the oestrogen receptor. Nature. 1997; 389(6652):753-758.
38. Masand VH, Al-Hussain SA, Alzahrani AY, Al-Mutairi AA, Hussien RA, Samad A, Zaki MEA. Estrogen Receptor Alpha Binders for Hormone-Dependent Forms of Breast Cancer: e-QSAR and Molecular Docking Supported by X-ray Resolved Structures. ACS Omega. 2024; 9(14):16759-16774.
39. Nursamsiar SM, Awaluddin A, Nurnahari N, Nur S, Febrina E, Asnawi A. Molecular docking and molecular dynamic simulation of the aglycone of curculigoside a and its derivatives as alpha-glucosidase inhibitors. Rasayan J Chem. 2020; 13(1):690-698.
40. Murugesan DK, Rajagopal K, Vijayakumar AR, Sundararajan G, Raman K, Byran G, Murugesan E, Gupta JK, Kankate RS, Nainu F, Barua R, Ahmad SF, Emran TB. Design and Synthesis of Pyrazole-Substituted 9-Anilinoacridine Derivatives and Evaluation against Breast Cancer. J. Biol Regul Homeost Agents. 2024; 38(4): 2845–2859
41. Nursamsiar NS, Febrina E, Asnawi A, Syafiie S. Synthesis and Inhibitory Activity of Curculigoside A Derivatives as Potential Anti-Diabetic Agents with β-Cell Apoptosis. J Mol Struct. 2022; 1265:133292.
42. Chen D, Oezguen N, Urvil P, Ferguson C, Dann SM, Savidge TC. Regulation of protein-ligand binding affinity by hydrogen bond pairing. Sci Adv. 2016; 2(3): e1501240.
43. Kalaimathi RV, Krishnaveni K, Murugan M, Basha AN, Gilles A, Kandeepan C, Senthilkumar N, Mathialagan B, Ramya S, Ramanathan L, Jayakumararaj R, Loganathan T, Pandiarajan G, Ram CJ. ADMET informatics of Tetradecanoic acid (Myristic Acid) from ethyl acetate fraction of Moringa oleifera leaves. J Drug Deliv Ther. 2022; 12(4-S):101-111.
44. Mak K, Kamal MB, Ayuba SB, Sakirolla R, Kang Y, Mohandas K, Balijepalli MK, Ahmad SH, Pichika MR. A Comprehensive Review on Eugenol’s Antimicrobial Properties and Industry Applications: A Transformation from Ethnomedicine to Industry. Pharmacogn Rev. 2019; 13:25.
45. Mehta S, Kadian V, Dalal S, Dalal P, Kumar S, Garg M, Rao R. A Fresh Look on Bergenin: Vision of Its Novel Drug Delivery Systems and Pharmacological Activities. Future Pharmacol. 2022; 2:64–91
46. Djeujo FM, Stablum V, Pangrazzi E, Ragazzi E, Froldi G. Luteolin and Vernodalol as Bioactive Compounds of Leaf and Root Vernonia amygdalina. Extracts: Effects on α-Glucosidase, Glycation, ROS, Cell Viability, and In Silico ADMET Parameters.Int J Pharm. 2023; 15:1541.
47. Wu W, Han X, Wu C, Wei G, Zheng G, Li Y, Yang Y, Yang L, He D, Zhao Y, Cai Z. Vernodalol mediates antitumor effects in acute promyelocytic leukemia cells. Oncol Lett. 2018; 15(2):2227-2235.
48. Mal R, Magner A, David J, Datta J, Vallabhaneni M, Kassem M, Manouchehri J, Willingham N, Stover D, Vandeusen J, Sardesai S, Williams N, Wesolowski R, Lustberg M, Ganju RK, Ramaswamy B, Cherian MA. Estrogen Receptor Beta (ERβ): A Ligand Activated Tumor Suppressor. Front. Oncol. 2020; 10:587386.
49. Choi Y, Kim H, Pollack S. ERβ Isoforms Have Differential Clinical Significance in Breast Cancer Subtypes and Subgroups. Curr Issues Mol Biol. 2022; 44: 1564–1586.
50. Choi, Y. Estrogen Receptor β Expression and Its Clinical Implication in Breast Cancers: Favorable or Unfavorable? J Breast Cancer. 2022; 25: 75
51. Lipinski CA, Lombardo F, Dominy BW, Feeney PJ. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev. 2001; 46(1-3):3-26.
52. Teague SJ, Davis AM, Leeson PD, Oprea T. The Design of Leadlike Combinatorial Libraries. Angew Chem Int Ed Engl. 1999; 38(24):3743-3748.
53. Ertl P, Schufenhauer A. Estimation of synthetic accessibility score of drug-like molecules based on molecular complexity and fragment contributions. J. Cheminf. 2009; 1:1–11
54. Rai M, Singh AV, Paudel N, Kanase A, Falletta E, Kerkar P, Heyda J, Barghash RF, Pratap SS, Soos M. Herbal concoction Unveiled: a computational analysis of phytochemicals’ pharmacokinetic and toxicological profiles using novel approach methodologies (NAMs). Curr Res Toxicol. 2023; 5:100118
55. Kerns EH, Di L. Drug-Like Properties: Concepts, Structure Design and Methods from ADME to Toxicity Optimization. New York, USA: Academic Press. 2008
56. Daina A, Michielin O, Zoete V. SwissADME: a free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules. Sci Rep. 2017; 7:42717.