Abstract

Barnûf (Pluchea dioscoridis L.) is a wild plant that grows in Egypt. Barnûf leaves are utilized as a folk medicine, as well as part of food and drink formulations. Their numerous biological benefits include anti-inflammatory and antioxidant properties. We examined the antioxidant, antidiabetic, anti-obesity, antithyroid, and anticancer activities of methanol, ethanol, and acetone extracts of barnûf leaves. The methanol extract exhibited the highest total phenolic (241.50 ± 3.71 mg GAE/g extract) and flavonoid (256.18 ± 3.19 mg QE/g extract) contents. All three extracts proved to possess good antioxidant, antimicrobial, antidiabetic, anti-obesity, antithyroid, and anticancer activities. Ellagic acid was the most abundant phenolic acid in the methanolic (30.33%) and ethanolic (24.71%) extracts. The antioxidant experiments revealed that the methanolic extract had potent DPPH• (IC50 = 18.21 μg/mL) and ABTS•+ (IC50 = 17.6 μg/mL) scavenging properties. The acetone extract demonstrated the highest antimicrobial activity against gramnegative bacteria. Regarding α-amylase and α-glucosidase inhibition, the methanolic extract showed the most potent activity with IC50 values of 104.28 ± 1.97 and 133.76 ± 2.09 μg/mL, respectively. The methanolic extract also proved to be the strongest inhibitor of lipase and thyroid peroxidase, with IC50 values of 127.35 and 211.2 μg/mL, respectively. In addition, the methanolic extract showed the strongest anticancer activity against MCF7-1 and H1299-1 lines with IC50 values of 29.3 and 18.4 μg/mL, respectively. The findings suggest that barnûf leaf extracts could be used in functional foods and pharmaceuticals.

Keywords

Pluchea dioscoridis L., medicinal properties, extraction, DPPH, ABTS, herbal medicine

REFERENCES

1. Hlila MB, Mosbah H, Zanina N, Ben Nejma A, Ben Jannet H, Aouni M, et al. Characterisation of phenolic antioxidants in Scabiosa arenaria flowers by LC-ESI-MS/MS and NMR. Journal of Pharmacy and Pharmacology. 2016;68(7):932–940. https://doi.org/10.1111/jphp.12561
2. Kumarasingha R, Preston S, Yeo T-C, Lim DSL, Tu C-L, Palombo EA, et al. Anthelmintic activity of selected ethno-medicinal plant extracts on parasitic stages of Haemonchus contortus. Parasites and Vectors. 2016;9:187. https://doi.org/10.1186/s13071-016-1458-9
3. Halliwell B. Drug antioxidant effects. A basis for drug selection? Drugs. 1991;42:569–605. https://doi.org/10.2165/00003495-199142040-00003
4. Shaltout KH, Slima DF. The biology of Egyptian woody perrenials. 3. Pluchea dioscoridis (L.) DC. Assuit University Bulletin for Environmental Researches. 2007;10(1):85–103.
5. Elsebaie EM, Essa RY. Application of barnûf (Pluchea dioscoridis) leaves extract as a natural antioxidant and antimicrobial agent for eggs quality and safety improvement during storage. Journal of Food Processing and Preservation. 2022;46:e16061. https://doi.org/10.1111/jfpp.16061
6. Buhmann A, Papenbrock J. An economic point of view of secondary compounds in halophytes. Functional Plant Biology. 2013;40(9):952–967. https://doi.org/10.1071/FP12342
7. Falleh H, Trabelsi N, Bonenfant-Magné M, Le Floch G, Abdelly C, Magné C, et al. Polyphenol content and biological activities of Mesembryanthemum edule organs after fractionation. Industrial Crops and Products. 2013;42:145–152. https://doi.org/10.1016/j.indcrop.2012.05.033
8. Harborne AJ. Phytochemical methods a guide to modern techniques of plant analysis. Dordrecht: Springer; 1998. 302 p.
9. Liu X, Dong M, Chen X, Jiang M, Lv X, Yan G. Antioxidant activity and phenolics of an endophytic Xylaria sp. from Ginkgo biloba. Food Chemistry. 2007;105(2):548–554. https://doi.org/10.1016/j.foodchem.2007.04.008
10. Dai J, Mumper RJ. Plant phenolics: Extraction, analysis and their antioxidant and anticancer properties. Molecules. 2010;15(10):7313–7352. https://doi.org/10.3390/molecules15107313
11. Rubio-Moraga Á, Argandoña J, Mota B, Pérez J, Verde A, Fajardo J, et al. Screening for polyphenols, antioxidant and antimicrobial activitiesof extracts from eleven Helianthemum taxa (Cistaceae) used in folk medicine in south-eastern Spain. Journal of Ethnopharmacology. 2013;148(1):287–296. https://doi.org/10.1016/j.jep.2013.04.028
12. Wang H, Helliwell K. Determination of flavonols in green and black tea leaves and green tea infusions by high-performance liquid chromatography. Food Research International. 2001;34(2–3):223–227. https://doi.org/10.1016/S0963-9969(00)00156-3
13. Hayouni EA, Abedrabba M, Bouix M, Hamdi M. The effects of solvents and extraction method on the phenolic contents and biological activities in vitro of Tunisian Quercus coccifera L. and Juniperus phoenicea L. fruit extracts. Food Chemistry. 2007;105(3):1126–1134. https://doi.org/10.1016/j.foodchem.2007.02.010
14. Prior RL, Wu X, Schaich K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. Journal of Agricultural and Food Chemistry. 2005;53(10):4290–4302. https://doi.org/10.1021/jf0502698
15. El-Hamouly MA, Ibraheim MT. GC/MS analysis of the volatile constituents of individual organs of Conyza dioscorides L. (Desf.), growing in Egypt. Alexandria Journal of Pharmaceutical Sciences. 2003;17:75–81.
16. Kamel EM, Ahemd S. Phenolic constituents and biological activity of the genus Pluchea. Der Pharma Chemica. 2013;5(5):109–114.
17. Uchiyama T, Miyase T, Ueno A, Usmanghani K. Terpene and lignan glycosides from Pluchea indica. Phytochemistry. 1991;30(2):655–657. https://doi.org/10.1016/0031-9422(91)83746-8
18. Bazzano LA, Serdula MK, Liu S. Dietary intake of fruits and vegetables and risk of cardiovascular disease. Current Atherosclerosis Reports. 2003;5:492–499. https://doi.org/10.1007/s11883-003-0040-z
19. Saravanan S, Parimelazhagan T. In vitro antioxidant, antimicrobial and anti-diabetic properties of polyphenols of Passiflora ligularis Juss. fruit pulp. Food Science and Human Wellness. 2014;3(2):56–64.
20. Shaw JE, Sicree RA, Zimmet PZ. Global estimates of the prevalence of diabetes for 2010 and 2030. Diabetes Research and Clinical Practice. 2010;87(1):4–14. https://doi.org/10.1016/j.diabres.2009.10.007
21. Zhang R, Zeng Q, Deng Y, Zhang M, Wei Z, Zhang Y, et al. Phenolic profiles and antioxidant activity of litchi pulp of different cultivars cultivated in Southern China. Food Chemistry. 2013;136(3–4):1169–1176. https://doi.org/10.1016/j.foodchem.2012.09.085
22. Boaventura BCB, Di Pietro PF, Klein GA, Stefanuto A, de Morais EC, de Andrade F, et al. Antioxidant potential of mate tea (Ilex paraguariensis) in type 2 diabetic mellitus and pre-diabetic individuals. Journal of Functional Foods. 2013;5(3):1057–1064. https://doi.org/10.1016/j.jff.2013.03.001
23. Ahmad LA, Crandall JP. Type 2 diabetes prevention: A review. Clinical Diabetes. 2010;28(2):53–59. https://doi.org/10.2337/diaclin.28.2.53
24. Yee HS, Fong NT. A review of the safety and efficacy of acarbose in diabetes mellitus. Pharmacotherapy. 1996;16(5):792–805. https://doi.org/10.1002/j.1875-9114.1996.tb02997.x
25. Padwal RS, Majumdar SR. Drug treatments for obesity: Orlistat, sibutramine, and rimonabant. The Lancet. 2007;369(9555):71–77. https://doi.org/10.1016/S0140-6736(07)60033-6
26. Tahrani AA, Piya MK, Kennedy A, Barnett AH. Glycaemic control in type 2 diabetes: Targets and new therapies. Pharmacology and Therapeutics. 2010;125(2):328–361. https://doi.org/10.1016/j.pharmthera.2009.11.001
27. Allouche N, Fki I, Sayadi S. Toward a high yield recovery of antioxidants and purified hydroxytyrosol from olive mill wastewaters. Journal of Agricultural and Food Chemistry. 2004;52(2):267–273. https://doi.org/10.1021/jf034944u
28. Pereira DF, Cazarolli LH, Lavado C, Mengatto V, Figueiredo MSRB, Guedes A, et al. Effects of flavonoids on α-glucosidase activity: Potential targets for glucose homeostasis. Nutrition. 2011;27(11–12):1161–1167. https://doi.org/10.1016/j.nut.2011.01.008
29. Doerge DR, Divi RL. Porphyrin π-cation and protein radicals in peroxidase catalysis and inhibition by anti-thyroid chemicals. Xenobiotica. 1995;25(7):761–767. https://doi.org/10.3109/00498259509061891
30. Gaitan E. Flavonoids and the thyroid. Nutrition. 1996;12(2):127–129. https://doi.org/10.1016/S0899-9007(97)85052-7
31. Divi RL, Doerge DR. Inhibition of thyroid peroxidase by dietary flavonoids. Chemical Research in Toxicology. 1996;9(1):16–23. https://doi.org/10.1021/tx950076m
32. Ferreira ACF, Lisboa PC, Oliveira KJ, Lima LP, Barros IA, Carvalho DP. Inhibition of thyroid type 1 deiodinase activity by flavonoids. Food and Chemical Toxicology. 2002;40(7):913–917. https://doi.org/10.1016/S0278-6915(02)00064-9
33. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians. 2018;68:394–424. https://doi.org/10.3322/caac.21492
34. Karpuz M, Silindir Gunay M, Ozer AY. Current and future approaches for effective cancer imaging and treatment. Cancer Biotherapy and Radiopharmaceuticals. 2018;33(2):39–51. https://doi.org/10.1089/cbr.2017.2378
35. Szablewski L. Diabetes mellitus: Influences on cancer risk. Diabetes/Metabolism Research and Reviews. 2014;30(7):543–553. https://doi.org/10.1002/dmrr.2573
36. Moglad EHO, Abdalla OM, Koko WS, Saadabi AM. In vitro anticancer activity and cytotoxicity of Solanum nigrum on cancers and normal cell lines. International Journal of Cancer Research. 2014;10(2):74–80. https://doi.org/10.3923/ijcr.2014.74.80
37. Waterhouse AL. Determination of total phenolics. In: Wrolstad RE, editor. Current protocols in food analytical chemistry. New York: John Wiley and Sons; 2002. Ppp. I1.1.1–I1.1.8.
38. Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry. 1999;64(4):555–559. https://doi.org/10.1016/S0308-8146(98)00102-2
39. Elsebaie EM, Essa RY. Microencapsulation of red onion peel polyphenols fractions by freeze drying technicality and its application in cake. Journal of Food Processing and Preservation. 2018:42:e13654. https://doi.org/10.1111/jfpp.13654
40. Fki I, Allouche N, Sayadi S. The use of polyphenolic extract, purified hydroxytyrosol and 3,4-dihydroxyphenyl acetic acid from olive mill wastewater for the stabilization of refined oils: A potential alternative to synthetic antioxidants. Food Chemistry. 2005;93(2):197–204. https://doi.org/10.1016/j.foodchem.2004.09.014
41. Sayah K, Marmouzi I, Naceiri Mrabti H, Cherrah Y, Faouzi MEA. Antioxidant activity and inhibitory potential of Cistus salviifolius (L.) and Cistus monspeliensis (L.) aerial parts extracts against key enzymes linked to hyperglycemia. BioMed Research International. 2017;2017:2789482. https://doi.org/10.1155/2017/2789482
42. Elsebaie EM, El-Wakeil NHM, Khalil AMM, Bahnasy RM, Asker GA, El-Hassnin MF, et al. Silver nanoparticle synthesis by Rumex vesicarius extract and its applicability against foodborne pathogens. Foods. 2023;12(9):1746. https://doi.org/10.3390/foods12091746
43. Ademiluyi AO, Oboh G. Aqueous extracts of Roselle (Hibiscus sabdariffa Linn.) varieties inhibit α-amylase and α-glucosidase activities in vitro. Journal of Medicinal Food. 2013;16(1):88–93. https://doi.org/10.1089/jmf.2012.0004
44. Telagari M, Hullatti K. In-vitro α-amylase and α-glucosidase inhibitory activity of Adiantum caudatum Linn. and Celosia argentea Linn. extracts and fractions. Indian Journal of Pharmacology. 2015;47(4):425–429. https://doi.org/10.4103/0253-7613.161270
45. Nakai M, Fukui Y, Asami S, Toyoda-Ono Y, Iwashita T, Shibata H, et al. Inhibitory effects of oolong tea polyphenols on pancreatic lipase in vitro. Journal of Agricultural and Food Chemistry. 2005;53(11):4593–4598. https://doi.org/10.1021/jf047814+
46. Jomaa B, de Haan LHJ, Peijnenburg AACM, Bovee TFH, Aarts JMMJG, Rietjens IMCM. Simple and rapid in vitro assay for detecting human thyroid peroxidase disruption. ALTEX – Alternatives to Animal Experimentation. 2015;32(3):191–200. https://doi.org/10.14573/altex.1412201
47. Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, et al. New colorimetric cytotoxicity assay for anticancer-drug screening. JNCI: Journal of the National Cancer Institute. 1990;82(13):1107–1112. https://doi.org/10.1093/jnci/82.13.1107
48. Qasim M, Aziz I, Rasheed M, Gul B, Khan MA. Effect of extraction solvents on polyphenols and antioxidant activity of medicinal halophytes. Pakistan Journal of Botany. 2016;48(2):621–627.
49. Asimi O, Sahu NP, Pal AK. Antioxidant activity and antimicrobial property of some Indian spices. International Journal of Scientific and Research Publications. 2013;3:1–8.
50. Howlader MSI, Rahman MM, Khalipha ABR, Ahmed F, Rahman MM. Antioxidant and antidiarrhoeal potentiality of Diospyros blancoi. International Journal of Pharmacology. 2012;8(5):403–409. https://doi.org/10.3923/ijp.2012.403.409
51. Sridhar K, Charles AL. In vitro antioxidant activity of Kyoho grape extracts in DPPH and ABTS assays: Estimation methods for EC50 using advanced statistical programs. Food Chemistry. 2019;275:41–49. https://doi.org/10.1016/j.foodchem.2018.09.040
52. Kala CP. Current status of medicinal plants used by traditional Vaidyas in Uttaranchal state of India. Ethnobotany Research and Applications. 2005;3:267–278. https://doi.org/10.17348/era.3.0.267-278
53. Aruoma OI, Cuppett SL. Antioxidant methodology: in vivo and in vitro concepts. Champaign: The American Oil Chemists Society; 1997. 241 p.
54. Tsai C-E, Lin L-H. DPPH scavenging capacity of extracts from Camellia seed dregs using polyol compounds as solvents. Heliyon. 2019;5(8):e02315. https://doi.org/10.1016/j.heliyon.2019.e02315
55. Helfand SL, Rogina B. Genetics of aging in the fruit fly, Drosophila melanogaster. Annual Review of Genetics. 2003;37:329–348. https://doi.org/10.1146/annurev.genet.37.040103.095211
56. Prashith KTR, Manasa M, Poornima G, Abhipsa V, Rekha C, Upashe SP, et al. Antibacterial, cytotoxic and antioxidant potential of Vitex negundo var. negundo and Vitex negundo var. purpurascens – A comparative study. Science, Technology and Arts Research Journal. 2013;2(3):59–68. https://doi.org/10.4314/star.v2i3.98737
57. Bonina F, Puglia C, Tomaino A, Saija A, Mulinacci N, Romani A, et al. In-vitro antioxidant and in-vivo photoprotective effect of three lyophilized extracts of Sedum telephium L. leaves. Journal of Pharmacy and Pharmacology. 2000;52(10):1279–1285. https://doi.org/10.1211/0022357001777261
58. Ghedadba N, Bousselsela H, Hambaba L, Benbia S, Mouloud Y. Evaluation of antioxidant and antimicrobial activity of leaves and flowering tops of Marrubium vulgare L. Phytothérapie. 2014;12:15–24. https://doi.org/10.1007/s10298-014-0832-z (In French.).
59. Fidrianny I, Rizkiya A, Ruslan K. Antioxidant activities of various fruit extracts from three solanum sp. using DPPH and ABTS method and correlation with phenolic, flavonoid and carotenoid content. Journal of Chemical and Pharmaceutical Research. 2015;7(5):666–672.
60. Sarr SO, Fall AD, Gueye R, Diop A, Diatta K, Diop N, et al. Study of the antioxidant activity of extracts from the leaves of Vitex doniana (Verbenacea). International Journal of Biological and Chemical Sciences. 2015;9(3):1263–1269.
61. Saber RA. Evaluation of antiurolithiatic and antioxidant activity of the Egyptian Pluchea dioscoridis L. leaves extracts in vitro. African Journal of Biological Sciences. 2021;17(1):233–249. https://doi.org/10.21608/ajbs.2021.201676
62. Cano A, Acosta M, Arnao MB. A method to measure antioxidant activity in organic media: Application to lipophilic vitamins. Redox Report. 2000;5(6):365–370. https://doi.org/10.1179/135100000101535933
63. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine. 1999;26(9–10):1231–1237. https://doi.org/10.1016/S0891-5849(98)00315-3
64. Ndhlala AR, Ncube B, Abdelgadir HA, Du Plooy CP, van Staden, J. Antioxidant potential of African medicinal plants. In: Al-Gubory KH, Laher I, editors. Nutritional antioxidant therapies: Treatments and perspectives. Cham: Springer; 2017. pp. 65–88. https://doi.org/10.1007/978-3-319-67625-8_3
65. Vongsak B, Kongkiatpaiboon S, Jaisamut S, Konsap K. Comparison of active constituents, antioxidant capacity, and α-glucosidase inhibition in Pluchea indica leaf extracts at different maturity stages. Food Bioscience. 2018;25:68–73. https://doi.org/10.1016/j.fbio.2018.08.006
66. Obeidat M, Shatnawi M, Al-alawi M, Al-Zu`bi E, Al-Dmoor H, Al-Qudah M, et al. Antimicrobial activity of crude extracts of some plant leaves. Research Journal of Microbiology. 2012;7(1):59–67. https://doi.org/10.3923/jm.2012.59.67
67. Zalabani SM, Hetta MH, Ismail AS. Anti-inflammatory and antimicrobial activity of the different Conyza dioscoridis L. Desf. Organs. Biosafety. 2013;2(1):1000106. https://doi.org/10.4172/2167-0331.1000106
68. Aruwa CE, Amoo S, Kudanga T. Phenolic compound profile and biological activities of Southern African Opuntia ficus-indica fruit pulp and peels. LWT. 2019;111:337–344. https://doi.org/10.1016/j.lwt.2019.05.028
69. El-Ghorab AH, Ramadan MM, Abd El-Moezc SI, Soliman A-MM. Essential oil, antioxidant, antimicrobial and anticancer activities of Egyptian Pluchea dioscoridis extract. Research Journal of Pharmaceutical, Biological and Chemical Sciences. 2015;6(2):1255
70. Jhong C-H, Riyaphan J, Lin S-H, Chia Y-C, Weng C-F. Screening alpha‐glucosidase and alpha‐amylase inhibitors from natural compounds by molecular docking in silico. BioFactors. 2015;41(4):242–251. https://doi.org/10.1002/biof.1219
71. Gowri PM, Tiwari AK, Ali AZ, Rao JM. Inhibition of α‐glucosidase and amylase by bartogenic acid isolated from Barringtonia racemosa Roxb. seeds. Phytotherapy Research. 2007;21(8):796–799. http://dx.doi.org/10.1002/ptr.2176
72. Tarling CA, Woods K, Zhang R, Brastianos HC, Brayer GD, Andersen RJ, et al. The search for novel human pancreatic α‐amylase inhibitors: High‐throughput screening of terrestrial and marine natural product extracts. ChemBioChem. 2008;9:433–438. https://doi.org/10.1002/cbic.200700470
73. Shobana S, Sreerama YN, Malleshi NG. Composition and enzyme inhibitory properties of finger millet (Eleusine coracana L.) seed coat phenolics: Mode of inhibition of α-glucosidase and pancreatic amylase. Food Chemistry. 2009;115(4):1268–1273. https://doi.org/10.1016/j.foodchem.2009.01.042
74. Apostolidis E, Lee CM. In vitro potential of Ascophyllum nodosum phenolic antioxidant‐mediated α‐glucosidase and α‐amylase inhibition. Journal of Food Science. 2010;75(3):H97–H102. https://doi.org/10.1111/j.1750-3841.2010.01544.x
75. Ramkumar KM, Thayumanavan B, Palvannan T, Rajaguru P. Inhibitory effect of Gymnema montanum leaves on α-glucosidase activity and α-amylase activity and their relationship with polyphenolic content. Medicinal Chemistry Research. 2010;19:948–961. https://doi.org/10.1007/s00044-009-9241-5
76. Ali MB, Mnafgui K, Feki A, Damak M, Allouche N. In vitro antidiabetic, anti-obesity and antioxidant proprities of Rosemary extracts. Journal of Advances in Chemistry. 2014;10(2):2305–2316. https://doi.org/10.24297/jac.v10i2.5497
77. McDougall GJ, Kulkarni NN, Stewart D. Berry polyphenols inhibit pancreatic lipase activity in vitro. Food Chemistry. 2009;115(1):193–199. https://doi.org/10.1016/j.foodchem.2008.11.093
78. Habza-Kowalska E, Kaczor AA, Żuk J, Matosiuk D, Gawlik-Dziki U. Thyroid peroxidase activity is inhibited by phenolic compounds – Impact of interaction. Molecules. 2019;24(15):2766. https://doi.org/10.3390/molecules24152766
79. Leonard JA, Tan Y-M, Gilbert M, Isaacs K, El-Masri H. Estimating margin of exposure to thyroid peroxidase inhibitors using high-throughput in vitro data, high-throughput exposure modeling, and physiologically based pharmacokinetic/pharmacodynamic modeling. Toxicological Sciences. 2016;151(1):57–70. https://doi.org/10.1093/toxsci/kfw022
80. Iawsipo P, Poonbud R, Somtragool N, Mutapat P, Meejom A. Pluchea indica tea-leaf extracts exert anti-cancer activity by inducing ROS-mediated cytotoxicity on breast and cervical cancer cells. British Food Journal. 2022;124(12):4769–4781. https://doi.org/10.1108/BFJ-05-2021-0497
81. Bibi Y, Nisa S, Zia M, Waheed A, Ahmed S, Chaudhary MF. In vitro cytotoxic activity of Aesculus indica against breast adenocarcinoma cell line (MCF-7) and phytochemical analysis. Pakistan Journal of Pharmaceutical Sciences. 2012;25(1):183–187.