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Antioxidant, anticoagulant and antihyperlipidemic activities of Thymus willdenowii Boiss. collected from three different harvesting sites in Eastern High Atlas

DOI
https://doi.org/10.14719/pst.10621
Submitted
13 July 2025
Published
14-05-2026
Versions

Abstract

Thyme species are commonly used in Moroccan folk medicine to prevent and treat cardiovascular diseases and their risk factors. In this respect, Thymus willdenowii Boiss. collected from 3 different harvesting site in Eastern High Atlas (Outerbate, Tissila and Aït Ali Ou Ikkou) was evaluated and compared for its antioxidant, anticoagulant and antihyperlipidemic activities.The antioxidant activity was assessed through 2,2-diphenyl-1-picrylhydrazyle (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and ferric reducing ability of plasma (FRAP) methods. The anticoagulant activity was evaluated based on prothrombin time (PT) and activated partial thromboplastin time (aPTT). While the antihyperlipidemic effect was examined using Triton-1339-induced hyperlipidemia in rats. The Tissila extract had the most phenolic (116.74 ± 0.52 mg GAE/g) and flavonoid (16.73 ± 0.9 mg RE/g) content, which was linked to better antioxidant activity, with IC50 values of 66.45 µg/mL for DPPH and 20.53 µg/mL for ABTS.  In coagulation assays, the Tissila extract exhibited the most pronounced effect, extending PT to 25.13 sec at 16.29 µg/mL, in contrast to 12.37 sec in the control.  Additionally, in hyperlipidemic rats, the Tissila extract (400 mg/kg) elicited the most significant biological response, decreasing total cholesterol by 63 %, LDL-C by 62 %  and the atherogenic index by 84 %, while elevating HDL-C levels by 121 %. On the other hand, the Outerbate extract did not have a strong effect on lowering blood lipids. Findings suggest the potential of T. willdenowii as a source of natural antioxidant and cardioprotective agents.

References

  1. 1. WHO. Cardiovascular diseases (CVDs); 2025. https://tinyurl.com/yta4hk93
  2. 2. Bays HE, Taub PR, Epstein E, Michos ED, Ferraro RA, Bailey AL, et al. Ten things to know about ten cardiovascular disease risk factors. Am J Prev Cardiol. 2021;5:100149. https://doi.org/10.1016/j.ajpc.2021.100149
  3. 3. Duraipandiyan V, Al-Dhabi NA, Irudayaraj SS, Sunil C. Hypolipidemic activity of friedelin isolated from Azima tetracantha in hyperlipidemic rats. Rev Bras Farmacogn. 2016;26(1):89–93. https://doi.org/10.1016/j.bjp.2015.07.025
  4. 4. Oduah EI, Linhardt RJ, Sharfstein ST. Heparin: Past, present and future. Pharmaceuticals (Basel). 2016;9(3). https://doi.org/10.3390/ph9030038
  5. 5. Jamshidikia F, Lorigooini Z, Amini-Khoei H. Medicinal plants: Past history and future perspective. J HerbMed Pharmacol. 2018;7:1–7. https://doi.org/10.15171/jhp.2018.01
  6. 6. Ekor M. The growing use of herbal medicines: Issues relating to adverse reactions and challenges in monitoring safety. Front Pharmacol. 2014;4:177. https://doi.org/10.3389/fphar.2013.00177
  7. 7. El-Hilaly J, Hmammouchi M, Lyoussi B. Ethnobotanical studies and economic evaluation of medicinal plants in Taounate province (Northern Morocco). J Ethnopharmacol. 2003;86(2–3):149–58. https://doi.org/10.1016/s0378-8741(03)00012-6
  8. 8. Riaz M, Khalid R, Afzal M, Anjum F, Fatima H, Zia S, et al. Phytobioactive compounds as therapeutic agents for human diseases: A review. Food Sci Nutr. 2023;11(6):2500–29. https://doi.org/10.1002/fsn3.3308
  9. 9. Hmidani A, Bouhlali EdT, Khouya T, Ramchoun M, Filali-Zegzouti Y, Benlyas M, et al. Effect of extraction methods on antioxidant and anticoagulant activities of Thymus atlanticus aerial part. Sci Afr.2019;5:e00143. https://doi.org/10.1016/j.sciaf.2019.e00143
  10. 10. Ramchoun M, Harnafi H, Alem C, Büchele B, Simmet T, Rouis M, et al. Hypolipidemic and antioxidant effect of polyphenol-rich extracts from Moroccan thyme varieties. e-SPEN J. 2012;7(3):e119–24. https://doi.org/10.1016/j.clnme.2012.02.005
  11. 11. Pietrzak W, Nowak R. Impact of harvest conditions and host tree species on chemical composition and antioxidant activity of extracts from Viscum album L. Molecules. 2021;26(12). https://doi.org/10.3390/molecules26123741
  12. 12. Hmidani A, Bouhlali EDT, Khouya T, Ramchoun M, Filali-Zegzouti Y, Alem C, et al. Antioxidant, anti-inflammatory and anticoagulant activities of three Thymus species grown in southeastern Morocco. Future J Pharm Sci. 2019;5(1):4. https://doi.org/ 10.1186/s43094-019-0005-x
  13. 13. Kim D-O, Jeong SW, Lee CY. Antioxidant capacity of phenolic phytochemicals from various cultivars of plums. Food Chem. 2003;81(3):321–6. https://doi.org/10.1016/S0308-8146(02)00423-5.
  14. 14. Blois MS. Antioxidant determinations by the use of a stable free radical. Nature. 1958;181(4617):1199–200. https://doi.org/10.1038/1811199a0
  15. 15. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999;26(9–10):1231–7. https://doi.org/10.1016/s0891-5849(98)00315-3
  16. 16. Benzie IF, Strain JJ. Ferric reducing/antioxidant power assay: Direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Methods Enzymol. 1999;299:15–27. https://doi.org/10.1016/s0076-6879(99)99005-5
  17. 17. Athukorala Y, Jung W-K, Vasanthan T, Jeon Y-J. An anticoagulative polysaccharide from an enzymatic hydrolysate of Ecklonia cava. Carbohydr Polym. 2006;66:184–91. https://doi.org/10.1016/j.carbpol.2006.03.002
  18. 18. Bouhlali EDT, Hmidani A, Bourkhis B, Khouya T, Harnafi H, Filali-Zegzouti Y, et al. Effect of Phoenix dactylifera seeds (dates) extract in triton WR-1339 and high fat diet induced hyperlipidaemia in rats: A comparison with simvastatin. J Ethnopharmacol. 2020;259:112961. https://doi.org/10.1016/j.jep.2020.112961
  19. 19. Eddouks M, Maghrani M, Lemhadri A, Ouahidi ML, Jouad H. Ethnopharmacological survey of medicinal plants used for the treatment of diabetes mellitus, hypertension and cardiac diseases in the south-east region of Morocco (Tafilalet). J Ethnopharmacol. 2002;82(2–3):97–103. https://doi.org/10.1016/s0378-8741(02)00164-2.
  20. 20. Radi FZ, Bencheikh N, Bouhrim M, Noman OM, Mothana RA, Ibrahim MN, et al. Phytochemical analysis, antioxidant, antimicrobial and antihyperglycemic activities and adverse effect assessment of Thymus zygis L. and Thymus willdenowii Bois. J Biol Regul Homeost Agents. 2023;37:6113–30. https://doi.org/10.23812/j.biol.regul.homeost.agents.20233711.584.
  21. 21. Zejli H, Metouekel A, Zouirech O, Maliki I, El Moussaoui A, Lfitat A, et al. Phytochemical analysis, antioxidant, analgesic, anti-inflammatory, hemagglutinin and hemolytic activities of chemically characterized extracts from Origanum grosii (L.) and Thymus pallidus (L.). Plants. 2024;13(3):385. https://doi.org/10.3390/plants13030385
  22. 22. Zeghib A, Calliste CA, Simon A, Charfeddine R, Aouni M, Duroux JL, et al. Chemical composition and biological potential of Thymus willdenowii Boiss. & Reut. essential oil. Nat Prod Res. 2021;35(5):845–8. https://doi.org/10.1080/14786419.2019.1602830
  23. 23. Taghouti M, Martins-Gomes C, Félix LM, Schäfer J, Santos JA, Bunzel M, et al. Polyphenol composition and biological activity of Thymus citriodorus and Thymus vulgaris: Comparison with endemic Iberian Thymus species. Food Chem. 2020;331:127362. https://doi.org/10.1016/j.foodchem.2020.127362
  24. 24. De Santis S, Clodoveo ML, Corbo F. Correlation between chemical characterization and biological activity: An urgent need for human studies using extra virgin olive oil. Antioxidants (Basel). 2022;11(2). https://doi.org/10.3390/antiox11020258
  25. 25. Endara M-J, Coley PD. The resource availability hypothesis revisited: A meta-analysis. Funct Ecol. 2011;25(2):389–98. https://doi.org/10.1111/j.1365-2435.2010.01803.x
  26. 26. Gouvea D, Gobbo-Neto L, Sakamoto H, Lopes N, Lopes J, Meloni F, et al. Seasonal variation of the major secondary metabolites present in the extract of Eremanthus mattogrossensis Less (Asteraceae: Vernonieae) leaves. Quim Nova. 2012;35:2139–45. https://doi.org/10.1590/S0100-40422012001100007
  27. 27. Rawat S, Bhatt ID, Rawal RS, Nandi SK. Geographical and environmental variation in chemical constituents and antioxidant properties in Roscoea procera Wall. J Food Biochem. 2017;41(2):e12302. https://doi.org/10.1111/jfbc.12302.
  28. 28. El-Jalel LFA, Elkady WM, Gonaid MH, El-Gareeb KA. Difference in chemical composition and antimicrobial activity of Thymus capitatus L. essential oil at different altitudes. Future J Pharm Sci. 2018;4(2):156–60. https://doi.org/10.1016/j.fjps.2017.12.004.
  29. 29. Ashraf MV, Khan S, Misri S, Gaira KS, Rawat S, Rawat B, et al. High-altitude medicinal plants as promising source of phytochemical antioxidants to combat lifestyle-associated oxidative stress-induced disorders. Pharmaceuticals (Basel). 2024;17(8). https://doi.org/10.3390/ph17080975.
  30. 30. Intharuksa A, Kuljarusnont S, Sasaki Y, Tungmunnithum D. Flavonoids and other polyphenols: Bioactive molecules from traditional medicine recipes/medicinal plants and their potential for phytopharmaceutical and medical application. Molecules. 2024;29(23):5760. https://doi.org/10.3390/molecules29235760.
  31. 31. Prior RL, Wu X, Schaich K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. J Agric Food Chem. 2005;53(10):4290–302. https://doi.org/10.1021/jf0502698.
  32. 32. Bahukhandi A, Pandey A, Sekar K, Bhatt I. Polyphenolics, nutrients and antioxidant activity of Gaultheria trichophylla Royle: A high value wild edible plant of Trans Himalaya. Horticult Int J. 2017;1:00007. https://doi.org/10.15406/hij.2017.01.00007
  33. 33. Gaber N, El-Dahy S, Shalaby E. Comparison of ABTS, DPPH, permanganate and methylene blue assays for determining antioxidant potential of successive extracts from pomegranate and guava residues. Biomass Convers Biorefin. 2021;13:1–10. https://doi.org/10.1007/s13399-021-01386-0.
  34. 34. Liao W, Chen L, Ma X, Jiao R, Li X, Wang Y. Protective effects of kaempferol against reactive oxygen species-induced hemolysis and its antiproliferative activity on human cancer cells. Eur J Med Chem. 2016;114:24–32. https://doi.org/10.1016/j.ejmech.2016.02.045
  35. 35. Xiang Z, Lin C, Zhu Y, Deng J, Xia C, Chen J. Phytochemical profiling of antioxidative polyphenols and anthocyanins in the wild plant Campanumoealancifolia (Roxb.) Merr. Int J Food Prop. 2021;24(1):105–14. https://doi.org/10.1080/10942912.2020.1867570
  36. 36. Loucif K, Hassiba B, Benchikh F, Mehlous S, Souici C, Amira S. Total phenolic contents, DPPH radical scavenging and β-carotene bleaching activities of aqueous extract from Ammoides atlantica. J Drug Deliv Ther. 2020;10:196–8. https://doi.org/10.22270/jddt.v10i3-s.4151
  37. 37. Matamane R, Manoharan K, Magama S. DPPH radical scavenging activity of extracts from Urtica urens (Urticaceae). J Med Plants Res. 2020;14:232–8. https://doi.org/10.5897/JMPR2019.6880
  38. 38. Rudrapal M, Khairnar SJ, Khan J, Dukhyil AB, Ansari MA, Alomary MN, et al. Dietary polyphenols and their role in oxidative stress-induced human diseases: Insights into protective effects, antioxidant potentials and mechanism(s) of action. Front Pharmacol. 2022;13:806470. https://doi.org/10.3389/fphar.2022.806470
  39. 39. Clemente-Suárez VJ, Martín-Rodríguez A, Beltrán-Velasco AI, Rubio-Zarapuz A, Martínez-Guardado I, Valcárcel-Martín R, et al. Functional and therapeutic roles of plant-derived antioxidants in type 2 diabetes mellitus: Mechanisms, challenges and considerations for special populations. Antioxidants. 2025;14(6):725. https://doi.org/10.3390/antiox14060725
  40. 40. Liu Y, Tang Q, Duan X, Tang T, Ke Y, Zhang L, et al. Antioxidant and anticoagulant activities of mycelia polysaccharides from Catathelasma ventricosum after sulfated modification. Ind Crops Prod. 2018;112:53–60. https://doi.org/10.1016/j.indcrop.2017.10.064
  41. 41. Ho TC, Kiddane AT, Sivagnanam SP, Park JS, Cho YJ, Getachew AT, et al. Green extraction of polyphenolic-polysaccharide conjugates from Pseuderanthemum palatiferum (Nees) Radlk.: Chemical profile and anticoagulant activity. Int J Biol Macromol. 2020;157:484–93. https://doi.org/10.1016/j.ijbiomac.2020.04.113
  42. 42. Leite PM, Miranda APN, Amorim JM, Duarte RCF, Bertolucci SKV, Carvalho MDG, et al. In vitro anticoagulant activity of Mikania laevigata: Deepening the study of the possible interaction between guaco and anticoagulants. J Cardiovasc Pharmacol. 2019;74(6):574–83. https://doi.org/10.1097/FJC.0000000000000745
  43. 43. Raha A, Mukherjee P, Paul M, Roy S, Mamud R, Sarkar S, et al. In vitro anticoagulant activity of Datura stramonium flower extracts on blood plasma of poultry bird. 2019;8:1146–8.
  44. 44. Mira A, Alkhiary W, Shimizu K. Antiplatelet and anticoagulant activities of Angelica shikokiana extract and its isolated compounds. Clin Appl ThrombHemost. 2017;23(1):91–9. https://doi.org/10.1177/1076029615595879
  45. 45. Bijak M, Ponczek MB, Nowak P. Polyphenol compounds belonging to flavonoids inhibit activity of coagulation factor X. Int J Biol Macromol. 2014;65:129–35. https://doi.org/10.1016/j.ijbiomac.2014.01.023
  46. 46. Khouya T, Ramchoun M, Abdelbassat H, Amrani S, Harnafi H, Benlyas M, et al. Anti-inflammatory, anticoagulant and antioxidant effects of aqueous extracts from Moroccan thyme varieties. Asian Pac J Trop Biomed. 2015;5. https://doi.org/10.1016/j.apjtb.2015.05.011
  47. 47. Koriem KMM. Antihyperlipidemic activity of the medicinal plants among Kadazan and Dusun communities in Sabah, Malaysia: A review. Asian Pac J Trop Biomed. 2014;4(10):768–79. https://doi.org/10.12980/APJTB.4.2014C1144
  48. 48. Otway S, Robinson DS. The use of a non-ionic detergent (Triton WR 1339) to determine rates of triglyceride entry into the circulation of the rat under different physiological conditions. J Physiol. 1967;190(2):321–32. https://doi.org/10.1113/jphysiol.1967.sp008211
  49. 49. Cai G, Shi G, Xue S, Lu W. The atherogenic index of plasma is a strong and independent predictor for coronary artery disease in the Chinese Han population. Medicine (Baltimore). 2017;96(37):e8058. https://doi.org/10.1097/MD.0000000000008058
  50. 50. Wadhwa D, Mahajan VK, Mehta KS, Chauhan PS, Yadav RS, Bhushan S, et al. Malondialdehyde, lipoprotein-a, lipoprotein ratios, comprehensive lipid tetrad index and atherogenic index as surrogate markers for cardiovascular disease in patients with psoriasis: A case-control study. Arch Dermatol Res. 2019;311(4):287–97. https://doi.org/10.1007/s00403-019-01896-y
  51. 51. Harnafi M, Bekkouch O, Touiss I, Khatib S, Mokhtari I, Milenkovic D, et al. Phenolic-rich extract from almond (Prunus dulcis) hulls improves lipid metabolism in Triton WR-1339 and high-fat, diet-induced hyperlipidemic mice and prevents lipoprotein oxidation: A comparison with fenofibrate and butylated hydroxyanisole. Prev Nutr Food Sci. 2020;25(3):254–62. https://doi.org/10.3746/pnf.2020.25.3.254
  52. 52. Zeni ALB, Moreira TD, Dalmagro AP, Camargo A, Bini LA, Simionatto EL, et al. Evaluation of phenolic compounds and lipid-lowering effect of Morus nigra leaves extract. An Acad Bras Cienc. 2017;89(4):2805–15. https://doi.org/10.1590/0001-3765201720160660

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