Inhibitory Effect of Major Terpenoids of Essential Oil from Agathosma betulina Leaves on Collagenase and Elastase Enzymes Involved in Skin Aging

Abstract

Collagenase and elastase enzymes that cause the degradation of collagen and elastase have both been implicated in skin aging. Agathosma betulina is a popular plant in South Africa used in aromatherapy and folk medicine but the scientific knowledge regarding its anti-aging activity is yet to be explored. This study aimed to investigate the roles of the essential oil of A. betulina leaves on skin aging and its inhibitory effect on collagenase and elastase activities. In this work, the chemical profiling of essential oils of A. betulina leaves was explored. The antioxidant activity of essential oil of A. betulina leaves at different concentrations (0.07–10 µg/mL) was assessed using standard assay methods for 1,1-dihenyl-2-picryhydrayl (DPPH) and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS). Standard procedures for elastase and collagenase assays were also employed to determine the anti-aging skin activity of the essential oil at the same concentrations. Gas chromatography-mass spectrometric analysis showed the presence of 43 constituents with menthone (29.2%), limonene (23.7%), and pulegone (8.4%) as the major compounds. The oil expressed strong radical scavenging activity against DPPH and ABTS radicals at higher concentrations, with over 90% inhibition, particularly at 2.5, 5, and 10 µg/mL compared to the control (butylated hydroxytoluene). The oil also demonstrated stronger inhibitory activity against elastase at higher concentrations (5 and 10 µg/mL) and concentration-dependent inhibitory activity against collagenase compared to the control (ursolic acid). The essential oil of A. betulina leaves could be a potential candidate for the cosmetic industry to retard skin wrinkles and other manifestations of skin aging particularly associated with extrinsic factors.

Keywords

anti-aging - antioxidant - essential oil - collagenase - elastase - Agathosma betulina

Publication History

Received: 27 September 2024

Accepted: 14 March 2025

Article published online:
28 April 2025

• References

• 1 Zhang S, Dong Z, Peng Z, Lu F. Anti-aging effect of adipose-derived stem cells in a mouse model of skin aging induced by D-galactose. PLoS One 2014; 9 (05) e97573
• 2 Thring TS, Hili P, Naughton DP. Anti-collagenase, anti-elastase and anti-oxidant activities of extracts from 21 plants. BMC Complement Altern Med 2009; 9: 27
• 3 Kim YH, Kim KS, Han CS. et al. Inhibitory effects of natural plants of Jeju Island on elastase and MMP-1 expression. J Cosmet Sci 2007; 58 (01) 19-33
• 4 Kammeyer A, Luiten RM. Oxidation events and skin aging. Ageing Res Rev 2015; 21: 16-29
• 5 Jagetia GC, Baliga MS. The evaluation of nitric oxide scavenging activity of certain Indian medicinal plants in vitro: a preliminary study. J Med Food 2004; 7 (03) 343-348
• 6 Ibrahim FA, Usman LA, Akolade JO, Idowu OA, Abdulazeez AT, Amuzat AO. Anti-diabetic potentials of Citrus aurantifolia leaf essential oil. Drug Res (Stuttg) 2019; 69 (04) 201-206
• 7 Gattefosse RM. Gattefosse's Aromatherapy. New York: Random House; 1993
• 8 Molla J, Van Vuuren SF, Van Zyl RL, Viljoen AM. Biological activity and toxicity profile of 17 Agathosma (Rutaceae) species. S Afr J Bot 2007; 73: 588-592
• 9 van Wyk BE, Gericke N. People's Plants. Pretoria, South Africa: Briza Publications; 2000
• 10 Simpson D. Buchu—South Africa's amazing herbal remedy. Scott Med J 1998; 43 (06) 189-191
• 11 Watt J, Breyer-Brandwijk M. The Medicinal and Poisonous Plants of Southern and Eastern Africa. 2nd ed. London: E. and S. Livingstone Ltd.; 1962
• 12 Campbell WE, Williamson BK. Composition of Agathosma capensis essential oil. Planta Med 1991; 57 (03) 291-292
• 13 Evans WC. Trease and Evans Pharmacognosy. 14th ed. London: W. B. Saunders Company Ltd.; 1996
• 14 Skosana B, Aboua G, du Plessis SS. Buchu—the multi-purpose ethnomedicinally important specie and its benefits in the reproductive system, antioxidant-antidiabetic agents and human health. Intech Open 2014; 13: 288-316
• 15 Viljeon MA, Molla A, van Vuuren FS. et al. The biological activity and essential oil composition of 17 Agathosma (Rutaceae) species. J Essent Oil Res 2006; 18: 2-16
• 16 Verma K, Singh A, Deep P, Srivastava V, Verma S. Pharmacognostical properties and medicinal uses of Agathosma betulina (Rutaceae): a review. Int J Pharm Sci Rev Res 2020; 64 (01) 92-95
• 17 Lis-Balchin M, Stanley GD, Eaglesham E. Relationship between bioactivity and chemical composition of commercial essential oils. Flavour Fragrance J 1998; 13: 98-104
• 18 Braca A, De Tommasi N, Di Bari L, Pizza C, Politi M, Morelli I. Antioxidant principles from Bauhinia tarapotensis. J Nat Prod 2001; 64 (07) 892-895
• 19 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): 1231-1237
• 20 Kong Y, Yoo D, Nam M, Kim H. Excellent anti-aging effects of ursoloc acid and oleanolic acid present in Ligustrum lucidum . J Soc Cosmet Sci Korea 2012; 38 (02) 181-187
• 21 Van Wart HE, Steinbrink DR. A continuous spectrophotometric assay for Clostridium histolyticum collagenase. Anal Biochem 1981; 113 (02) 356-365
• 22 Tu PT, Tawata S. Anti-oxidant, anti-aging, and anti melano genic properties of the essential oils from two varieties of Alpinia zerumbet. Molecules 2015; 20 (09) 16723-16740
• 23 Yang W, Chen X, Li Y, Guo S, Wang Z, Yu Z. Advances in pharmacological activities of terpenoids. Nat Prod Commun 2020; 15: 3
• 24 Collins NF, Graven EH, van Beek TA, Lelyveld GP. Chemotaxonomy of commercial buchu species (Agathosma betulina and A. crenulata). J Essent Oil Res 1996; 8: 229-235
• 25 Flamini G, Cioni PL, Puleio R, Morelli I, Panizzi L. Antimicrobial activity of the essential oil of Calamintha nepeta and its constituent pulegone against bacteria and fungi. Phytother Res 1999; 13 (04) 349-351
• 26 Khallouki F, Hmamouchi M, Younos C, Soulimani R, Bessiere JM, Essassi EM. Antibacterial and molluscicidal activities of the essential oil of Chrysanthemum viscidehirtum. Fitoterapia 2000; 71 (05) 544-546
• 27 Cobos MI, Rodriguez JL, Oliva ML, Demo M, Faillaci SM, Zygadlo JA. Composition and antimicrobial activity of the essential oil of Baccharis notosergila. Planta Med 2001; 67 (01) 84-86
• 28 Vieira AJ, Beserra FP, Souza MC, Totti BM, Rozza AL. Limonene: aroma of innovation in health and disease. Chem Biol Interact 2018; 283: 97-106
• 29 Baylac S, Racine P. Inhibition of 5-lipoxygenase by essential oils and other natural fragrant extracts. Inter J Aromatherapy 2003; 13: 138-144
• 30 Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999; 12 (04) 564-582
• 31 Saliu OA, Akanji AM, Idowu OA, Saliu BN. Free radical and reactive oxygen species scavenging potentials of Luffa cylindrica leaf extracts. J Cell Biol Biochem 2020; 4 (01) 013-019
• 32 Torres-Martínez R, García-Rodríguez YM, Ríos-Chávez P. et al. Antioxidant activity of the essential oil and its major terpenes of Satureja macrostema (Moc. and Sessé ex Benth.). Pharmacogn Mag 2018; 13 (Suppl. 04) S875-S880
• 33 Ben Hsouna A, Hamdi N, Ben Halima N, Abdelkafi S. Characterization of essential oil from Citrus aurantium L. flowers: antimicrobial and antioxidant activities. J Oleo Sci 2013; 62 (10) 763-772
• 34 Lin X, Cao S, Sun J, Lu D, Zhong B, Chun J. The chemical compositions, and antibacterial and antioxidant activities of four types of citrus essential oils. Molecules 2021; 26 (11) 3412
• 35 Oulebsir C, Mefti-Korteby H, Djazouli Z, Zebib B, Merah O. Essential oil of Citrus aurantium L. leaves: composition, antioxidant activity, elastase and collagenase inhibition. Agronomy (Basel) 2022; 12: 1466
• 36 Tzaphlidou M. The role of collagen and elastin in aged skin: an image processing approach. Micron 2004; 35 (03) 173-177
• 37 Solano F. Metabolism and functions of amino acids in the skin. In: Wu GY. eds. Amino Acids in Nutrition and Health. SpringerCham; Switzerland: 2020: 187-199
• 38 Kang J, Kim MJ, Hyun JM. et al. Antibacterial, whitening, and anti-wrinkling effects of essential oil from Curcuma aromatica leaves. Pharma Chem 2016; 8 (18) 95-99
• 39 Fraternale D, Flamini G, Ascrizzi R. In vitro anticollagenase and antielastase activities of essential oil of Helichrysum italicum subsp. italicum (Roth) G. Don. J Med Food 2019; 22 (10) 1041-1046
• 40 Ashmawy NS, Gad HA, El-Nashar HAS. Comparative study of essential oils from different organs of Syzygium cumini (pamposia) based on GC/MS chemical profiling and in vitro antiaging activity. Molecules 2023; 28 (23) 7861
• 41 Oboh G, Olasehinde TA, Ademosun AO. Essential oil from lemon peels inhibit key enzymes linked to neurodegenerative conditions and pro-oxidant induced lipid peroxidation. J Oleo Sci 2014; 63 (04) 373-381
• 42 Mori M, Ikeda N, Kato Y, Minamino M, Watabe K. Inhibition of elastase activity by essential oils in vitro . J Cosmet Dermatol 2002; 1 (04) 183-187
• 43 Franklin LU, Cunnington GD, Young D. Terpene based pesticide treatments for killing terrestrial arthropods including, amongst others, lice, lice eggs, mites, and ants. U.S. Patent 19990379268. August, 1999
• 44 Fahmy NM, Elhady SS, Bannan DF, Malatani RT, Gad HA. Citrus reticulata leaves essential oil as an antiaging agent: a comparative study between different cultivars and correlation with their chemical compositions. Plants 2022; 11 (23) 3335
• 45 Aumeeruddy-Elalfi Z, Lall N, Fibrich B, van Staden AB, Hosenally M, Mahomoodally MF. Selected essential oils inhibit key physiological enzymes and possess intracellular and extracellular antimelanogenic properties in vitro . Yao Wu Shi Pin Fen Xi 2018; 26 (01) 232-243
• 46 Gad HA, El Hassab MA, Elhady SS, Fahmy NM. Insights on Citrus clementina essential oil as a potential antiaging candidate with a comparative chemometric study on different cultivars. Ind Crops Prod 2023; 194: 116349
• 47 Prommaban A, Chaiyana W. Microemulsion of essential oils from citrus peels and leaves with anti-aging, whitening, and irritation reducing capacity. J Drug Deliv Sci Technol 2022; 69: 103188
• 48 Mangena T, Muyima NY. Comparative evaluation of the antimicrobial activities of essential oils of Artemisia afra, Pteronia incana and Rosmarinus officinalis on selected bacteria and yeast strains. Lett Appl Microbiol 1999; 28 (04) 291-296
• 49 Tzakou O, Pitarokili D, Chinou IB, Harvala C. Composition and antimicrobial activity of the essential oil of Salvia ringens. Planta Med 2001; 67 (01) 81-83