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Planta Med 2021; 87(10/11): 827-835
DOI: 10.1055/a-1538-5289
Biological and Pharmacological Activity
Original Papers

Cytotoxicity and Antiviral Activities of Haplophyllum tuberculatum Essential Oils, Pure Compounds, and Their Combinations against Coxsackievirus B3 and B4[ # ]

Assia Hamdi
1   Laboratory of Chemical, Pharmaceutical and Pharmacological Development of Drugs, Faculty of Pharmacy, University of Monastir, Tunisia
2   Department of Analytical Chemistry, Applied Chemometrics and Molecular Modelling (FABI), Center for Pharmaceutical Research (CePhaR), Vrije Universiteit Brussel (VUB), Belgium
,
Aymen Halouani
3   Laboratory of Transmissible Diseases and Biologically Active Substances LR99ES27, Faculty of Pharmacy, University of Monastir, Tunisia
,
Ines Aouf
3   Laboratory of Transmissible Diseases and Biologically Active Substances LR99ES27, Faculty of Pharmacy, University of Monastir, Tunisia
,
Johan Viaene
2   Department of Analytical Chemistry, Applied Chemometrics and Molecular Modelling (FABI), Center for Pharmaceutical Research (CePhaR), Vrije Universiteit Brussel (VUB), Belgium
,
Belsem Marzouk
1   Laboratory of Chemical, Pharmaceutical and Pharmacological Development of Drugs, Faculty of Pharmacy, University of Monastir, Tunisia
,
Jamil Kraiem
1   Laboratory of Chemical, Pharmaceutical and Pharmacological Development of Drugs, Faculty of Pharmacy, University of Monastir, Tunisia
,
Hela Jaïdane
3   Laboratory of Transmissible Diseases and Biologically Active Substances LR99ES27, Faculty of Pharmacy, University of Monastir, Tunisia
,
Yvan Vander Heyden
2   Department of Analytical Chemistry, Applied Chemometrics and Molecular Modelling (FABI), Center for Pharmaceutical Research (CePhaR), Vrije Universiteit Brussel (VUB), Belgium
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Abstract

Haplophyllum tuberculatum is a plant commonly used in folk medicine to treat several diseases including vomiting, nausea, infections, rheumatism, and gastric pains. In the current study, H. tuberculatum essential oils, hydrosols, the pure compounds R-(+)-limonene, S-(−)-limonene, and 1-octanol, as well as their combinations R-(+)-limonene/1-octanol and S-(−)-limonene/1-octanol, were screened for their cytotoxicity on HEp-2 cells after 24, 48, and 72 h, and then tested for their activity against Coxsackievirus B3 and B4 (CV-B3 and CV-B4) at 3 different moments: addition of the plant compounds before, after, or together with virus inoculation. Results showed that the samples were more cytotoxic after 72 h than after 24 h or 48 h cell contact. However, the combinations R-(+)-limonene/1-octanol and S-(−)-limonene/1-octanol showed less effect on HEp-2 cells than pure R-(+)-limonene and S-(−)-limonene after 24 h, 48 h, and 72 h. 1-octanol exhibited the highest concentration causing 50% cytotoxicity (CC50) on HEp-2 cells after 24 h (CC50 = 93 µg/mL) and 48 h (CC50 = 83 µg/mL). The antiviral assays showed that the tested samples exhibited potent inhibition of CV-B. IC50 values ranged from 0.66 µg/mL to 28.4 µg/mL. In addition, CV-B3 was more sensitive than CV-B4. Both CV-B strains are more inhibited when cells were pretreated with the plant compounds. The hydrosols have no effect, neither on HEp-2 cells nor on the virus. 1-octanol, S-(−), and R-(+)-limonene/1-octanol had important selectivity indexes over time. Although essential oils had potent antiviral activity, they can be considered for application in the pretreatment of cells. However, 1-octanol and the combinations are within the safety limits, and thus, they can be used as an active natural antiviral agent for CV-B3 and CV-B4 inhibition.

Key words

Haplophyllum tuberculatum - Rutaceae - essential oils - pure compounds - Coxsackievirus B - cytotoxicity - selectivity index

# Dedicated to Professor Arnold Vlietinck on the occasion of his 80th birthday.




Publication History

Received: 14 January 2021

Accepted after revision: 24 June 2021

Article published online:
22 July 2021

© 2021. Thieme. All rights reserved.

Georg Thieme Verlag KG
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  • References

  • 1 Jaïdane H, Sauter P, Sane F, Goffard A, Gharbi J, Hober D. Enteroviruses and type 1 diabetes: towards a better understanding of the relationship. Reviews Med Virol 2010; 20: 265-280
    CrossrefPubMedGoogle Scholar
  • 2 Bilal RM, AL-Zobaei MA, AL-Ani ZR. Relation of Coxsackie B3 and B4 viral infections for development of type 1 diabetes mellitus in children: a case-control study. Egypt Acad J Biolog Sci, G Microbiol 2019; 11: 1-12
    PubMedGoogle Scholar
  • 3 Burrell CJ, Howard CR, Murphy FA. Chapter 32–Picornaviruses. In: Burrell CJ, Howard CR, Murphy FA. eds. Fenner and Whiteʼs medical Virology (Fifth Edition). London: Academic Press; 2017: 447-463
    Google Scholar
  • 4 Cherry JD. Chapter 24–Enterovirus and Parechovirus Infections. In: Remington JS, Klein JO, Wilson CB, Baker CJ. eds. Infectious Diseases of the Fetus and newborn Infant (Sixth Edition). Philadelphia: W.B. Saunders; 2006: 783-822
    Google Scholar
  • 5 Tariq N, Kyriakopoulos C. Group B Coxsackie Virus. Treasure Island (FL): StatPearls Publishing; 2021. Accessed July 15 2021 at: https://www.ncbi.nlm.nih.gov/books/NBK560783/
    Google Scholar
  • 6 Jaïdane H, Halouani A, Jmii H, Elmastour F, Mokni M, Aouni M. Coxsackievirus B4 vertical transmission in a murine model. Virol 2017; 14: 1-11
    PubMedGoogle Scholar
  • 7 Taylor DJ, Hamid SM, Andres AM, Saadaeijahromi H, Piplani H, Germano JF, Song Y, Sawaged S, Feuer R, Pandol SJ. Antiviral effects of menthol on Coxsackievirus B. Viruses 2020; 12: 373
    CrossrefPubMedGoogle Scholar
  • 8 Rossi RC, da Rosa SR, Weimer P, Moura JGL, de Oliveira VR, de Castilhos J. Assessment of compounds and cytotoxicity of Citrus deliciosa Tenore essential oils: from an underexploited by-product to a rich source of high-value bioactive compounds. Food Biosci 2020; 38: 100779
    CrossrefPubMedGoogle Scholar
  • 9 Dzah CS, Duan Y, Zhang H, Wen C, Zhang J, Chen G, Ma H. The effects of ultrasound assisted extraction on yield, antioxidant, anticancer and antimicrobial activity of polyphenol extracts: a review. Food Biosci 2020; 35: 100547
    CrossrefPubMedGoogle Scholar
  • 10 Pottier-Alapetite G. Flore de la Tunisie. Angiospermes-Dicotyledones. Tunis: Ministère de lʼEnseignement supérieur et de la Recherche scientifique et ministère de lʼAgriculture ed.; 1979. – 1981
    Google Scholar
  • 11 Collenette S. An Illustrated Guide to the Flowers of Saudi Arabia. London: Scorpion Publishing; 1985
    Google Scholar
  • 12 Tackholm V. Students Flora of Egypt. 2nd ed.. ed. Beirut: Cairo University; 1974
    Google Scholar
  • 13 Acheuk F, Cusson M, Doumandji-Mitiche B. Effects of a methanolic extract of the plant Haplophyllum tuberculatum and of teflubenzuron on female reproduction in the migratory locust, Locusta migratoria (Orthoptera: Oedipodinae). J Insect Physiol 2012; 58: 335-341
    CrossrefPubMedGoogle Scholar
  • 14 Al Yousuf MH, Bashir AK, Veres K, Dobos Á, Nagy G, Máthé I, Blunden G, Vera JR. Essential oil of Haplophyllum tuberculatum (Forssk.) A. Juss. from the United Arab Emirates. J Essent Oil Res 2005; 17: 519-521
    CrossrefPubMedGoogle Scholar
  • 15 Al-Rehaily AJ, Al-Howiriny TA, Ahmad MS, Al-Yahya MA, El-Feraly FS, Hufford CD, McPhail AT. Alkaloids from Haplophyllum tuberculatum . Phytochemistry 2001; 57: 597-602
    CrossrefPubMedGoogle Scholar
  • 16 Townsend C, Guest E. Flora of Iraq (Volume Four, Part Two). Baghdad: Ministry of Agriculture and Agrarian Reform; 1980
    Google Scholar
  • 17 Vahdani M, Faridi P, Zarshenas MM, Javadpour S, Abolhassanzadeh Z, Moradi N, Bakzadeh Z, Karmostaji A, Mohagheghzadeh A, Ghasemi Y. Major compounds and antimicrobial activity of essential oils from five Iranian endemic medicinal plants. Pharmacogn J 2011; 3: 48-53
    CrossrefPubMedGoogle Scholar
  • 18 Abdelgaleil S, Saad M, Ariefta N, Shiono Y. Antimicrobial and phytotoxic activities of secondary metabolites from Haplophyllum tuberculatum and Chrysanthemum coronarium . S Afr J Bot 2020; 128: 35-41
    CrossrefPubMedGoogle Scholar
  • 19 Hamdi A, Bero J, Beaufay C, Flamini G, Marzouk Z, Vander Heyden Y, Quetin-Leclercq J. In vitro antileishmanial and cytotoxicity activities of essential oils from Haplophyllum tuberculatum A. Juss leaves, stems, and aerial parts. BMC Complement Altern Med 2018; 18: 60
    CrossrefPubMedGoogle Scholar
  • 20 Jaafari A, Mouse HA, Rakib EM, Tilaoui M, Benbakhta C, Boulli A, Abbad A, Zyad A. Chemical composition and antitumor activity of different wild varieties of Moroccan thyme. Rev Bras Farmacogn 2007; 17: 477-491
    CrossrefPubMedGoogle Scholar
  • 21 Stammati A, Bonsi P, Zucco F, Moezelaar R, Alakomi HL, von Wright A. Toxicity of selected plant volatiles in microbial and mammalian short-term assays. Food Chem Toxicol 1999; 37: 813-823
    CrossrefPubMedGoogle Scholar
  • 22 Tao L, Zhou L, Zheng L, Yao M. Elemene displays anti-cancer ability on laryngeal cancer cells in vitro and in vivo . Cancer Chemother Pharmacol 2006; 58: 24
    CrossrefPubMedGoogle Scholar
  • 23 Rooney S, Ryan M. Effects of alpha-hederin and thymoquinone, constituents of Nigella sativa, on human cancer cell lines. Anticancer Res 2005; 25: 2199-2204
    PubMedGoogle Scholar
  • 24 Jaafari A, Tilaoui M, Mouse HA, Mʼbark LA, Aboufatima R, Chait A, Lepoivre M, Zyad A. Comparative study of the antitumor effect of natural monoterpenes: relationship to cell cycle analysis. Rev Bras Farmacogn 2012; 22: 534-540
    CrossrefPubMedGoogle Scholar
  • 25 Sobral MV, Xavier AL, Lima TC, de Sousa DP. Antitumor activity of monoterpenes found in essential oils. Sci World J 2014; 2014: 953451
    CrossrefPubMedGoogle Scholar
  • 26 Haag JD, Lindstrom MJ, Gould MN. Limonene-induced regression of mammary carcinomas. Cancer Res 1992; 52: 4021-4026
    PubMedGoogle Scholar
  • 27 Yu X, Lin H, Wang Y, Lv W, Zhang S, Qian Y, Deng X, Feng N, Yu H, Qian B. D-limonene exhibits antitumor activity by inducing autophagy and apoptosis in lung cancer. Onco Targets Ther 2018; 11: 1833-1847
    CrossrefPubMedGoogle Scholar
  • 28 Satomi Y, Ohara K, Yazaki K, Ito M, Honda G, Nishino H. Production of the monoterpene limonene and modulation of apoptosis-related proteins in embryonic-mouse NIH 3T3 fibroblast cells by introduction of the limonene synthase gene isolated from Japanese catnip (Schizonepeta tenuifolia). Biotechnol Appl Biochem 2009; 52: 185-190
    CrossrefPubMedGoogle Scholar
  • 29 Chi G, Wei M, Xie X, Soromou L, Liu F, Zhao S. Suppression of MAPK and NF-κB pathways by limonene contributes to attenuation of lipopolysaccharide-induced inflammatory responses in acute lung injury. Inflammation 2013; 36: 501-511
    CrossrefPubMedGoogle Scholar
  • 30 Jia SS, Xi GP, Zhang M, Chen YB, Lei B, Dong XS, Yang YM. Induction of apoptosis by D-limonene is mediated by inactivation of Akt in LS174T human colon cancer cells. Oncol Rep 2013; 29: 349-354
    CrossrefPubMedGoogle Scholar
  • 31 Manassero CA, Girotti JR, Mijailovsky S, García de Bravo M, Polo M. In vitro comparative analysis of antiproliferative activity of essential oil from mandarin peel and its principal component limonene. Nat Prod Res 2013; 27: 1475-1478
    CrossrefPubMedGoogle Scholar
  • 32 Chaudhary S, Siddiqui M, Athar M, Alam MS. D-Limonene modulates inflammation, oxidative stress and Ras-ERK pathway to inhibit murine skin tumorigenesis. Human Exp Toxicol 2012; 31: 798-811
    CrossrefPubMedGoogle Scholar
  • 33 Chen TC, Cho HY, Wang W, Barath M, Sharma N, Hofman FM, Schönthal AH. A novel temozolomid-perillyl alcohol conjugate exhibits superior activity against breast cancer cells in vitro and intracranial triple-negative tumor growth in vivo . Mol Cancer Ther 2014; 13: 1181-1193
    CrossrefPubMedGoogle Scholar
  • 34 Dietrich DR, Swenberg JA. The presence of α2u-globulin is necessary for d-limonene promotion of male rat kidney tumors. Cancer Res 1991; 51: 3512-3521
    PubMedGoogle Scholar
  • 35 Santiago JVA, Jayachitra J, Shenbagam M, Nalini N. Dietary d-limonene alleviates insulin resistance and oxidative stress-induced liver injury in high-fat diet and L-NAME-treated rats. Eur J Nutr 2012; 51: 57-68
    CrossrefPubMedGoogle Scholar
  • 36 Dar MY, Shah WA, Rather MA, Qurishi Y, Hamid A, Qurishi M. Chemical composition, in vitro cytotoxic and antioxidant activities of the essential oil and major constituents of Cymbopogon jawarancusa (Kashmir). Food Chem 2011; 129: 1606-1611
    CrossrefPubMedGoogle Scholar
  • 37 Sakirigui A, Gbaguidi F, Kasséhin UC, Poupaert J, Accrombessi GC, Kotchoni SO. Structural and antitrypanosomal data of different carbasones of piperitone. Data Brief 2016; 9: 1039-1043
    CrossrefPubMedGoogle Scholar
  • 38 Hayes AJ, Leach DN, Markham JL, Markovic B. In vitro cytotoxicity of Australian tea tree oil using Human cell lines. J Essent Oil Res 1997; 9: 575-582
    CrossrefPubMedGoogle Scholar
  • 39 Ashrafizadeh M, Ahmadi Z, Mohammadinejad R, Kaviyani N, Tavakol S. Monoterpenes modulating autophagy: a review study. Basic Clin Pharmacol Toxicol 2020; 126: 9-20
    CrossrefPubMedGoogle Scholar
  • 40 Chander S, Lansdown A, Luqmani Y, Gomm J, Coope R, Gould N, Coombes R. Effectiveness of combined limonene and 4-hydroxyandrostenedione in the treatment of NMU-induced rat mammary tumours. Br J Cancer 1994; 69: 879-882
    CrossrefPubMedGoogle Scholar
  • 41 Jafri SH, Glass J, Shi R, Zhang S, Prince M, Kleiner-Hancock H. Thymoquinone and cisplatin as a therapeutic combination in lung cancer: in vitro and in vivo . J Exp Clin Cancer Res 2010; 29: 87
    CrossrefPubMedGoogle Scholar
  • 42 Astani A, Schnitzler P. Antiviral activity of monoterpenes beta-pinene and limonene against herpes simplex virus in vitro . Iran J Microbiol 2014; 6: 149
    PubMedGoogle Scholar
  • 43 Sharifi-Rad J, Salehi B, Schnitzler P, Ayatollahi S, Kobarfard F, Fathi M, Eisazadeh M, Sharifi-Rad M. Susceptibility of herpes simplex virus type 1 to monoterpenes thymol, carvacrol, p-cymene and essential oils of Sinapis arvensis L., Lallemantia royleana Benth. and Pulicaria vulgaris Gaertn. Cell Mol Biol 2017; 63: 42-47
    CrossrefPubMedGoogle Scholar
  • 44 Chiang LC, Ng LT, Cheng PW, Chiang W, Lin CC. Antiviral activities of extracts and selected pure constituents of Ocimum basilicum . Clin Exp Pharmacol P 2005; 32: 811-816
    CrossrefPubMedGoogle Scholar
  • 45 Samra RM, Soliman AF, Zaki AA, El-Gendy AN, Hassan MA, Zaghloul AM. Chemical composition, antiviral and cytotoxic activities of essential oil from Cyperus rotundus growing in Egypt: evidence from chemometrics analysis. J Essent Oil-Bear Plants 2020; 23: 648-659
    CrossrefPubMedGoogle Scholar
  • 46 Stránská R, Schuurman R, Nienhuis E, Goedegebuure IW, Polman M, Weel JF, Wertheim-Van Dillen PM, Berkhout RJ, van Loon AM. Survey of acyclovir-resistant herpes simplex virus in the Netherlands: prevalence and characterization. J Clin Virol 2005; 32: 7-18
    CrossrefPubMedGoogle Scholar
  • 47 Ma L, Yao L. Antiviral effects of plant-derived essential oils and their components: an updated review. Molecules 2020; 25: 2627
    CrossrefPubMedGoogle Scholar
  • 48 Hamdi A, Majouli K, Vander Heyden Y, Flamini G, Marzouk Z. Phytotoxic activities of essential oils and hydrosols of Haplophyllum tuberculatum . Ind Crops Prod 2017; 97: 440-447
    CrossrefPubMedGoogle Scholar
  • 49 Reed LJ, Muench H. A simple method of estimating fifty percent endpoints. Am J Epidemiol 1938; 27: 493-497
    CrossrefPubMedGoogle Scholar
  • 50 Bernas T, Dobrucki J. Mitochondrial and nonmitochondrial reduction of MTT: interaction of MTT with TMRE, JC-1, and NAO mitochondrial fluorescent probes. Cytometry 2002; 47: 236-242
    CrossrefPubMedGoogle Scholar
  • 51 Berridge MV, Tan AS. Characterization of the cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT): subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction. Arch Biochem Biophys 1993; 303: 474-482
    CrossrefPubMedGoogle Scholar