Top

BMC Complementary Medicine and Therapies

Published in:

Open Access 01-12-2021 | Cytokines | Research

Three chemotypes of thyme (Thymus vulgaris L.) essential oil and their main compounds affect differently the IL-6 and TNFα cytokine secretions of BV-2 microglia by modulating the NF-κB and C/EBPβ signalling pathways

Authors: Györgyi Horváth, Adrienn Horváth, Gréta Reichert, Andrea Böszörményi, Katalin Sipos, Edina Pandur

Published in: BMC Complementary Medicine and Therapies | Issue 1/2021

Abstract

Background

The essential oils possess both antimicrobial and anti-inflammatory effects, therefore they can provide an effective treatment against infections. Essential oils are widely used as supportive ingredients in many diseases, especially in the acute and chronic diseases of the respiratory tract. Neuroinflammation is responsible for several diseases of the central nervous system. Some plant-derived bioactive molecules have been shown to have role in attenuating neuroinflammation by regulating microglia, the immune cells of the CNS.

Methods

In this study, the anti-inflammatory effect of three chemotypes of thyme essential oil and their main compounds (geraniol, thujanol and linalool) were examined on lipopolysaccharide-induced BV-2 microglia. Three different experimental setups were used, LPS pretreatment, essential oil pretreatment and co-treatments of LPS and essential oils in order to determine whether essential oils are able to prevent inflammation and can decrease it. The concentrations of the secreted tumour necrosis factor α (TNFα) and interleukin-6 (IL-6) proinflammatory cytokines were measured and we analysed by Western blot the activity of the cell signalling pathways, NF-κB and CCAAT-enhancer binding protein β (C/EBPβ) regulating TNFα and IL-6 proinflammatory cytokine expressions in BV-2 cells.

Results

Our results showed definite alterations in the effects of essential oil chemotypes and their main compounds at the different experimental setups. Considering the changes of IL-6 and TNFα secretions the best reduction of inflammatory cytokines could be reached by the pretreatment with the essential oils. In addition, the main compounds exerted better effects than essential oil chemotypes in case of LPS pretreatment. At the essential oil pretreatment experiment, the effect of linalool and geraniol was outstanding but there was no major difference between the actions of chemotypes and standards. Main compounds could be seen to have large inhibitory effects on certain cell signalling components related to the activation of the expression of proinflammatory cytokines.

Conclusion

Thyme essential oils are good candidates to use in prevention of neuroinflammation and related neurodegeneration, but the exact ratio of the components has to be selected carefully.

Available only for authorised users

Aziz ZAA, Ahmad A, Setapar SHM, Karakucuk A, Azim MM, Lokhat D, et al. Essential oils: extraction techniques, pharmaceutical and therapeutic potential – A review. Curr Drug Metab. 2018;19(13):1100–10. https://doi.org/10.2174/1389200219666180723144850.CrossRefPubMed

Plant RM, Dinh L, Argo S, Shah M. The essentials of essential oils. Adv Pediatr Infect Dis. 2019;66:111–22.

Nazzaro F, Fratianni F, Coppola R, De Feo V. Essential oils and antifungal activity. Pharmaceuticals. 2017;10(4):86. https://doi.org/10.3390/ph10040086.CrossRefPubMedCentral

Gakuubi MM, Maina AW, Wagacha JM. Antifungal activity of essential oil of Eucalyptus camaldulensis Dehnh. against selected Fusarium spp. Int J Microbiol. 2017;2017:8761610.CrossRef

Cazella NL, Glamoclija J, Sokovic M, Gonçalves JE, Linde GA, Colauto NB, et al. Antimicrobial activity of essential oil Bacchalis dracunculifolia DC (Asteraceae) aerial parts at flowering period. Front Plant Sci. 2019;10:27. https://doi.org/10.3389/fpls.2019.00027.CrossRefPubMedPubMedCentral

Pérez G, Miguel Zavala S, Lucina Arias G, Miguel RL. Anti-inflammatory activity of some essential oils. J Essent Oil Res. 2011;23(5):38–44. https://doi.org/10.1080/10412905.2011.9700480.CrossRef

Shapouri-Moghaddam A, Mohammadian S, Vazini H, Taghadosi M, Esmaeili S-A, Mardani F, et al. Macrophage plasticity, polarization, and function in health and diseases. J Cell Physiol. 2019;233:6424–40.

Xing T, Wang Y, Ding W-J, Li Y-L, Hu X-D, Wang C, et al. Thrombospondin-1 production regulates the inflammatory cytokine secretion in THP-1 cells through NF-κB signaling pathway. Inflammation. 2017;40:1606–21.CrossRef

Sevenich L. Brain-resident microglia and blood-borne macrophages orchestrate central nervous system inflammation in neurodegenerative disorders and brain cancer. Front Immunol. 2018;9:697. https://doi.org/10.3389/fimmu.2018.00697.CrossRefPubMedPubMedCentral

10.

Wang J, Song Y, Chen Z, Leng XS. Connection between systemic inflammation and neuroinflammation underlies neuroprotective mechanism of several phytochemicals in neurodegenerative diseases. Oxid Med and Cell Longev. 2018;2018:1972714.

11.

Welcome MO. Cellular mechanisms and molecular signaling pathways in stress-induced anxiety, depression, and blood–brain barrier inflammation and leakage. Inflammopharmacol. 2020;28(3):643–65. https://doi.org/10.1007/s10787-020-00712-8.CrossRef

12.

Cherry JD, Olschowka JA, O’Banion MK. Neuroinflammation and M2 microglia: the good, the bad, and the inflamed. J Neuroinflammation. 2014;11(1):98. https://doi.org/10.1186/1742-2094-11-98.CrossRefPubMedPubMedCentral

13.

Oh Y-C, Jeong YH, Li W, Go Y. Angelicae gigantis radix regulates LPS-induced neuroinflammation in BV-2 microglia by inhibiting NF-κB and MAPK activity and inducing Nrf-2 activity. Molecules. 2019;24(20):3755. https://doi.org/10.3390/molecules24203755.CrossRefPubMedCentral

14.

Miao X-D, Zheng L-J, Zhao Z-Z, Su S-L, Zhu Y, Guo J-M, et al. Protective effect and mechanism of Boswellic acid and Myrrha sesquiterpenes with different proportions of compatibility on neuroinflammation by LPS-induced BV-2 cells combined with network pharmacology. Molecules. 2019;24(21):3946. https://doi.org/10.3390/molecules24213946.CrossRefPubMedCentral

15.

Weinberg RP, Koledova VV, Schneider K, Sambandan TG, Grayson A, Zeidman G, et al. Palm fruit bioactives modulate human astrocyte activity in vitro altering the cytokine secretome reducing levels of TNFα, RANTES and IP-10. Sci Rep. 2018;8(1):16423. https://doi.org/10.1038/s41598-018-34763-3.CrossRefPubMedPubMedCentral

16.

Agatonovic-Kustrin S, Kustrin E, Morton DW. Essential oils and functional herbs for healthy aging. Neural Regen Res. 2019;14(3):441–5. https://doi.org/10.4103/1673-5374.245467.CrossRefPubMedPubMedCentral

17.

Agatonovic-Kustrin S, Chan CKY, Gegechkori V, Morton DW. Models for skin and brain penetration of major components from essential oils used in aromatherapy for dementia patients. J Biomol Struct Dyn. 2020;38(8):2402–11. https://doi.org/10.1080/07391102.2019.1633408.CrossRefPubMed

18.

Vieira AI, Guerreiro A, Antunes MD, Miguel MDG, Faleiro ML. Edible coatings enriched with essential oils on apples impair the survival of bacterial pathogens through a simulated gastrointestinal system. Foods. 2019;8(2):57. https://doi.org/10.3390/foods8020057.CrossRefPubMedCentral

19.

Inouye S, Yamaguchi H, Takizawa T. Screening of the antibacterial effects of a variety of essential oils on respiratory tract pathogens, using a modified dilution assay method. J Infect Chemother. 2001;7(4):251–4. https://doi.org/10.1007/s101560170022.CrossRefPubMed

20.

Inouye S, Takizawa T, Yamaguchi H. Antibacterial activity of essential oils and their major constituents against respiratory tract pathogens by gaseous contact. J Antimicrob Chemother. 2001;47(5):565–73. https://doi.org/10.1093/jac/47.5.565.CrossRefPubMed

21.

Rota MC, Herrera A, Martínez RM, Sotomayor JA, Jordán MJ. Antimicrobial activity and chemical composition of Thymus vulgaris, Thymus zygis and Thymus hyemalis essential oils. Food Control. 2008;19(7):681–7. https://doi.org/10.1016/j.foodcont.2007.07.007.CrossRef

22.

De Cássia da Silveira e Sá R, Andrade L.N, Dos Reis Barreto de Oliveira R., De Sousa D.P. A review on anti-inflammatory activity of phenylpropanoids found in essential oils. Molecules 2014, 19, 1459–1480, 2, DOI: https://doi.org/10.3390/molecules19021459.

23.

Juhas Š, Bujňáková D, Rehák P, Čikoš Š, Czikková S, Veselá J, et al. Anti-inflammatory effects of thyme essential oil in mice. Acta Vet Brno. 2008;77(3):327–34. https://doi.org/10.2754/avb200877030327.CrossRef

24.

Fachini-Queiroz FC, Kummer R, Estevão-Silva CF, Dalva de Barros Carvalho M, Cunha JM, Grespan R, et al. Effects of thymol and carvacrol, constituents of Thymus vulgaris L. essential oil, on the inflammatory response. Evid Based Complement Alternat Med. 2012;2012:657026.CrossRef

25.

Mei-Lin Tsai M-L, Lin C-C, Lin W-C, Yang C-H. Antimicrobial, antioxidant, and anti-inflammatory activities of essential oils from five selected herbs. Biosci Biotechnol Biochem. 2011;75:1977–83.CrossRef

26.

Vigo E, Cepeda A, Gualillo O, Perez-Fernandez R. In-vitro anti-inflammatory effect of Eucalyptus globulus and Thymus vulgaris: nitric oxide inhibition in J774A.1 murine macrophages. J Pharm Pharmacol. 2004;56(2):257–63. https://doi.org/10.1211/0022357022665.CrossRefPubMed

27.

Ocaña A, Reglero G. Effects of thyme extract oils (from Thymus vulgaris, Thymus zygis, and Thymus hyemalis) on cytokine production and gene expression of oxLDL-stimulated THP-1-macrophages. J Obes. 2012;2012:104706.CrossRef

28.

Braga PC, Dal Sasso M, Culici M, Bianchi T, Bordoni L, Marabini L. Anti-inflammatory activity of thymol: inhibitory effect on the release of human neutrophil elastase. Pharmacology. 2006;77(3):130–6. https://doi.org/10.1159/000093790.CrossRefPubMed

29.

Chauhan AK, Kang SC. Thymol disrupts the membrane integrity of Salmonella ser. typhimurium in vitro and recovers infected macrophages from oxidative stress in an ex vivo model. Res Microbiol. 2014;165(7):559–65. https://doi.org/10.1016/j.resmic.2014.07.001.CrossRefPubMed

30.

Ács K, Balázs VL, Kocsis B, Bencsik T, Böszörményi A, Horváth G. Antibacterial activity evaluation of selected essential oils in liquid and vapor phase on respiratory tract pathogens. BMC Complement Altern Med. 2018;18(1):227. https://doi.org/10.1186/s12906-018-2291-9.CrossRefPubMedPubMedCentral

31.

Pandur E, Tamási K, Pap R, Varga E, Miseta A, Sipos K. Fractalkine induces Hepcidin expression of BV-2 microglia and causes Iron accumulation in SH-SY5Y cells. Cell Mol Neurobiol. 2019;39(7):985–1001. https://doi.org/10.1007/s10571-019-00694-4.CrossRefPubMedPubMedCentral

32.

Pandur E, Varga E, Tamási K, Pap R, Nagy J, Sipos K. Effect of inflammatory mediators lipopolysaccharide and lipoteichoic acid on iron metabolism of differentiated SH-SY5Y cells alters in the presence of BV-2 microglia. Int J Mol Sci. 2019;20:17.CrossRef

33.

Pap R, Montskó G, Jánosa G, Sipos K, Kovács GL, Pandur E. Fractalkine regulates HEC-1A/JEG-3 interaction by influencing the expression of implantation-related genes in an in vitro co-culture model. Int J Mol Sci. 2020;21(9):3175. https://doi.org/10.3390/ijms21093175.CrossRefPubMedCentral

34.

ImageJ. [(Accessed 23 Sept 1997)]; Available online: https://imagej.nih.gov/ij/

35.

Balázs VL, Horváth B, Kerekes E, Ács K, Kocsis B, Varga A, et al. Anti-Haemophilus activity of selected essential oils detected by TLC-direct bioautography and biofilm inhibition. Molecules. 2019;24(18):3301. https://doi.org/10.3390/molecules24183301.CrossRefPubMedCentral

36.

Ekpenyong C, Akpan E, Nyoh A. Ethnopharmacology, phytochemistry, and biological activities of Cymbopogon citratus (DC.) Stapf extracts. Chin J Nat Med. 2015;13(5):321–37. https://doi.org/10.1016/S1875-5364(15)30023-6.CrossRefPubMed

37.

Cavalleri R, Becker JS, Pavan AM, Bianchetti P, Goettert MI, Ethur EM, et al. Essential oils rich in monoterpenes are unsuitable as additives to boar semen extender. Andrologia. 2018;50(8):e13074. https://doi.org/10.1111/and.13074.CrossRefPubMed

38.

Bhatt D, Ghosh S. Regulation of the NF-κB-mediated transcription of inflammatory genes. Front Immunol. 2014;5:71.CrossRef

39.

Ho C, Li LH, Weng YC, Hua KF, Ju TC. Eucalyptus essential oils inhibit the lipopolysaccharide-induced inflammatory response in RAW264.7 macrophages through reducing MAPK and NF-κB pathways. BMC Complement Med Ther. 2020;20(1):200. https://doi.org/10.1186/s12906-020-02999-0.CrossRefPubMedPubMedCentral

40.

Pope RM, Leutz A, Ness SA. C/EBP beta regulation of the tumor necrosis factor alpha gene. J Clin Invest. 1994;94(4):1449–55. https://doi.org/10.1172/JCI117482.CrossRefPubMedPubMedCentral

41.

Schmidt E, Wanner J, Höferl M, Jirovetz L, Buchbauer G, Gochev V, et al. Chemical composition, olfactory analysis and antibacterial activity of Thymus vulgaris chemotypes geraniol, 4-thujanol/terpinen-4-ol, thymol and linalool cultivated in Southern France. NPC. 2012;7:1095–8.PubMed

42.

Satyal P, Murray BL, McFeeters RL, Setzer WN. Essential oil characterization of Thymus vulgaris from various geographical locations. Foods. 2016;5(4):70. https://doi.org/10.3390/foods5040070.CrossRefPubMedCentral

43.

Borugă O, Jianu C, Mişcă C, Goleţ I, Gruia AT, Horhat FG. Thymus vulgaris essential oil: chemical composition and antimicrobial activity. J Med Life. 2014;7:56–60.PubMedPubMedCentral

44.

Cheng BH, Sheen LY, Chang ST. Evaluation of anxiolytic potency of essential oil and S-(+)-linalool from Cinnamomum osmophloeum ct. Linalool leaves in mice. J Tradit Complement Med. 2014;16:27–34.

45.

Sabogal-Guáqueta AM, Osorio E, Cardona-Gómez GP. Linalool reverses neuropathological and behavioral impairments in old triple transgenic Alzheimer's mice. Neuropharmacology. 2016;102:111–20. https://doi.org/10.1016/j.neuropharm.2015.11.002.CrossRefPubMed

46.

Rekha KR, Selvakumar G, Sethupathy S, Santha K, Sivakamasundari RI. Geraniol ameliorates the motor behavior and neurotrophic factors inadequacy in MPTP-induced mice model of Parkinson's disease. J Mol Neurosci. 2013;51(3):851–62. https://doi.org/10.1007/s12031-013-0074-9.CrossRefPubMedPubMedCentral

47.

Abd Rashed A, Abd Rahman AZ, Rathi DNG. Essential oils as a potential Neuroprotective remedy for age-related neurodegenerative diseases: A review. Molecules. 2021;26(4):1107. https://doi.org/10.3390/molecules26041107.CrossRefPubMedPubMedCentral

48.

Chitnis T, Weiner LH. CNS inflammation and neurodegeneration. J Clin Invest. 2017;127(10):3577–87. https://doi.org/10.1172/JCI90609.CrossRefPubMedPubMedCentral

49.

Park B-K, Kim YH, Kim YR, Choi JJ, Yang C, Jang I-S, et al. Antineuroinflammatory and neuroprotective effects of Gyejibokryeong-Hwan in lipopolysaccharide-stimulated BV2 microglia. Evid Based Complementary Altern Med. 2019;2019:7585896.

50.

Ismail EN, Jantan I, Vidyadaran S, Jamal JA, Azmi N. Phyllanthus amarus prevents LPS-mediated BV2 microglial activation via MyD88 and NF-κB signaling pathways. BMC Complement Med Ther. 2020;20(1):202. https://doi.org/10.1186/s12906-020-02961-0.CrossRefPubMedPubMedCentral

51.

Elmann A, Mordechay S, Rindner M, Ravid U. Anti-neuroinflammatory effects of geranium oil in microglial cells. J Funct Foods. 2010;2(1):17–22. https://doi.org/10.1016/j.jff.2009.12.001.CrossRef

52.

Barrera-Sandoval AM, Osorio E, Cardona-Gómez GP. Microglial-targeting induced by intranasal linalool during neurological protection postischemia. Eur J Pharmacol. 2019;857:172420. https://doi.org/10.1016/j.ejphar.2019.172420.CrossRefPubMed

53.

Ackerman K, Fiddler J, Soh T, Clarke S. BV-2 microglial cells used in a model of neuroinflammation. FASEB J. 2015;29:608.2.

54.

Wang J, Su B, Zhu H, Chen C, Zhao G. Protective effect of geraniol inhibits inflammatory response, oxidative stress and apoptosis in traumatic injury of the spinal cord through modulation of NF-κB and p38 MAPK. Exp Ther Med. 2016;12(6):3607–13. https://doi.org/10.3892/etm.2016.3850.CrossRefPubMedPubMedCentral

55.

Smale TS. Dimer-specific regulatory mechanisms within the NF-κB family of transcription factors. Immunol Rev. 2012;246(1):193–204. https://doi.org/10.1111/j.1600-065X.2011.01091.x.CrossRef

56.

Bonjardim LR, Cunha ES, Guimarães AG, Santana MF, Oliveira MG, Serafini MR, et al. Evaluation of the anti-inflammatory and antinociceptive properties of p-cymene in mice. Z Naturforsch C J Biosci. 2012;67(1-2):15–21. https://doi.org/10.1515/znc-2012-1-203.CrossRefPubMed

57.

Ramalho TR, Oliveira MT, Lima AL, Bezerra-Santos CR, Piuvezam MR. Gamma-Terpinene modulates acute inflammatory response in mice. Planta Med. 2015;81(14):1248–54. https://doi.org/10.1055/s-0035-1546169.CrossRefPubMed

58.

Held S, Schieberle P, Somoza V. Characterization of alpha-terpineol as an anti-inflammatory component of orange juice by in vitro studies using oral buccal cells. J Agric Food Chem. 2007;55(20):8040–6. https://doi.org/10.1021/jf071691m.CrossRefPubMed

59.

Hart PH, Brand C, Carson CF, Riley TV, Prager RH, Finlay-Jones JJ. Terpinen-4-ol, the main component of the essential oil of Melaleuca alternifolia (tea tree oil), suppresses inflammatory mediator production by activated human monocytes. Inflamm Res. 2000;49(11):619–26. https://doi.org/10.1007/s000110050639.CrossRefPubMed

60.

de Cássia da Silveira e Sá R., Andrade L.N., de Sousa D.P. A review on anti-inflammatory activity of monoterpenes. Molecules 2013, 18, 1227–1254, A Review on Anti-Inflammatory Activity of Monoterpenes, 1, DOI: https://doi.org/10.3390/molecules18011227.

61.

de Cássia da Silveira E Sá R., Lima T.C., da Nóbrega F.R., de Brito A.E.M., de Sousa D.P. Analgesic-like activity of essential oil constituents: an update. Int J Mol Sci 2017, 18, 2392.

62.

Gonçalves MJ, Cruz MT, Tavares AC, Cavaleiro C, Lopes MC, Canhoto J, et al. Composition and biological activity of the essential oil from Thapsia minor, a new source of geranyl acetate. Ind Crops and Prod. 2012;35(1):166–71. https://doi.org/10.1016/j.indcrop.2011.06.030.CrossRef

63.

Brito, L.F., Oliveira, H.B.M., das Neves Selis N., E Souza C.L.S.; Júnior, M.N.S.; de Souza, E.P.; Silva, L.S.C.D.; de Souza Nascimento, F.; Amorim, A.T., Campos, G.B.; de Oliveira, M.V.; Yatsuda, R., Timenetsky, J.; marques, L.M. anti-inflammatory activity of β-caryophyllene combined with docosahexaenoic acid in a model of sepsis induced by Staphylococcus aureus in mice. J Sci Food Agric 2019, 99, 5870–5880, 13, DOI: https://doi.org/10.1002/jsfa.9861.

64.

Narnoliya, L.K.; Jadaun, J.S.; Singh, S.P. The Phytochemical Composition, Biological Effects and Biotechnological Approaches to the Production of High-Value Essential Oil from Geranium. Malik S. (eds). Essential Oil Research 2019, 327–352.

65.

Yang H, Jung EM, Ahn C, Lee GS, Lee SY, Kim SH, et al. Elemol from Chamaecyparis obtusa ameliorates 2,4-dinitrochlorobenzene-induced atopic dermatitis. Int J Mol Med. 2015;36(2):463–72. https://doi.org/10.3892/ijmm.2015.2228.CrossRefPubMed

Title: Three chemotypes of thyme (Thymus vulgaris L.) essential oil and their main compounds affect differently the IL-6 and TNFα cytokine secretions of BV-2 microglia by modulating the NF-κB and C/EBPβ signalling pathways
Authors: Györgyi Horváth
Adrienn Horváth
Gréta Reichert
Andrea Böszörményi
Katalin Sipos
Edina Pandur
Publication date: 01-12-2021
Publisher: BioMed Central
Keyword: Cytokines
Published in: BMC Complementary Medicine and Therapies / Issue 1/2021
Electronic ISSN: 2662-7671
DOI: https://doi.org/10.1186/s12906-021-03319-w

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Three chemotypes of thyme (Thymus vulgaris L.) essential oil and their main compounds affect differently the IL-6 and TNFα cytokine secretions of BV-2 microglia by modulating the NF-κB and C/EBPβ signalling pathways

Abstract

Background

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2021

Multiple facial candidal abscesses after self-administered acupuncture in a patient with undiagnosed diabetes mellitus: a case report

A randomized placebo-controlled phase I clinical trial to evaluate the immunomodulatory activities of Atractylodes lancea (Thunb) DC. in healthy Thai subjects

Ethanol extract of mulberry leaves partially restores the composition of intestinal microbiota and strengthens liver glycogen fragility in type 2 diabetic rats

Piperlongumine inhibits the growth of non-small cell lung cancer cells via the miR-34b-3p/TGFBR1 pathway

The use of complementary and alternative medicine during pregnancy: a cross-sectional study from Palestine

Molecular mechanism of Epicedium treatment for depression based on network pharmacology and molecular docking technology