Eucalyptus Citriodora Extract Regulates Cutaneous Homeostasis Including Immune Dysregulation and Skin Barrier Dysfunction Via the Modulation of Peroxisome Proliferator-Activated Receptor-β/δ (PPAR-β/δ) Pathway

Takuhiro Yamada, Akihiro Aioi

Article ID: 1130
Vol 4, Issue 2, 2020, Article identifier:69-80

VIEWS - 1199 (Abstract) 494 (PDF)


Perturbation of cutaneous homeostasis including immune dysregulation and skin barrier dysfunction evokes skin disorders. In this study, we examined the effect of Eucalyptus citriodora (Euc-c) extract on cytokine production, cell proliferation and cell differentiation in HaCaT cells to elucidate its influence on cutaneous homeostasis. Euc-c suppressed significantly LPS-induced IL-6 and TNF-a-induced IL-8 production from HaCaT cells. Conversely IL-1ra production was significantly enhanced by Euc-c. The expressions of IVL, CERS3 and CERS4, keratinocyte differentiation markers, were upregulated to 3.1, 2.8 and 2.7-fold respectively by Euc-c treatment, compared to the control, while the proliferation was downregulated. The lipid contents in Euc-c-treated cells tended to increase, compared with non-treated cells. To explore the underlying mechanism of these effect, we next performed siRNA experiments against PPAR-b/d. Euc-c enhanced PPAR-b/d mRNA expression to 3.25-fold, while PPAR-b/d mRNA expression in transfected cells was suppressed. The expressions of IVL, CERS3 and CERS4 in transfected cells were suppressed to 1.48, 0.82 and 0.72-fold respectively, concomitant with suppression of PPAR-b/d mRNA expression. These results indicated that Euc-c exerts anti-inflammatory effects and regulates keratinocyte differentiation via the modulation of PPAR-b/d pathway. Therefore, the application of Euc-c is expected to exert beneficial effect on skin disorders evoked by perturbation of skin homeostasis.


Eucalyptus citriodora; PPAR-β/δ; Inflammation; Barrier Function; Cutaneous Homeostasis

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Menona GK, Clearyb GW, Lanec ME. The structure and function of the stratum corneum. Intl J Pharm 2012; 435(1):3-9 doi: 10.1016/j.ijpharm.2012.06.005

Ansel J, Perry P, Brown J, et al. Cytokine modulation of keratinocyte cytokines. J Invest Dermatol 1990; 94(6):101s-107s

Elias PM. Stratum corneum defensive functions: An integrated view. J Invest Dermatol 2005; 125(2):183-200

Jungersted JM, Hellgren LI, Høgh JK, et al. Ceramides and barrier function in healthy skin. Acta Derm Venereol 2010; 90(4):350-353 doi: 10.2340/00015555-0894

Sandilands A, Sutherland C, Irvine AD, et al. Filaggrin in the frontline: role in skin barrier function and disease. J Cell Sci 2009; 122(9):1285-1294 doi:10.1242/jcs.033969

Koch PJ, de Viragh PA, Scharer E, et al. Lessons from loricrin-deficient mice: Compensatory mechanisms maintaining skin barrier function in the absence of a major cornified envelope protein. J Cell Biol 2000; 151(2):389-400 doi: 10.1083/jcb.151.2.389

Elsholz F, Harteneck C, Muller W, et al. Calcium - a central regulator of keratinocyte differentiation in health and disease. Eur J Dermatol 2014; 24(6): 650-661 doi:10.1684/ejd.2014.2452

Hänel KH, Cornelissen C, Lüscher B, et al. Cytokines and the Skin Barrier. Int. J Mol Sci 2013; 14(4): 6720-6745 doi:10.3390/ijms14046720

Grone A. Keratinocytes and cytokines. Vet Immunol Immunopathol 2002; 88(1-2): 1-12

Issemann I, Green S. Activation of a member of the steroid receptor superfamily by peroxisome proliferator. Nature 1990; 347(6294): 645-649 doi:10.1038/347645a0

Sher T, Hua-Fang Y, McBride O, et al. cDNA cloning, chromosomal mapping, and functional characterization of the human peroxisome proliferator activated receptor. Biochemistry 1993; 32(21): 5598-5604 doi:10.1021/bi00072a015

Kliewer SA, Umesono K, Nooman DJ, et al. Convergence of 9-cis retinoic acid and peroxisome proliferator signaling pathways through heterodimer formation of their receptors. Nature 1992; 358(6389): 771-774 doi:10.1038/358771a0

Lamichane S, Lamichane BD, Kwon SM. Pivotal roles of peroxisome proliferator-activated receptors (PPARs) and their signal cascade for cellular and whole-body energy homeostasis. Int J Mol Sci 2018; 19: 949, 2018 doi:10.3390/ijms19040949

Rivier M, Safonova I, Lebrun P, et al. Differential expression of peroxisome proliferator-activated receptor subtypes during the differentiation of human keratinocytes. J Invest Dermatol 1998; 111(6):1116-1121 doi:10.1046/j.1523-1747.1998.00439.x

Aioi A. Peroxisome proliferator-activated receptors (PPARs) activation as therapeutic targets in skin inflammation. Trends in Immunother 2018; 2(4): 1-14. doi:10.24294/ti.v2i4.1063

Sertznig P, Seifert M, Tilgen W, Reichrath J. Peroxisome proliferator-activated receptors (PPARs) and the human skin: Importance of PPARs in skin physiology and dermatologic diseases. Am J Clin Dermatol 2008; 9(1): 15-31

Sertznig P, Reichrath J. Peroxisome proliferator-activated receptors (PPARs) in dermatology. Dermato-Encrinol 2011; 3(3):130-135 doi:10.4161/derm.3.3.15025

Sadlon AE, Lamson DW. Immune-modifying and antimicrobial effects of eucalyptus oil and simple inhalation devices. Altern Med Rev 2010; 15(1):33-47

Gbenou JD, Ahounou JF, Akakpo HB, et al. Phytochemical composition of Cymbopogon citratus and Eucalyptus citriodora essential oils and their anti-inflammatory and analgesic properties on Wistar rats. Mol Biol Rep 2013; 40(2):1127-1134 doi: 10.1007/s11033-012-2155-1

Singh HP, Kaur S, Negi K, et al. Assessment of in vitro antioxidant activity of essential oil of Eucalyptus citriodora (lemon-scented Eucalypt; Myrtaceae) and its major constituents. Food Sci Tech 2012; 48 (2) 237-241 doi:10.1016/j.lwt.2012.03.019

Sepúlveda-Ariasa JC. Velozab A, Escobar LM, et al. Anti-inflammatory effects of the main constituents and epoxides derived from the essential oils obtained from Tagetes lucida, Cymbopogon citratus, Lippia alba and Eucalyptus citriodora. J Essential Oil Res 2013; 25(3):186-193 doi: 10.1080/10412905.2012.751556

Duh PD, Chen ZT, Lee SW, et al. Antiproliferative activity and apoptosis induction of Eucalyptus citriodora resin and its major bioactive compound in melanoma B16F10 cells. J Agric Food Chem 2012; 60(32):7866-7872 doi: 10.1021/jf301068z

Camp D, Davis RA, Campitelli M, et al. Drug-like Properties: Guiding Principles for the Design of Natural Product Libraries. J Nat Prod2012; 75(1):72-81 doi: 1.1021/np200687v

Camp D, Campitelli M, Carroll AR, et al. Front Loading Natural Product Screening Libraries for log P: Background, Development and Implementation. Chem Biodiversity, 2013; 10(4):524-537

Deyrieux AF, Wilson VG. In vitro culture conditions to study keratinocyte differentiation using the HaCaT cell line. Cytotechnology 2007; 54(2):77-83 doi: 10.1007/s10616-007-9076-1

Westergaard M, Henningsen J, Svendsen ML. et al. Modulation of keratinocyte gene expression and differentiation by PPAR-selective ligands and tetradecylthioacetic acid. J. Invest. Dermatol. 2001; 116(5):702–712 doi:10.1046/j.1523-1747.2001.01329.x

Boukamp P, Petrussevska RT, Breitkreutz D, et al. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol 1988; 106(3):761-771

Micallef L, Belaubre F, Pinon A, et al. Effects of extracellular calcium on the growth-differentiation switch in immortalized keratinocyte HaCaT cells compared with normal human keratinocytes. Exp Dermatol 2009; 18(2):143-151 doi:10.1111/j.1600-0625.2008.00775.x

Straus DS, Glass CK. Anti-inflammatory actions of PPAR ligands: new insights on cellular and molecular mechanisms. Trends Immunol 2007; 28(12):551-558 doi: 10.1016/

Devchand PR, Keller H, Peters JM, et al. The PPAR-leukotriene B4 pathway to inflammation control, Nature 1996; 384:39–43 doi:10.1038/384039a0

Jiang C, Ting AT, Sees B. PPAR- agonists inhibit production of monocyte inflammatory cytokines. Nature 1998; 391:83-86 doi:10.1038/34184

Staels B, Koenig W, Habib A, R. et al. Activation of human aortic smooth-muscle cells is inhibited by PPAR alpha but not by PPAR gamma activators. Nature 1998; 393:790–793 doi:10.1038/31701

Marx N, Sukhova GK, Collins T, et al. PPAR alpha activators inhibit cytokine-induced vascular cell adhesion molecule-1 expression in human endothelial cells. Circulation 1999; 99(24):3125–3131

Welch JS, Ricote M, Akiyama TE, et al. PPARγ and PPARγ negatively regulate specific subsets of lipopolysaccharide and IFNγ target genes in macrophages. Proc Natl Acad Sci USA 2003; 100(11):6712–6717 doi:10.1073/pnas.1031789100

Ding G, Cheng L, Qin Q, et al. PPAR modulates lipopolysaccharide-induced TNF- inflammation signaling in cultured cardiomyocytes. J Mol Cell Cardiol 2006; 40(6):821–828 doi:10.1016/j.yjmcc.2006.03.422

Natarajan C, Bright JJ. Peroxisome proliferator-activated receptor-gamma agonists inhibit experimental allergic encephalomyelitis by blocking IL-12 production, IL-12 signaling and Th1 differentiation. Genes and Immunity 2002; 3(2):59–70 doi:10.1038/sj/gene/6363832

Gervois P, Mansouri RM. PPARα as a therapeutic target in inflammation-associated diseases. Expert Opin Ther Targets 2012; 16(11):1113-1125 doi:10.1517/14728222.2012.715633

Kuribayashi S, Xu X, Ishii S, et al. A novel thiazolidinediones MCC-555 down-regulates tumor necrosis factor--induced expression of vascular adhesion molecule-1 in vascular endothelial cells. Atherosclerosis 2005; 182(1):71-77 doi: 10.1016/j.atherosclerosis.2005.02.004

Tyrone E, James PG, Zineh WI. Modulatory effect of fenofibrate on endothelia production of neutrophil chemokines IL-8 and ENA-78. Cardiovasc Drugs Ther 2012; 26(2):95-99 doi:10.1007/s10557-011-6368-7

Ricote M, Glass CK. PPARs and molecular mechanism of transcription. Biochim Biophys Acta 2007; 1771(8):926-936 doi:10.1016/j.bbalip.2007.02.013

Stienstra R, Mandard S, Tan NS, et al. The Interleukin-1 receptor antagonist is a direct target gene of PPAR- in liver. J Hepatol 2007; 46 (5):869-877 doi:10.1016/j.jhep.2006.11.019

Kim HJ, Kim MY, Hwang JS, et al. PPAR inhibits IL-1-stimulated proliferation and migration of vascular smooth muscle cells via up-regulation of IL-1Ra. Cell Mol Life Sci 2010; 67(12):2119-2130 doi:10.1007/s00018-010-0328-4

Romanowska M, Yacoub N, Seidel H, et al. PPAR- enhances keratinocyte proliferation in psoriasis and induces hparin-binding EGF-like growth factor. J Investigative Dermatol 2008; 128(1):110–124 doi:10.1038/sj.jid.5700943

Lianga P, Jiang B, Yanga X, et al. The role of peroxisome proliferator-activated receptor-/ in epidermal growth factor-induced HaCaT cell proliferation. Exp Cell Res 2008; 314(17):3142-3151 doi:10.1016/j.yexcr.2008.06.013

Kim DJ, Bility MT, Billin AN, et al. PPAR/ selectively induces differentiation and inhibits cell proliferation. Cell Death Differ 2006; 13(1):53-60 doi:10.1038/sj.cdd.4401713

Burdick AD, Bility MT, Girroir EE, et al. Ligand activation of peroxisome proliferator-activated receptor-/ (PPAR/) inhibits cell growth of human N/TERT-1 keratinocytes. Cell Signalling 2007; 19 (6) 1163-1171 doi: 10.1016/j.cellsig.2006.12.007

Levy M, Futerman AH. Mammalian ceramide synthases. IUBMB Life, 2010; 62(5):347-356 doi: 10.1002/iub.319

Mizutani Y, Kihara A, Chiba H, et al. 2-Hydroxy-ceramide synthesis by ceramide synthase family: enzymatic basis for the preference of FA chain length. J Lipid Res 49(11):2356-2364 doi:10.1194/jlr.M800158-JLR200

Kima B, Kima JE, Kim HS. Caffeic acid induces keratinocyte differentiation by activation of PPAR-. J Pharm Pharmacol 2014; 66(1):84-92 doi:10.1111/jphp.12159

Mao-Qiang M, Fowler AJ, Schmuth M, et al. Peroxisome-proliferator-activated receptor (PPAR)- activation stimulates keratinocyte differentiation. 2004; J Invest Dermatol 123(2):305 –312 doi:10.1111/j.0022-202X.2004.23235.x

Finegold KR. The role of epidermal lipids in cutaneous permeability barrier homeostasis. J Lipid Res 2007; 48(2):2531-2546 doi: 10.1194/jlr.R700013-JLR200

Man MQ, Choi EH, Schmuth M, et al. Basis for improved permeability barrier homeostasis induced by PPAR and LXR activators: Liposensors stimulate lipid synthesis, lamellar body secretion and post-secretory lipid processing. J Invest Dermatol 2006; 126(2):386-392 doi:10.1038/sj.jid.5700046

Schmuth M, Haqq CM, Cairns WJ, et al. Peroxisome proliferator-activated receptor (PPAR)-/ stimulates differentiation and lipid accumulation in keratinocytes. J Invest Dermatol 2004; 122(4):971-983 doi:10.1111/j.0022-202X.2004.22412.x

Chon SH, Tannahill R, Yao X, et al. Keratinocyte differentiation and upregulation of ceramide synthesis induced by an oat lipid extract via the activation of PPAR pathways. Exp Dermatol 2015; 24(4):290-295 doi:10.1111/exd.12658

Kima MS, Pyun HB, Hwanga JK. Panduratin A, an activator of PPAR- suppresses the development of oxazolone-induced atopic dermatitis-like symptoms in hairless mice. Life Sci 2014; 100(1):45-54 doi:10.1016/j.lfs.2014.01.076

Hatano Y, Man MQ, Uchida Y, et al. Murine atopic dermatitis responds to peroxisome proliferator-activated receptors α and β/δ (but not γγ) and liver X receptor activators. J Allergy Clin Immunol 2010; 125(1):160-169 doi:10.1016/j.jaci.2009.06.049

Michalik L, Desvergne B, Tan NS, et al. Impaired skin wound healing in peroxisome proliferator–activated receptor (PPAR)α and PPARβ mutant mice. J Cell Biol 2001; 154(4):799-814 doi:10.1083/jcb.200011148

Borland MG, Yao PL, Kehres EM, et al. PPAR and PPAR inhibit melanoma tumorigenicity by modulating inflammation and apoptosis. Toxicol Sci 2017; 159(2):436-448 doi: 10.1093/toxsci/kfx147

Ham SA. Yoo T, Hwang JS, et al. Ligand-activated PPAR modulates the migration and invasion of melanoma cells by regulating Snail expression. Am J Cancer Res 2014; 4(6):674-682

Jung YR, Lee EK, Kim DJ, et al. Upregulation of collagen expression via PPAR activation in aged skin by magnesium Lithospermate B from Salvia miltiorrhiza. J Nat Prod 2015; 78(8):2110-2115 doi:10.1021/acs.jnatprod.5b00348



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