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Phytosphingosine enhances moisture level in human skin barrier through stimulation of the filaggrin biosynthesis and degradation leading to NMF formation

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Phytosphingosine (PHS) is a sphingoid that is a key component of phytoceramides NP, AP and EOP. PHS has been known to have anti-inflammation and antimicrobial activities and to stimulate epidermal differentiation. In addition, it is reported that PHS treatment notably increased phytoceramide content in keratinocytes. In this study, we tried to investigate whether PHS has any effect on the maturation of corneocytes such as formation of cornified envelope and natural moisturizing factor (NMF) that is also an essential event during the formation of skin barrier, stratum corneum. Special focus was made on the filaggrin (FLG) metabolism that is directly responsible for NMF production. PHS increased the expression of essential keratinocyte differentiation genes such as involucrin and transglutaminase 1 in cultured human keratinocytes. Interestingly, the expressions of FLG, caspase 14 and bleomycin hydrolase, all of which involved in NMF production in corneocytes, were significantly induced by PHS treatment in vitro. The effect of PHS on FLG metabolism was manifested as the increase of pyrrolidone carboxylic acid and skin hydration in vivo human skin. Results showed PHS had skin moisturizing effect by modulating FLG metabolic pathways and suggested to be an essential role in coordinated formation of the corneocyte envelope and NMF within.
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Arch Dermatol Res (2017) 309:795–803
DOI 10.1007/s00403-017-1782-8
Phytosphingosine enhances moisture level inhuman skin barrier
throughstimulation ofthefilaggrin biosynthesis anddegradation
leading toNMF formation
HyunKyungChoi1· YoungHoonCho1· EunOkLee2· JinWookKim2·
Received: 8 March 2017 / Revised: 7 September 2017 / Accepted: 13 September 2017 / Published online: 21 September 2017
© Springer-Verlag GmbH Germany 2017
Keywords Phytosphingosine· Filaggrin· Natural
moisturizing factor· Pyrrolidone carboxylic acid· Skin
The epidermis provides physical and physiological protec-
tive functions such as permeability barrier to prevent water
loss from inside and invasion of harmful external factors
such as chemicals, microbial pathogens, allergens and UV
radiation [4, 9, 12]. The skin barrier function is located in the
outermost layer of the epidermis, the stratum corneum (SC),
which consists of corneocytes and intercellular lipid lamellar
organization. These two structural compartments have been
depicted as ‘bricks and mortar model’ [10, 32, 35]. Formation
of normal corneocytes (bricks) requires two separate events
to be completed: first, maturation of cornified envelope (CE)
that replaces cell membrane of viable keratinocytes with the
aggregates of proteins such as involucrin and loricrin tightly
cross-linked each other by transglutaminase-1 (TGase-1).
Subsequently, CE becomes corneocyte lipid envelope (CLE)
through covalent attachment of ω-hydroxyceramide. Second,
anucleated corneocytes, flattened by the compressive action
of filaggrin (FLG) on keratin intermediate filaments (KIF),
should be filled with natural moisturizing factors (NMF) such
as PCA, UCA and other amino acids derived from degrada-
tion of FLG through hydrolyzing processes involved calpain
1, caspase 14 and bleomycin hydrolase (BLMH) [4, 11, 40].
The importance of CE maturation has been well documented.
TGase-1 is essential to maintain epidermal homeostasis and
healthy skin conditions [4, 19]. Rawlings etal. have reported
that reduced TGase activity can lead to immaturity of CEs and
this has been specially represented in dry skin [8]. Further-
more, dysregulated differentiation process is closely connected
Abstract Phytosphingosine (PHS) is a sphingoid that is a
key component of phytoceramides NP, AP and EOP. PHS
has been known to have anti-inflammation and antimicro-
bial activities and to stimulate epidermal differentiation. In
addition, it is reported that PHS treatment notably increased
phytoceramide content in keratinocytes. In this study, we
tried to investigate whether PHS has any effect on the matu-
ration of corneocytes such as formation of cornified enve-
lope and natural moisturizing factor (NMF) that is also an
essential event during the formation of skin barrier, stratum
corneum. Special focus was made on the filaggrin (FLG)
metabolism that is directly responsible for NMF produc-
tion. PHS increased the expression of essential keratinocyte
differentiation genes such as involucrin and transglutami-
nase 1 in cultured human keratinocytes. Interestingly, the
expressions of FLG, caspase 14 and bleomycin hydrolase,
all of which involved in NMF production in corneocytes,
were significantly induced by PHS treatment invitro. The
effect of PHS on FLG metabolism was manifested as the
increase of pyrrolidone carboxylic acid and skin hydration
invivo human skin. Results showed PHS had skin mois-
turizing effect by modulating FLG metabolic pathways and
suggested to be an essential role in coordinated formation of
the corneocyte envelope and NMF within.
* Chang Seo Park
1 Department ofChemical andBiochemical Engineering,
Dongguk University, 3-26, Pil-dong, Chung-gu,
Seoul100-715, RepublicofKorea
2 LCS Biotech, Seodun-dong 103-2, Gwonseon-gu, Suwon-si,
Gyeonggi-do441-857, RepublicofKorea
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... Across the world, many cosmetic companies have tried to create products that include a cosmeceutical ingredient to address these aging factors. [46][47][48] However, the aging process can vary because of the exposome (external and lifestyle factors). 1,9,10,49 Although skin health and aging are affected by both the genome and exposome, an individual's intrinsic genes do not change. ...
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Sphingolipids (SPs) are structurally diverse and represent one of the most quantitatively abundant classes of lipids in mammalian cells. In addition to their structural roles, many SP species are known to be bioactive mediators of essential cellular processes. Historically, studies have focused on SP species that contain the canonical 18‑carbon, mono-unsaturated sphingoid backbone. However, increasingly sensitive analytical technologies, driven by advances in mass spectrometry, have facilitated the identification of previously under-appreciated, molecularly distinct SP species. Many of these less abundant species contain noncanonical backbones. Interestingly, a growing number of studies have identified clinical associations between these noncanonical SPs and disease, suggesting that there is functional significance to the alteration of SP backbone structure. For example, associations have been found between SP chain length and cardiovascular disease, pain, diabetes, and dementia. This review will provide an overview of the processes that are known to regulate noncanonical SP accumulation, describe the clinical correlations reported for these molecules, and review the experimental evidence for the potential functional implications of their dysregulation. It is likely that further scrutiny of noncanonical SPs may provide new insight into pathophysiological processes, serve as useful biomarkers for disease, and lead to the design of novel therapeutic strategies.
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Background: New ceramide (CER) NPs were prepared by linking fatty acids derived from oils of Korean traditional plants to phytosphingosine(PHS). The oils of Korean traditional plants were extracted from the seeds of Panax ginseng, Camellia sinensis, Glycine max napjakong, Glycine max seoritae and Camellia japonica as sources of diverse fatty acids AIMS: To investigate signaling bioactivities of HP-C. sinensis ceramide NP that was column purified to remove any residual PHS and to evaluate the skin barrier functions of the HP-C. sinensis ceramide NP in human skin. Methods: The expressions of genes related with epidermal differentiation was analyzed in vitro by qPCR. Human studies were also performed to determine the skin barrier functions with respect of TEWL and SC cohesion. Results: The HP-C. sinensis CER NP significantly enhanced the expressions of FLG, CASP14 and INV indicates that the signaling biological activities of oil-derived ceramide NPs could be different depend on the natural oils. The control ceramide, C18-CER NP had no effect on the expression of the three genes. HP-C. sinensis CER NP was selected for the in vivo human studies. Application of 0.5% HP-C. sinensis CER NP cream stimulated significantly faster recovery of a disrupted skin barrier than that of the control C18-CER NP. A significant enhancement of SC cohesion of the skin treated with 0.5% HP-C. sinensis CER NP was also observed. Conclusion: Taken all together, our results clearly demonstrate that HP-C. sinensis CER NP, P. ginseng CER NP and other oil-derived CER NP could be a better choice for developing moisturizers to improve skin barrier function as they more closely mimic the endogenous CER composition of the actual human skin barrier.
Introduction This study investigated the role of natural polymers as moisturizers with low toxicity and biodegradability in the cosmetic and pharmaceutical industries. We isolated a polysaccharide extract from Dendrobium candidum (D. candidum) and determined its efficacy in skin hydration when used as an active cosmetic ingredient. Methods The molecular weight distribution of D. candidum polysaccharides was analyzed via gel permeation chromatography (GPC). We performed real-time reverse transcription PCR (RT-PCR) and western blotting assays to investigate the physiological mechanism of the polysaccharides extracted from D. candidum (PDC). Based on in vitro data, the efficacy of PDC in improving skin condition was tested on the face of 21 volunteers. Results The expression of filaggrin (FLG), caspase-14, and bleomycin hydrolase, which are the major components contributing to skin hydration, was significantly increased in the PDC-treated group. Further, the PDC upregulated the mRNA expression of occludin and claudin-1, which play a key role in epidermal barrier function. In addition, a topical application of PDC markedly increased skin hydration and improved trans-epidermal water loss (TEWL) and skin elasticity after 2 weeks. Conclusions It is the first study reporting the efficacy of PDC-mediated FLG mechanism associated with positive skin hydration. PDC can be used as an active ingredient in moisturizers. Long-term application of PDC-based moisturizers may result in significant improvement in elasticity and barrier function.
Untargeted metabolomics approach based on ultra high performance liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS) was used to investigate the differences in cage duck eggs and sea duck eggs that from different poultry breeding system, which could help to combat fraud within the egg industry. High dimensions and complex data collected by UHPLC-HRMS were analyzed by multivariate statistical analysis. Identification model of sea duck eggs based on was established. After matching with the chemical databases, four potential markers were putatively matched. Further analysis showed that three of them were confirmed by reference standards. All these three markers (n-behenoyl-d-erythro-sphingosine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine and n-nervonoyl-d-erythro-sphingosine) have higher content in sea duck eggs. The quantitative analysis showed that the content difference of three markers in farm samples were in highly consistent with the concentration changes measured in experimental samples, which indicate that these three markers are reliable.
Galaxolide (HHCB), one of the most widely used synthetic musks in personal care products (PCPs), has been recognized as an emerging contaminant with potential human health concerns. To overcome such adverse effects, a systematic molecular design, screening and performance evaluation approach was developed to generate functionally improved and environmentally friendly HHCB derivatives. Among the 90 designed HHCB derivatives, 15 were screened with improved functional properties (i.e., odor stability and intensity) and less environmental impacts (i.e., lower bio-toxicity, bio-accumulation ability, and mobility) using 3D-QSAR models and density functional theory methods. Their human health risks were then assessed by toxicokinetic analysis, which narrowed the candidates to four. Derivative 7, the designed molecule with the least dermal adsorption potential, was evaluated for its interaction with other PCPs additives (i.e., anti-photosensitivity materials and moisturizer) and such impacts on human health risks using molecular docking and molecular dynamic simulation. The environmental fate of Derivative 7 after transformation (i.e., photodegradation, biotransformation, and chlorination) was also discussed. Biotransformation and chlorination were recognized as optimum options for Derivative 7 mitigation. This study provided the theoretical basis for the design of functionally improved and environmentally friendly HHCB alternatives and advanced the understanding of their environmental behaviors and health risks.
Introduction: Lactic fermentation products (LFPs) are thought to affect "good" bacteria in the gut. We previously reported that oral administration of LFPs has beneficial therapeutic effects in a mouse model of atopic dermatitis. However, it is unclear how LFPs affect human epidermal cell differentiation, ceramide (Cer), and amino acid production. Objective: The aim of this study was to determine the effects of LFPs on epidermal cell differentiation, by assessing amino acid and Cer production. Methods: A 3-dimensional cultured human epidermis model and normal human epidermal keratinocytes were used. Cytotoxicity tests were performed using alamar Blue. Transepidermal water loss (TEWL) was used as an index to assess barrier function. Keratin 1 (K1), keratin 5 (K5), keratin 10 (K10), involucrin (INV), calpain 1, and transglutaminase (TGase) (markers of differentiation) and profilaggrin (proFLG) and bleomycin hydrolase (amino acid synthesis-related genes) expression levels were quantified by RT-PCR. In addition, TGase protein levels were measured by Western blotting. The intercellular lipid content of the stratum corneum was measured by high-performance thin-layer chromatography. Amino acids were quantified using an amino acid analyzer. Finally, bound water content in the stratum corneum was measured by differential scanning calorimetry. Results: Cell viability did not change, but TEWL was significantly decreased in the cells treated with LFPs compared with the control cells. Treatment with LFPs significantly increased expression of the late-differentiation markers INV and TGase at the RNA level. Furthermore, TGase protein expression was significantly increased by treatment with LFPs. Treating a 3-dimensional cultured epidermis model with LFPs significantly increased the intercellular lipid content of the stratum corneum and production of the amino acid arginine (Arg). The amount of bound water in the stratum corneum was increased significantly in the LFP application group. Conclusion: Treatment with LFPs promotes human epidermal cell differentiation and increases the intercellular content of the free fatty acid, Chol, Cer [NS], Cer [AS], and Cer [AP]. This may result in improved skin barrier function. The increased amount of Arg observed in keratinocytes may help improve water retention.
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Xerosis affects up to 75% of older people and develops as a result of a skin barrier defect. Emollients are widely used to treat xerosis; however, there is limited understanding of the differences between them and their effects on the skin barrier in older people. This study aimed to compare the effect of a commercially available emollient containing 5% urea, ceramide NP and lactate (test emollient) to an alternative emollient without these additives (control emollient) on the properties of the skin barrier in older people. Two cohorts of 21 volunteers aged 60+ years with dry skin were recruited. The first applied the test emollient to one forearm and no treatment to the other for 28-days. The second compared the test emollient to the control emollient observing the same parameters. Effects on the skin barrier were determined by measuring skin barrier function, hydration, skin surface pH and by analyzing FTIR spectra before and after treatment. A third cohort of 6 young adults was recruited to investigate the effect of a single treatment with the test emollient on the molecular structure of the skin barrier at greater depths by employing the tape-stripping technique. The test emollient hydrated the skin to a significantly greater extent and for a longer period of time compared to the control emollient, an effect associated with a significant elevation of carboxylate groups (a marker of NMF content) within the stratum corneum. Furthermore, the test emollient imparted additional benefits to the structure and function of the skin barrier not exhibited by the control emollient. In conclusion the test emollient addressed the pathological features of xerotic aged skin, supporting its use as first-line therapy for xerotic skin conditions in this population.
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The skin plays a key role in protecting the body from the environment and from water loss. Cornified envelope (CE) and natural moisturizing factor (NMF) are considered as the primary regulators of skin hydration and barrier function. The CE prevents loss of water from the body and is formed by cross-linking of several proteins. Among these proteins, filaggrin is an important protein because NMF is produced by the degradation of filaggrin. Proteases, including matriptase and prostasin, stimulate the generation of filaggrin from profilaggrin and caspase-14 plays a role in the degradation of filaggrin. This study elucidated the effects of an ethanol extract of Boesenbergia pandurata (Roxb.) Schltr., known as fingerroot, and its active compound panduratin A on CE formation and filaggrin processing in HaCaT, human epidermal keratinocytes. B. pandurata extract (BPE) and panduratin A significantly stimulated not only CE formation but also the expression of CE proteins, such as loricrin, involucrin, and transglutaminase, which were associated with PPARα expression. The mRNA and protein levels of filaggrin and filaggrin-related enzymes, such as matriptase, prostasin, and caspase-14 were also up-regulated by BPE and panduratin A treatment. These results suggest that BPE and panduratin A are potential nutraceuticals which can enhance skin hydration and barrier function based on their CE formation and filaggrin processing.
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Background Maintenance of water balance in the stratum corneum (SC) is determined by the content of intercellular lipids and natural moisturizing factors (NMFs) in corneocytes.AimTo investigate the association between the NMFs and (pro)filaggrin and the proteases responsible for the processing of (pro)filaggrin to NMFs in the SC of hydrated and dry skin areas of healthy human subjects.Methods The SC hydration state and the transepidermal water loss (TEWL) were measured using a Corneometer and a Tewameter, respectively. Proteases, (pro)filaggrin and NMFs were extracted from SC samples obtained by tape-stripping of the tested skin. Expression levels of (pro)filaggrin were determined by dot blotting and western blotting, and total NMFs by ultra-high performance liquid chromatography. Expression of the proteases caspase-14, calpain-1 and bleomycin hydrolase was measured by western blotting.ResultsThe levels of (pro)filaggrin were not significantly different between hydrated and dry skin, whereas the level of total NMFs was significantly reduced in dry skin. A negative correlation between (pro)filaggrin and NMFs was found in dry skin (Pearson correlation coefficient r = − 0.57, *P < 0.05). Bleomycin hydrolase expression was significantly decreased in the SC of dry skin.Conclusions These results suggest that the low hydration state of dry skin may be due to the reduction in (pro)filaggrin degradation caused by decreased bleomycin hydrolase expression.
Ceramides based on phytosphingosine, sphingosine and dihydrosphingosine are essential constituents of the skin lipid barrier that protects the body from excessive water loss. The roles of the individual ceramide subclasses in regulating skin permeability and the reasons for C4-hydroxylation of these sphingolipids are not completely understood. We investigated the chain length-dependent effects of dihydroceramides, sphingosine ceramides (with C4-unsaturation) and phytoceramides (with C4-hydroxyl) on the permeability, lipid organization and thermotropic behavior of model stratum corneum lipid membranes composed of ceramide/lignoceric acid/cholesterol/cholesteryl sulfate. Phytoceramides with very long C24 acyl chains increased the permeability of the model lipid membranes compared to dihydroceramides or sphingosine ceramides with the same chain lengths. Either unsaturation or C4-hydroxylation of dihydroceramides induced chain length-dependent increases in membrane permeability. Infrared spectroscopy showed that C4-hydroxylation of the sphingoid base decreased the relative ratio of orthorhombic chain packing in the membrane and lowered the miscibility of C24 phytoceramide with lignoceric acid. The phase separation in phytoceramide membranes was confirmed by X-ray diffraction. In contrast, phytoceramides formed strong hydrogen bonds and highly thermostable domains. Thus, the large heterogeneity in ceramide structures and in their aggregation mechanisms may confer resistance towards the heterogeneous external stressors that are constantly faced by the skin barrier.
Background: Sensitive skin is a poorly understood skin condition. Defects in stratum corneum (SC) barrier function and/or extrasensory neuronal networks in the epidermis are believed to be involved in the problem. Objectives: This study aimed to unravel the relationships between bleomycin hydrolase (BH) and calpain-1 (C-1), pyrrolidone carboxylic acid (PCA) levels, corneocyte maturation, transglutaminase (TG) and plasmin activities on the cheeks of subjects with sensitive skin. Methods: Forty-eight female Caucasian subjects, Fitzpatrick skin phototypes II-III, with self - perceived sensitive facial skin were assessed and underwent a capsaicin reactivity test. Expert grading of skin condition was conducted as well as measurement of transepidermal water loss (TEWL), skin capacitance, SC cohesion and SC integrity. BH, C-1 and plasmin activities were measured as well as PCA levels, plasmin and TG activity. Differential Nile red and involucrin immunostaining was performed to assess corneocyte maturation and size. Results: 52% of the subjects reacted to capsaicin. There were no significant differences between the capsaicin-sensitive and non-capsaicin-sensitive subjects with reference to skin grading, TEWL, skin capacitance and SC cohesion. PCA levels and BH activity were lowest in the capsaicin-sensitive panel (p<0.05) and were correlated in non-capsaicin-sensitive subjects (r = 0.72). The activity of TG was significantly lower (48%) in the capsaicin-sensitive subjects (p<0.001) and their corneocytes were less mature and smaller (p ≤ 0.03). SC was estimated to be thinner (6.87 ± 0.28 vs. 8.68 ± 0.26 μm; p=0.001) in the capsaicin-sensitive subjects with a corresponding shorter SC path length (83.2± 4.4 μm and. 113.1 ± 4.5 μm; p=0.001). Conclusions: Despite the physiological similarities between the two groups of sensitive skin subjects, differences in their biochemistry were clearly evident. Lower levels of PCA, BH and TG activities together with a greater number of smaller and immature corneocytes indicate inferior SC maturation in the capsaicin-sensitive subjects. The reduced maturation of corneocytes and thinner SC likely contributes to a greater penetration of capsaicin and the associated increased skin sensitivity. This article is protected by copyright. All rights reserved.
Background: Knowledge of the ethnic differences and effects of photodamage on the relative amounts of natural moisturizing factor (NMF) together with filaggrin (FLG) processing enzymes in facial stratum corneum (SC) is limited. Our aim was to characterize the activities of calpain-1 (C-1), bleomycin hydrolase (BH) and the levels of pyrrolidone carboxylic acid (PCA) as a marker for total NMF levels and to relate them to plasmin activities and corneocyte maturation. Methods: Enzyme activities, PCA levels and corneocyte maturation were determined from facial tape strippings of photoexposed cheek and photoprotected post-auricular areas (PA) of healthy Caucasian (C), Black African (BA) and Albino African (AA) female subjects living in South Africa. Results: PCA concentration levels were of the order AA > BA > C subjects and the highest activities of BH were present in the AA subjects. BH activities were greater on the photoexposed sites for the BA and C subjects but they were only numerically elevated in the AA subjects. Photoprotected sites had an increase in C-1 activity in pigmented groups (C and BA) whereas in the AA subjects the opposite was measured. Plasmin activities were greater on the cheek compared with the PA site for the AA and C subjects but the activity was low in the BA subjects, In both sites. In both test sites, the AA, but not the BA and C subjects, had smaller, parakeratotic and less mature corneocytes. Conclusion: Variation in PCA levels has been found for different ethnic groups in this study (AA > BA > C subjects). The values in the AA subjects are surprising as one might expect that the lack of pigmentation, and thereby increased photodamage, might lead to lower levels. Increased BH, but not C1 activity, was observed in the AA subjects indicating that BH is associated with PCA production to a greater extent. Surprisingly, corneocyte maturation is still impaired with elevated PCA levels in AA subjects. The higher levels of plasmin and BH activities on the cheeks, especially for AA and C subjects, suggest that they can be used as markers for epidermal photodamage. This article is protected by copyright. All rights reserved.
Background: Therapeutic options for atopic dermatitis mostly address the symptoms but causal therapies are still missing. Peroxisome proliferator activated receptor (PPAR) agonists exert beneficial effects in patients suffering this disease, whereas the stimulation of PPARα and γ seemed most promising. Objectives: To elucidate the effects of the PPARα specific agonist WY14643, the PPARγ agonist ciglitazone, and the dual PPARα+γ agonist docosahexaenoic acid (DHA) on the homeostasis and barrier function of filaggrin deficient skin. Methods: The effects of the PPAR agonists on skin differentiation were evaluated via qPCR, Western blot, histological or immunofluorescence staining. Skin lipid organization was determined by ATR-FTIR and lipid composition was analyzed by HPTLC. Ultimately, the skin barrier function was assessed by skin absorption studies using the radioactively labeled compound testosterone. Results: Significant upregulation of filaggrin after DHA and WY14643 supplementation, but no effect of ciglitazone, on protein and mRNA level was detected. DHA and WY14643, but not ciglitazone, normalized the molar ratio of the main skin barrier lipids to 1:1:1 (free fatty acids:ceramides:cholesterol). Furthermore, DHA and WY14643 supplementation normalized the skin lipid profile in filaggrin deficient skin, but only WY14643 significantly improved the skin barrier function. Conclusion: Supplementation particularly with the PPARα agonist WY14643 improved the homeostasis and barrier function of filaggrin deficient skin models by normalization of the free fatty acid profile underlining the potential of PPAR agonists for the treatment of filaggrin-associated skin diseases.
The epidermis functions as a barrier against the environment by means of several layers of terminally differentiated, dead keratinocytes - the cornified layer, which forms the endpoint of epidermal differentiation and death. The cornified envelope replaces the plasma membrane of differentiating keratinocytes and consists of keratins that are enclosed within an insoluble amalgam of proteins, which are crosslinked by transglutaminases and surrounded by a lipid envelope. New insights into the molecular mechanisms and the physiological endpoints of cornification are increasing our understanding of the pathological defects of this unique form of programmed cell death, which is associated with barrier malfunctions and ichthyosis.
Activation of peroxisome proliferator-activated receptors (PPARs) has been shown to have an important role in skin barrier function by regulating differentiation and lipid synthesis in keratinocytes. Oat (Avena sativa) has long been used as a soothing agent to relieve skin irritations and the clinical benefits of topical oat formulations have been proven; however, the mechanistic understanding of oat's mode of action remains unknown. We investigated whether an oat lipid extract could activate PPARs and subsequently increase epidermal lipid synthesis and differentiation markers. Primary human epidermal keratinocytes and transformed cell lines were treated with PPAR agonists and oat lipid extracts to investigate the PPAR agonism. PPAR target genes and epidermal differentiation markers were analyzed by using quantitative real time PCR and HPTLC analysis. Oat lipid extract demonstrated robust dual agonism for PPARα and β/δ, and increased direct PPAR target gene induction in primary human keratinocytes. In addition, oat oil treatment increased both receptor expression and, consistent with the literature on PPARs, oat oil treatment resulted in a significant up-regulation of differentiation genes (Involucrin, SPRRs and transglutaminse-1) and ceramide processing genes (β-glucocerebrosidase, sphingomyelinases 3 and ABCA12). Further, oat oil treatment in keratinocytes significantly increased ceramide levels (70%), suggesting a functional translation of PPAR activation by oat oil in keratinocytes. Taken together, these results demonstrate that oat lipids possess robust dual agonistic activities for PPARα and β/δ, increase their gene expression, and induce differentiation and ceramide synthesis in keratinocytes, which can collectively improve skin barrier function.This article is protected by copyright. All rights reserved.
Peroxisome proliferator-activated receptors (PPARs) are potentially useful for treatment of skin diseases, because they stimulate keratinocyte differentiation, exert anti-inflammatory effects and improve barrier function. We examined five PPAR-γ agonists, including four thiazolidinediones (ciglitazone, troglitazone, rosiglitazone and pioglitazone) and an angiotensin-II receptor blocker (telmisartan), for their ability to up-regulate filaggrin and loricrin expression at both mRNA and protein levels in cultured normal human keratinocytes (NHKs). Troglitazone, rosiglitazone, pioglitazone and telmisartan significantly increased filaggrin expression at both mRNA and protein levels in calcium-induced differentiated NHKs. Rosiglitazone and pioglitazone, but not troglitazone nor telmisartan, also significantly increased loricrin expression at both mRNA and protein levels in differentiated NHKs. These effects were not found in undifferentiated NHKs nor differentiated NHKs treated with ciglitazone. This study revealed differential effects of various PPAR-γ agonists on epidermal differentiation, and the most potent of those are rosiglitazone and pioglitazone.This article is protected by copyright. All rights reserved.