CN116531274A

CN116531274A - Skin care composition derived from olive and application thereof

Info

Publication number: CN116531274A
Application number: CN202310271223.0A
Authority: CN
Inventors: 黄虎; 陶侃; 王静一; 郭莉莉; 黄云霞
Original assignee: Shanghai Zhongyi Daily Chemical Co ltd
Current assignee: Shanghai Zhongyi Daily Chemical Co ltd
Priority date: 2023-03-20
Filing date: 2023-03-20
Publication date: 2023-08-04

Abstract

The invention provides a skin care composition derived from olive and application thereof, wherein olive leaf extract and ceramide NP derived from olive oil are combined, the intracellular calcium ion steady state is maintained, inflammatory reaction is reduced, and the damage of ultraviolet rays to skin is reduced.

Description

Skin care composition derived from olive and application thereof

Technical Field

The invention relates to the field of cosmetics, and mainly relates to a skin care composition derived from olive.

Background

Olea europaea L.) is a evergreen tree belonging to Oleaceae, olea, and is native to the coast of Mediterranean, a well-known woody oil tree species in the world, and the cultivation history can be traced up to four thousand years ago. The olive oil is extracted from olive fruits by cold pressing, wherein the main component is triglyceride containing various unsaturated fatty acids, and contains various active substances such as olive polyphenol, squalene, carotene, vitamin E, etc., which has special nutritive value and is called as liquid gold. Besides the application in diet, the olive oil is widely applied to the fields of medicine and cosmetics, and researches show that the olive oil has certain effects of resisting cancer, improving cardiovascular system, resisting aging, promoting wound healing and the like.

Besides fruit oil, olive leaves also contain abundant active ingredients including oleuropein, hydroxytyrosol, flavonoids, polyphenols, etc. The research shows that the olive leaf extract has the functions of antioxidation, anti-inflammatory, antibacterial, antivirus and the like, and has wide application prospect in the fields of medicines, health-care foods and cosmetics.

The human skin Stratum Corneum (SC) is a stack of keratinocytes transformed into dead keratinocytes, with intercellular filling with lipids and natural moisturizing factors, forming the "brick wall structure" of the skin barrier. The main components of lipids are ceramide, cholesterol and free fatty acids. The structure of ceramide is composed of two parts, namely long-chain fatty acid is connected to sphingosine skeleton through amide bond. The ceramide with the highest content in the stratum corneum of human skin is NP, N represents non-hydroxy fatty acid, and P represents phytosphingosine. In addition, the human cuticle ceramide has broader fatty acid carbon chain distribution, can form various lamellar structure crystal domains in the cuticle, and is beneficial to maintaining the function of skin barrier. The ceramide derived from vegetable oil also has broader fatty acid carbon chain distribution, and has better compatibility with human stratum corneum lipid (Oh, M.J. et al, novel phytoceramides containing fatty acids of diverse chain lengths are better than asingle C-cephamide N-stearoyl phytosphingosine to improve the physiological properties of human stratum corn. Clin cosmetic invest Dermatol.2017; 10:363-371). Supplementing lipid to horny layer helps to improve horny layer structure, increase lipid content in horny layer, repair and strengthen skin barrier, and reduce skin water loss.

The invention comprises the following steps:

in order to make comprehensive use of the various active substances of olive origin, the present invention surprisingly found that two active substances of olive origin: the olive leaf extract and the composition of ceramide NP derived from olive oil can maintain the stability of skin barrier and has a certain repairing effect on skin injury induced by ultraviolet rays.

The olive leaf extract has a wide application in cosmetics, for example, patent CN111789799a mentions the effect of the olive leaf extract and the composition thereof on anti-inflammatory and soothing, and patent CN114469820a mentions that the olive leaf extract realizes the whitening effect by inhibiting tyrosinase activity. Ceramide NP is also widely used in cosmetics for repairing skin barrier, and for example, the nourishing and repairing emulsion containing ceramide NP is described in patent CN115006324A for repairing skin barrier, and the ceramide composition used in patent CN112402282A can reduce penetration of irritant substances, enhance moisture content of horny layer and water locking capacity of skin. Olive oil derived ceramides have good soothing and repairing efficacy, but there is no related patent application for a while. Both materials have many good effects on the skin, but there is no patent for a moment to protect the subject with a combination of both.

In the prior art, only the efficacy of olive oil-derived ceramide and olive leaf extract are studied, and the combination of the two is not studied, and the effect of olive oil-derived ceramide on the relief and repair is not specifically described.

In order to solve the problems, the invention provides a skin care composition derived from olive, which contains olive leaf extract and ceramide NP derived from olive oil.

Further, the oleuropein content in the olive leaf extract is more than 0.1%.

Further, the olive leaf extract in the composition may be extracted by extraction, ultrasonic-assisted extraction, microwave-assisted extraction, supercritical carbon dioxide extraction, etc., and the extraction mode is not particularly limited. The extract may be a solution or a solid, and the form thereof is not particularly limited. Olive leaves for extraction have oleuropein content of more than 0.1%, and are stored in cool and ventilated place to avoid degradation of active substances.

Preferably, the olive leaf extract is purified by a macroporous resin adsorption method.

Further, the ceramide is olive oil-derived ceramide NP, and the structure is shown as a formula (1). Wherein R represents a saturated or unsaturated alkyl group having 11 to 30 carbon atoms.

Specifically, the source of fatty acid in the olive oil-derived ceramide NP is olive oil.

Specifically, the olive oil-derived ceramide NP refers to a final product obtained by performing an amine transesterification reaction between phytosphingosine obtained by fermentation and triglyceride in olive oil.

Preferably, the olive leaf is from the same origin as the fatty acid donor olive oil in the olive oil derived ceramide NP.

In the composition provided by the invention, the ratio of oleuropein to olive oil source ceramide NP in the olive leaf extract is (0.1-20): 1.

Preferably, the ratio of oleuropein to olive oil derived ceramide NP in the olive leaf extract is (0.5-2): 1.

The composition provided by the invention can be applied to cosmetics, including but not limited to milk, cream, water and essence.

Further, the composition comprises olive leaf extract and olive oil derived ceramide NP.

Further, the composition is applied to cosmetics, and the content range is 0.01-30%.

Preferably, the composition is contained in the cosmetic in an amount ranging from 0.1 to 10%.

The composition provided by the invention can be used for reducing inflammatory reaction and maintaining skin barrier stability in cosmetics, and comprises olive leaf extract and olive oil-derived ceramide NP.

Further, the composition promotes VAPB and MICU1 gene expression to maintain calcium homeostasis within the skin barrier.

Further, the composition promotes CBR1 gene expression to reduce cell inflammation.

Further, the composition inhibits IFNA7 gene expression, alleviating uv-induced inflammatory processes.

Further, the composition inhibits WIF1 gene expression, affecting the proliferation and differentiation process of cells.

Further, the composition inhibits PMAIP1 gene expression and inhibits uv-induced apoptosis processes.

The invention has the remarkable advantages that:

1. the invention provides a skin care composition derived from olive, which comprises olive leaf extract and olive oil-derived ceramide NP.

2. The skin care composition comprises olive leaf extract and olive oil-derived ceramide NP, and can maintain intracellular calcium ion steady state by promoting barrier gene expression, reduce inflammatory reaction, reduce apoptosis, reduce damage of ultraviolet rays to skin and maintain stability of skin barrier.

3. The olive-derived skin care composition provided by the invention comprises olive leaf extract and olive oil-derived ceramide NP, and can be used in various skin care products, including but not limited to milk, cream, water and essence.

Description of the drawings:

in order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

Fig. 1: HPLC (high Performance liquid chromatography) spectrum of ceramide NP derived from Gansu Lonnan olive oil

Fig. 2: effect of olive oil derived ceramide NP on SLS stimulation-induced changes in epidermal model tissue morphology

Fig. 3: effect of olive oil derived ceramide NP on SLS stimulation-induced changes in the epidermal model silk-fibroin FLG content

Fig. 4: effect of olive oil derived ceramide NP on SLS stimulation-induced changes in the TGM1 content of the glutamine transaminase in the epidermis model

Fig. 5: structural formula of olive oil-derived ceramide NP

Fig. 6: GO enrichment analysis of expression upregulated genes in epidermis models following composition administration compared to negative controls

Fig. 7: specific embodiment of GO enrichment analysis of expression downregulated genes in epidermis model after composition administration compared to negative control

In the examples below, a commercially available ceramide NP, available from win-creation specialty chemicals, inc, characterized by the fatty acid moiety being stearic acid; the olive oil is from Gansu Lonnan and purchased from Gansu Lonnan Xiangyu Olive development Limited liability company; phytosphingosine from fermentation was purchased from korean soxhlet biotechnology limited; the olive leaf is from Gansu Lonnan, and the olive leaf extract is purified by macroporous resin adsorption method, wherein the extract is powder, and the oleuropein content is 32%. The preparation method of the olive leaf extract comprises the following steps: pulverizing dried olive leaf, adding into 70% ethanol water solution, refluxing for 3 times, mixing extractive solutions, and filtering to obtain concentrated extractive solution. The extract is adsorbed with macroporous resin column, and after the adsorption, the resin column is washed with water to remove impurities, and then is desorbed with 70% ethanol water solution. Collecting desorption solution, adding dextrin, and lyophilizing to obtain olive leaf extract powder.

Embodiment case one: preparation of olive oil derived ceramide NP

The ceramide is prepared by a conventional amine transesterification method. 3g of phytosphingosine is added into 10g of olive oil of Gansu Lonnan, and the mixture is stirred and heated to 100 ℃ under the protection of nitrogen, and the mixture is refluxed for 10 hours until the reaction is complete. And removing the heat source, cooling to room temperature under the nitrogen atmosphere, and recrystallizing with petroleum ether to obtain the final product of ceramide with 93 percent of yield. HPLC shows purity over 92%, and is shown in figure 1 of the specification.

The above synthesis method is only one of the possibilities, the transesterification reaction may be carried out under nitrogen, air or vacuum conditions by using an acid, a base, a catalyst or an enzyme to increase the reaction rate or reduce the degree of side reactions, and the present invention is not particularly limited to the synthesis method.

Implementation case two: distribution of fatty acid chains in olive oil-derived ceramide NP and raw olive oil

The method comprises the steps of determining ceramide NP from olive oil and raw material olive oil by adopting a transesterification method, precisely weighing about 0.3g of the olive oil sample, adding 5mL of sodium hydroxide-methanol solution, shaking uniformly, reacting at normal temperature for 40min, adding 5mL of petroleum ether, shaking uniformly, standing, and adding 5mL of distilled water. The upper organic phase is sucked into a centrifuge tube, dried by anhydrous sodium sulfate, centrifuged, filtered, diluted and analyzed by GC-MS, and compared with a database for searching for qualitative and quantitative analysis. For ceramide samples, about 0.2g of the sample is precisely weighed, 5mL of potassium hydroxide-methanol solution is added for shaking, the mixture is reacted for 1 hour in a water bath at 80 ℃, 5mL of BF3-methanol solution is added for shaking, and the mixture is reacted for 1 hour in a water bath at 80 ℃.20 mL of chloroform, 10mL of a 0.9% sodium chloride solution was added, the lower layer solution was removed after shaking and centrifugation, and the lower layer solution was washed once with 10mL of methanol and 10mL of a 0.9% sodium chloride solution, respectively, dried over anhydrous sodium sulfate, detected by GC-MS and analyzed by comparison with a database. The results of the experiment are shown in Table one.

Table one: comparison of carbon chain distribution of fatty acid chains

	Raw material olive oil (%)	Olive oil derived ceramide NP (%)
			C16:0	12.09	10.69
C16:1	1.59	1.63
			C18:0	1.89	2.03
C18:1	74.69	75.37
			C18:2	7.92	8.13
C18:3	0.79	0.80
			C20:0	0.34	0.32
C20:1	0.30	0.29
			Others	0.40	0.74

As can be seen from the data in Table I, the contents of C18:0, C18:1 and C18:2 in the raw olive oil are respectively 1.89%, 74.69% and 7.92%, and the contents of the raw olive oil source ceramide NP are respectively 2.03%, 75.37% and 8.13%. For C16:0 and C16:1, the content of the raw olive oil was 12.09% and 1.59%, respectively, and the content of the olive oil-derived ceramide NP was 10.69% and 1.63%, respectively. Namely, the ceramide NP from olive oil has similar fatty acid chain distribution with the raw material olive oil, and the unsaturated fatty acid content is higher, which indicates that the fatty acid in the ceramide NP prepared by the method of the invention is derived from olive oil.

Implementation case three: comparison of the soothing Effect of olive oil derived ceramide NP, other vegetable oil derived ceramide NP and commercially available ceramide NP

The effect of various ceramide NPs on the levels of inflammatory factors IL-1 alpha and inflammatory mediators PGE2 in UVB-stimulated keratinocytes was determined. The first sample group is the long-nan olive oil-derived ceramide, the second sample group is the Mediterranean olive oil-derived ceramide, the third sample group is the shea butter-derived ceramide, the fourth sample group is the mixed vegetable oil-derived ceramide (the vegetable oil comprises shea butter, white pool flower seed oil, moringa seed oil and macadamia nut seed oil), and the fifth sample group is the commercially available stearic acid-derived ceramide. The ceramide NP from other vegetable oils in the sample group is obtained by taking different vegetable oils as raw materials and adopting a synthesis and purification mode similar to that of the first embodiment. The test concentration for sample groups one to five was 0.003%, the positive control dexamethasone concentration was 0.01% and the UVB stimulation was 300mJ/cm2. The experimental results are shown in Table II.

And (II) table: effects of ceramide NP on changes in the levels of inflammatory factors IL-1 alpha and inflammatory mediators PGE2

From the data in Table II, the ceramides from sample group I to sample group five have inhibition effects on the content of keratinocyte inflammatory factors and inflammatory mediators caused by UVB stimulation, wherein the most inhibitory effect is olive oil-derived ceramides, and the least inhibitory effect is commercially available stearic acid-derived ceramides. Comparing the first, second and third and fourth sample groups, it can be seen that the ceramide with higher unsaturated fatty acid content has better relieving effect. Comparing sample group one and sample group two, it can be seen that the soothing effect of olive oil derived ceramide in the long and the south China is significantly better than that of olive oil derived ceramide in the middle sea (p < 0.01).

Implementation case four: comparison of anti-inflammatory Effect of olive oil derived ceramide NP and commercially available stearic acid derived ceramide NP

The effect of ceramide on SLS (sodium dodecyl sulfate, sodium lauryl sulfate) to stimulate skin barrier damage was studied using a 3D skin model (available from guangdong bosch biotechnology limited) and compared with commercially available ceramide NP.

The skin model was divided into blank, negative, positive and sample groups, each group containing 3 replicates. The blank control group was not subjected to any treatment, 25. Mu.L of 0.2% SLS solution was added to the surface of the negative control group, 12.5. Mu.L of 0.4% SLS solution and 12.5. Mu.L of 0.01% dexamethasone were added to the surface of the positive control group, 12.5. Mu.L of 0.4% SLS solution and 12.5. Mu.L of 0.05% olive oil-derived ceramide NP were added to the surface of the sample group, 12.5. Mu.L of 0.4% SLS solution and 12.5. Mu.L of 0.1% olive oil-derived ceramide NP were added to the surface of the sample group, and 12.5. Mu.L of 0.4% SLS solution and 12.5. Mu.L of 0.1% commercially available ceramide NP were added to the surface of the sample group

After the administration, the skin model is placed in an incubator for incubation for 24 hours, after the completion, the test substance remained on the surface of the model is washed by a sterile PBS solution, and the residual liquid inside and outside the model is wiped off by a sterile cotton swab. After the incubation is completed, collecting the epidermic skin model culture solution in a centrifuge tube, and measuring the contents of inflammatory factors IL-1 alpha and inflammatory mediators PGE2 by adopting an ELISA kit. The experimental results are shown in Table three.

Table three: content changes of inflammatory factor IL-1 alpha and inflammatory mediator PGE2

From the data in Table three, SLS stimulation resulted in increased inflammatory factor IL-1 alpha and inflammatory mediator PGE2 levels compared to the negative control and the blank. Compared with a negative control group, the olive oil source ceramide NP can obviously reduce the content of inflammatory factors IL-1 alpha and inflammatory mediators PGE2, and the p value is less than 0.01. The inhibition rates of the olive oil derived ceramide NP on IL-1α and PGE2 were 24.67% and 33.01%, respectively, and the inhibition rates of the olive oil derived ceramide NP on IL-1α and PGE2 were 40.71% and 34.39%, respectively. The commercial ceramide NP has a remarkable inhibition effect on inflammatory factor IL-1 alpha, the inhibition rate is 21.47%, but has no remarkable effect on inflammatory mediator PGE 2. Compared with the commercial ceramide NP, the inhibition effect of the olive oil-derived ceramide NP on the inflammatory factors IL-1 alpha and the inflammatory mediators PGE2 is obviously enhanced, and for the inflammatory factors IL-1 alpha, the p value of the sample group II to the sample group III is 0.045<0.05; for inflammatory mediator PGE2, sample group two versus sample group three, p-value was 0.002<0.01.

In the experimental data of the group, the inhibition effect of the olive oil source ceramide NP on inflammatory factors IL-1 alpha and inflammatory mediators PGE2 is obviously different from that of the commercial ceramide NP.

Implementation case five: comparison of the protective Barrier Effect of olive oil derived ceramide NP and commercially available ceramide NP

The skin model was divided into blank, negative, positive and sample groups, each group containing 3 replicates. The blank was not treated with any treatment, 25. Mu.L of 0.2% SLS solution was added to the surface of the negative control, 12.5. Mu.L of 0.4% SLS solution and 12.5. Mu.L of 50. Mu.M PPAR agonist WY14643 were added to the surface of the positive control, 12.5. Mu.L of 0.4% SLS solution and 12.5. Mu.L of 0.05% olive oil derived ceramide NP were added to the surface of one of the sample groups, 12.5. Mu.L of 0.4% SLS solution and 12.5. Mu.L of 0.1% olive oil derived ceramide NP were added to the surface of the two of the sample groups, and 12.5. Mu.L of 0.4% SLS solution and 12.5. Mu.L of 0.1% commercially available ceramide NP were added to the three surfaces of the sample groups.

Filaggrin (FLG) interacts with keratin fibres to form a dense keratin fibre bundle, which is linked to paphiopedin (LOR) or the like under the action of glutamine transaminase to form a stable, water-insoluble, keratinous envelope (Cornified Envelope, CE) which forms the basis of the skin barrier. In addition, the silk fibroin is gradually degraded into small molecular active substances such as natural moisturizing factors by proteases such as Caspase-14 at the end of the differentiation of keratinocytes, and the skin hydration is maintained. Various skin conditions such as ichthyosis vulgaris and atopic dermatitis may be associated with impaired skin barrier due to abnormal function of silk fibroin. Glutamine transaminase 1 (tgm 1) is mainly expressed in epidermal cells and participates in the formation of epsilon- (gamma-glutamyl) lysine isopeptide bond cross-links in the formation process of keratinocyte envelope, and the cross-links are very stable and are key steps for the terminal differentiation of keratinocyte to form the keratinocyte envelope. The levels of FLG and TGM1 enzymes are therefore one of the key indicators characterizing skin barrier function.

After the administration, the skin model is placed in an incubator for incubation for 24 hours, after the completion, the test substance remained on the surface of the model is washed by a sterile PBS solution, and the residual liquid inside and outside the model is wiped off by a sterile cotton swab. After the model for detection was fixed with 4% paraformaldehyde for 24 hours, the contents of silk fibroin and glutamine transaminase I were determined by immunofluorescence, and the changes in tissue morphology were analyzed after H & E staining. The results of the experiments are shown in Table IV and in the accompanying figures 2-4 of the description.

Table four: content variation of FLG and TGM1 enzymes

As can be seen from fig. 2, the SLS stimulation resulted in tissue morphology damage in the skin model, which is characterized by a reduced number of viable cell layers and cavitation, compared to the blank control and the negative control. The olive oil source ceramide NP has a certain repairing effect on tissue morphology damage, the quantity and the volume of cavitation bubbles are reduced by 0.05 percent of the olive oil source ceramide NP, the cavitation bubbles are reduced by 0.1 percent of the olive oil source ceramide NP, the arrangement of living cell layers is compact, and the phenomenon of living cell damage is obviously improved. However, commercially available ceramide NPs have no ameliorating effect on cavitation in tissues and on living cell layers.

As can be seen from table four and fig. 3, the SLS stimulation resulted in a decrease in the area of the green fluorescent portion (upper half of the image, lighter colored portions of the stratum corneum) in the skin model, i.e., a decrease in the silk fibroin content, compared to the blank and negative control groups. The content of filaggrin in the skin model is obviously increased by the olive oil-derived ceramide NP and the commercial ceramide NP, and the promotion effect of 0.05 percent, 0.1 percent of the olive oil-derived ceramide NP and 0.1 percent of the commercial ceramide NP on the filaggrin content is 225.58 percent, 281.40 percent and 262.79 percent respectively. The effect of the same concentration of olive oil derived ceramide NP on the increase of the silk fibroin content is significantly better than that of the commercial ceramide NP, and the p value is 0.041<0.05.

From table four and fig. 4, SLS stimulation resulted in a decrease in the green fluorescent portion area (upper half of the image, lighter colored portions of the stratum corneum), i.e., a decrease in TGM1 content, in the skin model. Both olive oil derived ceramide NP and commercially available ceramide NP significantly increased TGM1 content in skin models, with 0.05%, 0.1% olive oil derived ceramide NP and 0.1% commercially available ceramide NP promoting silk fibroin content by 491.67%, 558.33% and 366.67%, respectively. The effect of the same concentration of olive oil derived ceramide NP on the increase of TGM1 content was significantly better than that of the commercial ceramide NP, with a p-value of 0.012<0.05.

In this set of experimental data, olive oil derived ceramide NP and commercially available ceramide NP have significantly different promotion effects on filaggrin FLG and glutamine transaminase 1 (TGM 1).

The experimental conclusion shows that the olive oil source ceramide NP has obvious influence on the inflammatory response of an epidermis skin model and skin barrier damage caused by SLS stimulation, the reduction of the content of inflammatory factors and inflammatory mediators shows that the olive oil source ceramide NP has good relieving effect, the skin tissue structure is repaired, and the content of the related protein of the enhanced barrier shows that the olive oil source ceramide NP has good skin barrier repairing effect. In comparison with commercially available ceramide NPs, olive oil-derived ceramide NPs have been found to have more remarkable effects on both soothing and repairing effects.

Implementation case six: skin care composition derived from olive

The invention provides a skin care composition derived from olive, which adopts olive leaves from Gansu Longnan, and obtains olive leaf extract in powder form by a macroporous resin purification technology, wherein the oleuropein content is 32%. The ceramide NP is prepared by adopting olive oil from Gansu Lonnan, and the ceramide NP is prepared by mixing and heating phytosphingosine from fermentation sources and olive oil, so that the phytosphingosine and triglyceride undergo transesterification reaction to obtain the product, namely the ceramide NP from olive oil. The effect of olive leaf extract, olive oil-derived ceramide NP and composition (olive leaf extract+olive oil-derived ceramide NP) on the Ultraviolet (UVB) irradiation skin model was tested by using a gene microarray sequencing method, and the soothing and repairing effects of single components and compositions on skin damage caused by ultraviolet were verified.

Implementation case seven: GO enrichment analysis of epidermal model gene expression by administration of compositions

The full thickness epidermore model EpiDermFT (purchased from MatTek company, usa) was divided into 5 groups, namely a blank (no treatment), a negative control (UVB treatment only), olive leaf extract (0.2%), olive oil derived ceramide NP (0.05%)The compositions prepared according to the invention (olive leaf extract 0.2% + olive oil derived ceramide NP 0.05%) each comprise 3 replicates. The epidermis model was treated with 200. Mu.L of the corresponding test substance for 4 hours by gene microarray sequencing, UVB irradiation (280-340 nm,200mJ/cm ² ) And then the sample is treated by the corresponding sample for 24 hours, after incubation is finished, the sample remained on the surface of the model is washed by a sterile PBS solution, and the liquid remained inside and outside the model is wiped by a sterile cotton swab. Total RNA was extracted for microarray analysis (Affymetrix Human Clariom S, 21448 genes total). Genes with p-value less than 0.05 (significant change) compared to negative control group were screened for composition experimental group for GO enrichment analysis, and the results are shown in fig. 6 and 7.

The results show that: after administration of the combination of olive leaf extract and olive oil derived ceramide NP, the up-regulated expression genes are associated with processes including detoxification of copper ions, modulation of exogenous apoptosis-related signaling pathways, negative modulation of cysteine endopeptidase activity during apoptosis, mitochondrial membranes, etc. After administration of the combination of olive leaf extract and olive oil derived ceramide NP, the down-regulated expression genes are associated with processes including DNA damage sites, double stranded RNA responses, cell-matrix adhesion, modulation of DNA damage stimulation responses, and the like.

Implementation case eight: the composition promotes expression of barrier-related genes in an epidermis model

Calcium ions play an important role in maintaining the structure and stability of the skin barrier. The normal epidermis structure has a gradient of calcium ion concentration, the stratum corneum layer has the highest concentration of calcium ion, the stratum corneum layer gradually decreases with increasing skin depth, and the stratum basale layer has the lowest concentration of calcium ion. Lower calcium ion concentrations promote keratinocyte proliferation, higher calcium ion concentrations promote keratinocyte differentiation and stratification. The intracellular and extracellular calcium ion concentrations interact to cooperatively maintain the barrier effect. The elevation of ROS by uv irradiation affects the calcium ion concentration in the endoplasmic reticulum, causing endoplasmic reticulum stress (Endoplasmic Reticulum stress), and thus the skin barrier function. VAPB encodes a Vesicle-associated membrane protein binding protein (Vesicle-associated membrane protein-binding protein B) that is located on the surface of the endoplasmic reticulum membrane and interacts with the mitochondrial protein PTPIP51 to regulate calcium ion concentration. The loss of VAPB affects the process of mitochondrial uptake of calcium ions released by the endoplasmic reticulum (De Vos, K.J. et al VAPB interacts with the mitochondrial protein PTPIP, to regulate calcium Homeostasis. Hum. Mol. Genet 21,1299-1311 (2012)). MICU1 regulates the mitochondrial calcium transporter MCU (Antonny, A.N. et al, MICU1 regulation of mitochondrial Ca (2+) uptake dictates survival and tissue regeneration. Nat. Commun 7,10955 (2012)), affecting mitochondrial uptake of calcium ions. The absence of MICU1 results in excessive calcium uptake by mitochondria, inducing a cell death process that affects wound healing. The expression levels of the above two genes affect intracellular calcium ion homeostasis, and may also affect the calcium ion concentration in the stratum corneum of the skin, resulting in altered skin barrier function. CBR1 encodes an NADPH-dependent carbonyl reductase carbonyl reductase-1 that catalyzes the reduction of a variety of carbonyl compounds, including prostaglandins, and reduces the inflammatory response caused by oxidative stress. IFNA7 encodes interferon alpha 7, a low molecular weight glycoprotein produced by macrophages during the immune process, and is involved in JAK/STAT signaling pathways affecting immune modulating function. WIF1 encodes Wnt inhibitor 1, inhibits the anti-inflammatory canonical Wnt signaling pathway, inhibits keratinocyte proliferation and differentiation (schlouter, h. Et al, WIF1 Is Expressed by Stem Cells of the Human Interfollicular Epidermis and Acts to Suppress Keratinocyte production.j Invest Dermatol 133,1669-1673 (2013)). PMAIP1 encodes a pro-apoptotic protein belonging to the Bcl-2 protein family, also known as Noxa, involved in the p53 mediated apoptosis process.

The rate of change of the expression of the related genes is shown in Table six.

Table six: rate of change in gene expression

The VAPB, MICU1 and CBR1 expression levels were decreased and IFNA7, WIF1 and PMAIP1 expression levels were increased after UVB irradiation compared to the non-irradiated blank, i.e. calcium ion homeostasis in the skin barrier was disrupted (VAPB, MICU 1), skin inflammation was aggravated (CBR 1, IFNA 7), cell differentiation and apoptosis processes were affected (WIF 1, PMAIP 1). After the olive leaf extract, olive oil-derived ceramide NP and the combination of the olive leaf extract and the olive oil-derived ceramide NP are applied, the expression levels of VAPB, MICU1 and CBR1 are increased compared with the negative control group treated by UVB irradiation, and the expression levels of IFNA7, WIF1 and PMAIP1 are reduced, namely skin barrier injury, skin inflammation and cell differentiation apoptosis are all relieved to a certain extent. Compared with a single raw material group, the composition group has more remarkable change of gene expression and obvious synergistic effect. The above gene expression changes indicate that the composition of olive leaf extract and olive oil-derived ceramide NP has good repairing and relieving effects, and can reduce the damage of UVB irradiation to skin tissues.

The results of the microarray sequencing of the skin model show that the combination of olive leaf extract and olive oil-derived ceramide NP has a significant effect on maintaining the skin barrier function. Promoting VAPB and MICU1 gene expression, maintaining calcium ion homeostasis in skin barrier, promoting CBR1 gene expression, and reducing inflammation. Inhibiting IFNA7 gene expression, alleviating uv-induced inflammatory processes; inhibiting WIF1 gene expression, reducing the impact on anti-inflammatory Wnt pathway, and modulating cellular proliferation and differentiation processes; inhibit PMAIP1 gene expression and inhibit ultraviolet-induced apoptosis. The composition plays roles in relieving, repairing and maintaining skin barriers by regulating the expression of related genes, so that the skin care product using the composition also has the roles in relieving, repairing and maintaining skin barriers.

While the foregoing embodiments have been described in connection with the exemplary embodiments of the invention, it should be understood that the foregoing embodiments are merely illustrative of the invention, and that any modifications, additions, substitutions and the like made without departing from the scope of the invention.

Although the present invention is disclosed above, the present invention is not limited thereto. Various changes and modifications may be made by one skilled in the art without departing from the spirit and scope of the invention, and the scope of the invention should be assessed accordingly to that of the appended claims.

Claims

1. A skin care composition comprising olive leaf extract and olive oil derived ceramide NP.

2. The composition of claim 1, wherein the oleuropein content of the olive leaf extract is greater than 0.1%.

3. The composition of claim 2, wherein the olive oil derived ceramide NP in the composition is formed by linking phytosphingosine and fatty acids in olive oil through amide linkages.

4. A composition according to claim 3, wherein the olive oil derived ceramide NP in the composition is prepared by an amine transesterification reaction between a chiral phytosphingosine ((2 s,3s,4 r) -2-amino-1, 3, 4-octadecanol) of fermentative origin and a triglyceride.

5. The composition of claim 4, wherein the olive oil derived ceramide NP in the composition has a structure represented by formula (1), and R is a saturated or unsaturated alkyl group having 11 to 30 carbon atoms.

6. The composition according to claim 5, wherein the ratio of oleuropein to olive oil derived ceramide NP in the olive leaf extract is (0.1-20) 1 by weight.

7. Use of a composition for the preparation of a skin care formulation for maintaining stable skin barrier by reducing inflammatory response, maintaining intracellular calcium homeostasis, reducing uv damage to the skin.

8. The use according to claim 7, wherein the composition is olive leaf extract and olive oil derived ceramide NP.

9. The use according to claim 7, wherein the composition maintains calcium ion homeostasis in the skin barrier by modulating VAPB and MICU1 gene expression; regulating and controlling CBR1 and IFNA7 gene expression, and reducing inflammatory reaction in skin; regulating WIF1 and PMAIP1 gene expression, and affecting cell differentiation and apoptosis.

10. The use according to claim 7, wherein the composition is present in the skin care formulation in an amount ranging from 0.01 to 30%.