Emotional beautifying composition containing camellia nitidissima flower extract and application thereof

Abstract

The invention discloses an emotion beautifying composition containing a camellia nitidissima flower extract and application of the emotion beautifying composition, and relates to the technical field of cosmetics. The invention provides an emotional beautifying composition. The emotional beautifying composition is prepared from the following components in parts by weight: 0.1 to 5 parts of purple coneflower extract, 0.01 to 2 parts of tephros glauca seed extract, 0.01 to 3 parts of golden camellia flower extract and 0.1 to 3 parts of lactobacillus fermentation lysate. Aiming at the core problem of skin barrier gene (FLG / LOR) expression inhibition caused by cortisol rhythm disorder caused by emotional pressure, the invention provides a solution scheme of three-stage linkage of cortisol rhythm synchronization, barrier gene repair and nerve sensitivity regulation.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to an emotional beauty composition containing camellia nitidissima leaf extract and its application. Background Art

[0002] There is a very close relationship between emotions and skin health. In particular, negative emotions such as stress, anxiety, and depression will significantly exacerbate the problems of sensitive skin. As a key factor affecting the efficacy of traditional skin care products, stress further demonstrates its profound impact on the skin. Emotional stress can have a wide and deep impact on the skin through the neuro-endocrine-immune axis. Specifically, under stress, the skin barrier function is damaged more quickly, resulting in an increase in transepidermal water loss. At the same time, the skin becomes more sensitive to external stimuli such as ultraviolet rays, pollutants, and allergens. This barrier damage then triggers a series of sensitive symptoms, including stinging, tightness, and flushing. Among them, the excessive secretion of cortisol under stress is one of the core mechanisms leading to this series of problems. As a key hormone regulated under stress activation, cortisol has a certain anti-inflammatory effect in the short term. However, when it is at a high level for a long time, it will weaken the skin barrier function and make the skin more vulnerable by promoting inflammation and oxidative stress. In addition, excessive cortisol will also inhibit the self-repair ability of the skin, thus further aggravating the symptoms of sensitive skin.

[0003] Traditional skin care products for sensitive skin problems mostly use anti-inflammatory ingredients such as dipotassium glycyrrhizinate and bisabolol, as well as barrier lipid supplements such as ceramides and cholesterol. Their mechanism of action is mainly to inhibit mast cell degranulation or supplement intercellular lipids to relieve immediate sensitive symptoms, such as reducing skin stinging and tightness. However, such solutions have obvious limitations. They only intervene in local skin inflammation or physical barrier damage and cannot effectively cope with gene expression abnormalities caused by stress. Research shows that long-term stress will cause cortisol to continuously inhibit the expression of FLG (filaggrin) and LOR (loricrin) genes. Traditional barrier lipid supplements can only temporarily fill the lipid gap and cannot restore the autonomous synthesis ability of FLG and LOR genes. Some products on the market try to use aromatherapy, such as using lavender essential oil and bergamot volatiles, to indirectly improve the skin condition by regulating emotions. However, such technologies have obvious limitations. They ignore the rhythm existing in the human body, and such technologies cannot distinguish the circadian rhythm requirements. The normal secretion of cortisol in the human body has a circadian rhythm of high in the morning and low at night. At night, if cortisol abnormally increases, it will continuously inhibit the self-repair of skin cells. For example, the synthesis peak of the skin barrier protein FLG at night will be suppressed by the increase in cortisol, and existing technologies such as aromatherapy do not fully consider this circadian rhythm factor and are difficult to achieve precise and effective improvement of the skin condition.

[0004] As an important emotional factor affecting skin health, stress is gradually highlighting the shortcomings of traditional skin care products in dealing with complex skin sensitivity issues, which also leads to an in-depth discussion on the close connection between emotions and skin health. In view of this, this application is filed. Summary of the invention

[0005] Based on this, the purpose of the present invention is to overcome the shortcomings of the above-mentioned prior art and provide a mood beauty composition containing Camellia chrysantha extract and its application.

[0006] To achieve the above-mentioned purpose, the technical solution adopted by the present invention is: an emotional beauty composition, comprising the following components in parts by weight: 0.1-5 parts of purple coneflower extract, 0.01-2 parts of ginkgo seed extract, 0.01-3 parts of golden camellia extract, and 0.1-3 parts of lactic acid bacteria fermentation lysate.

[0007] Aiming at the core problem that cortisol rhythm disorder caused by emotional stress leads to the inhibition of skin barrier gene (FLG / LOR) expression, the present invention proposes a three-level linkage solution of "cortisol rhythm synchronization-barrier gene repair-neural sensitivity regulation".

[0008] The present invention uses echinacoside in purple coneflower extract to target and regulate the activity of the HPA axis, inhibit the release of hypothalamic CRH, reduce the nocturnal cortisol peak to the physiological rhythm level, and relieve the transcriptional inhibition of cortisol on FLG / LOR genes; at the same time, it combines with 5-hydroxytryptamine precursors in glycine max seed extract to regulate the amygdala stress response through the blood-brain barrier, blocking the vicious cycle of "skin sensitivity → increased anxiety → cortisol secretion". On the skin side, the lactic acid bacteria fermentation lysate activates the skin "clock" gene, rebuilds the circadian repair rhythm of epidermal cells, naturally misaligns FLG synthesis and cortisol secretion peaks, and cooperates with tea polyphenol derivatives in golden camellia extract to reshape the tight junction structure, achieving dual repair of physical-immune barriers. For the first time, the present invention integrates the regulation of emotion-induced cortisol rhythm with the repair of the skin biological clock-barrier gene network, breaking through the limitations of the traditional "treating the symptoms but not the root cause", and forming a full-link barrier repair system from intervention at the source of stress to skin physiological reconstruction.

[0009] Preferably, the mood beauty composition comprises the following components in parts by weight: 0.5-2 parts of Echinacea purpurea extract, 0.05-1 parts of Ganoderma lucidum seed extract, 0.05-1 parts of Camellia chrysantha extract, and 0.5-2 parts of lactic acid bacteria fermentation lysate.

[0010] Preferably, the mood beauty composition comprises the following components in parts by weight: 1-1.5 parts of Echinacea purpurea extract, 0.1-0.5 parts of Ganoderma lucidum seed extract, 0.5-0.8 parts of Camellia chrysantha extract, and 1-1.5 parts of lactic acid bacteria fermentation lysate.

[0011] In actual research, the inventors found that the weight parts of the components in the composition provided by the present invention can affect the performance of the product. When the weight parts of the components are further selected within the above range, the obtained product can better achieve the regulation of the emotional cortisol rhythm, activate the skin "clock" gene, reconstruct the circadian repair rhythm of epidermal cells, and form a full-link barrier repair system from the source of stress intervention to skin physiological reconstruction.

[0012] Preferably, based on the total weight of the emotional beauty composition, the weight percentage content of the lysate of lactic acid bacteria fermentation is 15-40%.

[0013] In actual research, the inventors found that when the weight percentage content of the lysate of lactic acid bacteria fermentation in the emotional beauty composition is within the above range, the obtained product has a more excellent effect of realizing the regulation of the emotional cortisol rhythm, activating the skin "clock" gene, and reconstructing the circadian repair rhythm of epidermal cells.

[0014] In actual research, the inventors found that the lysate of lactic acid bacteria fermentation can be a commercially available product or can be prepared by oneself. To prepare it by oneself, a conventional preparation method of the lysate of lactic acid bacteria fermentation can be used.

[0015] In one embodiment, the fermentation culture method of the lysate of lactic acid bacteria fermentation is as follows:

[0016] S1. Expand and culture the lactic acid bacteria strains in a seed medium respectively;

[0017] S2. Inoculate the expanded lactic acid bacteria seeds into a fermentation medium respectively for fermentation to obtain a fermentation broth;

[0018] S3. Centrifuge the fermentation broth to obtain a bacterial sludge;

[0019] S4. Mix the bacterial sludge, add a protective agent according to the mass of the bacterial sludge, and stir to mix evenly to obtain a mixed solution;

[0020] S5. Sterilize the mixed solution at a high temperature to obtain the lysate of lactic acid bacteria fermentation.

[0021] Preferably, in S1, the lactic acid bacteria strains are at least one of Lactobacillus plantarum and Lactobacillus brevis.

[0022] In the fermentation culture method of the lysate of lactic acid bacteria fermentation of the present application, when the lactic acid bacteria strains are not limited to one kind, in S1, the expansion and culture are carried out for each lactic acid bacteria strain in different seed media respectively; in S2, the fermentation is to inoculate each lactic acid bacteria strain into different fermentation media respectively, and the bacterial sludge is mixed after each lactic acid bacteria strain is separately expanded and cultured and fermented. Among them, the components of the expanded seed media are the same, the components of the fermentation media are the same, and the subsequent fermentation culture conditions are the same.

[0023] Preferably, in S2, at least one of microaerobic fermentation and anaerobic fermentation is used for the fermentation.

[0024] Preferably, the lactic acid bacteria fermentation lysate is obtained by microaerobic fermentation, and in the microaerobic fermentation, the dissolved oxygen content in the fermentation system is 0.2 - 0.5 mg / L; or, the lactic acid bacteria fermentation lysate is obtained by anaerobic fermentation, and in the anaerobic fermentation, the dissolved oxygen content in the fermentation system < 0.1 mg / L.

[0025] More preferably, in S2, a combination of microaerobic fermentation and anaerobic fermentation is used for the fermentation. First, microaerobic fermentation is carried out, and then anaerobic fermentation is carried out; in the microaerobic fermentation, the dissolved oxygen content in the fermentation system is 0.2 - 0.5 mg / L, and in the anaerobic fermentation, the dissolved oxygen content in the fermentation system < 0.1 mg / L.

[0026] In one embodiment, in S2, microaerobic fermentation is carried out first, and the conditions are as follows: 5 - 10% (volume ratio, that is, the volume of the bacterial solution accounts for the volume of the fermentation medium) of the bacterial solution is inoculated into the fermentation medium, and under a stirring rate of 100 - 200 rpm and an aeration rate of 0.3 - 0.5 vvm (volume ratio aeration rate), the dissolved oxygen content in the culture system is stabilized within the range of 0.2 - 0.5 mg / L, and the culture is carried out at 34 - 37 °C for 24 - 30 hours; then anaerobic fermentation is carried out, and the conditions are as follows: aeration is stopped and the stirring rate is reduced to 50 - 100 rpm, maintaining the dissolved oxygen content < 0.1 mg / L, and the culture is carried out at 30 - 34 °C for 36 - 48 hours.

[0027] Preferably, in S4, mixing the bacterial sludge means that when there is more than one kind of lactic acid bacteria strain, if there is only one kind of lactic acid bacteria strain, there is no need to mix, and the protective agent can be directly added.

[0028] When the lactic acid bacteria strains are a mixed strain of Lactobacillus plantarum and Lactobacillus brevis, the weight ratio of the Lactobacillus plantarum bacterial sludge to the Lactobacillus brevis bacterial sludge is (1 - 3):(2 - 5).

[0029] More preferably, in S4, the mass ratio of the bacterial sludge to the protective agent is 1:(1 - 10).

[0030] Preferably, the protective agent includes trehalose and water, and the mass percentage content of trehalose is 5 - 50%; preferably, the mass percentage content of trehalose is 28%.

[0031] Preferably, in S5, the sterilization temperature is 95 - 121 °C, and the sterilization time is 15 - 30 min.

[0032] In actual research, the inventors found that the performance of lactic acid bacteria fermentation lysate obtained by using different lactic acid bacteria fermentation cultures and different fermentation culture methods is different. Furthermore, when the lactic acid bacteria fermentation lysate prepared by different fermentation culture methods is added to the emotional beauty composition, the efficacy of the obtained emotional beauty composition will vary. When the above method is selected, the comprehensive effect of the obtained emotional beauty composition is better.

[0033] In addition, the present invention provides an application of the above-mentioned emotional beauty composition in the preparation of skin care products.

[0034] Furthermore, the present invention provides a skin care product, which includes the above-mentioned emotional beauty composition.

[0035] Preferably, the skin care product includes the following components in mass percentage: 0.1-5% of the above-mentioned emotional beauty composition, 2-40% of a cosmetic matrix, and the balance being water.

[0036] Preferably, the skin care product includes any one of lotion, emulsion, cream, mask, essence, essential oil, spray.

[0037] In one embodiment, the present invention provides an emulsion, which includes the following components in mass percentage: 0.1-5% of the above-mentioned emotional beauty composition, 0.1-0.5% of a thickener, 3-10% of a moisturizer, 0.1-10% of an emulsifier, 0.5-5% of a preservative, 0.05-0.5% of a pH regulator, 0.05-0.1% of a fragrance, and the balance being water.

[0038] In one embodiment, the thickener is one or more of polyacrylate crosspolymer-6, carbomer (including various configurations and different molecular weights), carrageenan, gellan gum, microcrystalline cellulose, cellulose gum, ethyl cellulose, tara gum, guar gum; the moisturizer is one or more of glycerin, D-panthenol, vitamin B5, 1,3-butanediol, 1,2-hexanediol, 1,3-propanediol, sodium hyaluronate, tremella polysaccharide, trehalose, betaine, allantoin, ceramide; the emulsifier is one or more of triglyceride caprylate / caprate, C14-22 alcohol, C12-20 alkyl glucoside, cetearyl glucoside, isononyl isononanoate, pentaerythritol tetra(ethylhexanoate), polydimethylsiloxane, stearyl alcohol, sucrose stearate, hydroxystearic acid, polymethylsilsesquioxane, pentaerythritol distearate, sucrose stearate; the preservative is p-hydroxyacetophenone and / or polyol, avoiding the use of traditional preservatives; the pH regulator is one or more of disodium ethylenediaminetetraacetate, arginine.

[0039] Compared with the prior art, the beneficial effects of the present invention are as follows: In view of the core problem that the cortisol rhythm disorder caused by emotional stress inhibits the expression of skin barrier genes (FLG / LOR), the present invention proposes a solution through a three-level linkage of "cortisol rhythm synchronization-barrier gene repair-neuro-sensitivity regulation".

[0040] The present invention targets and regulates the activity of the HPA axis through echinacoside in Echinacea purpurea extract, inhibits the release of hypothalamic CRH, reduces the nocturnal cortisol peak to the physiological rhythm level, and relieves the transcriptional inhibition of FLG / LOR genes by cortisol; at the same time, in combination with the 5-hydroxytryptamine precursor in Tephrosia purpurea seed extract, it regulates the amygdala stress response through the blood-brain barrier and blocks the vicious cycle of "skin sensitivity→increased anxiety→cortisol secretion". At the skin end, the lysate of Lactobacillus fermentation activates the skin "clock" genes, reconstructs the circadian repair rhythm of epidermal cells, makes the synthesis of FLG naturally misaligned with the cortisol secretion peak, and synergistically remodels the tight junction structure with the tea polyphenol derivatives in Camellia nitidissima extract to achieve dual repair of the physical-immune barrier. The present invention integrates for the first time the regulation of emotion-derived cortisol rhythm and the repair of the skin biological clock-barrier gene network, breaks through the limitation of the traditional "treating the symptoms rather than the root cause", and forms a full-link barrier repair system from the intervention of the stress source to the physiological reconstruction of the skin. Brief Description of the Drawings

[0041] Figure 1 It is a schematic diagram of improving the skin circadian rhythm of the normal group, the model group and Example 1;

[0042] Figure 2 It is the human efficacy diagram of Application Example 1 and Comparative Application Example 1. Detailed Embodiments

[0043] To better illustrate the purpose, technical solution and advantages of the present invention, the present invention will be further described below in conjunction with the drawings and specific embodiments. The purpose is to understand the content of the present invention in detail, rather than to limit the present invention. All other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention. The experimental reagents and instruments designed in the embodiments and comparative examples of the present invention are all common ordinary reagents and instruments unless otherwise specified, and can be obtained from commercial channels. In the embodiments and comparative examples, the experimental methods used are all conventional methods unless otherwise specified; and unless otherwise specified, the raw materials used in parallel experiments are of the same batch.

[0044] The raw materials used in this application are introduced as follows, but are not limited to the following raw materials:

[0045] Echinacea purpurea extract: purchased from Symrise AG, trade name SymFinity;

[0046] Glechoma longituba seed extract: purchased from Givaudan under the trade name Neurophroline;

[0047] Camellia chrysantha extract: Ximuyuan (Xiamen) Biotechnology Co., Ltd.;

[0048] Calendula officinalis extract: purchased from Zhenghe (Guangzhou) Biotechnology Co., Ltd.;

[0049] Soybean seed extract: purchased from Quzhou Zhanhong Biotechnology Co., Ltd.

[0050] Green tea extract: purchased from Shanyang Lianfeng Biotechnology;

[0051] Bacillus fermentation products: purchased from Guangzhou Youke Biotechnology Co., Ltd.;

[0052] Lactobacillus plantarum: the collection number is CGMCC 1.12934, purchased from China General Microbiological Culture Collection Center;

[0053] Lactobacillus brevis: deposit number CGMCC 1.2028, purchased from China General Microbiological Culture Collection Center;

[0054] Lactic acid bacteria fermentation lysate-1: Homemade, preparation method is as follows:

[0055] S1. The lactic acid bacteria strains are cultured in seed culture medium respectively; wherein the seed culture medium includes: 20g glucose, 10g peptone, 10g beef extract powder, 5g yeast extract, 5g anhydrous sodium acetate, 1ml Tween-80, 2g diammonium hydrogen citrate, 2g dipotassium hydrogen phosphate, 0.58g magnesium sulfate heptahydrate, 0.19g manganese sulfate monohydrate, add water to 1000ml, adjust pH to 6.8, sterilize at 118°C for 20min; the lactic acid bacteria strains are Lactobacillus plantarum and Lactobacillus brevis;

[0056] S2, inoculating the cultured lactic acid bacteria seeds into fermentation medium to obtain fermentation liquid;

[0057] First, microaerobic fermentation was carried out under the following conditions: 5% (volume ratio, i.e., the ratio of the volume of bacterial liquid to the volume of fermentation medium) of bacterial liquid was inoculated into a fermentation medium with a pH of 6.0, and the dissolved oxygen content in the culture system was stabilized within the range of 0.5 mg / L at a stirring rate of 150 rpm and a ventilation volume of 0.3 vvm (volume ratio ventilation rate), and cultured at 37° C. for 24 hours; then, anaerobic fermentation was carried out under the following conditions: ventilation was stopped and the stirring rate was reduced to 100 rpm, the dissolved oxygen content was maintained at <0.1 mg / L, and cultured at 30° C. for 48 hours.

[0058] The fermentation medium includes: 2% glucose, 1.2% peptone, 0.5% beef extract powder, 1% yeast extract, 0.5% anhydrous sodium acetate, 0.1% Tween-80, 0.2% diammonium hydrogen citrate, 0.2% dipotassium hydrogen phosphate, 0.058% magnesium sulfate heptahydrate, 0.05% manganese sulfate monohydrate, 0.1% L-cysteine ​​hydrochloride, and the rest is water;

[0059] S3, centrifuging the fermentation broth at a speed of 6500 r / min for 15 min, pouring out the supernatant to collect the bacterial sludge; washing with physiological saline 2-3 times to remove the residual culture medium;

[0060] S4. Mix the bacterial mud, add a protective agent according to the mass of the mixed bacterial mud at a ratio of 1:8, the protective agent composition: 28% trehalose by mass, and fully mix with a high-speed stirrer to obtain a mixed solution; the lactic acid bacteria strain is a mixed strain of Lactobacillus plantarum and Lactobacillus brevis, and the weight ratio of Lactobacillus plantarum mud to Lactobacillus brevis mud is 1:2.

[0061] S5. Sterilize the mixed solution by high temperature moist heat at 121° C. for 20 min to obtain a lactic acid bacteria fermentation lysate.

[0062] Lactic acid bacteria fermentation lysate-2: homemade, the preparation method is different from that of lactic acid bacteria fermentation lysate-1, only the weight ratio of the bacterial mud is different, the total weight of the bacterial mud is the same, the lactic acid bacteria species are Lactobacillus plantarum and Lactobacillus brevis, and in S4, the weight ratio of the Lactobacillus plantarum mud to the Lactobacillus brevis mud is 3:5.

[0063] Lactic acid bacteria fermentation lysate-3: homemade, the preparation method is different from that of lactic acid bacteria fermentation lysate-1, only the weight ratio of bacterial mud is different, the total weight of bacterial mud is the same, the lactic acid bacteria species are Lactobacillus plantarum and Lactobacillus brevis, and in S4, the weight ratio of Lactobacillus plantarum mud to Lactobacillus brevis mud is 1:10.

[0064] Lactic acid bacteria fermentation lysate-4: homemade, the preparation method is different from that of lactic acid bacteria fermentation lysate-1, only the lactic acid bacteria species are selected differently, and only Lactobacillus plantarum is selected as the lactic acid bacteria species. The weight of Lactobacillus plantarum mud in S4 is the same as the total weight of Lactobacillus plantarum mud and Lactobacillus brevis mud in Example 1.

[0065] Lactic acid bacteria fermentation lysate-5: homemade, the preparation method is different from that of lactic acid bacteria fermentation lysate-1, only the selection of lactic acid bacteria species is different, and only Lactobacillus brevis is selected as the lactic acid bacteria species. The weight of Lactobacillus plantarum brevis mud in S4 is the same as the total weight of Lactobacillus plantarum mud and Lactobacillus brevis mud in Example 1.

[0066] Lactic acid bacteria fermented lysate - 6: Self - made. Compared with lactic acid bacteria fermented lysate - 1, in the S2 step only, the fermentation method of the strain is different, and only micro - aerobic fermentation is carried out.

[0067] Lactic acid bacteria fermented lysate - 7: Self - made. Compared with lactic acid bacteria fermented lysate - 1, in the S2 step only, the fermentation method of the strain is different, and only anaerobic fermentation is carried out.

[0068] Lactic acid bacteria fermented lysate - 8: Commercially available product, purchased from CLR Company in Germany, trade name ProRenewComplex CLR TM 。

[0069] Examples 1 - 13 and Comparative Examples 1 - 12

[0070] The examples and comparative examples of the present invention provide an emotional beauty composition. The components and parts by weight of the composition are selected as shown in Table 1 - Table 2. W1 represents the weight percentage of the lactic acid bacteria fermented lysate based on the total weight of the composition. The preparation method of the composition includes the following steps: Mix each component evenly to obtain the emotional beauty composition.

[0071] Table 1

[0072]

[0073] Table 2

[0074]

[0075] Application Examples

[0076] The application examples of the present invention provide an emulsion. The components (mass percentage) of the emulsion are shown in Table 3. Among them, the emotional beauty compositions in Application Examples 1 - 13 are respectively the emotional beauty compositions prepared in Examples 1 - 13, and the emotional beauty compositions in Comparative Application Examples 1 - 12 are respectively the emotional beauty compositions prepared in Comparative Examples 1 - 12. For example, the emotional beauty composition used in Application Example 1 is the emotional beauty composition prepared in Example 1, the emotional beauty composition used in Application Example 2 is the emotional beauty composition prepared in Example 2, the emotional beauty composition used in Application Example 14 is the emotional beauty composition prepared in Example 1, the emotional beauty composition used in Comparative Application Example 1 is the emotional beauty composition prepared in Comparative Example 1, and so on.

[0077] Table 3

[0078]

[0079]

[0080] The preparation method of the emulsion is as follows: Put the components in Phase A into water in sequence. After the Carbopol U30 is fully swollen, homogenize and mix evenly. Heat Phase A and Phase B separately until the solid substances melt and become uniform. Mix and homogenize and emulsify at 80 °C. Heat Phase C until the p-hydroxyacetophenone melts, add it to the emulsion and stir evenly. After the temperature drops to 40 °C, add Phase D and stir to form a uniform emulsion. Finally, add arginine to adjust the pH to obtain the emulsion.

[0081] Performance test - 1 Regulation of the clock gene PER1 by the composition.

[0082] Skin cells regulate the circadian repair rhythm through "clock genes" (such as PER1). The disruption of the cortisol rhythm caused by stress inhibits the expression of clock genes and destroys the skin's nocturnal repair ability. In this experiment, the function of restoring the skin biological clock was verified by detecting the regulatory effect of the composition on the clock gene.

[0083] The cell line used was human keratinocyte HaCaT (Shanghai Yaji Biotechnology Co., Ltd.). The test conditions were: incubator temperature 37 ± 1 °C, humidity 90 ± 5%, carbon dioxide 5 ± 1%; cell culture and treatment were carried out according to the grouping, and then the test was carried out. The test method was as follows:

[0084] (1) Sample preparation: The compositions prepared in each example and comparative example were dissolved in 0.1% DMSO (volume fraction of DMSO 0.1%) to form a sample solution, and the mass percentage content of the composition in the sample solution was 0.1%.

[0085] (2) Resuscitation of HaCaT cells: Inoculate the cell suspension into a 96-well cell culture plate at a density of 2000 cells / well, add 100 μL of DMEM + 10% fetal bovine serum (FBS) + 1% penicillin / streptomycin culture medium to each well, and resuscitate and culture for 24 h;

[0086] (3) Cell rhythm synchronization: When the cells were fused to 70%, they were treated with a medium containing 50% FBS for 2 hours to cause cell serum shock to induce circadian synchronization.

[0087] (4) Induction and sample addition: Discard the supernatant of the cells. The normal group (blank group) was added with 100 μL of DMEM medium (Gibco, 210534). The model group was added with 100 μL of DMEM medium containing 100 nM cortisol (MedChemExpress, HY-N0583). The sample groups of the examples and comparative examples were added with 100 μL of medium containing 100 nM cortisol and 0.1% corresponding sample solution, cultured for 24 h, and the cells of each group were collected at 6 h, 12 h, 18 h, and 24 h respectively. Discard the medium, wash twice with PBS. Add 1 mL of TRIzol lysis solution, scrape the cells, transfer them to an EP tube, and store at -80 °C for later use.

[0088] (4) Gene content determination: The expression level of PER1 gene in HaCaT cells of each group was detected by real-time fluorescence quantitative PCR method, and β-actin was used as the internal reference gene. Total RNA of cells in each sample group was extracted by TRIzol method, and then cDNA was synthesized using PrimeScript RT kit.

[0089] Primers were synthesized according to the following primer information. For PER1 gene: forward primer: 5'-CAAGGACTCAGAAGGA-ACTCATGACAG-3', reverse primer: 5'-GTACCGAGGCCCTGGATCCCG-GTCAG-3';

[0090] For the internal reference gene (β-actin): forward primer: 5'-CAT GTA CGT TGC TAT CCA GGC-3', reverse primer: 5'-CTC CTT AAT GTC ACG CAC GAT-3'.

[0091] Then, relative quantitative analysis of mRNA expression was performed using the CFX96 Touch Real-Time PCR detection system (Bio-Rad, Hercules, CA, USA). After normalizing the expression levels of each clock gene to the β-actin transcript level, the 2 -ΔΔCt method was used to calculate the gene level; software such as CircaCompare was used to fit the circadian positive sine curve. The mean square error (MSE) was used to calculate the overall difference in the PER1 expression levels of the model group and each sample group relative to the normal group at 4 time points, so as to judge the regulation of the clock gene "circadian rhythm" by the model group and each sample group. The smaller the MSE, the closer the gene expression level is to that of the normal group, and the stronger the ability of the sample to improve the skin circadian rhythm. The MSE calculation method is as follows:

[0092]

[0093] The expression levels of genes at each time point in the normal group, model group and Example 1 are as Figure 1 shown. From the data of the model group, it can be seen that the model of cortisol interfering with the skin cell rhythm was successfully established, and the subsequent data of sample treatment are credible.

[0094] The effects of each group on the skin keratinocyte rhythm are shown in Table 4, showing the gene expression levels at 6h, 12h, 18h and 24h, as well as the mean square error.

[0095] Table 4

[0096] Group 6h 12h 18h 24h MSE (compared with the normal group) Normal group 0.81 1.22 0.92 1.15 0 (baseline) Model group 1.21 2.30 0.37 1.73 0.491 Example 1 0.83 1.16 0.75 1.17 0.008 Example 2 0.82 1.03 0.93 1.13 0.009 Example 3 0.62 1.04 0.83 1.00 0.025 Example 4 0.67 1.13 0.77 0.85 0.035 Example 5 0.58 0.99 0.81 0.98 0.037 Example 6 0.53 1.02 0.83 0.92 0.045 Example 7 0.61 0.99 0.82 0.88 0.044 Example 8 0.52 1.05 0.76 0.91 0.049 Example 9 0.78 1.12 0.87 0.92 0.017 Example 10 0.68 1.12 0.69 0.95 0.030 Example 11 0.67 1.15 0.70 0.97 0.026 Example 12 0.54 0.83 0.82 1.03 0.062 Example 13 0.63 0.83 0.92 0.92 0.059 Comparative Example 1 0.93 1.73 0.42 1.63 0.189 Comparative Example 2 0.93 1.63 0.32 1.33 0.144 Comparative Example 3 0.10 1.42 0.32 1.23 0.228 Comparative Example 4 0.73 1.53 0.26 1.55 0.175 Comparative Example 5 1.21 1.74 0.41 1.74 0.260 Comparative Example 6 0.72 0.83 0.32 1.29 0.135 Comparative Example 7 0.53 0.93 0.32 1.34 0.140 Comparative Example 8 0.62 0.83 0.51 0.91 0.103 Comparative Example 9 0.71 0.69 0.53 1.24 0.113 Comparative Example 10 0.74 0.82 0.42 1.11 0.104 Comparative Example 11 0.72 0.99 0.37 1.43 0.110 Comparative Example 12 0.82 1.23 0.61 1.72 0.105

[0097] As can be seen from the above table, compared with the normal group, the mean square error MSE of the gene expression level of the emotional beauty composition prepared in the embodiments of the present invention is less than 0.07. The emotional beauty composition prepared in the embodiments of the present invention can well regulate the circadian repair rhythm and restore the function of the skin biological clock.

[0098] It can be seen from the comparison of Examples 1-8 that when the lysate of lactic acid bacteria prepared by different fermentation and culture methods is added to the emotional beauty composition, it will affect the effect of the obtained emotional beauty composition in regulating the circadian repair rhythm and restoring the skin biological clock.

[0099] It can be seen from the comparison of Example 1 and Examples 9-13 that the weight parts of the components in the composition will affect the performance of the product. When the weight parts of the components are further selected within the preferred range, the obtained product can better achieve the regulation of the cortisol rhythm of emotional origin, activate the skin "clock" gene, and reconstruct the circadian repair rhythm of epidermal cells.

[0100] It can be seen from the comparison of Example 1 and Comparative Examples 1-7 that when one or two of the components are not added, or when other component parts are supplemented while not adding, the effects of the present invention cannot be achieved; it can be seen from the comparison of Example 1 and Comparative Examples 8-11 that when other similar components are used for substitution, the corresponding effects of the present invention cannot be obtained. It can be seen from the comparison of Example 1 and Comparative Example 12 that when the weight parts of the components in the composition are not within the scope of the present application, good synergistic effects cannot be exerted.

[0101] Performance test - 2 Promotion effect of the composition on FLG and LOR barrier proteins.

[0102] FLG (filaggrin) and LOR (loricrin) are the core proteins of the skin barrier, and their expression is inhibited by cortisol for a long time. This experiment verifies whether the compositions prepared in the examples and comparative examples can reverse the inhibition of cortisol on the expression of barrier genes.

[0103] The cell line used is human keratinocyte HaCaT (Shanghai Yaji Biotechnology Co., Ltd.). The test conditions are: the temperature of the incubator is 37±1°C, the humidity is 90±5%, and the carbon dioxide is 5±1%; according to the grouping, cell culture and treatment are carried out, and then the test is carried out. The test method is as follows:

[0104] (1) Sample preparation: The compositions prepared in each example and comparative example are dissolved in 0.1% DMSO (the volume fraction of DMSO is 0.1%) to form a sample solution, and the mass percentage content of the composition in the sample solution is 0.1%.

[0105] (2) Resuscitate HaCaT cells: Inoculate the cell suspension into a 96-well cell culture plate at a density of 2000 cells / well, add 100 μL of DMEM + 10% fetal bovine serum (FBS) + 1% penicillin / streptomycin culture medium to each well, and resuscitate and culture for 24 h;

[0106] (3) Induction and sample administration: Discard the supernatant of the cells. Add 100 μL of DMEM medium containing 100 nM cortisol (MedChemExpress, HY-N0583) to the control group. Add 100 μL of medium containing 100 nM cortisol and 0.1% corresponding sample solution to the examples and comparative examples, culture for 24 h, collect the cells of each group, discard the medium, and wash twice with PBS. Add 1 mL of TRIzol lysis solution, scrape the cells, transfer them to an EP tube, and store at -80 °C for later use.

[0107] (4) Gene content determination: Gene content determination: Use real-time fluorescence quantitative PCR to detect the expression levels of FLG and LOR genes in HaCaT cells of each group, and use β-actin as an internal reference gene.

[0108] Use the TRIzol method to extract the total RNA of the cells in each sample group respectively, then use the PrimeScript RT kit to synthesize cDNA, and synthesize primers according to the following primer information. FLG gene: Forward primer: 5'-ATG GCT AAG ACC CCT AGT GAC CA-3', Reverse primer: 5'-TCA GTC AGG CCC CTG ACC ACC A-3'; LOR gene: Forward primer: 5'-CAG CCA GCT GTC TCA GCT TC-3', Reverse primer: 5'-TGC TGC TGA TGA TGG TGA AG-3'; Internal reference gene (β-actin): Forward primer: 5'-CAT GTA CGT TGC TAT CCA GGC-3', Reverse primer: 5'-CTC CTT AAT GTC ACG CAC GAT-3'.

[0109] Then use the CFX96 Touch Real-Time PCR detection system (Bio-Rad, Hercules, CA, USA) to perform relative quantitative analysis of mRNA expression levels. After normalizing the expression levels of each gene to the β-actin transcript level, use 2 -ΔΔCt methods to calculate the gene expression levels of FLG and LOR.

[0110] Gene expression improvement rate = (gene expression level 样品组 - gene expression level 对照组 ) / gene expression level 对照组 × 100%;

[0111] Among them, the higher the gene improvement rates of FLG and LOR, the stronger the ability to combine and repair the barrier. The calculated results are shown in Table 5 below;

[0112] Table 5

[0113]

[0114]

[0115] As can be seen from the above table, the emotional beauty composition prepared in the embodiment of the present invention can reverse the inhibition of cortisol on the expression of barrier genes. Specifically, after using the emotional beauty composition prepared in the embodiment of the present invention, the improvement rate of the FLG gene reaches more than 120%, and the improvement rate of the LOR gene is more than 110%. The emotional beauty composition prepared in the embodiment of the present invention can well repair the barrier genes.

[0116] It can be seen from the comparison of Examples 1-8 that when the lactic acid bacteria fermentation lysate prepared by different fermentation and culture methods is added to the emotional beauty composition, it will affect the effect of the obtained emotional beauty composition on repairing barrier genes.

[0117] It can be seen from the comparison of Example 1 and Examples 9-13 that the weight parts of the components in the composition will affect the performance of the product. When the weight parts of the components are further selected within the preferred range, the obtained product can better achieve the effect of repairing barrier genes.

[0118] It can be seen from the comparison of Example 1 and Comparative Examples 1-7 that when one or two of the components are not added, or when other component parts are supplemented while not adding, the effects of the present invention cannot be achieved; it can be seen from the comparison of Example 1 and Comparative Examples 8-11 that when replaced with other similar components, the corresponding effects of the present invention cannot be obtained. It can be seen from the comparison of Example 1 and Comparative Example 12 that when the weight parts of the components in the composition are not within the scope of the present application, good synergistic effects cannot be exerted.

[0119] Performance Test - 3 Improvement of Facial Skin of Sensitive Skin Population by the Composition

[0120] The effect examples of the present invention explore the application effects of the emulsions prepared in Application Examples 1-14, Comparative Application Examples 1-12 and Blank Application Example on the human body, including the following steps:

[0121] 18 - 45-year-old sensitive skin subjects were selected according to the tests in the "Technical Specifications for Cosmetic Safety" (2015). The screening method was self-sensitivity assessment questionnaire + lactic acid stinging ≥ 3 points. The subjects were randomly divided into 27 groups, with 5 people in each group. The volunteers used the lotion on the whole face once in the morning and once in the evening every day. Data were collected on the 0th day and the 28th day of use. After the volunteers arrived, they washed their faces with facial cleanser, and after sitting still for 30 minutes in an air-conditioned room at a temperature of 21 ± 1°C and a humidity of 50 ± 10%, the trans-epidermal water loss value (TEWL) of the cheekbones was measured using the skin water loss test probe TewaMeter, and the improvement rate of facial TEWL was calculated. Before and after use, the skin redness degree of the subjects was measured using the skin color test probe Colorimeter CL440, the improvement rate of facial erythema was calculated, and the facial erythema situation was photographed using the VISIA-CR instrument. At the same time, an appropriate amount of saliva was collected each time the volunteers arrived, and the content of the stress hormone cortisol in the saliva was detected using the human salivary cortisol (SC) ELISA detection kit (Jianglai Company, JL10796-96T).

[0122] The soothing ability is represented by the improvement of facial erythema, the skin barrier repair ability is represented by the improvement of the TEWL value, and the reduction of stress is represented by the improvement rate of cortisol:

[0123] Improvement rate = |(T0 测量值 - T28 测量值 )| / T0 测量值 * 100%.

[0124] The data are shown in Table 6.

[0125] Table 6

[0126]

[0127]

[0128] As can be seen from the above table, when the emotional beauty composition prepared in the examples of the present invention is prepared into a lotion, it has good effects of improving skin erythema, enhancing skin barrier ability and relieving stress. From Figure 1 it can be seen that when the emotional beauty composition prepared in the examples of the present invention is prepared into a lotion, it has good effects of improving skin erythema and enhancing skin barrier ability.

[0129] By comparing Application Examples 1 - 8, it can be seen that using different fermentation and culture methods to prepare the lactic acid bacteria fermented lysate will affect the effects of the emotional beauty composition prepared into a lotion in improving skin erythema, enhancing skin barrier ability and relieving stress.

[0130] From the comparison between Application Example 1 and Application Examples 9 - 13, it can be seen that the parts by weight of the components in the emotional beauty composition added to the emulsion will affect the performance of the product. When the parts by weight of the components are further selected within the preferred range, the obtained product can better achieve the effects of improving skin erythema, enhancing skin barrier ability, and relieving stress.

[0131] From the comparison between Application Example 1 and Comparative Application Examples 1 - 7, it can be seen that when one or two of the components are not added, or when other component parts are used to make up for the non - addition, the effects of the present invention cannot be achieved; from the comparison between Application Example 1 and Comparative Application Examples 8 - 11, it can be seen that when other similar components are used for substitution, the corresponding effects of the present invention cannot be obtained. From the comparison between Application Example 1 and Comparative Application Example 12, it can be seen that when the parts by weight of the components in the composition are not within the scope of this application, good synergistic effects cannot be exerted.

[0132] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. An emotional beauty composition, characterized in that, It comprises the following components in parts by weight: 0.1 - 5 parts of Echinacea purpurea extract, 0.01 - 2 parts of Tephrosia purpurea seed extract, 0.01 - 3 parts of Camellia nitidissima extract, and 0.1 - 3 parts of lactic acid bacteria fermentation lysate.

2. The emotional beauty composition according to claim 1, wherein It comprises the following components in parts by weight: 0.5 - 2 parts of Echinacea purpurea extract, 0.05 - 1 part of Tephrosia purpurea seed extract, 0.05 - 1 part of Camellia nitidissima extract, and 0.5 - 2 parts of lactic acid bacteria fermentation lysate.

3. The emotional beauty composition according to claim 2, characterized in that, It comprises the following components in parts by weight: 1 - 1.5 parts of Echinacea purpurea extract, 0.1 - 0.5 part of Tephrosia purpurea seed extract, 0.5 - 0.8 part of Camellia nitidissima extract, and 1 - 1.5 parts of lactic acid bacteria fermentation lysate.

4. The emotional beauty composition according to claim 1, characterized in that, Based on the total weight of the emotional beauty composition, the weight percentage content of the lactic acid bacteria fermentation lysate is 15 - 40%.

5. The emotional beauty composition according to claim 1, characterized in that, The lactic acid bacteria fermentation lysate is obtained by fermentation of at least one of Lactobacillus plantarum and Lactobacillus brevis; And / or, the fermentation method of the lactic acid bacteria fermentation lysate is at least one of microaerobic fermentation and anaerobic fermentation.

6. The emotional beauty composition according to claim 5, wherein The lactic acid bacteria fermentation lysate is obtained by microaerobic fermentation, and in the microaerobic fermentation, the dissolved oxygen content in the fermentation system is 0.2 - 0.5 mg / L; Or, the lactic acid bacteria fermentation lysate is obtained by anaerobic fermentation, and in the anaerobic fermentation, the dissolved oxygen content in the fermentation system < 0.1 mg / L; Or, the lactic acid bacteria fermentation lysate is obtained by first performing microaerobic fermentation and then anaerobic fermentation. In the microaerobic fermentation, the dissolved oxygen content in the fermentation system is 0.2 - 0.5 mg / L, and in the anaerobic fermentation, the dissolved oxygen content in the fermentation system < 0.1 mg / L.

7. Use of an emotional beauty composition according to any one of claims 1 - 6 in the preparation of skin care products.

8. A skin care product, characterized in that, The skin care product comprises an emotional beauty composition according to any one of claims 1 - 6.

9. The skin care product according to claim 8, characterized in that, The skin care product comprises the following components in mass percentage: 0.1 - 5% of an emotional beauty composition according to any one of claims 1 - 6, 2 - 40% of a cosmetic matrix, and the balance being water.

10. The skin care product according to claim 9, wherein, The skin care product includes any one of lotion, emulsion, cream, facial mask, essence, essential oil, and spray.

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