A mood-enhancing cosmetic composition containing Camellia chrysantha flower extract and its application

Abstract

This invention discloses an emotional beauty composition containing Camellia chrysantha extract and its application, relating to the field of cosmetic technology. The invention provides an emotional beauty composition comprising the following components in parts by weight: 0.1-5 parts of Echinacea purpurea extract, 0.01-2 parts of Edamame seed extract, 0.01-3 parts of Camellia chrysantha extract, and 0.1-3 parts of lactic acid bacteria fermentation lysate. This invention addresses the core problem of cortisol rhythm disorder caused by emotional stress leading to the inhibition of skin barrier gene (FLG / LOR) expression, proposing a three-tiered solution involving "cortisol rhythm synchronization - barrier gene repair - neurosensitivity regulation."

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and in particular to a mood-enhancing composition containing Camellia chrysantha extract and its application. Background Technology

[0002] There is a very close relationship between emotions and skin health, especially negative emotions such as stress, anxiety, and depression, which can significantly exacerbate problems with sensitive skin. Stress, as a key factor affecting the effectiveness of traditional skincare products, 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, stress accelerates damage to the skin barrier function, leading to increased transepidermal water loss and making the skin's reaction to external stimuli, such as ultraviolet rays, pollutants, and allergens, more intense. This barrier damage then triggers a series of sensitivity symptoms, including stinging, tightness, and redness. Excessive secretion of cortisol under stress is one of the core mechanisms leading to these problems. While cortisol, a key hormone regulated under stress, has some anti-inflammatory effects in the short term, prolonged high levels weaken the skin barrier function and make the skin more fragile by promoting inflammation and oxidative stress. Furthermore, excessive cortisol also inhibits the skin's self-repair ability, further aggravating the symptoms of sensitive skin.

[0003] Traditional skincare products targeting sensitive skin often employ anti-inflammatory ingredients such as dipotassium glycyrrhizate and bisabolol, as well as barrier lipid supplements like ceramides and cholesterol. Their mechanism of action primarily involves inhibiting mast cell degranulation or replenishing intercellular lipids to alleviate immediate sensitivity symptoms, such as reducing stinging and tightness. However, these approaches have significant limitations. They only address localized inflammation or damage to the physical skin barrier and cannot effectively address gene expression abnormalities caused by stress. Research shows that chronic stress causes cortisol to persistently suppress FLG (filaggrin) and LOR (lobeline) gene expression, and traditional barrier lipid supplements can only temporarily fill the lipid gap without restoring the autonomous synthesis capacity of FLG and LOR genes. Some products on the market attempt to use aromatherapy, such as lavender essential oil and bergamot volatiles, to indirectly improve skin condition by regulating emotions. However, these techniques have significant limitations. They ignore the body's circadian rhythms and cannot differentiate between circadian rhythm needs. The normal secretion of cortisol in the human body has a circadian rhythm, with higher levels in the morning and lower levels at night. If cortisol levels rise abnormally at night, it will continuously inhibit the self-repair of skin cells. For example, the peak synthesis of skin barrier protein FLG at night will be suppressed by the rise in cortisol. Existing techniques such as aromatherapy have not fully considered this circadian rhythm factor, making it difficult to achieve precise and effective improvement of skin condition.

[0004] Stress, as a significant emotional factor affecting skin health, is increasingly highlighting the shortcomings of traditional skincare products in addressing complex skin sensitivity issues. This has led to in-depth discussions on the close link between emotions and skin health. Therefore, this application is submitted. Summary of the Invention

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

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows: an emotional beauty composition comprising the following components in parts by weight: 0.1-5 parts of Echinacea purpurea extract, 0.01-2 parts of Edamame seed extract, 0.01-3 parts of Camellia chrysantha extract, and 0.1-3 parts of lactic acid bacteria fermentation lysate.

[0007] This invention addresses the core issue of cortisol rhythm disorder caused by emotional stress leading to the inhibition of skin barrier gene (FLG / LOR) expression, and proposes a three-level linkage solution involving "cortisol rhythm synchronization - barrier gene repair - neurosensitivity regulation".

[0008] This invention targets and regulates HPA axis activity through echinacea purpurea extract, inhibiting hypothalamic CRH release, reducing nighttime cortisol peaks to circadian rhythm levels, and relieving cortisol's transcriptional repression of FLG / LOR genes. Simultaneously, it combines with serotonin precursors from soybean seed extract to regulate amygdala stress response across the blood-brain barrier, blocking the vicious cycle of "skin sensitivity → increased anxiety → cortisol secretion." At the skin level, lactic acid bacteria fermentation lysates activate skin "clock" genes, reconstructing the diurnal repair rhythm of epidermal cells, naturally misaligning FLG synthesis and cortisol secretion peaks. This, in conjunction with tea polyphenol derivatives from camellia oleifera extract, remodels tight junction structures, achieving dual repair of the physical and immune barriers. This invention is the first to integrate emotionally driven cortisol rhythm regulation with skin biological clock-barrier gene network repair, overcoming the limitations of traditional "treating the symptoms but not the root cause" approaches, and forming a comprehensive barrier repair system from stress source intervention to skin physiological reconstruction.

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

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

[0011] The inventors discovered in their actual research that the weight proportions of the components in the composition provided by this invention affect the performance of the product. When the weight proportions of the components are further selected within the above range, the resulting product can better achieve the regulation of the cortisol rhythm of mood, activate the skin's "clock" gene, rebuild the diurnal repair rhythm of epidermal cells, and form a full-link barrier repair system from stress source intervention to skin physiological reconstruction.

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

[0013] In their actual research, the inventors discovered that when the weight percentage of lactic acid bacteria fermentation lysate in the emotional beauty composition is within the above-mentioned range, the resulting product has a superior effect in achieving emotional cortisol rhythm regulation, activating skin "clock" genes, and rebuilding the diurnal repair rhythm of epidermal cells.

[0014] The inventors discovered in their actual research that the lactic acid bacteria fermentation lysate can be a commercially available product or can be prepared in-house. In-house preparation simply requires the use of conventional methods for preparing fermentation lysates.

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

[0016] S1. Propagate the lactic acid bacteria strains separately in seed culture medium;

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

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

[0019] S4. Mix the mushroom sludge, add a preservative according to the quality of the mushroom sludge, and stir to mix thoroughly to obtain a mixture;

[0020] S5. Sterilize the mixture at high temperature to obtain the lactic acid bacteria fermentation lysate.

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

[0022] In the fermentation culture method of the lactic acid bacteria fermentation lysate of this application, when the lactic acid bacteria strain is not limited to one type, in step S1, the expansion culture involves expanding each type of lactic acid bacteria in different seed culture media; in step S2, the fermentation involves inoculating each type of lactic acid bacteria into different fermentation culture media, and then mixing the fermented bacteria sludge. The seed culture media used for expansion have the same composition, the fermentation culture media have the same composition, and the subsequent fermentation culture conditions are the same.

[0023] Preferably, in step S2, the fermentation is carried out using at least one of microaerobic fermentation and anaerobic fermentation.

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

[0025] More preferably, in step S2, the fermentation adopts a combination of microaerobic fermentation and anaerobic fermentation, with microaerobic fermentation performed first and anaerobic fermentation performed later; 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 is <0.1 mg / L.

[0026] In one embodiment, in step S2, microaerobic fermentation is first performed under the following conditions: 5-10% (volume ratio, i.e., the ratio of bacterial volume to fermentation medium volume) of bacterial culture is inoculated into the fermentation medium; the dissolved oxygen content in the culture system is stabilized at 0.2-0.5 mg / L at a stirring rate of 100-200 rpm and an aeration rate of 0.3-0.5 vvm (volume ratio aeration rate); and the culture is carried out at 34-37℃ for 24-30 hours. Then, anaerobic fermentation is performed under the following conditions: aeration is stopped and the stirring rate is reduced to 50-100 rpm to maintain a dissolved oxygen content <0.1 mg / L; and the culture is carried out at 30-34℃ for 36-48 hours.

[0027] Preferably, in step S4, mixing the bacterial sludge refers to the case where there is more than one type of lactic acid bacteria. If there is only one type of lactic acid bacteria, then there is no need to mix them, and a protectant can be added directly.

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

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

[0030] Preferably, the protective agent comprises trehalose and water, wherein the trehalose has a mass percentage of 5-50%; preferably, the trehalose has a mass percentage of 28%.

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

[0032] The inventors discovered in their actual research that the properties of lactic acid bacteria fermentation lysates obtained by using different lactic acid bacteria fermentation cultures and different fermentation culture methods are different. Therefore, when lactic acid bacteria fermentation lysates prepared by different fermentation culture methods are added to the mood beauty composition, the efficacy of the resulting mood beauty composition will vary. When the above method is selected, the overall effect of the resulting mood beauty composition is better.

[0033] Furthermore, the present invention provides an application of the aforementioned mood-enhancing composition in the preparation of skin care products.

[0034] Furthermore, the present invention provides a skin product comprising the aforementioned mood-enhancing composition.

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

[0036] Preferably, the skin care products include any one of toner, lotion, cream, mask, serum, essential oil, and spray.

[0037] In one embodiment, the present invention provides an emulsion comprising the following components in weight percentages: 0.1-5% of the 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 adjuster, 0.05-0.1% of a fragrance, and the balance being water.

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

[0039] Compared with the prior art, the beneficial effects of the present invention are as follows: The present invention addresses the core problem of the inhibition of skin barrier gene (FLG / LOR) expression caused by cortisol rhythm disorder induced by emotional stress, and proposes a three-level linkage solution of "cortisol rhythm synchronization - barrier gene repair - neurosensitivity regulation".

[0040] This invention targets and regulates HPA axis activity through echinacea purpurea extract, inhibiting hypothalamic CRH release, reducing nighttime cortisol peaks to circadian rhythm levels, and relieving cortisol's transcriptional repression of FLG / LOR genes. Simultaneously, it combines with serotonin precursors from soybean seed extract to regulate amygdala stress response across the blood-brain barrier, blocking the vicious cycle of "skin sensitivity → increased anxiety → cortisol secretion." At the skin level, lactic acid bacteria fermentation lysates activate skin "clock" genes, reconstructing the diurnal repair rhythm of epidermal cells, naturally misaligning FLG synthesis and cortisol secretion peaks. This, in conjunction with tea polyphenol derivatives from camellia oleifera extract, remodels tight junction structures, achieving dual repair of the physical and immune barriers. This invention is the first to integrate emotionally driven cortisol rhythm regulation with skin biological clock-barrier gene network repair, overcoming the limitations of traditional "treating the symptoms but not the root cause" approaches, and forming a comprehensive barrier repair system from stress source intervention to skin physiological reconstruction. Attached Figure Description

[0041] Figure 1 A schematic diagram illustrating the improvement of skin circadian rhythm in the normal group, model group, and Example 1;

[0042] Figure 2 The diagram shows the human effects of application example 1 and the comparison application example 1. Detailed Implementation

[0043] To better illustrate the purpose, technical solution, and advantages of this invention, the invention will be further described below in conjunction with the accompanying drawings and specific embodiments. The purpose is to provide a detailed understanding of the invention, not to limit it. All other embodiments obtained by those skilled in the art without inventive effort are within the scope of protection of this invention. Unless otherwise specified, the experimental reagents and instruments designed in the embodiments and comparative examples of this invention are commonly used reagents and instruments, all of which are commercially available. Unless otherwise specified, the experimental methods used in the embodiments and comparative examples are conventional methods; and unless otherwise specified, the raw materials used in parallel experiments are from the same batch.

[0044] The raw materials used in this application are described below, but are not limited to the following:

[0045] Purple coneflower extract: purchased from SymFinity, a company specializing in purple coneflower extract.

[0046] Edamame seed extract: purchased from Givaudan, brand name Neurophroline;

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

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

[0049] Wild soybean seed extract: purchased from Quzhou Zhanhong Biotechnology Co., Ltd.;

[0050] Green tea extract: purchased from Shanyang Lianfeng Biotechnology Co., Ltd.

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

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

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

[0054] Lactic acid bacteria fermentation lysate-1: prepared in-house, the preparation method is as follows:

[0055] S1. The lactic acid bacteria strains were cultured separately in seed culture media; wherein, the seed culture media consisted of: 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, water added to 1000ml, pH adjusted to 6.8, and sterilized at 118℃ for 20min; the lactic acid bacteria strains were Lactobacillus plantarum and Lactobacillus brevis.

[0056] S2. Inoculate the expanded lactic acid bacteria seeds into the fermentation medium for fermentation to obtain fermentation broth;

[0057] Microaerobic fermentation was first performed under the following conditions: 5% (volume ratio, i.e., the ratio of bacterial volume to fermentation medium volume) of bacterial culture was inoculated into fermentation medium with a pH of 6.0. The dissolved oxygen level in the culture system was stabilized within the range of 0.5 mg / L at a stirring speed of 150 rpm and an aeration rate of 0.3 vvm (volume ratio aeration rate). The culture was then incubated at 37°C for 24 hours. Anaerobic fermentation was then performed under the following conditions: aeration was stopped and the stirring speed was reduced to 100 rpm to maintain a dissolved oxygen level of <0.1 mg / L. The culture was then incubated at 30°C for 48 hours.

[0058] The fermentation medium consisted of: 2% glucose, 1.2% peptone, 0.5% beef extract, 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 remainder being water.

[0059] S3. Centrifuge the fermentation broth at 6500 r / min for 15 min, then decant the supernatant to collect the bacterial sludge; wash with physiological saline 2-3 times to remove residual culture medium.

[0060] S4. Mix the bacterial sludge, and add a protective agent at a ratio of 1:8 according to the mass of the mixed bacterial sludge. The protective agent consists of 28% trehalose by mass. Mix thoroughly with a high-speed mixer to obtain a mixture. The lactic acid bacteria strain is a mixture of Lactobacillus plantarum and Lactobacillus brevis, and the weight ratio of Lactobacillus plantarum bacterial sludge to Lactobacillus brevis bacterial sludge is 1:2.

[0061] S5. Sterilize the mixture with high temperature and humidity at 121℃ for 20 minutes to obtain the lactic acid bacteria fermentation lysate.

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

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

[0064] Lactic acid bacteria fermentation lysate-4: self-made, the preparation method is the same as that of lactic acid bacteria fermentation lysate-1, only the selection of lactic acid bacteria species is different, only Lactobacillus plantarum is selected as the lactic acid bacteria species, and the weight of Lactobacillus plantarum sludge in S4 is the same as the total weight of Lactobacillus plantarum sludge and Lactobacillus brevis sludge in Example 1.

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

[0066] Lactic acid bacteria fermentation lysate-6: self-made, the preparation method is the same as that of lactic acid bacteria fermentation lysate-1, only the fermentation method of the strain in S2 is different, only microaerobic fermentation is carried out.

[0067] Lactic acid bacteria fermentation lysate-7: self-made, the preparation method is the same as that of lactic acid bacteria fermentation lysate-1, only the fermentation method of the strain in S2 is different, only anaerobic fermentation is carried out.

[0068] Lactic acid bacteria fermentation lysate-8: A commercially available product, purchased from CLR GmbH, Germany, under the trade name ProRenewComplex CLR. TM .

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

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

[0071] Table 1

[0072]

[0073] Table 2

[0074]

[0075] Application examples

[0076] This invention provides an emulsion whose components (mass percentage) are shown in Table 3. The emotional beauty compositions used in Application Examples 1-13 are those prepared in Examples 1-13, and the emotional beauty compositions used in Comparative Application Examples 1-12 are those prepared in Comparative Examples 1-12. For example, the emotional beauty composition used in Application Example 1 is the one prepared in Example 1; the emotional beauty composition used in Application Example 2 is the one prepared in Example 2; the emotional beauty composition used in Application Example 14 is the one prepared in Example 1; the emotional beauty composition used in Comparative Application Example 1 is the one prepared in Comparative Example 1, and so on.

[0077] Table 3

[0078]

[0079]

[0080] The emulsion is prepared as follows: Each component in phase A is added to water in sequence. After the carbomer U30 is fully swollen, it is homogenized and mixed. Phase A and phase B are heated separately until the solid substances are melted and homogenized. They are then mixed and homogenized at 80°C. Phase C is heated until p-hydroxyacetophenone is melted. It is then added to the emulsion and stirred evenly. After the temperature drops to 40°C, phase D is added and stirred to form a homogeneous emulsion. Finally, arginine is added 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 their circadian rhythm of repair through "clock genes" (such as PER1). Stress-induced cortisol rhythm disruption inhibits clock gene expression, impairing the skin's nighttime repair capabilities. This experiment verifies the skin's ability to restore its biological clock by examining the regulatory effect of the composition on clock genes.

[0083] The cell line used was human keratinocytes (HaCaT, Shanghai Yaji Biotechnology Co., Ltd.). The test conditions were: incubator temperature 37±1℃, humidity 90±5%, carbon dioxide 5±1%. Cells were cultured and treated according to groups, followed by testing. The test methods are as follows:

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

[0085] (2) Resuscitating HaCaT cells: The cell suspension was seeded into a 96-well cell culture plate at a density of 2000 cells / well. 100 μL of DMEM + 10% fetal bovine serum (FBS) + 1% penicillin / streptomycin culture medium was added to each well and the cells were revived and cultured for 24 h.

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

[0087] (4) Induction and Sample Delivery: The supernatant was discarded from the cells. The normal control group (blank group) was given 100 μL of DMEM medium (Gibco, 210534). The model group was given 100 μL of DMEM medium containing 100 nM cortisol (MedChemExpress, HY-N0583). The example and comparative sample groups were given 100 μL of medium containing 100 nM cortisol and 0.1% of the corresponding sample solution. Cells were cultured for 24 h, and collected at 6 h, 12 h, 18 h, and 24 h. The medium was discarded, and the cells were washed twice with PBS. 1 mL of TRIzol lysis buffer was added, cells were scraped, transferred to EP tubes, and stored at -80℃ 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 quantitative PCR, with β-actin as an internal reference gene. Total RNA was extracted from cells of each group using the TRIzol method, and then cDNA was synthesized using the PrimeScript RT kit.

[0089] Primers were synthesized based on the following primer information for the PER1 gene: forward primer: 5'-CAAGGACTCAGAAGGA-ACTCATGACAG-3', reverse primer: 5'-GTACCGAGGCCCTGGATCCCG-GTCAG-3';

[0090] 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 levels was performed using a CFX96 Touch Real-Time PCR detection system (Bio-Rad, Hercules, CA, USA). After normalizing the expression levels of each clock gene to β-actin transcript levels, 2... -ΔΔCt Methods: Gene levels were calculated; circadian rhythm sine and cosine curves were fitted using software such as CircaCompare. Mean squared error (MSE) was used to calculate the overall difference in PER1 expression levels at four time points between the model group and each sample group and the normal group, to assess the regulatory effect of the model group and each sample group on the circadian rhythm of clock genes. A smaller MSE indicates that the gene expression levels are closer to those of the normal group, and thus the stronger the sample's ability to improve the skin's circadian rhythm. The MSE calculation method is as follows:

[0092]

[0093] The expression levels of genes at various time points in the normal group, model group, and Example 1 are as follows: Figure 1 As shown in the data from the model group, the model of cortisol interfering with skin cell rhythm has been successfully established, and the data from subsequent sample processing are reliable.

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

[0095] Table 4

[0096] Group 6h 12h 18h 24h MSE (compared to normal group) normal group 0.81 1.22 0.92 1.15 0 (benchmark) 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 shown in the table above, the gene expression level of the mood-enhancing composition prepared in this embodiment of the invention is less than 0.07 compared with the normal group. The mood-enhancing composition prepared in this embodiment of the invention can effectively regulate the diurnal repair rhythm and restore the function of the skin's biological clock.

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

[0099] As can be seen from the comparison of Examples 1 and Examples 9-13, 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 resulting product can better achieve the regulation of mood-related cortisol rhythm, activate the skin's "clock" gene, and rebuild the diurnal repair rhythm of epidermal cells.

[0100] As can be seen from the comparison of Example 1 and Comparative Examples 1-7, the effects of the present invention cannot be achieved when one or two of the components are not added, or when other components are used to make up the difference while the components are not added. As can be seen from the comparison of Example 1 and Comparative Examples 8-11, the corresponding effects of the present invention cannot be obtained when other similar components are substituted. As can be seen from the comparison of Example 1 and Comparative Example 12, when the weight proportions of the components in the composition are not within the scope of this application, a good synergistic effect cannot be achieved.

[0101] Performance Test - 2: The promoting effect of the composition on FLG and LOR barrier proteins.

[0102] FLG (figrin) and LOR (lobelin) are core proteins of the skin barrier, and their expression is suppressed by cortisol over a long period of time. This experiment verifies whether the compositions prepared in the examples and comparative examples can reverse the inhibition of barrier gene expression by cortisol.

[0103] The cell line used was human keratinocytes (HaCaT, Shanghai Yaji Biotechnology Co., Ltd.). The test conditions were: incubator temperature 37±1℃, humidity 90±5%, carbon dioxide 5±1%. Cells were cultured and treated according to groups, followed by testing. The test methods are as follows:

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

[0105] (2) Resuscitating HaCaT cells: The cell suspension was seeded into a 96-well cell culture plate at a density of 2000 cells / well. 100 μL of DMEM + 10% fetal bovine serum (FBS) + 1% penicillin / streptomycin culture medium was added to each well and the cells were revived and cultured for 24 h.

[0106] (3) Induction and Sample Delivery: The supernatant of the cells was discarded. For the control group, 100 μL of DMEM medium containing 100 nM cortisol (MedChemExpress, HY-N0583) was added. For the examples and comparative groups, 100 μL of medium containing 100 nM cortisol and 0.1% of the corresponding sample solution was added. Cells were cultured for 24 h, and cells from each group were collected, the medium was discarded, and the cells were washed twice with PBS. 1 mL of TRIzol lysis buffer was added, cells were scraped, transferred to EP tubes, and stored at -80℃ for later use.

[0107] (4) Gene content determination: The expression levels of FLG and LOR genes in HaCaT cells of each group were detected by real-time fluorescence quantitative PCR, and β-actin was used as an internal reference gene.

[0108] Total RNA was extracted from cells in each group using the TRIzol method, and cDNA was synthesized using the PrimeScript RT kit. Primers were synthesized according to the following primer information: FLG gene: forward primer: 5'-ATG GCT AAG ACC CCT AGTGAC 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 CTTAAT GTC ACG CAC GAT-3'.

[0109] Then, relative quantitative analysis of mRNA expression levels was performed using a CFX96 Touch Real-Time PCR detection system (Bio-Rad, Hercules, CA, USA). After normalizing the expression levels of each gene to β-actin transcript levels, 2... -ΔΔCt The method calculates the gene expression levels of FLG and LOR.

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

[0111] The higher the gene improvement rate of FLG and LOR, the stronger the combined ability to repair the barrier. The calculated results are shown in Table 5.

[0112] Table 5

[0113]

[0114]

[0115] As shown in the table above, the mood-enhancing composition prepared in the embodiments of the present invention can reverse the inhibition of barrier gene expression by cortisol. Specifically, after use, the mood-enhancing composition prepared in the embodiments of the present invention improves the FLG gene by more than 120% and the LOR gene by more than 110%. The mood-enhancing composition prepared in the embodiments of the present invention can effectively repair barrier genes.

[0116] As can be seen from the comparison of Examples 1-8, when lactic acid bacteria fermentation lysate prepared by different fermentation culture methods is added to the mood beauty composition, it will affect the effect of the obtained mood beauty composition on barrier gene repair.

[0117] As can be seen from the comparison of Examples 1 and Examples 9-13, 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 resulting product can better achieve the effect of barrier gene repair.

[0118] As can be seen from the comparison of Example 1 and Comparative Examples 1-7, the effects of the present invention cannot be achieved when one or two of the components are not added, or when other components are used to make up the difference while the components are not added. As can be seen from the comparison of Example 1 and Comparative Examples 8-11, the corresponding effects of the present invention cannot be obtained when other similar components are substituted. As can be seen from the comparison of Example 1 and Comparative Example 12, when the weight proportions of the components in the composition are not within the scope of this application, a good synergistic effect cannot be achieved.

[0119] Performance Test - 3: Improvement of Facial Skin in Individuals with Sensitive Skin

[0120] The application effects of the emulsions prepared in Application Examples 1-14, Comparative Application Examples 1-12, and Blank Application Examples on the human body are investigated through the following steps:

[0121] According to the "Cosmetic Safety Technical Specifications" (2015), subjects aged 18-45 with sensitive skin were selected for the trial. The screening method was a self-sensitivity assessment questionnaire plus a lactic acid stinging score of ≥3. Subjects were randomly divided into 27 groups of 5 people each. Volunteers applied the lotion to their entire face twice daily, morning and evening. Data was collected on day 0 and day 28. After the trial, volunteers washed their faces with facial cleanser and sat quietly for 30 minutes in an air-conditioned room with a temperature of 21±1℃ and humidity of 50±10%. The transepidermal water loss (TEWL) value of the cheekbone was measured using a TewaMeter probe, and the facial TEWL improvement rate was calculated. Before and after application, the degree of skin redness was measured using a Colorimeter CL440 probe, and the facial erythema improvement rate was calculated. Facial erythema was also photographed using a VISIA-CR instrument. At the same time, a suitable amount of saliva was collected each time a volunteer visited, and the content of the stress hormone cortisol in the saliva was detected using a human salivary cortisol (SC) ELISA kit (Jianglai Company, JL10796-96T).

[0122] Soothing ability was represented by the improvement in facial erythema, skin barrier repair ability was represented by the improvement in TEWL value, and stress reduction was represented by the improvement rate of cortisol levels.

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

[0124] The data is shown in Table 6.

[0125] Table 6

[0126]

[0127]

[0128] As shown in the table above, the mood-enhancing composition prepared in the embodiments of the present invention, when formulated into an emulsion, has excellent effects in improving skin erythema, enhancing skin barrier function, and relieving stress. Figure 1 It is evident that the emotional beauty composition prepared in the embodiments of the present invention, when formulated into an emulsion, has a very good effect on improving skin erythema and enhancing skin barrier function.

[0129] As can be seen from the comparison of application examples 1-8, the lactic acid bacteria fermentation lysate prepared by different fermentation culture methods will affect the effect of the obtained mood beauty composition on improving skin erythema, enhancing skin barrier function and relieving stress when it is prepared into an emulsion.

[0130] As can be seen from the comparison of Application Examples 1 and 9-13, the weight parts of the components in the mood-enhancing composition added to the emulsion will affect the performance of the product. When the weight parts of the components are further selected within the preferred range, the resulting product can better achieve the effects of improving skin erythema, enhancing skin barrier function and relieving stress.

[0131] As can be seen from the comparison between Application Example 1 and Comparative Application Examples 1-7, the effects of the present invention cannot be achieved when one or two components are not added, or when other components are used to make up the difference while the components are not added. As can be seen from the comparison between Application Example 1 and Comparative Application Examples 8-11, the corresponding effects of the present invention cannot be achieved when other similar components are substituted. As can be seen from the comparison between Application Example 1 and Comparative Application Example 12, when the weight proportions of the components in the composition are not within the scope of this application, a good synergistic effect cannot be achieved.

[0132] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A mood-enhancing composition, characterized in that, The product comprises the following components in parts by weight: 0.5-2 parts of Echinacea purpurea extract, 0.05-1 part of Edamame seed extract, 0.05-1 part of Camellia chrysantha extract, and 0.5-2 parts of lactic acid bacteria fermentation lysate; wherein the lactic acid bacteria fermentation lysate is obtained by fermentation of Lactobacillus plantarum and Lactobacillus brevis; wherein the lactic acid bacteria fermentation lysate is obtained by first performing microaerobic fermentation and then anaerobic fermentation.

2. The mood-enhancing composition according to claim 1, characterized in that, It includes the following components in parts by weight: 1-1.5 parts of Echinacea purpurea extract, 0.1-0.5 parts of Edamame seed extract, 0.5-0.8 parts of Camellia chrysantha flower extract, and 1-1.5 parts of lactic acid bacteria fermentation lysate.

3. The mood-enhancing composition as described in claim 1, characterized in that, The weight percentage of lactic acid bacteria fermentation lysate is 15-40% based on the total weight of the emotional beauty composition.

4. The mood-enhancing composition as described in claim 1, characterized in that, 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 is <0.1 mg / L.

5. The use of the mood-enhancing composition as described in any one of claims 1-4 in the preparation of skin care products.

6. A skin care product, characterized in that, The skin care products include the mood-enhancing composition as described in any one of claims 1-4.

7. The skin product as described in claim 6, characterized in that, The skin care product comprises the following components in weight percentage: 0.1-5% of the emotional beauty composition according to any one of claims 1-4, 2-40% of the cosmetic base, and the balance being water.

8. The skin product as described in claim 7, characterized in that, The skin care products include any one of the following: toner, lotion, cream, mask, serum, facial oil, and spray.

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