Composition for adjusting skin micro-ecological flora structure and application of composition in improving red skin

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to a composition for adjusting a skin micro-ecological flora structure and application of the composition in improving red skin. Specifically, aiming at the problem of skin flora unbalance, skin physiological characteristics and skin flora characteristics of the red crowd are selected for research, key bacteria which influence flora composition of the red crowd are screened, and common cosmetic raw materials are screened by using characteristic bacteria propionibacterium acnes. On the basis, formula matching is carried out based on the proliferation rule of the raw materials on skin characteristic bacteria, the formula is further applied to the formula, the safety of the formula and the skin redness improvement condition after people use the formula are verified, and a product suitable for the skin characteristics of the redness people is obtained. Tests prove that the composition can remarkably improve the diversity of florae of the reddish skin, reduce the sebum content, the percutaneous moisture loss rate and the a value, recover the skin barrier function and improve the reddish symptom, so that the composition has good practical application value.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a composition for adjusting the structure of the skin's microecological flora and its application in improving redness in the skin. Background Technology

[0002] The information disclosed in this background section is intended only to enhance understanding of the overall background of the invention and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] The skin barrier function encompasses physical, chemical, immune, and microbial barriers, with the microbial barrier being crucial for skin health. Bacteria on the skin surface are divided into resident and transient bacteria. An imbalance in resident bacteria can easily lead to various skin problems. For example, the excessive proliferation of opportunistic pathogens such as Staphylococcus aureus is a significant factor causing skin redness. When the skin barrier function is impaired, such as due to dryness, trauma, or excessive cleansing, Staphylococcus aureus, which is normally colonized at low levels, will proliferate rapidly. Its secreted toxins and enzymes can stimulate the skin's immune system, triggering an inflammatory response, leading to vasodilation and subsequent redness. Furthermore, Propionibacterium acnes plays a vital role in the redness of acne patients. This bacterium can break down triglycerides in the skin, producing free fatty acids. These free fatty acids stimulate hair follicles and surrounding tissues, inducing inflammation and causing symptoms such as redness and papules. Simultaneously, abnormal interactions between microorganisms and the skin's immune system can exacerbate redness. For instance, when certain microbial antigens bind to receptors on the surface of skin immune cells, they can over-activate the immune response, continuously releasing inflammatory factors, making the redness persistent and difficult to resolve. Clinical studies have shown that approximately 30%-50% of adults experience varying degrees of skin redness, which severely impacts their appearance and quality of life.

[0004] However, the inventors discovered that, in the current technology, no one has analyzed the causes of skin redness and sensitivity from the perspective of microecology, nor has anyone systematically studied the characteristics of the microbial composition and skin physiological parameters of people with redness, nor has anyone studied the correlation between the skin microbial community and physiological parameters of people with redness, nor has anyone selected suitable cosmetic raw materials based on the characteristics of the microbial community of people with redness, in order to maintain the balance of the skin microecology and improve the skin microecological condition of people with redness as indicators to solve the problem of skin redness. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the present invention aims to provide a composition for adjusting the skin's microbiome structure and its application in improving redness in skin. Specifically, the present invention provides a composition screened based on skin physiological parameters and microbiome characteristics of individuals with redness, which can effectively adjust the skin's microbiome structure and improve skin redness. The present invention is based on the above research findings.

[0006] To achieve the above-mentioned technical objectives, the present invention provides the following technical solution: A first aspect of the present invention provides a composition for adjusting the structure of the skin microbiota, the composition comprising the following components in parts by weight: 0.1-2 parts panthenol, 0.1-0.5 parts dipotassium glycyrrhizate, 0.1-2 parts horse chestnut extract, 1-3 parts dioctyl carbonate and 1-3 parts diisostearyl malate.

[0007] Among them, the extract of horse chestnut is preferably prepared by supercritical CO2 extraction. The extract prepared by this method has better inhibitory and anti-inflammatory effects (reducing IL-6 expression) on Propionibacterium acnes than the extract prepared by ultrasonic extraction with ethanol.

[0008] A second aspect of this invention provides the application of the above-described composition for adjusting the skin's microecological flora structure in the preparation of cosmetics. Through experimental verification, this invention demonstrates that the above-described composition can effectively improve the microecology of reddened skin, restore flora balance, alleviate redness, and enhance the skin barrier function.

[0009] Therefore, in a third aspect, the present invention provides a cosmetic product comprising at least the above-described composition for adjusting the structure of the skin's microbiome.

[0010] The cosmetics mentioned can be skincare products.

[0011] A fourth aspect of the present invention provides a method for preparing the above-mentioned cosmetic, the method comprising mixing the raw material components of the composition for adjusting the skin microecological flora structure; or mixing the composition for adjusting the skin microecological flora structure with other raw material components.

[0012] A fifth aspect of the present invention provides the use of the above-described compositions and / or cosmetics for adjusting the structure of the skin microbiome in any one or more of the following: (a) Improves the microecology of reddened skin and restores the balance of flora; (b) Improves skin redness; (c) Enhance skin barrier function.

[0013] The beneficial technical effects of one or more of the above technical solutions are as follows: The above technical solution addresses the problem of imbalanced skin flora in problematic skin. It studies the skin physiological characteristics and skin flora features of individuals with redness, screens key bacterial genera affecting the composition of the skin flora in these individuals, and uses the characteristic genus *Propionibacterium acnes* to screen commonly used cosmetic raw materials. Based on this, it formulates a product according to the proliferation patterns of characteristic skin bacteria in these raw materials. This formulation is then applied to a formula to verify its safety and the improvement of skin redness after use in individuals, resulting in a product suitable for the skin characteristics of individuals with redness. Experimental verification shows that this composition can significantly improve the diversity of the skin flora in reddened skin, reduce sebum content, transepidermal water loss rate and α value, restore skin barrier function, and improve redness symptoms, thus possessing good practical application value. Attached Figure Description

[0014] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the following description of the embodiments will be briefly introduced. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0015] Figure 1 Changes in skin physiological parameters in healthy individuals and individuals with flushed skin; Figure 2 Changes in gut microbiota diversity in healthy individuals and those with flushed skin; Figure 3 The composition of skin flora in healthy individuals and those with reddened skin; Figure 4 Correlation analysis between skin physiological parameters and skin microbiota; Figure 5 The effect of oil-based raw materials on the growth of characteristic bacteria (Propionibacterium acnes) on erythematous skin; Figure 6 The effect of active ingredient raw materials on the growth of characteristic bacteria (Propionibacterium acnes) in erythematous skin; Figure 7 Effects of different concentrations of horse chestnut extract on keratinocyte viability; Figure 8 The effect of different concentrations of horse chestnut extract on LPS-induced IL-6 expression in keratinocytes; Figure 9 The inhibitory effect of fermentation filtrate obtained by co-culturing and fermenting active raw materials with Staphylococcus epidermidis on Staphylococcus aureus biofilm; Figure 10 Changes in skin microbiota characteristics before and after product use in volunteers; Figure 11 Changes in facial redness before and after volunteers used the product. Detailed Implementation

[0016] It should be noted that the following detailed description is illustrative and intended to provide further explanation of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0017] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the exemplary embodiments according to this application. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0018] As mentioned earlier, in the current technology, no one has analyzed the causes of skin redness and sensitivity from the perspective of microecology, nor has anyone systematically studied the characteristics of the microbial composition and skin physiological parameters of people with redness, nor has anyone studied the correlation between the skin microbial community and physiological parameters of people with redness, nor has anyone selected suitable cosmetic raw materials based on the microbial characteristics of people with redness, in order to maintain the skin microecological balance and improve the skin microecological status of people with redness as indicators to solve the problem of skin redness.

[0019] In view of this, the present invention studies the skin physiological parameters and microbial composition of women with redness through questionnaire surveys, skin physiological parameter measurement, and 16S rRNA sequencing technology. Furthermore, it combines traditional microbial culture methods to select relevant strains based on the microbial characteristics of the redness population for screening commonly used active ingredients in cosmetics. Finally, a set of compositions that can effectively improve skin problems in people with redness and are microecologically friendly were obtained.

[0020] This invention selected women aged 18-35 as the research subjects. The study grouped volunteers based on their skin redness using questionnaires and physiological parameter measurements. Volunteers were divided into two groups according to their self-assessment of redness and TEWL value. Volunteers who self-assessed no redness and had TEWL < 20 were classified as healthy individuals, while those who self-assessed redness and had TEWL ≥ 20 were classified as redness-prone individuals. Further research was conducted using 16S rRNA sequencing to study the distribution of gut microbiota in the two groups and the correlation between physiological parameters and gut microbiota.

[0021] Studies have found that compared to healthy individuals, people with flushed skin have significantly lower skin hydration, significantly increased transepidermal water loss, and significantly higher levels of sebum, heme, pH, and a value. Skin elasticity and luminance (L value) are also significantly reduced, indicating severely impaired barrier function, resulting in oily, flushed, and dull skin. Furthermore, 16S rRNA sequencing of collected microorganisms revealed significantly reduced microbial diversity and richness in the flushed individuals, with a significantly increased abundance of Propionibacterium spp. (…). P <0.05), with an increase rate of 22.3%, and a decrease in Staphylococcus spp.; Spearman correlation studies on skin physiological parameters and skin microbiota in individuals with redness revealed that Propionibacterium spp. showed a significant positive correlation with TEWL, sebum content, heme content, and α-value, and a significant negative correlation with L-value and b-value; Staphylococcus spp. showed a significant positive correlation with glossiness; the heat map showed that sebum content and heme content always showed the same correlation with skin microbiota, indicating that redness is accompanied by oiliness. In summary, the above analysis indicates that Propionibacterium spp. has undergone significant changes in the microbial composition of individuals with redness, and is a key spp. causing dysbiosis in this population. Changes in the abundance of Propionibacterium spp. lead to a decrease in overall species diversity, a decline in skin barrier function, and skin that becomes red, sensitive, and oily.

[0022] This invention, through analysis of changes in skin microbiota characteristics, screened *Propionibacterium acnes*, a characteristic genus of *Propionibacterium* in people with redness, as a characteristic strain for screening commonly used raw materials in skincare products. Addressing the issue of excessive proliferation of pathogenic bacteria such as *Staphylococcus aureus* in people with redness, a suitable raw material formulation was selected and co-cultured with *Staphylococcus epidermidis*, a sentinel bacterium abundant in the skin, to obtain a fermentation filtrate that inhibited the formation of *Staphylococcus aureus* biofilm. This filtrate was then further processed into a simple serum for human trials. This invention addresses the problem of imbalanced skin microbiota by studying the skin physiological characteristics and skin microbiota features of people with redness. Key bacterial genera affecting the microbiota composition of people with redness were screened, and commonly used cosmetic raw materials were screened using the characteristic genus *Propionibacterium acnes*. Based on this, a formulation was developed based on the proliferation patterns of characteristic skin bacteria in the raw materials. This formulation was then applied to a product, and the safety of the formula and the improvement of skin redness after use were verified, resulting in a composition suitable for the skin characteristics of people with redness.

[0023] Specifically, the present invention provides a composition for adjusting the structure of the skin microbiome, the composition comprising the following components in parts by weight: 0.1-2 parts panthenol, 0.1-0.5 parts dipotassium glycyrrhizate, 0.1-2 parts horse chestnut extract, 1-3 parts dioctyl carbonate and 1-3 parts diisostearyl malate.

[0024] Among them, the extract of horse chestnut is preferably prepared by supercritical CO2 extraction. The extract prepared by this method has better inhibitory and anti-inflammatory effects (reducing IL-6 expression) on Propionibacterium acnes than the extract prepared by ultrasonic extraction with ethanol.

[0025] Furthermore, the method for preparing the horse chestnut extract includes: Dry horse chestnut seeds are crushed, sieved, dried, and then subjected to supercritical CO2 extraction to obtain a crude extract. After concentration, filtration, and secondary drying, the final product is obtained.

[0026] The sieve mesh size is 40-60 mesh (preferably 50 mesh), and the specific drying conditions are: drying at 50-70℃ (preferably 60℃) for 1-3 hours (preferably 2 hours). The supercritical CO2 extraction conditions are as follows: the extraction temperature is controlled at 30-50℃ (preferably 40℃), the pressure is controlled at 20-40 MPa (preferably 30 MPa), and the extraction time is controlled at 2-6 hours (preferably 3 hours); furthermore, ethanol is used as an entrainer during extraction to improve the extraction efficiency, and the ethanol is high-concentration ethanol, more preferably 70% or higher ethanol.

[0027] The specific conditions for concentration, filtration, and secondary drying are as follows: concentration for 30-60 minutes at 45-55℃ and a vacuum of 0.08-0.1MPa to remove ethanol and water; filtration of the concentrated extract using a 0.45μm organic phase filter membrane to remove trace solid impurities; and drying of the filtered extract in a vacuum drying oven at 50℃ and under vacuum for 4-6 hours until the moisture content is ≤5% to obtain a powdered horse chestnut extract.

[0028] In another specific embodiment of the present invention, the application of the above-mentioned composition for adjusting the skin microecological flora structure is provided in the preparation of cosmetics. Through experimental verification, the present invention demonstrates that the above-mentioned composition can effectively improve the skin microecology of reddened skin, restore flora balance, alleviate redness, and enhance skin barrier function.

[0029] Therefore, in another specific embodiment of the present invention, a cosmetic is provided, which at least comprises the above-mentioned composition for adjusting the structure of the skin microecological flora.

[0030] The cosmetics mentioned can be skincare products.

[0031] The cosmetics may also contain other raw materials permitted in any cosmetic field, such as antioxidants, emulsifiers, thickeners, dispersants, pH adjusters, preservatives, etc., without specific limitations.

[0032] Furthermore, by rationally adding the above-mentioned raw material components, the present invention can also be used to prepare different cosmetic dosage forms, such as aqueous solutions, emulsions, gels, and creams. In addition, other cosmetic categories that are further derived and prepared based on the above-mentioned basic cosmetic categories are also within the scope of protection of this application.

[0033] In another specific embodiment of the present invention, the cosmetic comprises the following components by mass fraction: 0.1-2% panthenol, 0.1-0.5% dipotassium glycyrrhizate, 0.1-2% horse chestnut extract, 1-3% dioctyl carbonate, 1-3% diisostearyl malate, 0.1-1% ZEN, 1-10% glycerin, balance water.

[0034] In another specific embodiment of the present invention, a method for preparing the above-mentioned cosmetic is provided, the method comprising mixing the raw material components of the composition for adjusting the skin microecological flora structure; or mixing the composition for adjusting the skin microecological flora structure with other raw material components.

[0035] In another specific embodiment of the present invention, the above-described composition and / or cosmetic for adjusting the skin microbiota structure are provided for use in any one or more of the following: (a) Improves the microecology of reddened skin and restores the balance of flora; (b) Improves skin redness; (c) Enhance skin barrier function.

[0036] The present invention will be further described below with reference to embodiments. The present invention will be further illustrated by the embodiments described below, but this does not limit the present invention to the scope of the embodiments described. Based on the embodiments of the present invention, any changes to the present invention made by those skilled in the art without inventive step are within the scope of protection of the present invention. Furthermore, in the embodiments of the present invention, unless otherwise specified, all raw materials used in preparation are commercially available products well known to those skilled in the art. The Propionibacterium acnes CCSM0254 (hereinafter referred to as 254) and Staphylococcus epidermidis CCSM0283 (hereinafter referred to as 283) used in the present invention were isolated from the facial skin of individuals with rosacea and healthy individuals, respectively, by Shandong Freda Biotechnology Co., Ltd., and Staphylococcus aureus was purchased from Beina Biotechnology.

[0037] Culture medium used in the experiment TSA medium (g / L): tryptone 15.0, soybean papain hydrolysate 5.0, sodium chloride 5.0, agar 15.0, autoclaved at 121℃ for 15 min, ready for use.

[0038] TSB medium (g / L): tryptone 17, soybean peptone 3, sodium chloride 5, dipotassium hydrogen phosphate 2.5, glucose 2.5, autoclaved at 121℃ for 15 min, ready for use.

[0039] Example 1: Analysis of skin physiological parameters and skin microbiota characteristics in individuals with flushed skin A questionnaire survey was used to allow volunteers to self-assess their skin redness. Then, their TEWL (transient endothelial wound healing) values ​​were measured. Volunteers who self-assessed no redness and had a TEWL < 20 were classified as healthy skin (H1), while those who self-assessed redness and had a TEWL ≥ 20 were classified as reddened skin (H2). Compared to the healthy group, if... Figure 1 As shown, the skin moisture content of people with reddened skin is significantly reduced. p <0.05), the transdermal water loss rate increased significantly ( p <0.001), sebum content ( p <0.05), heme content ( p <0.001), pH value ( p <0.05) and the value of a ( p <0.05) significantly increased, while skin elasticity and luminance (L value) significantly decreased; skin microbiota diversity indicated ( Figure 2 Compared to healthy individuals, people with reddish skin have reduced gut microbiota diversity; species composition indicates ( Figure 3 Propionibacterium spp. increased ( p <0.01), Staphylococcus spp. decreased, Neisseria spp., Pseudomonas spp., Rhodococcus spp. and Streptococcus spp. ( p <0.05) decreased; through Spearman correlation studies on skin physiological parameters and skin microbiota in individuals with erythema ( Figure 4 The study found that Propionibacterium spp. was significantly positively correlated with TEWL, sebum content, heme content, and a value, and significantly negatively correlated with L value and b value; Staphylococcus spp. was significantly positively correlated with gloss; and Streptococcus spp. was significantly negatively correlated with sebum content and heme content.

[0040] Example 2: Effect of oil-soluble raw materials on the growth of characteristic bacteria Strain activation: P. acnes strain CCSM0254 was streaked onto a TSA plate and placed in a 37°C incubator for anaerobic incubation for 48-72 h for later use.

[0041] Seed culture: The activated Propionibacterium acnes strain CCSM0254 was inoculated into TSB liquid medium and placed in a 37℃ constant temperature incubator for anaerobic culture for 48~72 h for later use.

[0042] Bacterial cell proliferation rate determination (using 50mL centrifuge tubes, each containing 40mL of solution for the experiment). Control group: 40 mL TSB medium; Control group 1: 39.2 mL TSB medium + 0.8 mL seed culture; Control group 2: Different types of oil-soluble raw materials + 0.8 mL seed culture + the remainder of TSB medium; Experimental group: different types of oil-soluble raw materials + the remainder of TSB culture medium.

[0043] The samples from the different groups were placed in a 37°C incubator and anaerobically cultured in test tubes inoculated with Propionibacterium acnes seed culture for 48–72 h. After the culture was completed, the OD of the culture medium was measured. 600 The value was calculated according to Equation 1 to determine the effect of different compositions on the proliferation of the two strains. [(Experimental Group - Control 2) - (Control 1 - Blank Group)] / (Control 1 - Blank Group) --------------- Equation 1 The results are as follows Figure 5 As shown, perilla seed oil, grape seed oil, and avocado oil have a proliferative effect on the growth of strain 254, and the proliferative effect is stronger with increasing concentration. Dioctyl carbonate showed a certain degree of proliferative effect at a concentration of 1%, but it showed an inhibitory effect on strain 254 with increasing concentration, and the inhibitory effect was stronger at higher concentrations. Diisostearyl malate showed an inhibitory effect on the growth of strain 254, and the inhibitory effect was strongest at a concentration of 3%.

[0044] Example 3: Effects of water-soluble active ingredients on the growth of characteristic bacteria According to the method in Example 2, Propionibacterium acnes seed culture CCSM0254 was inoculated into TSB liquid medium at an inoculum volume of 2%, and different concentrations of active ingredients were tested, including 0.01~0.1% ectoine, 0.1~2% panthenol, 0.1~2% horse chestnut extract, 0.1~0.5% dipotassium glycyrrhizate, 0.05~0.2% sodium hyaluronate, and 0.01~0.5% allantoin.

[0045] The preparation method of horse chestnut extract 1 is as follows: Select dried horse chestnut seeds, remove impurities, pulverize, pass through a 50-mesh sieve, and dry in a vacuum drying oven at 60℃ for 2 hours. Use a supercritical CO2 extraction device for extraction. Weigh 150g of pretreated horse chestnut seed powder and inject 70% ethanol at a material-to-liquid ratio of 1:10 (g / mL). Evenly fill the extraction vessel, close the vessel lid to ensure good sealing, start the equipment, and start timing when the temperature inside the extraction vessel reaches 40℃ and the pressure reaches 30MPa and stabilizes. Maintain the extraction state for 3 hours, then open the bottom valve to collect the crude extract. Transfer the crude extract to a rotary evaporator and concentrate at 50℃ and a vacuum of 0.08 MPa for 45 minutes to remove ethanol and water. Filter the concentrated extract through a 0.45μm organic phase filter membrane to remove trace solid impurities. Send the filtered extract to a vacuum drying oven and dry at 50℃ and a vacuum for 3 hours until the moisture content is ≤5%, obtaining a powdered horse chestnut extract.

[0046] The preparation method of horse chestnut extract 2 is as follows: Remove the outer shell of fresh horse chestnut fruit, take the kernel, and dry it in a vacuum drying oven at 50℃ until the moisture content is below 8%. Crush the dried raw material using a pulverizer, pass it through a 50-mesh sieve, and collect the sieve-passing powder. If there are large particles, they need to be re-crushed to ensure uniform particle size for subsequent extraction. Weigh horse chestnut powder and 70% ethanol aqueous solution according to a material-to-liquid ratio of 1:15 (g / mL), add them to a stoppered conical flask, stir to fully wet the powder, place the conical flask in a constant temperature ultrasonic extractor, set the extraction temperature to 45℃, the ultrasonic power to 400W, and the extraction time to 70 minutes. Stir once every 30 minutes during ultrasonic extraction to ensure uniform extraction. After extraction, the extract was filtered while hot using a Buchner funnel to remove most of the solid residue, yielding a preliminary filtrate. This preliminary filtrate was cooled to room temperature and then precisely filtered through a 0.45 μm filter membrane to further remove fine impurities and colloidal substances, preventing precipitation during subsequent concentration and ensuring the purity of the extract. The precisely filtered filtrate was then transferred to an evaporation flask in a rotary evaporator. The water bath temperature was set to 50°C, and the vacuum degree to 0.08 MPa for reduced pressure concentration. During concentration, the liquid state in the evaporation flask was carefully observed to avoid bumping. Concentration was stopped when the volume of the concentrated extract decreased to 1 / 5 of the original filtrate volume, yielding the concentrated horse chestnut extract.

[0047] The results are as follows Figure 6As shown: Different concentrations of active ingredients have different effects on the growth of Propionibacterium acnes. Ectoin slightly promotes the growth of Propionibacterium acnes at a concentration of 0.01%, and inhibits growth at concentrations of 0.05% and 0.1%, but the inhibition rate is low. Panthenol and horse chestnut extract can inhibit the growth of Propionibacterium acnes, with the best inhibitory effects at concentrations of 1% and 2%, and the inhibitory effect of horse chestnut extract 1 is better than that of horse chestnut extract 2. Dipotassium glycyrrhizate also shows an inhibitory effect, with better results at a concentration of 0.5%. Sodium hyaluronate has almost no effect on Propionibacterium acnes. Allantoin only has a certain inhibitory effect at a concentration of 0.5%, and its effect is not significant at low concentrations.

[0048] Example 4: Effects of different horse chestnut extracts on IL-6 expression in keratinocytes Keratinocytes were divided into 1×10 4 Cells were seeded at a density of [number] cells / well in 96-well plates and cultured for 24 hours until adherence. The culture medium was discarded, and 100 mL of different concentrations (0.1 mg / mL to 2.0 mg / mL) of *Aesculus hippocastanum* extract 1 and extract 2 were added to each well, with three replicates per group. After 24 hours of culture, cell viability was assessed using the MTT assay. The concentration with ≥90% cell viability was selected as the experimental concentration. The results are shown below. Figure 7 As shown in the figure, different concentrations of horse chestnut extract 1 did not show toxicity to keratinocytes and had a certain proliferative effect. Horse chestnut extract 2 showed a certain degree of toxicity to keratinocytes at 2 mg / mL, but no toxicity at concentrations below 1 mg / mL. To ensure the consistency of subsequent experiments, a concentration of 1 mg / mL was finally selected for subsequent experiments to inhibit the expression of IL-6 cytokine.

[0049] The experiment included a blank control group: 100 ml of serum-free culture medium; Model control group: 100 ml of serum-free culture medium containing LPS; Positive control group: 50 ml serum-free culture medium containing LPS + 50 ml dexamethasone; Experimental group 1: 50 ml serum-free culture medium containing LPS + 50 ml horse chestnut extract 1 (1 mg / mL); Experimental group 2: 50 ml serum-free culture medium containing LPS + 50 ml horse chestnut extract 2 (1 mg / mL).

[0050] The IL-6 cytokine levels in each group were measured using the ELISA kit according to the instructions. The results are as follows: Figure 8 As shown, 1 mg / mL of horse chestnut extract 1 significantly reduced the expression of the inflammatory factor IL-6 compared to horse chestnut extract 2 (52.1% vs 17.7%), indicating that horse chestnut extract 1 has better anti-inflammatory function than horse chestnut extract 2.

[0051] Example 5: Effect of fermentation filtrate obtained from co-culturing and fermenting active raw materials with Staphylococcus epidermidis on Staphylococcus aureus biofilm. Preparation of fermentation filtrate: Staphylococcus epidermidis strain CCSM0283 was streaked onto TSA plates and incubated at 37℃ for 14-16 h. After two generations of activation, single colonies were picked and cultured with TSB liquid containing 0.01-0.1% ectoine, 0.1-2% panthenol, 0.1-2% horse chestnut extract, 0.1-0.5% dipotassium glycyrrhizate, 0.05-0.2% sodium hyaluronate, and 0.01-0.1% allantoin on a shaker for 14-16 h. After the culture was completed, the filtrate obtained by centrifugation at 10000 r / min for 10 min was the fermentation filtrate after co-culturing Staphylococcus epidermidis with different active ingredient compositions. The fermentation filtrate obtained without the addition of any active ingredient composition was used as a comparison with the active ingredient fermentation filtrate.

[0052] Staphylococcus aureus activation: Staphylococcus aureus powder purchased from Beina Biotechnology was dissolved in sterile water. A disposable sterile inoculation loop was used to pick up the bacterial suspension and streak it onto a TSA solid plate. Activation was performed twice. Single colonies from the plate were picked and aerobically cultured in TSB liquid medium for 18-24 hours to obtain a bacterial suspension. The concentration of the bacterial suspension was adjusted to OD0.05. 600 =0.5~0.8 is seed solution, for later use.

[0053] Inhibition of Staphylococcus aureus growth (experiment conducted using 96-well plates) Experimental group: 156 mL TSB medium + 40 mL fermentation filtrate + 4 mL Staphylococcus aureus seed culture; Control group: 156 mL TSB medium + 40 mL sterile water + 4 mL Staphylococcus aureus seed culture.

[0054] After incubating the 96-well plate in an aerobic environment for 24 h, the OD of each well was measured. 600 The growth inhibition rate of the fermentation filtrate against Staphylococcus aureus was calculated according to Equation 2: (Control group - Experimental group) / Control group * 100% ------------ Equation 2 The results are as follows Figure 9 As shown, the fermentation supernatant of *Staphylococcus epidermidis* CCSM0283 obtained without the addition of active ingredients has a certain inhibitory effect on the biofilm of *Staphylococcus aureus*. Among the fermentation supernatants obtained by co-culturing with strain 283 with the addition of active ingredients, 1% and 2% panthenol and horse chestnut extract, as well as 0.25% and 0.5% dipotassium glycyrrhizate and 0.5% allantoin showed the best inhibitory effects, and horse chestnut extract 1 had a better inhibitory effect than horse chestnut extract 2.

[0055] Examples 6-8: A composition formulation that can effectively improve skin redness. Based on the effects of the aforementioned oil-soluble and water-soluble raw materials on the growth of characteristic bacteria (Propionibacterium acnes) and the screening results regarding the inhibition of Staphylococcus epidermidis biofilm by the Staphylococcus epidermidis fermentation filtrate obtained through fermentation with water-soluble active ingredients, a composition was formulated. The composition is as follows: Table 1. Combinations for improving redness

[0056] The skincare serum was formulated according to the ingredient combinations in Table 1. To ensure the scientific rigor of the experiment, in addition to the ingredients mentioned above, ZEN (Cepic, Inc.: Polyacrylate Crosspolymer-6) was used as a suspending and thickening agent throughout the serum. The serum preparation method is as follows: (1) Phase A: Disperse the suspension thickener ZEN in glycerol, heat water to disperse it, and then place it in an 85℃ water bath to keep it warm until dissolved. Turn on the homogenizer at 4500r / min to accelerate dispersion. (2) Phase B: Mix the oil-soluble raw materials (dioctyl carbonate and diisostearyl malate) evenly according to the proportions in Table 1 above, and keep warm at 85°C; (3) Phase mixing: Place phase A under a homogenizer, then slowly add phase B into the phase A container, turn on the homogenizer at 4500 r / min and homogenize for 4 min to obtain emulsion-type oil suspension essence; (4) Subsequent phase: After dissolving the water-soluble active ingredients (panthenol, dipotassium glycyrrhizate, horse chestnut extract, allantoin), add them to the essence according to the proportions in Table 1, mix well, and name the essences containing the compositions of Examples 5-7 and Comparative Examples 1-5 in Table 1 as H1-H8 respectively.

[0057] Example 8: Recruiting volunteers to conduct pre- and post-use evaluations of the serum product. 180 female volunteers aged 18-35 with reddened skin were recruited to test a skincare serum. The volunteers were randomly divided into 8 groups of 20 each. Each group used this serum instead of their usual serum, twice daily, morning and evening. Microbial samples were collected from the volunteers' faces using sterile cotton swabs according to the method described in Example 1. Changes in the facial flora of the volunteers after using the 8 different serums were compared. Figure 10 ); VISIA was used to take facial photos of volunteers after 14 consecutive days of use to assess the improvement in facial redness. Figure 11The changes in facial sebum content of volunteers were measured using a Sebumeter SM 815 sebum analyzer, the transdermal water loss rate of volunteers' faces was monitored using a Tewamater TM 300 DNA transdermal water loss analyzer, and the changes in facial a-value of volunteers were measured using a Colorimeter CL 400 skin Lab value analyzer (Table 2).

[0058] The results showed that all eight serums could improve skin microbiome diversity to some extent, as well as inhibit sebum secretion, reduce transepidermal water loss, lower a value, improve skin redness, and enhance the skin barrier. Serum 2 showed the best improvement effect, indicating that the formulation in Example 6 can effectively improve the microecology of reddened skin, restore microbiome balance, improve redness, and enhance skin barrier function.

[0059] Table 2: Changes in skin physiological parameters before and after using 8 different serums

[0060] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A composition for adjusting the structure of the skin's microecological flora, characterized in that, The composition for adjusting the skin's microecological flora structure comprises the following components in parts by weight: 0.1-2 parts panthenol, 0.1-0.5 parts dipotassium glycyrrhizate, 0.1-2 parts horse chestnut extract, 1-3 parts dioctyl carbonate and 1-3 parts diisostearyl malate; Among them, the horse chestnut extract is an extract prepared by supercritical CO2 extraction.

2. The composition according to claim 1, characterized in that, The preparation method of the horse chestnut extract includes: Dry horse chestnut seeds are crushed, sieved, dried, and then subjected to supercritical CO2 extraction to obtain a crude extract. After concentration, filtration, and secondary drying, the final product is obtained.

3. The composition according to claim 2, characterized in that, The sieve mesh size is 40-60 mesh, and the specific drying conditions are: drying at 50-70℃ for 1-3 hours.

4. The composition according to claim 2, characterized in that, The supercritical CO2 extraction conditions are as follows: extraction temperature controlled at 30-50℃, pressure controlled at 20-40 MPa, and extraction time controlled at 2-6 hours. Furthermore, ethanol is used as an entrainer during extraction, and the ethanol is high-concentration ethanol, more specifically 70% or higher ethanol.

5. The composition according to claim 2, characterized in that, The specific conditions for concentration, filtration, and secondary drying are as follows: concentrate at 45-55℃ and vacuum degree 0.08-0.1MPa for 30-60 minutes, filter the concentrated extract with a 0.45μm organic phase filter membrane, and dry the filtered extract at 50℃ and vacuum for 4-6 hours until the moisture content is ≤5%.

6. The use of the composition according to any one of claims 1-5 for adjusting the structure of the skin microbiome in the preparation of cosmetics.

7. A cosmetic product, characterized in that, The cosmetic product comprises at least the composition according to any one of claims 1-5 that modulates the structure of the skin's microecological flora; Furthermore, the cosmetics in question are skincare products.

8. The cosmetic product as described in claim 7, characterized in that, The cosmetics also contain other ingredients permitted to be added in any cosmetic field, further including antioxidants, emulsifiers, thickeners, dispersants, pH adjusters, and preservatives.

9. A method for preparing the cosmetic according to claim 7 or 8, wherein the method comprises mixing the raw material components of the composition for adjusting the skin microecological flora structure; or mixing the composition for adjusting the skin microecological flora structure with other raw material components.

10. The use of the composition for adjusting the skin microbiome structure according to any one of claims 1-5 and / or the cosmetic according to any one of claims 7-8 in any one or more of the following: (a) Improves the microecology of reddened skin and restores the balance of flora; (b) Improves skin redness; (c) Enhance skin barrier function.

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