Staphylococcus epidermidis CCSM009 for repairing skin barrier and metagen of staphylococcus epidermidis CCSM009

Abstract

The invention discloses staphylococcus epidermidis CCSM009 for repairing a skin barrier and a metagen of the staphylococcus epidermidis CCSM009, and belongs to the field of microbial technologies and medicines. The staphylococcus epidermidis CCSM009 and the metagen thereof provided by the invention have a good barrier function repairing effect and application prospect in the aspect of external use, and are specifically shown as follows: the activity of damaged HaCaT cells is improved in vitro, and the expression of gene levels and protein levels in the aspects of functional proteins and structural proteins after damage is improved; meanwhile, sphingomyelinase 1 and tissue kininolase 7 (KLK7) are improved, and it is clear that the bacterium performs up-regulation of skin barrier related proteins and enzymes through an AHR-OVOL1 pathway. Therefore, the metagen of the staphylococcus epidermidis CCSM009 has a huge application prospect in preparation of cosmetics or medicines for repairing the skin barrier.

Description

Technical Field

[0001] This invention relates to a Staphylococcus epidermidis strain CCSM009 that repairs the skin barrier and its post-genes, belonging to the fields of microbial technology and pharmaceutical technology. Background Technology

[0002] Maintaining a healthy skin barrier is crucial for overall health. The skin barrier is the body's first line of defense against the external environment. It not only prevents the invasion of harmful substances, pathogens, and allergens but also maintains the body's water balance, preventing excessive water loss. A healthy skin barrier effectively protects the body from external stimuli, reducing the risk of infection and inflammation. Conversely, when the skin barrier is damaged, the skin becomes dry and sensitive, making it more susceptible to bacterial and other harmful substances, leading to various skin problems such as eczema, dermatitis, and allergic reactions. Furthermore, the integrity of the skin barrier is closely related to the health of the overall immune system. Therefore, maintaining a healthy skin barrier not only contributes to the appearance and comfort of the skin but is also a vital factor in maintaining overall health.

[0003] The skin's surface microbiome consists of bacteria, fungi, and viruses, but existing known symbiotic microorganisms still have significant functional limitations. For example, many common skin strains (such as Staphylococcus epidermidis and Propionibacterium) possess certain symbiotic and protective functions, but their anti-inflammatory, barrier repair-promoting, or metabolic activities are weak, and they are unstable in different individuals and under different skin conditions. Furthermore, some microorganisms are easily affected by changes in the environment, pH, and sebum composition, making it difficult to maintain stable function. Therefore, existing strains cannot fully meet the needs for efficient skin barrier repair, precise immune regulation, or the provision of controllable active metabolites. Thus, it is necessary to screen for novel functional strains from the skin microecology with stronger efficacy, more stable metabolic characteristics, and higher safety to overcome the limitations of traditional microorganisms in skin barrier repair. Summary of the Invention

[0004] This invention provides a Staphylococcus epidermidis (Staphylococcus epidermidis) Staphylococcus epidermidis The application of CCSM009 and its post-genes in the preparation of products that repair the skin barrier.

[0005] This invention provides a strain of Staphylococcus epidermidis (Staphylococcus epidermidis) Staphylococcus epidermidis Accession number CCSM009 was deposited at the Guangdong Provincial Center for Microbial Culture Collection on July 22, 2024, with accession number GDMCC NO: 648903.

[0006] In one embodiment, the Staphylococcus epidermidis CCSM009 is derived from the skin of a healthy person; the colonies of the Staphylococcus epidermidis CCSM009 on TSB solid medium are raised, pale yellow, smooth, and round.

[0007] The present invention also provides a metabiotic prepared using the aforementioned Staphylococcus epidermidis CCSM009.

[0008] In one embodiment, the metabiotic includes cell lysate, inactivated or dead cells, fermentation supernatant, or any of the above-mentioned powders prepared by drying.

[0009] In one embodiment, the metagener contains oleamide, D-erythrosine, (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid, and (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid.

[0010] In one embodiment, the inactivated or deadened cells are prepared by culturing the Staphylococcus epidermidis CCSM009 in a culture medium for a period of time, collecting the bacterial cells in the cell culture medium, and obtaining inactivated bacterial cells after heat treatment. In one embodiment, the heat treatment conditions are: 62~65℃, 30min.

[0011] In one embodiment, the method for preparing the bacterial lysate is as follows: Staphylococcus epidermidis CCSM009 is cultured in a culture medium for a period of time, bacterial cells are collected, homogenized under high pressure, and the supernatant obtained by centrifugation is the bacterial lysate.

[0012] In one embodiment, the fermentation supernatant is the supernatant obtained by centrifuging Staphylococcus epidermidis CCSM009 after culturing it in a culture medium for a period of time.

[0013] The present invention also provides a composition containing the aforementioned Staphylococcus epidermidis CCSM009 and / or its postgenes.

[0014] In one embodiment, the composition includes, but is not limited to, pharmaceuticals or daily chemical products.

[0015] In one embodiment, the pharmaceutical product includes the composition and conventional excipients.

[0016] In one embodiment, the conventional excipients include one or more of fillers, flavoring agents, binders, disintegrants, lubricants, antacids, and nutritional fortifiers.

[0017] In one embodiment, the composition is a probiotic powder.

[0018] In one embodiment, the bacterial powder is a solid powder prepared by drying the prepared liquid postbiotic Staphylococcus epidermidis CCSM009.

[0019] In one embodiment, the drying includes, but is not limited to, spray drying, vacuum freeze drying, fluidized bed drying, or vacuum drying.

[0020] The present invention also provides the use of the composition in the preparation of pharmaceuticals or cosmetics for repairing the skin barrier.

[0021] In one implementation, the application includes at least one of the following functions: (1) Enhance the cell activity of skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro; (2) In vitro, the expression of barrier function protein genes in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage was increased; (3) In vitro, the expression of barrier structure protein genes in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage was increased; (4) Increase the expression of barrier function proteins in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro; (5) Reduced the expression of structural proteins in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro; (6) Increase the expression of sphingomyelinase 1 (SMPD1) in keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro; (7) Reduced tissue kallikrein 7 (KLK7) expression in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro.

[0022] In one embodiment, the drug or cosmetic is applied topically.

[0023] In one embodiment, the amount of Staphylococcus epidermidis CCSM009 and the prepared postbiotic in the drug or cosmetic is not less than 1 × 10⁻⁶ of the corresponding viable bacterial count. 7 CFU / mL.

[0024] In one embodiment, the drug comprises the Staphylococcus epidermidis CCSM009, a drug carrier, and / or pharmaceutical excipients.

[0025] In one embodiment, the pharmaceutical excipient comprises excipients and additives.

[0026] In one embodiment, the pharmaceutical excipients include solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, integrators, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants and anti-flocculation agents, filter aids, and release inhibitors.

[0027] In one embodiment, the cosmetic contains Staphylococcus epidermidis CCSM009 postbiotic, matrix ingredients, and / or conventional excipients.

[0028] In one embodiment, the matrix raw materials include oil-based raw materials, wax-based raw materials, synthetic oil-based raw materials, powder-based raw materials, gel-based raw materials, coagulants, and surfactants.

[0029] In one embodiment, the conventional excipients include one or more of the following: moisturizers, whitening agents, flavoring agents, adhesives, lubricants, preservatives, film-forming agents, antioxidants, emulsifiers, and cosmetic nutritional additives.

[0030] The present invention also provides the use of the aforementioned Staphylococcus epidermidis CCSM009 in the preparation of products containing oleic acid amide, D-erythrosine, (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid or (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid.

[0031] Beneficial effects: Staphylococcus epidermidis of the present invention ( Staphylococcus epidermidis The post-biotic prepared with CCSM009 has the ability to alleviate skin barrier damage and increase the expression of related proteins when applied topically, specifically in the following ways: (1) Enhance the cell activity of skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage; (2) Increase the mRNA expression of barrier function proteins in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro; (3) Increase the mRNA expression of barrier structural proteins in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro; (4) Increase the expression of barrier function proteins in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro; (5) Reduces the expression of barrier structural proteins in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro; (6) Increase the expression of SMPD1 in keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) injury in vitro; (7) Reduced KLK7 expression in skin keratinocytes (HaCaT) after sodium dodecyl sulfate (SDS) damage in vitro; (8) Regulate the AHR-OVOL1 pathway receptor to regulate the expression of related proteins and thus restore skin barrier function.

[0032] Therefore, Staphylococcus epidermidis ( Staphylococcus epidermidis The post-biotic prepared by CCSM009 has great application potential in products that alleviate damage to the host skin barrier.

[0033] Preservation of biological materials Staphylococcus epidermidis ( Staphylococcus epidermidis CCSM009, taxonomically named Staphylococcus epidermidis It was deposited on July 22, 2024, at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC No: 64903, located at Building 59, No. 100 Xianlie Middle Road, Guangzhou. Attached Figure Description

[0034] Figure 1 To evaluate the effects of different concentrations of SDS on HaCaT cell damage.

[0035] Figure 2 The effect of post-genetics on HaCaT cell proliferation.

[0036] Figure 3 The effect of post-genetic repair on HaCaT cell viability after SDS damage.

[0037] Figure 4 Gene expression of HaCaT cell barrier function proteins that promote post-serogenetic repair of SDS-damaged cells.

[0038] Figure 5 Gene expression of HaCaT cell barrier structural proteins that promote the repair of SDS damage by metagenes.

[0039] Figure 6 The expression of HaCaT cell barrier function proteins for post-genetic repair of SDS-damaged cells.

[0040] Figure 7 The expression of barrier structural proteins in HaCaT cells after SDS damage is improved by metagenesis.

[0041] Figure 8 To promote the expression of SMPD1 in HaCaT cells after SDS damage by metagenesis.

[0042] Figure 9To promote the repair of KLK7 expression in HaCaT cells after SDS damage by metagenes.

[0043] Figure 10 The metagener used AHR-OVOL1 to repair HaCaT cells damaged by SDS.

[0044] Figure 11 This is a liquid chromatography-mass spectrometry (LC-MS) diagram of oleamide standard.

[0045] Figure 12 The liquid chromatography-mass spectrometry (LC-MS) plot shows the oleic acid amide content in the fermentation supernatant sample.

[0046] Figure 13 This is a liquid chromatography-mass spectrometry (LC-MS) graph showing the oleic acid amide content in the bacterial cell sample.

[0047] Figure 14 A comparison chart showing the retention times of L-glycine-valine standard, fermentation supernatant, and cell samples.

[0048] Figure 15 The image shows the liquid chromatography-mass spectrometry (LC-MS) of D-erythrosine disphingosine standard.

[0049] Figure 16 Liquid chromatography-mass spectrometry (LC-MS) plot of D-erythrosine content in fermentation supernatant sample.

[0050] Figure 17 This is a liquid chromatography-mass spectrometry (LC-MS) plot showing the D-erythrosine content in the bacterial cell sample.

[0051] Figure 18 A comparison chart showing the retention times of D-erythrosine standard, fermentation supernatant, and cell samples.

[0052] Figure 19 The LC-MS / MS chromatogram of (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid standard is shown.

[0053] Figure 20 Liquid chromatography-mass spectrometry (LC-MS) plot of (2-oxo-2,3-dihydro-1H-indole-3-yl)-acetic acid content in fermentation supernatant sample.

[0054] Figure 21 A comparison chart of retention times for (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid standards, fermentation supernatants, and cell samples.

[0055] Figure 22 This is a liquid chromatography-mass spectrometry (LC-MS) image of the L-glycine-valine dipeptide standard.

[0056] Figure 23 Liquid chromatography-mass spectrometry (LC-MS) plot of L-glycine-valve dipeptide content in fermentation supernatant sample.

[0057] Figure 24A comparison chart showing the retention times of L-glycine-valine standard, fermentation supernatant, and cell samples. Detailed Implementation

[0058] The present invention will be further described below with reference to specific embodiments.

[0059] The human keratinocytes (HaCaT) involved in the following examples were purchased from the Shanghai Cell Bank.

[0060] The culture media involved in the following examples are as follows: TSB liquid medium: tryptone: 17.0 g / L; soybean peptone: 3.0 g / L; sodium chloride: 5.0 g / L; potassium dihydrogen phosphate: 2.5 g / L and glucose: 2.5 g / L.

[0061] TSB solid medium: tryptone: 17.0 g / L; soybean peptone: 3.0 g / L; sodium chloride: 5.0 g / L; potassium dihydrogen phosphate: 2.5 g / L; glucose: 2.5 g / L and agar powder 20 g / L.

[0062] Modified BHI screening solid culture medium: bovine brain extract: 4.0 g / L, bovine heart extract: 4.0 g / L, peptone: 5.0 g / L, casein peptone: 16.0 g / L, sodium chloride: 5.0 g / L, glucose: 2.0 g / L, disodium hydrogen phosphate: 2.5 g / L, nicotinic acid: 0.3 g / L, neomycin: 0.4 g / L, and agar: 20 g / L.

[0063] Cell culture medium: 89% (v / v) DMEM medium + 10% (v / v) fetal bovine serum + 1% (v / v) 100× penicillin and streptomycin mixed solution (the mixed solution contains 10,000 U / mL penicillin and 10 mg / mL streptomycin).

[0064] Example 1: Cell resuscitation and culture First, remove the frozen human keratinocyte cell line (HaCaT) from the -80℃ freezer, thaw it rapidly in a 37℃ water bath, then add 5 mL of cell culture medium and centrifuge at 1000 r / min for 3 min, discarding the supernatant. Resuspend the cells in 1 mL of cell culture medium and place them in a culture dish, then incubate in a 37 °C incubator containing 5% CO2. When the cells regain viability and reach 70%-80% confluence after 1-2 days, passage the cells.

[0065] Example 2: Isolation and screening of Staphylococcus epidermidis The bacterial strains were selected from healthy human skin. Sterile swabs were applied to the back of the hand three times with physiological saline, and then streaked directly onto a modified BHI solid medium. The culture was incubated at 37°C for 48 hours. Three pale yellow, round colonies were selected from the resulting colonies and enriched in BHI liquid medium for 16 hours. The bacterial culture was then transferred to BHI solid plates for purification and incubation for 48 hours. Finally, single colonies were transferred to BHI liquid medium for enrichment. The strains were preserved in 30% glycerol. The genome of the strains was extracted, and 16S rDNA was amplified and sequenced (performed by Suzhou Genewiz Biotechnology Co., Ltd.). The results, confirmed by NCBI sequence alignment, identified them as Staphylococcus epidermidis, and they were named Staphylococcus epidermidis CCSM009, Staphylococcus epidermidis D16, and Staphylococcus epidermidis C13, respectively.

[0066] Example 3: Preparation of Staphylococcus epidermidis metabiotic Staphylococcus epidermidis CCSM009 was streaked from the preservation tube and revived. It was then cultured on TSB solid medium in a 37°C water-jacketed incubator for 48 h to obtain single colonies. Single colonies were picked and inoculated into TSB liquid medium and cultured at 37°C for 12-18 h to obtain culture solution 1. Culture solution 1 was inoculated into TSB liquid medium at an inoculation rate of 2% (v / v) and cultured at 37°C for 12 h to obtain seed culture; The seed culture was inoculated into TSB liquid medium at 2% (v / v) for expansion culture, and cultured at 37℃ for 18h. The number of viable bacteria was recorded and bacterial culture a was obtained.

[0067] The bacterial suspension a was centrifuged at 8000 r / min for 30 min to collect the bacterial sludge. The bacterial sludge was resuspended in double-distilled water at 75% of the original bacterial suspension volume. The resuspended suspension was heat-treated (65 ℃, 30 min) and then homogenized under high pressure (1000 MPa, 10 times) in a high-pressure homogenizer. After homogenization, the supernatant was collected by centrifugation at 8000 r / min for 30 min to obtain the bacterial cell lysate, which was designated CCSM009-S.

[0068] The same method was used to prepare the cell lysate of Staphylococcus epidermidis D16, denoted as D16-S, and the cell lysate of Staphylococcus epidermidis C13, denoted as C13-S.

[0069] Optionally, the supernatant or lysis buffer prepared above is freeze-dried at -10℃ to -50℃ to obtain Staphylococcus epidermidis postbiotic freeze-dried powder.

[0070] Example 4: Establishment of an in vitro model of keratinocyte damage caused by sodium dodecyl sulfate (SDS) (HaCaT) (1) Take 100 μL of HaCaT cells in the logarithmic growth phase and use 1.2 × 10⁻⁶ ppm.4 Cells were seeded at a concentration of 100 cells / well in 96-well plates, with the outermost ring filled with PBS solution to prevent edge effects. After culturing for 36 h to allow the cells to adhere, blank control, control and SDS treatment groups were set up. Control group: Contains only cell culture medium and does not contain HaCaT cells; Control group: Contains cell culture medium and HaCaT cells, but does not contain SDS; Treatment groups: containing cell culture medium, HaCaT cells, and SDS at final concentrations of 6.25 μg / mL, 12.5 μg / mL, 25 μg / mL, and 50 μg / mL, respectively.

[0071] (2) The above well plates were incubated in an incubator at 37 °C for 6 h, 12 h and 24 h respectively. After incubation, 10 μL of CCK8 solution was added to each well and incubated for 1.5 h to measure the absorbance (OD) at 450 nm.

[0072] Cell viability is calculated using the following formula: Cell viability (%) = (OD value of treatment group - OD value of blank group) / (OD value of control group - OD value of blank group) × 100%.

[0073] (3) The effect on cell activity after nonlinear regression fitting is as follows: Figure 1 As shown, compared with the control group (cell proliferation rate 100%), the cell viability of the model group treated with 15 μg / mL SDS for 6 h was 60.6%, and SDS modeling caused significant damage to HaCaT cells.

[0074] Example 5: Safety verification of postbiotics prepared from Staphylococcus epidermidis in HaCaT cells (1) Take 100 μL of HaCaT cells in the logarithmic growth phase and use 1.2 × 10⁻⁶ ppm. 4 Cells per well were seeded at a concentration of 100 cells / well in 96-well plates, with the outermost ring filled with PBS solution to prevent edge effects. After culturing for 36 h to allow them to adhere, blank control group, control group, and post-genetic treatment group were set up. Control group: Contains only cell culture medium and does not contain HaCaT cells; Control group: Contains cell culture medium and HaCaT cells, but does not contain post-genetics; The post-genetic treatment group contained cell culture medium and HaCaT cells, as well as post-genetic agents.

[0075] The metabiotic was resuspended in cell culture medium until the concentration reached 1.0 × 10⁻⁶. 7The amount of postbiotic prepared from bacterial culture with CFU / ml is equivalent. Add 100 μL of postbiotic CCSM009-S prepared from Staphylococcus epidermidis CCSM009, or postbiotic D16-S or C13-S prepared from strains D16 and C13-S.

[0076] (2) The above well plates were incubated in an incubator at 37 °C for 24 h. After incubation, 10 μL of CCK8 solution was added to each well and incubated for 1.5 h. The absorbance (OD) at 450 nm was measured.

[0077] Cell viability was calculated using the following formula: Cell viability (%) = (OD value of treated group - OD value of blank group) / (OD value of control group - OD value of blank group) × 100% Effects on cell viability, such as Figure 2 As shown, compared with the control group (cell proliferation rate 99.14%), the addition of post-genetic agents CCSM009-S, D16-S, and C13-S composed of Staphylococcus epidermidis CCSM009, D16, or C13 at an inactivated bacterial concentration of 1×10⁻⁶ showed a significant increase in cell proliferation. 7 The cell proliferation rates at CFU / ml were 100.89%, 98.99%, and 101.45%, respectively.

[0078] According to the ISO 10993-5:2009 toxicity classification evaluation method, cells with a viability greater than 70% can be considered non-toxic. The above results indicate that an inactivated bacterial concentration of 1.0 × 10⁻⁶ is appropriate. 7 HaCaT cells at a postbiotic concentration of CFU / ml showed high viability, exceeding 95%. Considering their lack of cytotoxicity, an inactivated cell concentration of 1.0 × 10⁻⁶ was selected. 7 CFU / ml is a suitable postbiotic concentration for subsequent cell experiments.

[0079] Example 6: Effect of post-biotics prepared from Staphylococcus epidermidis on SDS-induced damage to HaCaT cells (1) Take 100 μL of HaCaT cells in the logarithmic growth phase and use 1.2 × 10⁻⁶ ppm. 4 A concentration of 10 cells / well was seeded into a 96-well plate, with the outermost ring filled with PBS solution to prevent edge effects, and cultured for 36 h to allow the cells to adhere. (2) The well plates prepared in step (1) were incubated in an incubator at 37 °C for 6 h, and control group, model group and treatment group were set up respectively; Control group: After changing the medium of cells in step (1), the cells contained cell culture medium and HaCaT cells, without post-genetic treatment, and without SDS modeling agent; Model group: The cells from step (1) were replaced with a cell culture medium containing 15 μg / mL SDS, containing the original HaCaT cells, without post-genetic treatment; the cell culture medium containing SDS modeling agent was prepared by mixing SDS evenly in the culture medium and passing it through a 0.22 μm aqueous filter membrane for sterilization, so that the final concentration of SDS in the cell culture medium was 15 μg / mL. Postgenetic treatment group: The treatment method is the same as that of the model group.

[0080] (3) After incubation, discard the original culture medium and wash once with PBS. Add the corresponding metagenic sample to the metagenic treatment group, and add cell culture medium to the control group and model group. Incubate again for 24 hours.

[0081] In the metabiotic treatment group, the metabiotics were resuspended in cell culture medium, and the amount of resuspended metabiotics was adjusted to a fermentation concentration of 1.0 × 10⁻⁶. 7 The amount of metabiotics prepared from bacterial cultures of CFU / ml was equivalent, and 100 μL of metabiotics CCSM009-S, D16-S, or C13-S prepared from Staphylococcus epidermidis CCSM009, D16, and C13 were added respectively.

[0082] (4) Add 10 μL of CCK8 solution to each well and incubate for 1.5 h to measure the absorbance (OD) at 450 nm.

[0083] Cell viability was calculated using the following formulas: Model group cell viability (%) = (Model group OD value - Blank group OD value) / (Control group OD value - Blank group OD value) × 100%; Treatment group cell viability (%) = (Treatment group OD value - Blank group OD value) / (Control group OD value - Blank group OD value) × 100%.

[0084] The results of the effect of post-genetic agents on SDS-induced HaCaT cell damage are as follows: Figure 3 Compared with the control group (cell viability 102.28%), the cell viability of the model group was 66.53%, indicating that SDS modeling caused significant damage to HaCaT cells.

[0085] The cell viability of the treatment groups after adding CCSM009-S, D16-S, and C13-S were 85.8%, 71.61%, and 92.42%, respectively. Compared with the model group (66.53%), CCSM009-S can improve the viability of HaCaT cells, indicating that the metabiotics (cell lysate fractions) of Staphylococcus epidermidis CCSM009 and C13-S can effectively alleviate the damage to HaCaT cells caused by SDS. Therefore, subsequent investigations will only explore the efficacy of Staphylococcus epidermidis CCSM009 and C13-S.

[0086] Example 7: Effect of postbiotic preparation of Staphylococcus epidermidis CCSM009 on the expression of barrier function protein genes in SDS-damaged HaCaT cells.

[0087] (1) HaCaT cells were injected at a concentration of 1.8 × 10⁻⁶. 4 10 cells / well were seeded into a 6-well plate and cultured for 36 hours until the cells adhered. (2) The well plates prepared in step (1) were incubated in an incubator at 37 °C for 6 h, and control group, model group and treatment group were set up respectively: Control group: After changing the medium of cells in step (1), the cells contained cell culture medium and HaCaT cells, without post-genetic treatment and without SDS modeling agent; Model group: The cells from step (1) were replaced with a cell culture medium containing 15 μg / mL SDS, containing the original HaCaT cells, without post-genetic treatment; the cell culture medium containing SDS modeling agent was prepared by mixing SDS evenly in the culture medium and passing it through a 0.22 μm aqueous filter membrane for sterilization, so that the final concentration of SDS in the cell culture medium was 15 μg / mL. Postgenetic treatment group: The treatment method is the same as that of the model group.

[0088] (3) After incubation, discard the original culture medium and wash once with PBS. Add the corresponding metagenic sample to the metagenic treatment group, and add cell culture medium to the control group and model group. Incubate again for 24 hours.

[0089] In the epigenetic treatment group, the epigenetic elements were resuspended in cell culture medium (the amount of resuspended epigenetic elements was the same as that fermented to a concentration of 1×10⁻⁶). 7 (The amount of postbiotic prepared from bacterial culture of CFU / ml is equivalent), and 2 mL of postbiotic CCSM009-S and C16-S prepared from Staphylococcus epidermidis CCSM009 were added respectively.

[0090] (4) After incubation, discard the culture supernatant, wash each well three times rapidly with PBS, add 1 mL of cell lysis buffer to each well, repeatedly pipette, extract RNA from the cell lysis buffer, and reverse transcribe it into cDNA using an RT-PCR reverse transcription kit. Detect gene expression in HaCaT cells using real-time quantitative PCR. -△△Ct Formula calculation FLG, IVL and AQP3 The mRNA expression levels were measured, with GAPDH as the internal reference. The primers are described in Table 1 below, and the results are as follows: Figure 4 As shown.

[0091] Table 1 Primer Sequences

[0092] Filamentin (FLG) plays a crucial role in the skin barrier function. It is an important molecule connecting keratin fibers in the stratum corneum, forming a robust physical barrier on the outermost layer of the epidermis by helping keratin fibers aggregate in an orderly manner, preventing moisture loss and blocking the invasion of external allergens. FLG deficiency is closely associated with various skin diseases, such as eczema (atopic dermatitis) and ichthyosis. In these diseases, FLG deficiency or absence weakens the skin barrier function, making the skin more sensitive to external stimuli. Therefore, reducing SDS-induced damage to keratinocytes by increasing FLG gene expression has become a therapeutic target. FLG expression results are obtained from… Figure 4 It was found that the expression level of FLG mRNA in the control group was 1, while the expression level in the model group decreased to 0.22 after SDS intervention. The postbiotic (cell lysate) prepared from Staphylococcus epidermidis CCSM009 significantly upregulated the expression level of FLG mRNA to 4.8, while Staphylococcus epidermidis C13 could not increase the expression level of FLG mRNA, which was 0.45.

[0093] In keratinocytes, inner lamina protein (IVL) cross-links with laminarin via transglutaminase catalysis, forming an insoluble keratinized capsule that constitutes the unique stratum corneum barrier structure of the epidermis. Simultaneously, it interacts with other keratinocyte differentiation proteins such as filaggrin (FLG) to jointly maintain the structure and function of the skin barrier. IVL expression results are derived from… Figure 4 It can be seen that the expression level of IVL mRNA in the control group was about 1, while the expression level in the model group decreased to 0.17 after SDS intervention. The lysate of Staphylococcus epidermidis CCSM009 could increase the expression level of IVL to 3.36, while Staphylococcus epidermidis C13 could not increase the expression level of FLG mRNA, which was 0.89.

[0094] Aquaporins (AQPs) play a crucial role in skin barrier function, especially AQP3. AQPs regulate the transport and distribution of water in the skin, maintaining skin hydration and elasticity by controlling the water content within the stratum corneum. AQP3 not only promotes water transport but also regulates glycerol transport, playing a vital role in maintaining skin suppleness and barrier function. The expression of AQP3 is determined by… Figure 6 It can be seen that the expression level of AQP3 mRNA in the control group was about 1, while the expression level in the model group decreased to 0.54 after SDS intervention. After repair by Staphylococcus epidermidis CCSM009 bacterial lysate, the expression level of AQP3 was restored to 1.32, while Staphylococcus epidermidis C13 could not significantly increase the expression of FLG mRNA, with an expression level of 0.73.

[0095] Example 8: Effect of postbiotic preparation of Staphylococcus epidermidis CCSM009 on the expression of barrier structural protein genes in SDS-damaged HaCaT cells.

[0096] The specific implementation method is the same as in Example 6, except that after step (3) incubation, the original culture medium is discarded and washed once with PBS. The corresponding metagenic sample is added to the metagenic treatment group, and cell culture medium is added to the control group and the model group. The cells are incubated again for 24 hours.

[0097] In the metabiotic treatment group, the metabiotics were resuspended in cell culture medium, and the amount of resuspended metabiotics was adjusted to a concentration of 1×10⁻⁶ after fermentation. 7 The amounts of postbiotics prepared from bacterial cultures of CFU / ml were equal, and 2 mL of postbiotics CCSM009 prepared from Staphylococcus epidermidis and postbiotic C13-S prepared from C13 were added respectively.

[0098] After incubation, discard the culture supernatant, wash each well three times rapidly with PBS, add 1 mL of cell lysis buffer to each well, repeatedly pipette, extract RNA from the cell lysis buffer, and reverse transcribe it into cDNA using an RT-PCR reverse transcription kit. Gene expression in HaCaT cells is detected using real-time quantitative PCR. -△△Ct Formula calculation ZO-1, Occludin and CLDN The mRNA expression levels were measured, with GAPDH as the internal reference. The primers are described in Table 2 below, and the results are as follows: Figure 5 As shown.

[0099] Table 2 Primer Sequences

[0100] Tight junction proteins are essential components for maintaining skin barrier function. Located between keratinocytes, they form sealed junctions that prevent the permeation of external substances and water through the intercellular spaces, thus maintaining the integrity of the skin barrier. ZO-1, Occludin, and CLDN are core components of tight junctions and are important structural proteins of the skin barrier. They play a crucial role in regulating the transport of substances through the intercellular spaces, maintaining the selective permeability of the skin barrier. Furthermore, these proteins not only participate in physical cell-cell connections but also function in intracellular signal transduction, influencing cell proliferation, differentiation, and migration. The expression of ZO-1, CLDN, and Occludin is influenced by… Figure 5As shown, the mRNA expression of these three proteins in the model group decreased to 0.48, 0.50 and 0.34, respectively. After repair by Staphylococcus epidermidis CCSM009 bacterial lysate, the expression of these three proteins could be upregulated to varying degrees, reaching 8.22, 8.45 and 6.38, respectively. However, C13-S had no upregulating effect on the expression of tight junction protein genes, with expression levels of 0.53, 0.49 and 0.37, respectively.

[0101] Based on the results of Examples 7 and 8, C13-S has no positive effect on the gene expression of proteins related to skin barrier function and structure. Therefore, subsequent examples will only describe the efficacy of CCSM009-S.

[0102] Example 9: Effect of postbiotic preparation of Staphylococcus epidermidis CCSM009 on the expression of barrier function proteins in SDS-damaged HaCaT cells.

[0103] The specific implementation method is the same as in Example 6, except that after step (3) the original culture medium is discarded and washed once with PBS, the corresponding metagenic sample is added to the metagenic treatment group, and cell culture medium is added to the control group and the model group, and incubated again for 24h.

[0104] In the metabiotic treatment group, the metabiotics were resuspended in cell culture medium, and the amount of resuspended metabiotics was adjusted to a concentration of 1×10⁻⁶ after fermentation. 7 The amount of metabiotic prepared from the bacterial culture of CFU / ml is equivalent to that of the metabiotic CCSM009-S prepared from Staphylococcus epidermidis CCSM009.

[0105] After incubation, the culture supernatant was collected, and the expression levels of FLG, IVL, and LOR were calculated using the Sempercap ELISA kit via enzyme-linked adsorption reaction. The results are as follows: Figure 6 As shown.

[0106] The levels of FLG protein in the control group were 6610.62 pg / mL, while those in the model group were significantly reduced to 5548.12 pg / mL. After treatment with CCSM009-S, the levels increased to 6145.62 pg / mL. The levels of IVL protein in the control group were 1242.91 pg / mL, while those in the model group were significantly reduced to 1036.71 pg / mL. After treatment with CCSM009-S, the levels increased to 1117.54 pg / mL. The levels of LOR protein in the control group were 833.9 μg / mL, while those in the model group were significantly reduced to 736.62 μg / mL. After treatment with CCSM009-S, the levels increased to 789.25 μg / mL.

[0107] Example 10: Effect of postbiotic preparation of Staphylococcus epidermidis CCSM009 on the expression of barrier structural proteins in SDS-damaged HaCaT cells.

[0108] The specific implementation method is the same as in Example 6, except that after step (3) incubation, the original culture medium is discarded and washed once with PBS. The corresponding metagenic sample is added to the metagenic treatment group, and cell culture medium is added to the control group and the model group. The cells are incubated again for 24 hours.

[0109] In the metabiotic treatment group, the metabiotics were resuspended in cell culture medium, and the amount of resuspended metabiotics was adjusted to a concentration of 1×10⁻⁶ after fermentation. 7 The amount of metabiotic prepared from the bacterial culture of CFU / ml is equivalent to that of the metabiotic CCSM009-S prepared from Staphylococcus epidermidis CCSM009.

[0110] After incubation, the culture supernatant was collected, and the expression level of ZO-1 was calculated using the Sempercapto ELISA kit via enzyme-linked adsorption reaction. The results are as follows: Figure 7 As shown.

[0111] The ZO-1 protein content in the control group was 96.9 pg / mL, while the content in the model group decreased significantly to 78.7 pg / mL. After treatment with CCSM009-S, the content increased to 97.01 pg / mL, which was 1.23 times higher than that in the model group.

[0112] Example 11: Effect of postbiotic prepared from Staphylococcus epidermidis CCSM009 on the repair of SMPD1 expression in SDS-damaged HaCaT cells.

[0113] The specific implementation method is the same as in Example 6, except that after step (3) incubation, the original culture medium is discarded and washed once with PBS. The corresponding metagenic sample is added to the metagenic treatment group, and cell culture medium is added to the control group and the model group. The cells are incubated again for 24 hours.

[0114] In the metabiotic treatment group, the metabiotics were resuspended in cell culture medium, and the amount of resuspended metabiotics was adjusted to a concentration of 1×10⁻⁶ after fermentation. 7 The amount of metabiotic prepared from the bacterial culture of CFU / ml is equivalent to that of the metabiotic CCSM009-S prepared from Staphylococcus epidermidis CCSM009.

[0115] After incubation, the culture supernatant was collected, and the expression level of SMPD1 was calculated using the Smbega ELISA kit via enzyme-linked adsorption reaction. The results are as follows: Figure 8 As shown.

[0116] Sphingomyelinase 1 (SMPD1) plays a crucial role in maintaining skin barrier function. It breaks down sphingomyelin to generate ceramides, a lipid molecule that functions vitally in the skin barrier. In this study, the SMPD1 level in the control group was 264.27 μg / mL, while in the model group it was significantly reduced to 208.38 μg / mL. Treatment with CCSM009-S increased the level to 264.93 μg / mL, a 1.27-fold increase compared to the model group.

[0117] Example 12: Effect of postbiotic prepared from Staphylococcus epidermidis CCSM009 on the repair of KLK7 expression in SDS-damaged HaCaT cells.

[0118] The specific implementation method is the same as in Example 6, except that after step (3) incubation, the original culture medium is discarded and washed once with PBS. The corresponding metagenic sample is added to the metagenic treatment group, and cell culture medium is added to the control group and the model group. The cells are incubated again for 24 hours.

[0119] In the metabiotic treatment group, the metabiotics were resuspended in cell culture medium, and the amount of resuspended metabiotics was adjusted to a concentration of 1×10⁻⁶ after fermentation. 7 The amount of metabiotic prepared from the bacterial culture of CFU / ml is equivalent to that of the metabiotic CCSM009-S prepared from Staphylococcus epidermidis CCSM009.

[0120] After incubation, the culture supernatant was collected, and the expression level of SMPD1 was calculated using the Smbega ELISA kit via enzyme-linked adsorption reaction. The results are as follows: Figure 9 As shown.

[0121] Tissue kallikrein 7 (KLK7) is a serine protease that plays a crucial role in skin barrier function. It primarily participates in the degradation of intercellular junctions in the stratum corneum, a process essential for stratum corneum shedding and renewal. Excessive KLK7 activity leads to excessive stratum corneum degradation, weakening the skin barrier and resulting in increased skin moisture loss and sensitivity to external stimuli. In this study, the KLK7 level in the control group was 16.02 ng / mL, while the level in the model group significantly increased to 19.27 ng / mL. Treatment with CCSM009-S reduced the level to 14.09 g / mL, a 1.37-fold increase compared to the model group.

[0122] Example 13: Effect of postbiotic prepared from Staphylococcus epidermidis CCSM009 on the expression of AHR and OVOL1 receptors in SDS-damaged HaCaT cells.

[0123] The specific implementation method is the same as in Example 6, except that after step (3) incubation, the original culture medium is discarded and washed once with PBS. The corresponding metagenic sample is added to the metagenic treatment group, and cell culture medium is added to the control group. The cells are incubated again for 24 hours.

[0124] In the metabiotic treatment group, the metabiotics were resuspended in cell culture medium, and the amount of resuspended metabiotics was adjusted to a concentration of 1×10⁻⁶ after fermentation. 7 The amount of metabiotic prepared from the bacterial culture of CFU / ml is equivalent to that of the metabiotic CCSM009-S prepared from Staphylococcus epidermidis CCSM009.

[0125] After incubation, discard the culture supernatant, wash each well three times rapidly with PBS, add 1 mL of cell lysis buffer to each well, repeatedly pipette, extract RNA from the cell lysis buffer, and reverse transcribe it into cDNA using an RT-PCR reverse transcription kit. Gene expression in HaCaT cells is detected using real-time quantitative PCR. -△△Ct Formula calculation AHR and OVOL1 The mRNA expression levels were measured, with GAPDH as the internal reference. The primers are described in Table 3 below, and the results are as follows: Figure 10 As shown.

[0126] Table 3 Primer Sequences

[0127] The AHR signaling pathway plays a crucial role in maintaining the integrity of the skin barrier. Decreased AHR expression levels increase skin barrier permeability, leading to a higher risk of infection and more pronounced pathological features in atopic dermatitis models. OVOL1 is a transcription factor responsible for regulating the expression of genes related to skin barrier function. Studies have shown that OVOL1 is a direct transcriptional target of AHR, and its expression affects AHR's ability to regulate related genes in keratinocytes. The absence of OVOL1 in keratinocytes weakens the promoting effect of AHR on skin barrier function. Figure 9 The results show that the bacterial lysate of CCSM009 can significantly upregulate the gene expression of AHR and OVOL1, up to 1.65 and 1.7, respectively, thereby activating the expression of this pathway.

[0128] Example 14: Effective substance analysis of fermentation supernatant of Staphylococcus epidermidis CCSM009 1. Substance identification based on non-targeted metabolomics: The steps for metabolomics analysis of CCSM009 fermentation supernatant are as follows: (1) After culturing the strain in TSB medium for 12 h according to the method in Example 1, take 1 mL of bacterial solution and incubate at 4℃ for 10,000 hours. g Centrifuge for 5 minutes and collect the supernatant. (2) Transfer 100 μL of supernatant into a 1.5 mL centrifuge tube; (3) Add 400 μL of methanol:acetonitrile = (1:1, v / v) (pre-cooled at -20℃ in advance) to precipitate the protein; (4) Vortex for 30 s, followed by ice bath ultrasound for 10 min; (5) Place the sample in a -20℃ refrigerator for 1 h to increase the protein precipitation rate (secondary precipitation removes protein). (6) Centrifuge at 15000 rpm for 15 min at 4℃. (7) Take the supernatant and concentrate it under vacuum; (8) Redissolve by adding 100 μL of acetonitrile:water (1:1) and vortexing for 30 s; (9) Centrifuge at 15,000 rpm for 15 min at 4℃, take the supernatant, transfer the appropriate volume into a vial for instrument testing; (10) For polar metabolites, this project used a Vanquish (Thermo Fisher Scientific) ultra-high performance liquid chromatograph (UPLC) with a Waters ACQUITY UPLC BEH Amide (2.1 mm × 50 mm, 1.7 μm) column for chromatographic separation of the target compounds. Phase A of the HPLC was aqueous, containing 25 mmol / L ammonium acetate and 25 mmol / L ammonia, and phase B was acetonitrile. Sample tray temperature: 4℃, injection volume: 2 μL. The Orbitrap Exploris 120 mass spectrometer was able to acquire primary and secondary mass spectrometry data under the control of the control software (Xcalibur, version 4.4, Thermo). Detailed parameters are as follows: Sheath gasflow rate: 50 Arb, Aux gas flow rate: 15 Arb, Capillary temperature: 320℃, Full ms resolution: 60000, MS / MS resolution: 15000, Collision energy: SNCE 20 / 30 / 40, Spray voltage: 3.8 kV (positive) or -3.4 kV (negative).

[0129] 2. Metabolomics analysis of CCSM006 bacterial cell structure: After culturing for 12 hours using the aforementioned method, take 1 mL of bacterial culture and incubate at 4°C with 10,000... g Centrifuge for 5 minutes to collect the bacterial sludge; (1) Wash once with pre-cooled 0.9% physiological saline to remove residual culture medium; (2) After centrifuging to remove physiological saline, 1.5 mL centrifuge tubes containing bacterial sludge are placed into liquid nitrogen for quenching to stop metabolic activity; (3) Remove the sample from the liquid nitrogen and add 500 μL of methanol:acetonitrile = (1:1, v / v) (pre-cooled at -20℃ in advance); (4) After vortexing for 30 seconds, the cells were repeatedly frozen and thawed three times with liquid nitrogen to lyse the cells and release intracellular metabolites. (5) Place the sample in a -20℃ refrigerator for 1 h to increase the protein precipitation rate (secondary precipitation removes protein). (6) Centrifuge at 15,000 rpm for 15 min at 4℃; (7) Take the supernatant and evaporate it to dryness using a rotary evaporator; (8) Redissolve by adding 100 μL of acetonitrile:water (1:1) and vortexing for 30 s; (9) Centrifuge at 15,000 rpm for 15 min at 4℃, take the supernatant, transfer an appropriate volume into a vial for instrument testing; (10) For polar metabolites, this project used a Vanquish (Thermo Fisher Scientific) ultra-high performance liquid chromatograph (UPLC) with a Waters ACQUITY UPLC BEH Amide (2.1 mm × 50 mm, 1.7 μm) column for chromatographic separation of the target compounds. Phase A of the HPLC was aqueous, containing 25 mmol / L ammonium acetate and 25 mmol / L ammonia, and phase B was acetonitrile. Sample tray temperature: 4℃, injection volume: 2 μL. The Orbitrap Exploris 120 mass spectrometer was able to acquire primary and secondary mass spectrometry data under the control of the control software (Xcalibur, version 4.4, Thermo). Detailed parameters are as follows: Sheath gasflow rate: 50 Arb, Aux gas flow rate: 15 Arb, Capillary temperature: 320℃, Full ms resolution: 60000, MS / MS resolution: 15000, Collision energy: SNCE 20 / 30 / 40, Spray voltage: 3.8 kV (positive) or -3.4 kV (negative).

[0130] 3. Analysis of active ingredients in CCSM009: The raw data was converted into mzXML format using ProteoWizard software, and then metabolite identification was performed using a collaboratively developed R package. The database used was BiotreeDB (V3.0), and then visualization analysis was performed using a self-developed R package.

[0131] Literature review was conducted to collect the chemical formulas, molecular formulas, and molecular weights of potential effective substances in CCSM002. Based on non-target metabolomics, and according to substances with a P-value < 0.05 and differences compared to those before fermentation, as well as research on skin barrier repair, potential effective substances were screened. The substances are shown in Table 4.

[0132] Table 4 Potentially effective substances related to the skin barrier in CCSM009

[0133] Example 15: Quantitative detection of potential active substances in Staphylococcus epidermidis CCSM009 based on non-target metabolomics results 1. Liquid chromatography-mass spectrometry was used to detect substances in CCSM009, and the detection method was the same as in Example 13.

[0134] 2. Based on the results of non-targeted metabolomics, the following substances were detected in the sample after targeted quantitative detection: (1) Oleamide like Figures 11-14 As shown, oleamide was detected in both the fermentation supernatant and bacterial cells, with concentrations of 2.62 ppm and 4.46 ppm, respectively. Oleamide is a type of endogenous fatty amide with significant advantages in lipid homeostasis regulation and immune balance. It can directly activate the PPAR pathway and CB2 receptor, enhance the skin's tolerance to inflammatory stimuli, and inhibit the excessive release of inflammatory factors such as TNF-α and IL-1β, thereby rapidly restoring the damaged barrier environment. Simultaneously, oleamide can promote the production of key barrier lipids—ceramides, free fatty acids, and cholesterol—improving the fluidity and structure of the sebum film, thus enhancing the skin's water-locking and defense capabilities. As an endogenous regulatory molecule, oleamide possesses advantages such as high safety, well-defined target, and rapid barrier repair.

[0135] (2) D-erythrosine disphingosine like Figure 15-18As shown, this substance was detected in both the fermentation supernatant and bacterial cells, with concentrations of 2.27 ppm and 15 ppm, respectively. Sphingosine is an important naturally occurring sphingolipid molecule in the skin surface, possessing irreplaceable biological advantages in barrier repair. Firstly, sphingosine possesses natural broad-spectrum antibacterial properties, inhibiting opportunistic pathogens such as Staphylococcus aureus and Propionibacterium acnes, which are prone to proliferation when the skin barrier is damaged, thereby maintaining skin microecological stability. Secondly, sphingosine can significantly inhibit the TLR2 / TLR4-NF-κB inflammatory pathway, reducing the release of inflammatory mediators and providing a favorable low-inflammatory environment for barrier repair. Thirdly, sphingosine is a key intermediate in the sphingomyelin-ceramide metabolic pathway, promoting ceramide production and enhancing the lipid structure of the stratum corneum and overall barrier strength. As an inherent bioactive lipid of the skin, sphingosine combines high safety, strong microecological regulation capabilities, and a clearly defined barrier repair target.

[0136] (3) (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid like Figure 19-21 As shown, this substance was detected only in the fermentation supernatant, at a concentration of 19 ppb. (2-oxo-2,3-dihydro-1H-indole-3-yl)-acetic acid is a class of small, reactive molecules with an indole structure, possessing excellent antioxidant and cellular homeostasis regulation advantages. It can effectively scavenge reactive oxygen species (ROS) induced by ultraviolet radiation, pollution, or inflammation, reducing oxidative damage to stratum corneum structural proteins and lipids. Simultaneously, these indole derivatives can regulate the proliferation and differentiation of keratinocytes, promote the expression of barrier proteins (such as FLG and IVL), and accelerate the renewal of damaged barriers. Furthermore, this molecule also has the ability to inhibit inflammatory signals (such as NF-κB), maintaining a low-inflammatory environment in the skin during repair, thereby achieving more efficient and stable barrier reconstruction.

[0137] (4) L-gamma-valpy dipeptide like Figure 22-24 As shown, this substance was detected only in the fermentation supernatant, at a concentration of 2.99 ppm. L-Glyceryl dipeptide, as an active small-molecule dipeptide, can directly participate in the metabolic processes of keratinocytes in the skin. Its advantage lies in its ability to promote the expression of structural proteins (such as filaggrin and loricrin) and tight junction proteins (claudin and occludin) within keratinocytes, thereby improving the integrity of the skin barrier structure from its source. Simultaneously, this dipeptide can also enhance the ability of keratinocytes to synthesize natural moisturizing factors (NMF), improving the water content of the stratum corneum and significantly reducing transepidermal water loss (TEWL). Due to its small molecular weight and good transdermal permeability, L-Glyceryl dipeptide can achieve rapid and precise barrier repair, effectively improving dry and sensitive skin conditions.

[0138] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. Staphylococcus epidermidis ( Staphylococcus epidermidis Accession number CCSM009 was deposited at the Guangdong Provincial Center for Microbial Culture Collection on July 22, 2024, with accession number GDMCC NO: 648903.

2. The metabiotic prepared using the Staphylococcus epidermidis CCSM009 as described in claim 1.

3. The epigenetic agent according to claim 2, characterized in that, The metabiotic includes bacterial lysate, inactivated or dead cells, fermentation supernatant, or any of the above-mentioned powders prepared by drying.

4. The epigenetic agent according to claim 2 or 3, characterized in that, The post-gene contains oleamide, D-erythrosine, (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid, and (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid.

5. The epigenetic agent according to claim 3, characterized in that, The inactivated or deactivated cells are prepared as follows: the Staphylococcus epidermidis CCSM009 is cultured in a culture medium for a period of time, the bacterial cells in the cell culture medium are collected, and inactivated bacterial cells are obtained after heat treatment.

6. The epigenetic agent according to claim 3, characterized in that, The method for preparing the bacterial lysate is as follows: Staphylococcus epidermidis CCSM009 is cultured in a culture medium for a period of time, bacterial cells are collected, homogenized under high pressure, and the supernatant obtained by centrifugation is the bacterial lysate.

7. A composition containing Staphylococcus epidermidis CCSM009 as described in claim 1 and / or its metabiotic, characterized in that, The composition includes, but is not limited to, pharmaceuticals or daily chemical products.

8. The use of the post-genetic agent as described in claim 2 in the preparation of pharmaceuticals or daily chemical products for repairing the skin barrier.

9. The application according to claim 8, characterized in that, The application includes at least one of the following functions: (1) Enhance the cell activity of keratinocytes in the skin after injury; (2) Increase the expression of barrier function protein genes in keratinocytes of the skin after injury; (3) Increase the expression of barrier structure protein genes in keratinocytes of the skin after injury; (4) Increase the expression of barrier function proteins in keratinocytes of the skin after injury; (5) Reduces the expression of structural proteins in keratinocytes of the skin after injury; (6) Increase the expression of sphingomyelinase 1 in keratinocytes of damaged skin; (7) Reduces tissue kininase 7 expression in keratinocytes of the skin after injury.

10. The use of the Staphylococcus epidermidis CCSM009 of claim 1 in the preparation of products containing oleic acid amide, D-erythrosine, (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid or (2-oxo-2,3-dihydro-1H-indol-3-yl)-acetic acid.

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