Bifidobacterium adolescentis YG3912, exosome and extract thereof and application of bifidobacterium adolescentis YG3912 in skin care

Abstract

The invention relates to the field of microorganisms, in particular to bifidobacterium adolescentis YG3912, inactivated thalli, an extract, a culture, a culture supernatant, a fermentation product and / or a fermentation supernatant of the bifidobacterium adolescentis YG3912, an application of the bifidobacterium adolescentis YG3912 in preparation of a composition for promoting skin repair, reducing skin inflammation or allergy, improving skin quality and improving skin quality. The invention also relates to application of the composition in products for resisting skin aging and enhancing skin immune function. The bifidobacterium adolescentis YG3912 and the related products thereof have the advantages that the bifidobacterium adolescentis YG3912 and the related products of the bifidobacterium adolescentis YG3912 are high in skin cell repairing capacity, skin inflammation relieving, inhibiting or eliminating capacity and remarkable in skin aging resisting capacity, so that the bifidobacterium adolescentis YG3912 and the related products of the bifidobacterium adolescentis YG3912 are expected to be used for skin health care such as skin repairing, inflammation relieving and light aging resisting, and the bifidobacterium adolescentis YG3912 and the related products thereof have remarkable application value and market potential.

Description

Technical Field

[0001] The present invention relates to the field of microorganisms, and specifically to a strain of Bifidobacterium adolescentis YG3912, its inactivated bacteria, extracts, cultures, culture supernatants, fermentations and / or fermentation supernatants, compositions comprising the foregoing, and use of Bifidobacterium adolescentis or its inactivated bacteria, extracts, cultures and compositions comprising the foregoing in the preparation of products for repairing skin, resisting dermatitis, resisting skin aging and enhancing skin immune function. Background Art

[0002] Traditional skin care products rely on chemical ingredients and may have potential risks. With the advancement of biotechnology, especially the in-depth study of microbiome, the important role of microorganisms and their metabolites in skin health has been revealed. Exosomes, as specific membrane vesicles for intercellular communication, are becoming a research hotspot in the field of skin care and repair due to their unique biological properties. However, the exosomes currently on the market are mainly derived from human stem cells, which has limitations in terms of ethics, acquisition costs and large-scale production.

[0003] Bifidobacterium adolescentis is a Gram-positive, anaerobic bacterium belonging to the genus Bifidobacterium and is an important representative of probiotics. Existing technologies mainly focus on the health effects of Bifidobacterium adolescentis on the host, including immunomodulation, anti-diarrhea, anti-inflammatory and hypoglycemic effects, and there are no reports on its application in skin care.

[0004] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0005] Purpose of the Invention

[0006] The object of the present invention is to provide a strain of Bifidobacterium adolescentis YG3912 with the functions of repairing skin, resisting dermatitis, resisting skin aging and enhancing skin immunity, its inactivated bacteria, extracts, cultures, culture supernatants, fermentations and / or fermentation supernatants, compositions containing the aforementioned, and the use of Bifidobacterium adolescentis or its inactivated bacteria, extracts, cultures and compositions containing the aforementioned in the preparation of products for repairing skin, resisting dermatitis, resisting skin aging and enhancing skin immunity.

[0007] Solution

[0008] To achieve the purpose of the present invention, the present invention adopts the following technical scheme.

[0009] In the first aspect, the present invention provides a strain of Bifidobacterium adolescentis YG3912, which is classified and named Bifidobacterium adolescentis and is deposited in the General Microbiology Center of China Microbiological Culture Collection Administration. The address of the depository is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China. The deposit date is June 20, 2024, and the deposit number is CGMCC No. 31020.

[0010] The Bifidobacterium adolescentis YG3912 of the present invention is isolated from fecal samples of healthy adolescents and is identified as Bifidobacterium adolescentis through 16S rRNA sequencing and combined with its morphological characteristics.

[0011] The 16S rRNA sequence of Bifidobacterium adolescentis YG3912 of the present invention is as follows:

[0012]

[0013] The colony characteristics of this strain include: after 24 to 48 hours of culture on MRS agar medium, the diameter is between 0.5 and 2.0 mm, the colony edges are smooth, the surface is smooth, and it is opaque and milky white.

[0014] In vitro experiments have confirmed that the Bifidobacterium adolescentis YG3912 of the present invention has a good repair ability for human skin fibroblasts; specifically, the inventors found that the exosomes of the Bifidobacterium adolescentis YG3912 of the present invention can promote the production of collagens such as COL1A2 and COL3A1, and its fermentation broth and lysate can enhance the production of collagens such as COL1A1, COL17A1 and hyaluronic acid (HAS1, HAS3), thereby facilitating the deep repair and regeneration of skin fibroblasts.

[0015] The inventors also found that the exosomes, fermentation broth and lysate of Bifidobacterium adolescentis YG3912 of the present invention can promote the proliferation and survival of macrophages at appropriate concentrations, and can stimulate macrophage cells and enhance their ability to produce cytokines such as TNF-α, IL-6, IL-1β and iNOS, thereby promoting immune regulation and enhancing the skin immune barrier function.

[0016] In addition, the inventors also confirmed that the Bifidobacterium adolescentis YG3912 of the present invention also has obvious inflammation inhibition and anti-skin aging effects; specifically, the inventors confirmed through experiments that the exosomes, fermentation broth and lysate of Bifidobacterium adolescentis YG3912 at appropriate concentrations can significantly inhibit the expression of UVB-induced inflammatory factors (such as IL-1β, IL-6 and IL-8), inhibit the generation of matrix metalloproteinases (MMP-2, MMP-9) and ROS, thereby achieving the anti-photoaging effect.

[0017] In a second aspect, the present invention provides the inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of Bifidobacterium adolescentis YG3912 as described in the first aspect above.

[0018] Preferably, the extract of Bifidobacterium adolescentis YG3912 is a lysate and / or exosomes extracted therefrom.

[0019] In a feasible embodiment, the method for extracting the lysate comprises the following:

[0020] Bifidobacterium adolescentis YG3912 is fermented and cultured, the fermentation broth is centrifuged, the bacteria are collected, the bacteria are resuspended in PBS and the cell walls are broken by ultrasound or enzymatic hydrolysis to obtain a lysate; preferably, the obtained lysate is sterilized.

[0021] In a feasible embodiment, the method for extracting exosomes comprises the following:

[0022] Fermenting Bifidobacterium adolescentis YG3912, centrifuging the fermentation liquid, and collecting the fermentation supernatant; centrifuging, filtering, and concentrating the obtained fermentation supernatant to obtain exosomes;

[0023] Preferably, the centrifugation procedure of the fermentation supernatant is: first centrifuge at 200-400g, preferably 300g for 8-15min, preferably 10min, then centrifuge at 1500-3000g, preferably 2000g for 15-30min, preferably 20min, and finally centrifuge at 8000-12000g, preferably 10000g for 20-40min, preferably 30min;

[0024] Preferably, the filtration of the fermentation supernatant is: filtering with a filter membrane having a pore size of 0.2-2.0 μm, preferably, filtering with filter membranes having pore sizes of 0.8 μm and 0.45 μm respectively;

[0025] Preferably, the fermentation supernatant is concentrated by using a hollow fiber of 400-600 kd, preferably 500 kd, to concentrate the fermentation clarified liquid.

[0026] In a third aspect, the present invention provides a composition comprising the Bifidobacterium adolescentis YG3912 as described in the first aspect above, or the inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of the Bifidobacterium adolescentis YG3912 as described in the second aspect above, as an active ingredient.

[0027] In some feasible embodiments, the composition is a pharmaceutical composition, which further includes a pharmaceutically acceptable carrier and / or excipient.

[0028] In other possible embodiments, the composition is a cosmetic composition, which further comprises a cosmetically acceptable carrier and / or excipient.

[0029] Preferably, the composition is in the form of an external dosage form;

[0030] More preferably, the external dosage form is selected from the following dosage forms: aqueous solution, emulsion, ointment, cream, gel, powder, oil.

[0031] In a feasible embodiment, the composition further comprises other active agents;

[0032] Preferably, the other active agents include any one or more of antioxidants, cell active agents, moisturizing agents, anti-aging agents and anti-dermatitis agents.

[0033] In a fourth aspect, the present invention provides the use of the inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of Bifidobacterium adolescentis YG3912 as described in the first aspect, or the inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of Bifidobacterium adolescentis YG3912 as described in the second aspect, or a composition comprising one or more of the foregoing substances in the preparation of a product for the following purposes:

[0034] 1) Promote skin repair;

[0035] 2) Reduce, inhibit or eliminate skin inflammation or allergies;

[0036] 3) Anti-skin aging;

[0037] 4) Enhance skin immune function.

[0038] In a feasible embodiment, the promoting skin repair is promoting the repair of sensitive skin;

[0039] In a feasible embodiment, the skin inflammation is sterile skin inflammation or inflammation caused by external stimulation;

[0040] In a feasible embodiment, the skin allergy is a skin allergy symptom caused by stimulation from external factors;

[0041] In a feasible embodiment, the anti-skin aging is anti-skin photoaging.

[0042] In a fifth aspect, the present invention provides a method for repairing skin, reducing, inhibiting or eliminating skin inflammation or allergy, resisting skin aging, and / or enhancing skin immune function, which comprises: administering to a subject in need an effective amount of Bifidobacterium adolescentis YG3912 as described in the first aspect above, or the inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of Bifidobacterium adolescentis YG3912 as described in the second aspect above, or a composition comprising one or more of the foregoing substances.

[0043] In a feasible embodiment, the promoting skin repair is promoting the repair of sensitive skin;

[0044] In a feasible embodiment, the skin inflammation is sterile skin inflammation or inflammation caused by external stimulation;

[0045] In a feasible embodiment, the skin allergy is a skin allergy symptom caused by stimulation from external factors;

[0046] In a feasible embodiment, the anti-skin aging is anti-skin photoaging.

[0047] Beneficial Effects

[0048] Compared with the prior art, the Bifidobacterium adolescentis YG3912 of the present invention has the following obvious advantages:

[0049] (1) Its exosomes can promote the production of collagens such as COL1A2 and COL3A1 at appropriate concentrations, and its fermentation broth and lysate can enhance the production of collagens such as COL1A1 and COL17A1 and hyaluronic acid (HAS1, HAS3), thereby facilitating the deep repair and regeneration of skin fibroblasts;

[0050] (2) Its exosomes, fermentation broth and lysate can promote the proliferation and survival of macrophages at appropriate concentrations, and can stimulate macrophages and enhance their ability to produce cytokines such as TNF-α, IL-6, IL-1β and iNOS, thereby promoting immune regulation and enhancing the skin immune barrier function;

[0051] (3) Its exosomes, fermentation broth and lysate can significantly inhibit the expression of UVB-induced inflammatory factors (such as IL-1β, IL-6 and IL-8), inhibit the generation of matrix metalloproteinases (MMP-2, MMP-9) and ROS at appropriate concentrations, thereby achieving an anti-photoaging effect.

[0052] In summary, the Bifidobacterium adolescentis YG3912 and its related products of the present invention have good ability to repair skin cells, reduce, inhibit or eliminate skin inflammation, and resist skin aging. Therefore, they are expected to be used for skin repair, inflammation relief, and skin health care such as anti-photoaging, and have significant application value and market potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] One or more embodiments are exemplarily described by the pictures in the accompanying drawings, and these exemplary descriptions do not constitute limitations on the embodiments. The special word "exemplary" here means "used as an example, embodiment or illustrative". Any embodiment described as "exemplary" here is not necessarily interpreted as being superior or better than other embodiments.

[0054] Figure 1 This is the growth curve of Bifidobacterium adolescentis YG3912 of the present invention.

[0055] Figure 2 The particle concentration and particle size distribution of exosomes extracted from the fermentation broth of the strain YG3912 of the present invention measured by NTA technology are shown.

[0056] Figure 3 Heatmap showing HSF cell repair rates of fermentation filtrates and lysates of different Bifidobacterium adolescentis strains.

[0057] Figure 4 The effect of the exosomes of Bifidobacterium adolescentis YG3912 of the present invention on the fusion rate of HSF cells is shown, and * indicates P<0.05.

[0058] Figure 5 The results show the effects of the fermentation filtrate (Figure A), lysate (Figure B) and exosomes (Figure C) of Bifidobacterium adolescentis YG3912 of the present invention on the expression of HSF cell repair-related factors (including collagen COL1A1, COL1A2, COL3A1, COL17A and hyaluronic acid HAS1, HAS3); wherein the abscissa is the type of repair-related factors tested, and the ordinate is the relative mRNA expression level of each repair-related factor; * indicates P<0.05, ** indicates P<0.01, *** indicates P<0.001, and **** indicates P<0.0001.

[0059] Figure 6 The effects of the fermentation filtrate, exosomes, and lysates of Bifidobacterium adolescentis YG3912 of the present invention on the viability of RAW264.7 cells were shown.

[0060] Figure 7 The effects of the fermentation filtrate (Figure A), lysates (Figure B), and exosomes (Figure C) of Bifidobacterium adolescentis YG3912 of the present invention on the secretion of TNF-α by RAW264.7 cells were shown; where *, **, ***, and **** indicate P < 0.05, P < 0.01, P < 0.001, and P < 0.0001, respectively, compared with the blank control group (i.e., the Con group).

[0061] Figure 8 The effects of the fermentation filtrate (Figures A - D), lysates (Figures E - H), and exosomes (Figures I - L) of Bifidobacterium adolescentis YG3912 of the present invention on the secretion of inflammatory factors TNF-α, IL-6, IL-1β, and iNOS by RAW264.7 cells were shown; where *, **, ***, and **** indicate P < 0.05, P < 0.01, P < 0.001, and P < 0.0001, respectively, compared with the blank control group (i.e., the Con group).

[0062] Fig. 9 The effects of the fermentation filtrate, lysates, and exosomes of Bifidobacterium adolescentis YG3912 of the present invention on the cell morphology and fusion degree of UVB-damaged cells were shown.

[0063] Fig.10 The effects of the fermentation filtrate, lysates, and exosomes of Bifidobacterium adolescentis YG3912 of the present invention on the expression levels of cell inflammatory factors and matrix metalloproteinases in UVB-damaged cells were shown; where Figures A, B, and C show the protein levels of IL-6, IL-8, and IL-1β in each treatment group, and Figure D shows the relative mRNA levels of IL-1β, MMP2, and MMP9 in each treatment group; *, **, and *** indicate 0.01 < P < 0.05, 0.001 < P < 0.01, and P < 0.001, respectively, compared with the UVB group. ###Indicates: P < 0.001 compared with the blank control group (i.e., Con group).

[0064] Fig.11 Shows the effects of the fermentation filtrate, lysate, and exosomes of Bifidobacterium adolescentis YG3912 of the present invention on the cell morphology, confluence, and cellular ROS level of UVB damage; among them, Figure A shows the cell morphology and confluence of each treatment group (upper figure) and the fluorescence staining pictures of ROS (lower figure), and Figure B shows the relative fluorescence intensity (%) of ROS in each treatment group, where * indicates: 0.01 < P < 0.05 compared with the UVB group, ** indicates: 0.001 < P < 0.01 compared with the UVB group, *** indicates: P < 0.001 compared with the UVB group; ### Indicates: P < 0.001 compared with the blank control group (i.e., Con group).

[0065] Bifidobacterium adolescentis YG3912 of the present invention is classified and named as Bifidobacterium adolescentis, deposit date: June 20, 2024; deposit unit: General Microbiology Center of China Committee for Culture Collection of Microorganisms; deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China; deposit number is CGMCC NO. 31020. Detailed implementation manners

[0066] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts belong to the scope of protection of the present invention. Unless otherwise clearly stated, throughout the specification and claims, the term "comprise" or its variations such as "comprises" or "including" etc. will be understood to include the stated elements or components, without excluding other elements or other components.

[0067] In addition, for better illustration of the present invention, numerous specific details are given in the following detailed implementation manners. Those skilled in the art should understand that the present invention can also be implemented without some specific details. In some embodiments, details of raw materials, components, methods, means, etc. well-known to those skilled in the art are not described in detail to highlight the gist of the present invention.

[0068] In the following examples, unless otherwise specified, other reagents or raw materials used are commercially available.

[0069] In addition, in the following embodiments, the "concentration" of the strain fermentation filtrate, lysate, and exosomes refers to the volume dilution multiple of the original solution of the corresponding substance diluted with a complete culture medium (i.e., DMEM culture medium containing serum); for example, 1 / 1000, 1 / 3000, 1 / 10000 and 1 / 30000 (V / V) concentrations of the strain fermentation filtrate or lysate refer to the original solution of the strain fermentation filtrate or lysate diluted 1000 times, 3000 times, 10000 times and 30000 times by volume, respectively.

[0070] Example 1: Isolation and identification of Bifidobacterium adolescentis YG3912

[0071] (1) Strain isolation

[0072] Take 1g of healthy adolescent fecal sample and put it into 9mL PBS buffer. Vortex mix and then dilute it tenfold with sterile saline. -6 , 10 -7 , 10 -8 Three dilution gradients were spread on MRS agar medium respectively, and the solid plates were placed in an anaerobic constant temperature incubator at 37°C for 24 to 48 hours. Colonies were picked and purified on MRS agar medium for more than 3 times until the colony morphology on the plate was consistent. Several clones were picked and named YGMCC2531, YGMCC3246, YGMCC3487, YGMCC3492, YGMCC3390, YGMCC3533, YGMCC3625, YGMCC0568, YGMCC0809, YGMCC0896, YGMCC0879, YGMCC1108, YGMCC1129, YGMCC1777 and YGMCC3912 respectively. The strains were identified by strain morphology and 16S rRNA sequencing results, and they were all Bifidobacterium adolescentis. Moreover, after efficacy screening, the Bifidobacterium adolescentis of the present invention, namely strain YGMCC3912, also known as the Bifidobacterium adolescentis strain YG3912 of the present invention, was obtained (herein, strain YGMCC3912 is equivalent to strain YG3912, and the two can be used interchangeably).

[0073] (2) Identification of the strains of the present invention

[0074] A. Colony morphology

[0075] After the strain clone YG3912 of the present invention is cultured in MRS agar medium for 24 to 48 hours, the diameter is between 0.5 and 2.0 mm, the colony has a smooth edge, a smooth surface, and is opaque and milky white.

[0076] B. 16S rRNA sequencing

[0077] The strain YG3912 was sent to Sangon Biotech (Shanghai) Co., Ltd. for 16S rRNA gene sequencing, and its 16S rRNA sequence is shown in SEQ ID NO: 1; the 16S rRNA sequencing results were compared with the NCBI database by BLAST (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi), and combined with its colony morphological characteristics, the isolated strain YG3912 was determined to be Bifidobacterium adolescentis.

[0078] Example 2: Preparation of fermentation filtrate and lysate of strain YG3912, and extraction and characterization of exosomes

[0079] The culture medium involved in this embodiment is as follows:

[0080] MRS medium (g / L): peptone 10.0g / L, beef powder 5.0g / L, Tween-80 1mL / L, dipotassium hydrogen phosphate 2.0g / L, yeast powder 4.0g / L, magnesium sulfate 0.2g / L, triammonium citrate 2.0g / L, glucose 20.0g / L, manganese sulfate 0.05g / L, sodium acetate 5.0g / L, L-cysteine ​​hydrochloride 0.5g / L, solvent is water; the preparation method is: mix all the ingredients evenly and sterilize them under high pressure at 121℃ for 15min.

[0081] 2.1 Cultivation of strain YG3912 and preparation of its fermentation broth

[0082] The seed solution of strain YG3912 was transferred to MRS liquid medium at a 2% inoculation rate and cultured anaerobically at 37°C for 24 h. During the culture process, the OD600 value of the culture solution was measured by an ELISA instrument at regular intervals to obtain the growth curve of strain YG3912. Figure 1 shown. Figure 1 The results showed that strain YG3912 reached a stable growth phase (OD600=1.22) after 12 hours of culture.

[0083] The fermentation process parameters of strain YG3912 were optimized to obtain the best fermentation parameters as follows: pH 6.25, inoculation volume 2.0%, and culture time 48h. According to the above optimized fermentation parameters, strain YG3912 was fermented to obtain fermentation broth for later use.

[0084] 2.2 Preparation of fermentation filtrate and lysate of strain YG3912 and extraction and characterization of exosomes

[0085] (1) Preparation of fermentation filtrate:

[0086] The fermentation broth of strain YG3912 prepared above was centrifuged at 9000 r / min×4°C for 10 min to obtain the fermentation supernatant, which was filtered using a 0.22 μm polyethersulfone filter membrane to obtain the cell-free fermentation filtrate of strain YG3912, which was frozen at -80°C for later use.

[0087] (2) Lysate preparation:

[0088] The cells after centrifugation in (1) were harvested, and pre-cooled sterile PBS was added to 2-fold concentration and resuspended, and the cell wall was broken by ultrasound. The specific ultrasound program was as follows: ultrasound for 2 seconds, stop for 2 seconds, and the total ultrasound time was 60 minutes, and the entire ultrasound program was performed under ice bath conditions at -4 to 0°C. Then, the ultrasound product was heat-treated at 80°C for 30 minutes using an autoclave to obtain a sterilized lysate of the fermentation broth of strain YG3912, which was frozen at -80°C for later use.

[0089] (3) Exosome extraction and characterization:

[0090] The fermentation broth of strain YG3912 was centrifuged at 9000r / min×4℃ for 10min to obtain the fermentation supernatant. The fermentation supernatant was first centrifuged at 300g×4℃ for 10min, then at 2000g×4℃ for 20min, and finally at 10000g×4℃ for 30min, and then filtered with 0.8μm and 0.45μm polyethersulfone filter membranes in sequence to obtain the fermentation clarified liquid. The obtained fermentation clarified liquid was concentrated to 50mL using a 500kd hollow fiber, and 200mL of PBS was used to replace the liquid. The concentrated liquid was purified using a protein purifier to obtain the exosome solution of the fermentation broth of strain YG3912. The particle number and diameter distribution of exosomes in the obtained exosome solution were measured using NTA technology.

[0091] Through the above procedures, we obtained the fermentation filtrate, lysate and exosome solution of the stable and high-yield strain YG3912. The number and diameter range of exosome particles in the YG3912 exosome solution were measured based on the NTA technology. The results are shown in Figure 2 ; Figure 2 The results showed that the diameter of exosomes in the YG3912 exosome solution ranged from 100 to 500 nm, with an average of 178.4 nm (median diameter: 152.2 nm), which means that although the size of the main population was about 150 nm, it had a wide size distribution; and the number of particles per milliliter of exosome solution was 1.8*1010 indivual.

[0092] Using the above procedure, fermentation filtrates, lysates, and exosome solutions of strains YGMCC2531, YGMCC3246, YGMCC3487, YGMCC3492, YGMCC3390, YGMCC3533, YGMCC3625, YGMCC0568, YGMCC0809, YGMCC0896, YGMCC0879, YGMCC1108, YGMCC1129, and YGMCC1777 were also prepared, respectively.

[0093] Example 3: Detection of the ability of the fermentation filtrate and lysate of Bifidobacterium adolescentis YG3912 to promote skin cell repair

[0094] Human skin fibroblasts, as key cells in the dermis, are crucial to maintaining skin elasticity and tension. They can efficiently synthesize matrices such as elastin and collagen, generate elastic fibers and collagen fibers, secrete repair factors, and give the skin the ability to self-repair and renew.

[0095] In this example, human fibroblasts (hereinafter referred to as HSF) were used as the research object, and the repair abilities of fermentation filtrates and lysates of various strains were tested.

[0096] The culture method of HSF cells is as follows:

[0097] The HSF cells were cultured at 37°C and 5% CO 2 The cells were cultured in a cell culture incubator using a high glucose medium (DMEM) containing 15% special grade fetal bovine serum (FBS) and 1% 100× penicillin-streptomycin mixture (S / P).

[0098] The detection procedure of cell repair ability (cell repair rate) is as follows:

[0099] Select HSF cells that are growing well and in the logarithmic growth phase within 10 generations and culture them at a rate of 2×10 5 The cells were seeded into 6-well plates at a density of 1000 cells / well and placed in an incubator (37°C, 5% CO 2) after incubation for 24 hours, the fusion rate reached 90%; use a 200μL pipette tip to streak on the cell culture plate, and use PBS to wash the streaked culture plate 2 to 3 times to remove floating cells. Take a photo immediately after the scratch at 0h, and add 2mL of serum-free DMEM culture medium containing the sample after the photo is taken. Specifically, the control group is the DMEM treatment group, and the experimental group is different concentrations (including 1 / 1000, 1 / 3000, 1 / 10000 and 1 / 30000 (V / V) concentrations), fermentation filtrates of different strains and lysate treatment groups, each group has three replicates. Take pictures at 0 hours and 48 hours, and use Image J software to calculate the scratch area and cell repair rate. The calculation formula of cell repair rate is as follows:

[0100]

[0101] M--Migration, expressed as a percentage (%), B 0h --0h scratch area, B48 h --48h scratch area.

[0102] The cell repair rates of each experimental group and control group were counted and a cell repair rate heat map was made. The results are as follows Figure 3 As shown; Figure 3 The results showed that the repair ability of Bifidobacterium adolescentis showed strain selectivity, and the cell repair ability of the strain YG3912 of the present invention was significantly better than that of the control group and other strains, whether in the fermentation filtrate (32.3%-34.0%) or the lysate (41.4%-46.9%). Therefore, the strain YG3912 has become an ideal candidate strain due to its excellent cell repair ability.

[0103] Furthermore, HSF cells with good growth and within 10 generations in the logarithmic growth phase were selected and cultured at 2×10 5 Cells were seeded into 6-well plates at a seeding density of 100 cells / well and incubated in an incubator (37°C, 5% CO 2 ) for 24 h, the scratch was taken at 0 h, and then different concentrations of YG3912 exosomes were added. After 48 h of continuous culture, the scratch area was calculated using Image J software, and the cell repair rate was calculated. The calculation formula of the cell repair rate is the same as above.

[0104] The cell repair rate results of the YG3912 exosome groups at different concentrations and the control group are shown in Figure 4 shown. Figure 4 The results showed that YG3912 exosomes at concentrations of 1 / 30000, 1 / 10000 and 1 / 3000 could enhance the repair ability of HSF cells, among which the enhancement of YG3912 exosomes at a concentration of 1 / 10000 was significant (P<0.05).

[0105] Effects on the expression of cell repair-related factors

[0106] Select HSF cells that are growing well and in the logarithmic growth phase within 10 generations and culture them at a rate of 2×10 5 Cells were seeded into 6-well plates at a seeding density of 100 cells / well and incubated in an incubator (37°C, 5% CO 2 ) for 24 h, take a picture of the scratch at 0 h, and then add the test substances (different concentrations of YG3912 fermentation filtrate, lysate, exosomes), and continue to culture for 48 h before collecting the cells. Use TransZol (Quanshijin, R61225) to lyse the cells and extract RNA for the determination of the relative expression of cell repair-related factor mRNA.

[0107] The results are as follows Figure 5 shown.

[0108] Figure 5 Figure A is the test result of the YG3912 fermentation filtrate treatment group, which shows that the YGMCC03912 fermentation filtrate concentration of 1 / 10000 and 1 / 3000 can significantly increase the mRNA expression of COL1A1 in HSF cells, among which the concentration of 1 / 10000 has the most obvious effect (P<0.001), and the concentration of 1 / 10000 can significantly increase the mRNA expression of HAS3 in HSF cells (P<0.05).

[0109] Figure 5 Figure B is the test result of the YG3912 lysate treatment group, which shows that when the concentration of YGMCC03912 lysate is 1 / 10000, it can significantly increase the mRNA expression of COL17A1 in HSF cells (P<0.01), when it is 1 / 1000, it can significantly increase the mRNA expression of HAS1 in HSF cells (P<0.05), and when it is 1 / 30000, it can significantly increase the mRNA expression of HAS3 in HSF cells (P<0.05).

[0110] Figure 5 Figure C is the test result of the YG3912 exosome treatment group, which shows that the YGMCC03912 exosome concentration of 1 / 30000 can significantly increase the mRNA expression of COL1A2 and COL3A1 in HSF cells (P<0.01).

[0111] In summary, YG3912 fermentation filtrate, lysate and exosomes can enhance the production of collagens such as COL1A1, COL1A2, COL3A1, COL17A1 and hyaluronic acid such as HAS1 and HAS3 at appropriate concentrations, which are proteins that are beneficial to cell repair. Therefore, YG3912 fermentation filtrate, lysate and exosomes may promote the repair of damaged cells by increasing the expression of cell repair-related proteins.

[0112] Example 4: Detection of the fermentation filtrate, lysate and exosomes of Bifidobacterium adolescentis YG3912 in enhancing the skin immune barrier function

[0113] Macrophages are professional phagocytes that can remove dying or dead cells and cell fragments in a highly specialized manner. In addition, macrophages are the most important antigen-presenting cells and play a vital role in initiating immune responses. After receiving antigen stimulation, macrophages will present the antigen to the corresponding Th cells. Antigen presentation leads to the production of antibodies and completes the immune regulatory response.

[0114] In this example, macrophages RAW264.7 were used as the subjects to explore the effect of the YG3912 strain extract on the survival rate of RAW264.7 cells and the secretion of cytokines by RAW264.7 cells to explore its effect on the skin immune barrier function.

[0115] (1) Effects of YG3912 fermentation filtrate, exosomes, and lysate on the survival rate of RAW264.7 cells

[0116] RAW264.7 cells (Pnosai, CL-0190) in the logarithmic growth phase were resuspended and diluted to 1×10 5 100 μl of cell suspension was added to each well of a 96-well plate, and 200 μl of PBS was added to each well around the plate as a moisturizing well. The plate was placed at 37°C and 5% CO. 2 Incubate incubator for 24 h. Afterwards, discard the cell culture medium, add different concentrations of YG3912 fermentation filtrate, exosomes and lysate dilutions, 100 μl per well, set up 6 replicates, and place in 37°C, 5% CO 2The culture was continued in the incubator for 24 hours. Among them, the fermentation filtrate, lysate and exosome treatment groups included Con group (complete culture medium group), LPS group (LPS working solution, 1 μg / ml), 1 / 100 (100-fold lysate dilution), 1 / 300 group (300-fold lysate dilution), 1 / 1000 group (1000-fold lysate dilution), 1 / 3000 group (3000-fold lysate dilution), 1 / 10000 (10000-fold lysate dilution), and 1 / 30000 (30000-fold lysate dilution).

[0117] Then, add 50 μl of MTT solution (1×MTT) to each well and place in a 37°C, 5% CO2 incubator for 4 hours. All the above operations are performed under sterile conditions. After discarding the liquid in the culture plate, add 150 μl of DMSO to each well, mix well, measure the absorbance at a wavelength of 570 nm with an enzyme marker, and record the measurement results. The experimental results are analyzed using Graphpad prism. Select a concentration that is non-toxic to cells to continue the subsequent experiments.

[0118] The results are as follows Figure 6 shown. Figure 6 show:

[0119] 1) The cell survival rate of the LPS group (76.15%) was lower than that of the Con group, while that of the YG3912 extract treatment groups was higher than that of the LPS group;

[0120] 2) For the YG3912 fermentation filtrate treatment group, there was no significant change in the cell survival rate of the 1 / 30000, 1 / 10000, 1 / 3000 and 1 / 1000 concentration groups, while there was a decrease in the 1 / 100 and 1 / 300 concentration groups.

[0121] 3) For the YG3912 exosome-treated group, except for the 1 / 1000 concentration group whose cell survival rate was slightly lower than that of the Con group, the cell survival rates of other concentration groups were significantly improved, especially the 1 / 10000 concentration group whose cell survival rate even reached 121.74%.

[0122] 4) For the YG3912 lysate treatment groups, the cell survival rates of the 1 / 30000, 1 / 300 and 1 / 100 concentration groups were all improved compared to the Con group.

[0123] In summary, YG3912 exosomes and lysates can promote the proliferation and survival of macrophages at appropriate concentrations.

[0124] (2) Effects of YG3912 fermentation filtrate, exosomes, and lysates on the levels of cytokine secretion by RAW264.7 cells

[0125] Take RAW264.7 cells in the logarithmic growth phase, resuspend them and dilute them to 1×10 5 2 ml of cell suspension was added to each well of a 12-well plate and placed in an incubator at 37°C and 5% CO 2 Incubate incubator for 24 h. Afterwards, discard the culture medium, add complete culture medium, different concentrations of YG3912 fermentation filtrate, lysate and exosome dilution, 2 ml per well, set up 3 replicates, and place at 37°C, 5% CO 2 The culture was continued in the incubator for 24 hours. After 24 hours, the cell culture fermentation filtrate was collected and the level of cytokine TNF-α was detected by enzyme-linked immunosorbent assay (ELISA) detection kit (Enzyme-linked Biology, IC50325-1). The operation steps were carried out according to the product instructions provided by the kit company.

[0126] On the other hand, after collecting the cell fermentation filtrate, the cells were washed twice with PBS and then the PBS was discarded. The cells were lysed with TransZol (Full Formula Gold, R61225) to extract RNA for subsequent detection of the gene expression levels of inflammatory factors TNF-α, IL-6, IL-1β and iNOS (after obtaining the Ct value, 2 -△△Ct formula for calculation).

[0127] The ELISA test results of TNF-α in the YG3912 fermentation filtrate, lysate and exosome treatment groups were as follows Figure 7 As shown in Figures A, B, and C; Figure 7 As shown in Figures A, B, and C, the TNF-α content in the LPS group was 11-15 times that in the control group, which indicates that LPS treatment can stimulate macrophages to secrete the cytokine TNF-α.

[0128] Figure 7 Figure A shows that the TNF-α secretion was increased in the YG3912 fermentation filtrate low concentration (1 / 300 and 1 / 100) treatment groups compared with the control group, and there was a significant difference in the 1 / 300 treatment group (P < 0.05). Figure 7 Figure B shows that in the YG3912 lysate-treated group, compared with the control group, the TNF-α secretion in the 1 / 10000 and 1 / 1000 concentration groups was significantly increased (P<0.01), and the TNF-α secretion in the 1 / 300 and 1 / 100 concentration groups increased more (P<0.0001), increasing by 13.03 and 20.24 times, respectively. Figure 7 Figure C shows that the low-concentration treatment group of YG3912 exosomes reduced the secretion of TNF-α, and only the 1 / 100 concentration group significantly increased the secretion of TNF-α (P<0.001). These results show that the fermentation filtrate, lysate and exosomes of YG3912 can stimulate macrophages to secrete TNF-α at appropriate concentrations, thereby inducing cellular immune regulation.

[0129] The gene expression results of TNF-α, IL-6, IL-1β and iNOS in the YG3912 fermentation filtrate, lysate and exosome treatment groups are shown in Figure 8 shown.

[0130] Figure 8 Figures AD in the figure respectively show the gene expression levels of TNF-α, IL-6, IL-1β and iNOS in the YG3912 fermentation filtrate treatment group, which show that: compared with the control group, the TNF-α gene level of the YG3912 fermentation filtrate treatment group at each concentration did not change significantly; compared with the control group, the IL-6 gene level of the 1 / 100 concentration group increased significantly (P<0.0001), with an increase of up to 17.89 times; compared with the control group, the IL-1β gene level of the 1 / 100 and 1 / 300 concentration groups increased significantly (P<0.0001), with an increase of up to 8.21 and 36.81 times, respectively; compared with the control group, the iNOS gene level of all concentration groups increased significantly.

[0131] Figure 8 Figures EH in the figure show the gene expression levels of TNF-α, IL-6, IL-1β and iNOS in the YG3912 lysate treatment group, respectively. It shows that compared with the control group, except for the 1 / 30000 concentration group, the TNF-α gene level of each lysate treatment group increased significantly, and this increase had a certain dose-dependent effect, with the highest increase reaching 3.75 times; compared with the control group, the IL-6 gene level of the 1 / 1000 (P<0.001), 1 / 300 (P<0.0001) and 1 / 100 (P<0.0001) concentration groups increased significantly, In particular, the increase in the 1 / 100 concentration group was as high as 176.20 times; compared with the control group, the IL-1β gene levels in the 1 / 1000, 1 / 300 and 1 / 100 concentration groups were significantly increased (P<0.0001), in particular, the increase in the 1 / 100 concentration group was as high as 1003.59 times; compared with the control group, the iNOS gene levels in the 1 / 1000 (P<0.05), 1 / 300 (P<0.0001) and 1 / 100 (P<0.0001) concentration groups were significantly increased, in particular, the increase in the 1 / 100 concentration group was as high as 41.92 times.

[0132] Figure 8The IL graph in the figure shows the gene expression levels of TNF-α, IL-6, IL-1β and iNOS in the YG3912 exosome-treated group, respectively, which shows that: compared with the control group, the TNF-α gene level in the 1 / 300 (P<0.01) and 1 / 100 (P<0.001) concentration groups was significantly increased; compared with the control group, the IL-6 gene level in all concentration groups was decreased; compared with the control group, the IL-1β gene level in the 1 / 300 and 1 / 100 concentration groups was significantly increased (P<0.0001), especially, the increase in the 1 / 100 concentration group was as high as 11.17 times; compared with the control group, the iNOS gene level in the 1 / 300 and 1 / 100 concentration groups was significantly increased (P<0.0001), especially, the increase in the 1 / 100 concentration group was as high as 12.32 times.

[0133] Example 5: Evaluation of the anti-photoaging efficacy of Bifidobacterium adolescentis YG3912

[0134] (1) Establishment of UVB-irradiated HaCaT photoaging cell model

[0135] For the construction method of the cell photoaging model, please refer to the article Protective Effect of Bifidobacterium animalis subs. lactis MG741 as Probiotics against UVB-Exposed Fibroblasts and Hairless Mice (DOI: 10.3390 / microorganisms10122343).

[0136] Specifically, HaCaT cells were cultured in a high-glucose medium (DMEM) containing 10% special fetal bovine serum (FBS) and 1% 100× penicillin-streptomycin mixture (S / P) at 37°C and 5% CO. 2 The cells were cultured in a cell culture incubator; when the cell confluence reached about 80%, the culture medium was removed, the cells were washed with sterile PBS, sterile PBS was added to the cells, and UVB irradiation was started; the UVB irradiation conditions were: using a four-purpose ultraviolet analyzer (Qilin Bell, ZF-2, UVB312nm, 30W) for 40s to construct a cell photoaging model.

[0137] (2) YG3912 fermentation filtrate, lysate and exosomes alleviate UVB-induced photoaging of HaCaT cells

[0138] HaCaT cells were cultured to the logarithmic phase and the cells were collected. A normal control group (Con), a model group (UVB), and treatment groups (S-10^4, R-3*10^4, and W-3000) were set up in a 12-well plate. The concentration of the cell suspension was adjusted to 3×10 5 Each well was inoculated with 1 mL of cell suspension. The cell culture plate was placed at 37°C and 5% CO 2 Culture in incubator for 24 hours.

[0139] After 24 hours of culture, the cells grew to a monolayer, the culture medium was removed, and the cells were washed twice with sterile PBS, and 1 mL of sterile PBS was added to each well. The control group was wrapped with tin foil to avoid UVB irradiation. The model group and the treatment group were irradiated with UVB for 40 seconds. After irradiation, 1 mL of complete culture medium was added to the control group and the model group, and 1 mL of complete culture medium containing 10,000-fold diluted YG3912 fermentation filtrate (i.e., the aforementioned "S-10^4" group), 30,000-fold diluted lysate (i.e., the aforementioned "R-3*10^4" group) and 3,000-fold diluted exosomes (i.e., the aforementioned "W-3000" group) was added to the treatment group. After 24 hours of culture, the changes in cell morphology were observed under a microscope (20×), and the cell supernatant was collected. The protein levels of inflammatory factors IL-1β, IL-6, and IL-8 in the cell supernatant were determined using the MQ60 PLUS fully automatic chemiluminescence immunoassay system (Thermogene Bio). TransZol (Full Gold, R61225) was used to lyse cells and extract RNA for subsequent RT-PCR. The inflammatory factor IL-1β and matrix metalloproteinases MMP-2 and MMP-9 in the cell supernatant were detected at the gene level (after obtaining the Ct value, 2 -△△Ct formula for calculation).

[0140] The results of observing cell morphology under a microscope (20×) are as follows Fig. 9 As shown; Fig. 9 The results showed that under a 20× microscope field of view, UVB caused large-area cell detachment and poor fusion, while treatment with YG3912 fermentation filtrate, lysate and exosomes restored the cell state, proving that YG3912 fermentation filtrate, lysate and exosomes have the function of repairing cell damage.

[0141] The protein levels of inflammatory factors IL-1β, IL-6 and IL-8 were measured as follows: Fig.10 As shown in Figures A, B, and C; Fig.10As shown in Figures A, B, and C, after UVB irradiation, the levels of IL-1β, IL-6, and IL-8 increased significantly compared with the control group. However, YG3912 fermentation filtrate, lysate, and exosome treatment significantly reduced the levels of these factors, with inhibition rates of 28%, 38%, and 40% for IL-6, 43%, 39%, and 26% for IL-8, and 43%, 44%, and 32% for IL-1β, respectively.

[0142] The results of gene level determination of inflammatory factor IL-1β and matrix metalloproteinases MMP2 and MMP9 are as follows Fig.10 As shown in Figure D. Fig.10 As shown in Figure D, the trend of changes in the gene level of the inflammatory factor IL-1β is consistent with its protein level. This further confirms that the YG3912 fermentation filtrate, lysate and exosomes can inhibit the increased expression of the inflammatory factor IL-1β induced by UVB. Fig.10 It can also be seen from Figure D that after UVB irradiation, the gene expression levels of matrix metalloproteinases MMP2 and MMP9 increased significantly, while YG3912 fermentation filtrate, lysate and exosome treatment significantly downregulated their gene expression levels, indicating that YG3912 fermentation filtrate, lysate and exosomes can significantly inhibit the UVB-induced increase in MMP2 and MMP9 gene expression.

[0143] Judging from the above results, YG3912 fermentation filtrate, lysate and exosomes can significantly inhibit the increased expression of inflammatory factors IL-6, IL-8, and IL-1β caused by UVB irradiation, and can alleviate the abnormal increase in the gene levels of matrix metalloproteinases MMP2 and MMP9 caused by UVB, thereby alleviating UVB-induced cellular photoaging.

[0144] (3) YG3912 fermentation filtrate, lysate and exosomes alleviate UVB-induced ROS content in HaCaT cells

[0145] HaCaT cells were cultured to the logarithmic phase and the cells were collected. A normal control group (Con), a model group (UVB), and treatment groups (S-10^4, R-3*10^4, and W-3000) were set up in a 12-well plate. The concentration of the cell suspension was adjusted to 1.5×10 5 Each well was inoculated with 1 mL of cell suspension. The cell culture plate was placed in a 5% CO 2 , culture in a 37°C incubator for 24 hours.

[0146] After 24 hours of culture, the cells grew to a monolayer, the culture medium was removed, and the cells were washed twice with sterile PBS, and 1 mL of sterile PBS was added to each well. The control group was wrapped in tin foil to avoid UVB irradiation. The model group and the treatment group were irradiated with UVB for 40 seconds. After irradiation, 1 mL of complete culture medium was added to the control group and the model group, and 1 mL of complete culture medium containing 10^4 times diluted YG3912 fermentation filtrate (i.e., the aforementioned "S-10^4" group) or 3*10^4 times YG3912 lysate (i.e., the aforementioned "R-3*10^4" group) or 3000 times YG3912 exosomes (i.e., the aforementioned "W-3000" group) was added to the treatment group. After culturing for 24 h, the changes in cell morphology were observed under a microscope (20×). After washing twice with sterile PBS, the intracellular ROS fluorescence level was determined by DCFH-DA fluorescence using a reactive oxygen species detection kit (purchased from S0033S, Bio-Tech), and the relative fluorescence intensity of intracellular ROS was quantified using Image J to evaluate the effects of YG3912 fermentation filtrate, lysate and exosomes on UVB-induced cellular ROS levels.

[0147] The results are as follows Fig.11 shown.

[0148] Depend on Fig.11 As can be seen from Figure A:

[0149] 1) Under a 20× microscope field of view, UVB caused a large area of ​​cells to fall off and the degree of fusion to deteriorate. Treatment with YG3912 fermentation filtrate, lysate and exosomes restored the cell state to some extent, indicating that YG3912 fermentation filtrate, lysate and exosomes have the function of repairing cell damage;

[0150] 2) Under a 20× microscope field of view, UVB can cause a significant increase in intracellular ROS levels, while YG3912 fermentation filtrate, lysate, and exosome treatment can significantly inhibit the UVB-induced increase in cellular ROS levels. This conclusion can also be Fig.11 The statistical results of Figure B are obtained.

[0151] Anti-photoaging is an important aspect of skin aging, which involves the damage of ultraviolet rays to skin cells, leading to skin pigmentation, reduced elasticity, increased wrinkles and other problems. Ultraviolet rays can destroy the natural barrier of the skin, trigger the production of free radicals, and further accelerate the skin aging process. The extract of strain YG3912 (including fermentation filtrate, exosomes and lysates) can significantly improve the cell morphological disorder, poor fusion, and abnormal increase of inflammatory factors and matrix metalloproteinases caused by UVB irradiation, thereby having an anti-photoaging effect. It can be seen that the YG3912 strain or its extract can be used to develop economically effective natural anti-photoaging skin care products.

[0152] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A strain of Bifidobacterium adolescentis YG3912, characterized in that It is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit date being June 20, 2024 and the deposit number being CGMCC No.31020.

2. The inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of Bifidobacterium adolescentis YG3912 as claimed in claim 1.

3. The inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of Bifidobacterium adolescentis YG3912 according to claim 2, characterized in that: The extract of Bifidobacterium adolescentis YG3912 is a lysate and / or exosomes extracted therefrom.

4. The inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of Bifidobacterium adolescentis YG3912 according to claim 3, characterized in that: The method for extracting the lysate comprises the following: The Bifidobacterium adolescentis YG3912 is fermented and cultured, the fermentation broth is centrifuged, the cells are collected, the cells are resuspended with PBS, and the cell walls are broken by ultrasound or enzymatic hydrolysis to obtain a lysate; preferably, the obtained lysate is sterilized; And / or, the method for extracting exosomes comprises the following: The Bifidobacterium adolescentis YG3912 is fermented and cultured, the fermentation broth is centrifuged, and the fermentation supernatant is collected; the obtained fermentation supernatant is centrifuged, filtered, and concentrated to obtain exosomes.

5. The inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of Bifidobacterium adolescentis YG3912 according to claim 4, characterized in that: The centrifugation procedure of the fermentation supernatant is: first centrifuge at 200-400g for 8-15min, then centrifuge at 1500-3000g for 15-30min, and finally centrifuge at 8000-12000g for 20-40min; And / or, the filtration of the fermentation supernatant is: filtering with a filter membrane having a pore size of 0.2-2.0 μm; And / or, the fermentation supernatant is concentrated by using a 400-600 kd hollow fiber to concentrate the fermentation clarified liquid.

6. A composition, characterized in that The composition comprises the Bifidobacterium adolescentis YG3912 as described in claim 1 and / or the inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of Bifidobacterium adolescentis YG3912 as described in any one of claims 2-5 as active ingredients.

7. The composition according to claim 6, characterized in that The composition is a pharmaceutical composition or a cosmetic composition, which further comprises a pharmaceutically or cosmetically acceptable carrier and / or excipient.

8. The composition according to claim 6 or 7, characterized in that The composition is an external dosage form; the external dosage form is a dosage form selected from the following: aqueous solution, emulsion, ointment, cream, gel, powder, oil.

9. The composition according to claim 6 or 7, characterized in that The composition further comprises other active agents; the other active agents comprise any one or more of antioxidants, cell active agents, moisturizing agents, anti-aging agents and anti-dermatitis agents.

10. Use of the Bifidobacterium adolescentis YG3912 according to claim 1, or the inactivated bacteria, extract, culture, culture supernatant, fermentation product and / or fermentation supernatant of the Bifidobacterium adolescentis YG3912 according to any one of claims 2 to 5, or a composition comprising one or more of the foregoing substances in the preparation of a product for the following purposes: 1) Promote skin repair; 2) Reduce, inhibit or eliminate skin inflammation or allergies; 3) Anti-skin aging; 4) Enhance skin immune function.

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