A strain of Lactobacillus helveticus and its application

Abstract

This invention discloses a strain of *Lactobacillus helveticus* and its applications, relating to the field of microbial technology. The *Lactobacillus helveticus* strain disclosed in this invention (… Lactobacillus helveticus The sample, ProfMIC-226, was deposited at the China Center for Type Culture Collection on August 3, 2022, with accession number CCTCCNO:M20221224. Experiments have shown that ProfMIC-226 possesses anti-aging, cell proliferation-promoting, moisturizing, and skin barrier-repairing functions, and can be used in pharmaceutical preparations.

Description

Technical Field

[0001] This invention belongs to the technical field of microorganisms, specifically relating to a strain of Lactobacillus helveticus and its applications. Background Technology

[0002] The skin barrier is a structural barrier formed by the epidermal morphology cells of the stratum corneum and the lipids between the keratinocytes. The skin barrier prevents excessive water loss from the body and protects against harmful substances such as chemicals or microorganisms from entering our bodies. The keratinocyte epithelium, which makes up the surface of dead keratinocytes, plays an important role in the stability of intercellular lipids.

[0003] The structural lipid ceramides in the stratum corneum gradually increase in content during the differentiation of the basal layer into keratinocytes. Reaching the stratum corneum, they are excreted into the intercellular spaces, forming a barrier to prevent moisture loss. The stratum corneum's hydrophilic and barrier functions, along with the natural moisturizing factors it contains (amino acids, lactates, and sugars), ensure that it typically contains 10-30% water. This environment provides a cradle for the growth of the skin's own microbial community. However, with age, the water content of the stratum corneum gradually decreases. When the water content falls below 10%, various skin problems arise. Damage to the skin barrier leads to dry skin, skin aging, and inflammation, ultimately resulting in skin aging.

[0004] Recent research has demonstrated that the skin microbiome regulates the expression of various innate factors, and that resident bacteria on the skin are not merely passively residing organisms; they actively participate in the host's immune system through the intact skin barrier. The skin's innate immune system, in conjunction with the skin microbiome, forms a barrier against pathogenic and opportunistic pathogens. The skin possesses self-renewal capabilities, and the breakdown of phospholipids, sterols, and keratin by skin microbes allows skin cells to absorb these substances, promoting cell growth, delaying aging, and reducing wrinkles. Therefore, developing probiotic-related products utilizing microecological technology has significant practical implications. Summary of the Invention

[0005] In view of this, the object of the present invention is to provide a strain of Lactobacillus helveticus and its applications.

[0006] This invention provides Lactobacillus helveticus ( Lactobacillus helveticus The strain is Lactobacillus helveticus ProfMIC-226, which was deposited on August 3, 2022, at the China Center for Type Culture Collection (CCTCC, address: No. 299 Bayi Road, Wuchang District, Wuhan, 430072, China, Wuhan University, with accession number CCTCCNO: M20221224).

[0007] Another object of the present invention is to provide the use of the above-mentioned Lactobacillus helveticus in the preparation of products that improve the condition of skin cells.

[0008] Preferably, the improvement in skin cell condition includes at least one of anti-aging, cell proliferation promotion, moisturizing, and skin barrier repair.

[0009] In some implementations, the anti-aging effect refers to upregulating the expression of extracellular matrix-related genes and upregulating autophagy-related genes. LC3B Expression and upregulation of immune regulatory factor-related genes MOR The expression and / or upregulation of cell growth factor-related genes, including extracellular matrix-related genes, are discussed. SPTSSA and / or COL1A1 The cell growth factor-related genes include FGF2 and / or FGF21 .

[0010] In some embodiments, the anti-aging effect involves downregulating the expression of genes related to extracellular matrix degradation and / or downregulating apoptosis-related factors. BAX The expression of the genes related to the degradation of the extracellular matrix includes MMP At least one member of the family.

[0011] In some implementations, the promotion of cell proliferation includes promoting the proliferation of skin keratinocytes and / or promoting the proliferation of fibroblasts.

[0012] In some implementations, the moisturizing includes upregulating moisturizing-related genes. GBA The expression.

[0013] In some implementations, the skin barrier repair includes at least one of restoring cell vitality and upregulating the expression of barrier repair-related genes.

[0014] In some implementation schemes, the product is a pharmaceutical product.

[0015] In some implementations, the *Lactobacillus helveticus* in the product includes one or two of the following:

[0016] (1) Live and / or inactivated Lactobacillus helveticus;

[0017] (2) Cultures, exosomes, lysates and / or extracts of Lactobacillus helveticus.

[0018] The Lactobacillus helveticus disclosed in this invention ( Lactobacillus helveticus ProfMIC-226, with accession number CCTCCNO: M20221224, has been shown in experiments to promote cell proliferation, repair the skin barrier, moisturize, and have anti-aging functions, and can be used in pharmaceutical preparations.

[0019] Biological deposit description

[0020] Lactobacillus helveticus ( Lactobacillus helveticus ProfMIC-226 was deposited on August 3, 2022, at the China Center for Type Culture Collection (CCTCC, No. 299 Bayi Road, Wuchang District, Wuhan, 430072, China), with accession number CCTCCNO: M20221224. Detailed Implementation

[0021] This invention provides a strain of *Lactobacillus helveticus* and its applications. Those skilled in the art can refer to this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can clearly modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to implement and apply the technology of this invention.

[0022] This invention provides a sample of *Lactobacillus helveticus* ProfMIC-226 from the feces of a seven-year-old healthy boy, identified as *Lactobacillus helveticus* by 16S rDNA analysis. Lactobacillus helveticus This strain is Gram-positive and appears as rods under a microscope. When grown on MRS plates, it forms smooth, translucent, round colonies that are white with regular edges. In MRS liquid medium, it grows in a uniformly turbid manner, and the cells form a white precipitate after prolonged storage. The optimal growth temperature is 37°C.

[0023] Lactobacillus helveticus ( Lactobacillus helveticus (), depositary institution: China Center for Type Culture Collection, address: No. 299 Bayi Road, Wuchang District, Wuhan, Wuhan University, deposit date: August 3, 2022, accession number: CCTCCNO: M20221224.

[0024] In order to investigate the anti-aging effect of the aforementioned Lactobacillus helveticus, this invention measured indicators related to cell aging, including at least one of extracellular matrix synthesis, extracellular matrix degradation, apoptosis, and autophagy.

[0025] This invention uses HaCaT keratinocytes as test subjects to study the effects of *Lactobacillus helveticus* on genes related to extracellular matrix synthesis and degradation in HaCaT keratinocytes. The results show that *Lactobacillus helveticus* can upregulate genes related to extracellular matrix synthesis. COL1A1 The expression of these genes promotes the synthesis of extracellular matrix and downregulates genes related to the degradation of extracellular matrix. MMP The expression of 1 inhibits the degradation of the extracellular matrix.

[0026] This invention uses HaCaT keratinocytes as test subjects to study the effect of *Lactobacillus helveticus* on autophagy in HaCaT keratinocytes. The results show that *Lactobacillus helveticus* can upregulate autophagy-related genes. LC3B The expression of [a substance] promotes autophagy to clear senescent cells.

[0027] This invention uses HFF fibroblasts as the test subject to study the effects of *Lactobacillus helveticus* on genes related to extracellular matrix synthesis in HFF cells. The results show that *Lactobacillus helveticus* can upregulate genes related to extracellular matrix synthesis. SPTSSA The expression of [a substance] promotes the synthesis of extracellular matrix.

[0028] This invention uses HFF fibroblasts as the test subject to study the effect of *Lactobacillus helveticus* on HFF cell apoptosis. The results show that *Lactobacillus helveticus* can downregulate apoptosis-related genes. BAX The expression of [a substance] inhibits apoptosis.

[0029] This invention uses HFF fibroblasts as the test subject to study the effect of *Lactobacillus helveticus* on the expression of immunomodulatory factor-related genes in HFF keratinocytes. The results show that *Lactobacillus helveticus* can upregulate the expression of immunomodulatory factor-related genes and promote cell growth.

[0030] This invention uses HFF fibroblasts as the test subject to study the effect of *Lactobacillus helveticus* on the expression of keratinocyte growth factor-related genes in HFF. The results show that *Lactobacillus helveticus* can upregulate genes related to immune regulatory factors. MOR The expression of [something] enhances the cell's immune regulatory capacity.

[0031] Furthermore, in this invention, the promotion of cell proliferation includes promoting the proliferation of keratinocytes and / or promoting the proliferation of fibroblasts. This invention uses HaCaT keratinocytes as test subjects to study the cell proliferation-promoting effect of the aforementioned *Lactobacillus helveticus*. The results show that *Lactobacillus helveticus* can promote the repair of SDS-induced HaCaT keratinocyte damage, increasing the cell proliferation rate by 127.25%~150.75%. This invention uses HFF fibroblasts as test subjects to study the cell proliferation-promoting effect of the aforementioned *Lactobacillus helveticus*. The results show that *Lactobacillus helveticus* can promote the proliferation of HFF fibroblasts by 107.44%~155.48%.

[0032] Furthermore, in this invention, the moisturizing effect includes, but is not limited to, upregulating the expression of moisturizing-related genes, which include, but are not limited to, those related to moisturizing. GBAThis invention uses HaCaT keratinocytes as test subjects to study the moisturizing effect of *Lactobacillus helveticus* on the skin. The results show that *Lactobacillus helveticus* can upregulate moisturizing-related genes. GBA The expression of this substance promotes skin cell hydration.

[0033] Furthermore, in this invention, the repair of the skin barrier includes at least one of restoring cell vitality and upregulating the expression of barrier repair-related genes; the restoration of cell vitality is to repair SDS-induced cell damage, and the barrier repair-related genes include... FLG, IVL, OCLN and OVOL1 At least one of the following. Furthermore, this invention used HaCaT keratinocytes as test subjects to study the SDS damage repair ability of the aforementioned *Lactobacillus helveticus*. The results showed that *Lactobacillus helveticus* can repair cell damage caused by SDS, that is, *Lactobacillus helveticus* can restore cell vitality, increase cell proliferation rate, and upregulate the expression of barrier repair-related genes in undamaged cells.

[0034] The present invention also provides the application of *Lactobacillus helveticus* ProfMIC-226 described in the above technical solution in the preparation of products that improve the condition of skin cells. The *Lactobacillus helveticus* in the application or product exists in the form of live, dead, or intermittently sterilized bacteria, or in the form of lysates and / or extracts, or in the form of bacterial products, supernatants, or derivatives. The derivatives are preferably selected from: metabolites, metabolic biological products, exosomes, probiotics, cell walls and their components, extracellular polysaccharides, and compounds containing immunogenic components, preferably selected from: supernatants and inactivated bacterial cells.

[0035] Furthermore, the product includes the Lactobacillus helveticus;

[0036] Furthermore, the product includes lysates, extracts, and metabolites of the Lactobacillus helveticus;

[0037] Furthermore, the product includes a bacterial flora containing the aforementioned Lactobacillus helveticus;

[0038] Furthermore, the product includes a microbial agent containing *Lactobacillus helveticus* and its lysates, extracts, and metabolites, or a microbial agent containing a flora of *Lactobacillus helveticus*. This invention does not limit the scope of the invention.

[0039] The dosage form of the product includes, but is not limited to, at least one of the following: creams, lotions, oils, liquids, gels, powders, and freeze-dried products; the present invention does not limit this to any particular type.

[0040] The present invention also provides a method for improving skin condition, including applying, steaming or injecting the products described in the above technical solutions, without limiting the scope of the invention.

[0041] The test materials used in this invention are all commercially available products. The invention will be further illustrated below with reference to the embodiments:

[0042] Example 1: Separation of ProfMIC-226

[0043] Sampling was performed on the feces of a healthy seven-year-old boy. After appropriate processing, the samples were shaken and mixed in physiological saline. The supernatant was streaked onto MRS agar plates and incubated at 37°C for 48 hours. White colonies were picked and repeatedly inoculated until uniform single colonies were obtained, which were named ProfMIC-226.

[0044] Gram staining microscopy: strain ProfMIC-226 is Gram positive and appears as rod-shaped under a microscope; when grown on MRS plates, it forms small, white, smooth, round, semi-transparent colonies with neat edges; in MRS liquid medium, it grows in a uniformly turbid manner, and the bacterial cells form a white precipitate after prolonged storage.

[0045] Example 2: Nucleic acid identification of ProfMIC-226

[0046] 1. 16S rDNA gene sequence analysis

[0047] Single colonies were picked and placed in MRS liquid medium, incubated overnight at 37°C, and then centrifuged at 12,000 rpm for 1 min to collect the bacterial cells. The DNA extraction kit was then followed. Universal bacterial primers 27F and 1492R were used. The PCR amplification system was 50 μL, with pre-denaturation at 95°C for 5 min; followed by 35 cycles of 94°C for 15 s, 57°C for 15 s, and 72°C for 40 s; and a final extension at 72°C for 10 min.

[0048] 2. Results

[0049] After comparing the sequencing results of the PCR product with the published standard sequences in GenBank (BLASTN), the ProfMIC-226 strain was identified as *Lactobacillus helveticus*. Lactobacillus helveticus ).

[0050] Example 3: ProfMIC-226 promotes SDS-induced HaCaT damage repair in immortalized human keratinocytes.

[0051] 1. Preparation of ProfMIC-226 supernatant

[0052] Single colonies of *Lactobacillus helveticus* ProfMIC-226 were picked and cultured on MRS liquid medium at 37°C for 16–18 hours. The culture was then analyzed using a microplate reader, and the OD value was adjusted by dilution with PBS. 600=0.2, 121℃, autoclave for 30 min, centrifuge at 12000 rpm for 2 min, and filter through a 0.22 μm filter membrane to obtain the supernatant.

[0053] 2. Experiments promoting HaCaT cell repair

[0054] Seed HaCaT cells (5×10) 4 Cells / well were added to 96-well plates and cultured overnight until cell adhesion occurred. 100 μl of 50 μg / ml SDS was added to each well, and the plates were incubated at 37°C for 8 h in a 5% CO2 incubator. 5% supernatant was added to each well (for the control group, an equal volume of PBS was used instead of supernatant), and the plates were incubated for 24 h. 10 μl of CCK-8 solution was added to each well, and the plates were incubated for 4 h. The absorbance at 450 nm was then measured.

[0055] The formula and results for calculating cell proliferation rate are shown in the table below:

[0056]

[0057] The results in the table above show that ProfMIC-226 supernatant can repair HaCaT keratinocyte damage caused by SDS, that is, promote the proliferation of skin keratinocytes, with a proliferation rate of 127.25~150.75%.

[0058] Example 4: Experiment on ProfMIC-226 Upregulation of HaCaT Moisturizing-Related Gene Expression

[0059] 1. Preparation of ProfMIC-226 supernatant

[0060] The preparation method is the same as in Example 3.

[0061] 2. Experiment on upregulating the expression of HaCaT moisturizing-related genes

[0062] Inoculate with human immortalized keratinocytes HaCaT (2 ml / well, containing 5 × 10⁶ cells). 5 Cells were transferred to 6-well plates and cultured overnight at 37°C with 5% CO2 until cell adhesion. 5% (v / v) supernatant was added (the control group was replaced with an equal volume of PBS), and after 24 h of culture, lysis buffer was added to extract total RNA. RNA concentration and purity were measured, and then reverse transcribed into cDNA. GAPDH This is an internal reference gene, detected using real-time qPCR. GBA Gene expression. Using an equal volume of PBS treatment group as a control (relative gene expression fold F=1), 2... -ΔΔCT The F-value of each sample was calculated using the method described above.

[0063] Formula: F=2 -ΔΔCT ,in:

[0064] △CT 实验=CT 实验 -CT 内参（实验） ;

[0065] △CT 对照 =CT 对照 -CT 内参（对照） ;

[0066] △△CT=△CT 实验 -△CT 对照 .

[0067] The results are shown in the table below:

[0068]

[0069] In vitro cell experiments showed that the *Lactobacillus helveticus* ProfMIC-226 of this invention upregulates the hydration-related glucocerebrosidase gene. GBA The expression level was upregulated by 1.31–1.62, indicating that ProfMIC-226 promotes skin hydration.

[0070] Example 5: Experiment on ProfMIC-226 promoting the expression of HaCaT barrier repair-related genes

[0071] 1. Preparation of ProfMIC-226 supernatant and inactivated bacterial cells

[0072] Single colonies of *Lactobacillus helveticus* ProfMIC-226 were picked and cultured on MRS liquid medium at 37°C for 16–18 hours. The culture was then analyzed using a microplate reader, and the OD value was adjusted by dilution with PBS. 600 =0.2, autoclaved at 121℃ for 30 min, centrifuged at 12000 rpm for 2 min, and filtered through a 0.22 μm filter to obtain the supernatant. The centrifuged precipitate was resuspended in an appropriate amount of PBS and diluted to adjust the OD. 600 =0.2, indicating inactivated bacteria.

[0073] 2. Experiments to promote the expression of genes related to HaCaT barrier repair

[0074] Inoculate with human immortalized keratinocytes HaCaT (2 ml / well, containing 5 × 10⁶ cells). 5 Cells were transferred to 6-well plates and incubated overnight at 37°C with 5% CO2 until cell adhesion. 5% (v / v) supernatant and 10% (v / v) inactivated bacterial cells were added (for the control group, equal volumes of PBS were used instead of supernatant / inactivated cells). After 24 h of incubation, lysis buffer was added, total RNA was extracted, and the RNA concentration and purity were measured before reverse transcription to cDNA. GAPDH This is an internal reference gene, detected using real-time qPCR. FLG , IVL , OCLN andOVOL1 Gene expression was measured. A control group treated with an equal volume of PBS was used (relative gene expression fold F=1). Two... -ΔΔCT The F-value of each sample was calculated using the method described above.

[0075] The results are shown in the table below:

[0076]

[0077]

[0078] In vitro cell experiments showed that the supernatant and inactivated cells of *Lactobacillus helveticus* ProfMIC-226 of this invention upregulated the filaggrin gene, a skin barrier repair-related factor. FLG outer skin protein gene IVL Tight junction proteins OCLN and OVO-like transcription factor 1 gene OVOL1 The expression level was upregulated by 1.12 to 2.69 times, indicating that ProfMIC-226 promotes skin barrier repair.

[0079] Example 6: Experiment on the regulation of extracellular matrix / autophagy-related gene expression in photoaging HaCaT cells by ProfMIC-226

[0080] 1. Preparation of ProfMIC-226 supernatant and inactivated bacterial cells

[0081] The preparation method is the same as in Example 5.

[0082] 2. HaCaT cell preparation and ultraviolet damage

[0083] HaCaT cells were digested and then dispensed into 0.5 ml / well (containing 2 × 10⁶ cells). 5 Cells were seeded into 24-well plates and incubated overnight at 37°C with 5% CO2. The total dose of cells in the wells was 2 J / cm². 2 UVB radiation damage.

[0084] 3. Add ProfMIC-226

[0085] 5% (v / v) of supernatant and 10% (v / v) of inactivated bacterial cells were added to stimulated HaCaT cells (the control group was treated with an equal volume of PBS instead of supernatant / inactivated bacterial cells). Each group was in triplicate and incubated overnight at 37°C.

[0086] 4. Detection of the relative fold increase in extracellular matrix / autophagy-related gene expression using qPCR.

[0087] After discarding the culture medium from the cells, lysis buffer was added, total RNA was extracted, and the RNA concentration and purity were measured before reverse transcription into cDNA. GAPDH As internal reference genes, extracellular matrix-related genes were detected using real-time qPCR. COL1A1 Autophagy-related genes LC3B and genes related to the degradation of the extracellular matrix MMP1 The expression of genes was measured. Using a control group gene expression fold of F=1, 2... -ΔΔCT The F-value of each sample was calculated using the method described above.

[0088] Supernatant upregulates autophagy-related genes LC3B Downregulation of genes related to extracellular matrix degradation MMP1 The results are shown in the table below:

[0089]

[0090] Inactivated bacteria upregulate extracellular matrix genes COL1A1 The results are shown in the table below:

[0091]

[0092] In vitro cell experiments showed that the *Lactobacillus helveticus* ProfMIC-226 of this invention upregulates the type I collagen α1 chain gene. COL1A1 Genes related to autophagy LC3B The expression of the gene was relatively high, ranging from 1.27 to 3.97 fold; it has the function of downregulating matrix metalloproteinase family genes associated with the degradation of the extracellular matrix. MMP1 The role of expression, the relative fold increase of gene expression is 0.63~0.84.

[0093] Example 7: ProfMIC-226 promotes the proliferation of human fibroblasts (HFF).

[0094] 1. Preparation of ProfMIC-226 supernatant and inactivated bacterial cells

[0095] The preparation method is the same as in Example 5.

[0096] 2. HFF cell preparation and ProfMIC-226 addition

[0097] HFF cells cultured in DMEM were seeded at 100 μL / well (each well containing 3 × 10⁶ cells). 4Cells were transferred to 96-well plates and cultured overnight until adherence. 200 μM H₂O₂ was prepared, with 100 μl added to each well, and the cells were incubated at 37°C for 1 h in a 5% CO₂ incubator. The original culture medium was discarded, and the cells were washed twice with PBS. 100 μl of fresh culture medium was added to each well, followed by the addition of 5% (v / v) supernatant and 10% (v / v) inactivated bacterial cells (for the control group, equal volumes of PBS were used instead of supernatant / inactivated bacterial cells). The cells were incubated for 24 h. 10 μl of CCK-8 solution was added to each well, and the cells were incubated for 4 h. The absorbance A at 450 nm was measured, and the calculation formula and results are shown in the table below.

[0098]

[0099] In vitro cell experiments showed that the Lactobacillus helveticus ProfMIC-226 of the present invention promoted the proliferation of human fibroblasts (HFF) with a proliferation rate of 107.44%~155.48%.

[0100] Example 8: Experiment on the regulation of extracellular matrix / immunomodulatory factors / cell growth factors / apoptosis-related gene expression by ProfMIC-226 in HFF cells affected by oxidative damage

[0101] 1. Preparation of ProfMIC-226 supernatant and inactivated bacterial cells

[0102] The preparation method is the same as in Example 5.

[0103] 2. Preparation of HFF cells and H2O2-induced oxidative damage

[0104] HFF cells cultured in DMEM were digested and then injected into wells at a concentration of 0.5 ml (containing 2 × 10⁶ cells / well). 5 Cells were seeded into 24-well plates and incubated overnight at 37°C with 5% CO2. Each well was then stimulated with 200 μM H2O2 and incubated at 37°C for 1 hour.

[0105] 3. Add ProfMIC-226

[0106] 5% (v / v) of supernatant and 10% (v / v) of inactivated bacterial cells were added to the stimulated HFF cells (the control group was replaced with an equal volume of PBS instead of supernatant / inactivated bacterial cells). Each group was in triplicate and incubated overnight at 37°C.

[0107] 4. qPCR method for detecting the relative fold increase in expression of extracellular matrix / immunomodulatory factors / cell growth factors / apoptosis-related genes

[0108] After discarding the culture medium from the cells, lysis buffer was added, total RNA was extracted, and the RNA concentration and purity were measured before reverse transcription into cDNA. GAPDH As internal reference genes, extracellular matrix-related genes were detected using real-time qPCR.SPTSSA Immunomodulatory factor-related genes MOR Cell growth factor-related genes FGF2 and FGF21 ; and apoptosis-related genes BAX The expression of genes was measured. Using a control group gene expression fold of F=1, 2... -ΔΔCT The F-value of each sample was calculated using the method described above.

[0109] Supernatant upregulates genes related to immunomodulatory factors MOR Cell growth factor-related genes FGF21 Downregulation of apoptosis-related genes BAX The results are shown in the table below:

[0110]

[0111] Inactivated bacteria upregulate genes that inhibit extracellular matrix. SPTSSA Immunomodulatory factor-related genes MOR Cell growth factor-related genes FGF2 The results are shown in the table below:

[0112]

[0113] In vitro cell experiments showed that the *Lactobacillus helveticus* ProfMIC-226 of this invention possesses a gene that upregulates the HFF extracellular matrix-related serine palmitoyltransferase gene. SPTSSA β-endorphin receptor gene related to immune regulatory factors MOR Cell growth factor-related fibroblast growth factor genes FGF2 and FGF21 The gene expression function was assessed, with relative fold increases ranging from 1.13 to 8.12; it also downregulated the BCL2-Associated X protein gene, which is associated with apoptosis. BAX The role of expression, with a relative gene expression fold of 0.22~0.74.

[0114] Example 9: Preparation of Lactobacillus helveticus ProfMIC-226 supernatant powder

[0115] The preparation of ProfMIC-226 supernatant granules is shown in the table below:

[0116]

[0117] Preparation process: The supernatant of ProfMIC-226 prepared in Example 3 was spray-dried to make ProfMIC-226 baking powder. Red bean and barley powder, jujube powder, and silicon dioxide were added and stirred until evenly dispersed to complete the powder. The resulting ProfMIC-226 powder is convenient to carry. After being brewed with water, the powder has a natural taste and good palatability, and can be used as an oral beverage for skin care.

[0118] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A strain of Lactobacillus helveticus ( Lactobacillus helveticus The strain is Lactobacillus helveticus ProfMIC-226, which was deposited at the China Center for Type Culture Collection on August 3, 2022, with accession number CCTCCNO: M20221224.

2. The application of *Lactobacillus helveticus* with accession number CCTCCNO: M20221224 in the preparation of products that improve skin cell condition, characterized in that... The product is a pharmaceutical product. The improvement of skin cell condition is described as anti-aging, promoting cell proliferation, moisturizing and repairing the skin barrier. The promotion of cell proliferation is described as promoting the proliferation of keratinocytes and / or promoting the proliferation of fibroblasts. The Lactobacillus helveticus in the product is a live bacterium or a supernatant.

3. The application according to claim 2, characterized in that, When the improvement of skin cell condition is for anti-aging purposes, the drug can upregulate the expression of extracellular matrix-related genes and upregulate autophagy-related genes. LC3B Expression and upregulation of immune regulatory factor-related genes MOR The expression of and / or upregulation of cell growth factor-related genes, wherein the extracellular matrix-related genes are... SPTSSA and / or COL1A1 The cell growth factor-related gene is FGF2 and / or FGF21 .

4. The application according to claim 2, characterized in that, When the improvement of skin cell condition is for anti-aging purposes, the drug may downregulate the expression of genes related to extracellular matrix degradation and / or downregulate apoptosis-related factors. BAX The expression of the extracellular matrix degradation-related genes is a matrix metalloproteinase family gene. MMP1 .

5. The application according to claim 2, characterized in that, When the improvement in skin cell condition is hydration, the drug can upregulate hydration-related genes. GBA The expression.

6. The application according to claim 2, characterized in that, When the improvement in skin cell condition refers to repairing the skin barrier, the drug can restore keratinocyte vitality and upregulate filaggrin genes. FLG outer skin protein gene IVL Tight junction proteins OCLN OVO-like transcription factor 1 gene OVOL1 The expression.

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