CN115678807A - Bifidobacterium longum and application thereof - Google Patents
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- CN115678807A CN115678807A CN202211404065.3A CN202211404065A CN115678807A CN 115678807 A CN115678807 A CN 115678807A CN 202211404065 A CN202211404065 A CN 202211404065A CN 115678807 A CN115678807 A CN 115678807A
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Abstract
The invention discloses bifidobacterium longum and application thereof, and relates to the technical field of microorganisms. The strain of Bifidobacterium longum (Bifidobacterium longum) disclosed by the invention is Bifidobacterium longum ProfMIC-231, which is preserved in China center for type culture Collection in 9 months and 13 days in 2022, and the preservation number is CCTCCNO: M20221386. Experiments show that ProfMIC-231 has the functions of resisting aging, improving the cell antibacterial ability and improving alopecia, and can be used for preparing foods, medicines, cosmetics and the like.
Description
Technical Field
The invention relates to the technical field of microorganisms, and particularly relates to bifidobacterium longum and application thereof.
Background
Photoaging is the predominant form of extrinsic aging of the skin, and the primary cause of photoaging of the skin is ultraviolet radiation. In the aging process of skin, cell proliferation is reduced, apoptosis is increased, and extracellular matrix components (collagen, elastin, glycosaminoglycan, etc.) are significantly reduced with skin aging. In addition, oxidative stress is increased due to increased reactive oxygen species generated by various factors such as mitochondrial damage, inflammatory reaction, etc., and aging-damaged cells cannot be removed in time, thereby causing skin aging.
In the research of aging mechanism, the autophagy of cells is found to delay the function degradation of organs, clear damaged proteins and lipids in cells, repair DNA, help cells to restore metabolic homeostasis and provide protection for photoaging damaged skin. In addition, the reduction of apoptosis, the reduction of cell inflammation, the increase of extracellular matrix synthesis and degradation, the improvement of cell antioxidant capacity and the like are proved to be capable of slowing down skin cell aging, and the search for multi-layer and multi-dimensional anti-aging substances is a current research hotspot.
In recent years, the skin has been found to secrete proteins and polypeptides with antibacterial action, which are called antimicrobial peptides (AMPs), and are effector molecules of the natural immune system, capable of contacting microbial membranes and dissolving cells, and having broad-spectrum antimicrobial action. In addition, they are involved in interfering with cellular proliferation, immune response, wound healing, cytokine release, leukocyte chemotaxis, protease antiprotease balance, and other processes.
Skin microorganisms play an important role in the maturation and homeostasis of skin immunity, and most of the microorganisms living on the skin behave symbiotically or synergetically under steady-state conditions, and recent studies have demonstrated that the skin microflora regulates the expression of various innate factors. Skin microbial decomposition of phospholipids, sterols and keratin also allows skin cells to absorb and promote cell growth, delay aging and reduce wrinkle production. When the skin is damaged by external causes such as air pollution, excessive use of cosmetics, or use of cosmetics containing hormones, it may cause disorders in the flora on the surface of the skin, thereby causing various skin problems. Studies have shown that changes in the skin's micro-ecology are associated with the development and progression of a variety of inflammatory and infectious diseases, such as psoriasis, atopic dermatitis, acne and chronic wounds. When the probiotics are reduced or even eliminated, the skin has various problems of reduced resistance, easy inflammation, infection, allergy, acne growth, imbalance of the water and the oil and the like. Therefore, the method has important practical significance for providing the probiotic related product developed by utilizing the skin microbial technology.
Disclosure of Invention
In view of the above, the present invention provides a bifidobacterium longum and applications thereof.
The invention provides Bifidobacterium longum (Bifidobacterium longum), which is Bifidobacterium longum ProfMIC-231 which is preserved in China center for type culture Collection (CCTCC for short, with the address of No. 299 in Wuchang district, wuhan university, zip code 430072) in 13 months at 2022, and the preservation number of the Bifidobacterium longum is CCTCC NO: M20221386;
the invention also aims to provide the application of the bifidobacterium longum in preparing products for improving skin conditions.
In some embodiments, the improvement in skin condition is at least one of anti-aging, increasing cellular antibacterial ability, improving hair loss.
In some embodiments, the anti-aging is upregulating expression of extracellular matrix-associated genes; the extracellular matrix-related gene is at least one of COL1A1, COL13A1, SPTSSA, TIMP1 and/or SMAD 3;
the anti-aging is the expression of up-regulated apoptosis-inhibiting related gene BCL-2;
the anti-aging is the expression of up-regulated cell autophagy-related gene LC3B;
the anti-aging is the expression of up-regulated immune regulatory factor related gene MOR;
the anti-aging is the up-regulation of expression of an oxidation-resistant related gene PTEN.
In some embodiments, the anti-aging is down-regulation of expression of extracellular matrix-associated genes; the extracellular matrix-associated degrading gene is at least one of the MMP family;
the anti-aging is the expression of down-regulation apoptosis related genes; the apoptosis-related gene is expression of at least one of BAX and/or Caspase families;
the anti-aging is the down-regulation of the expression of the genes IL-6 and/or TNF-alpha related to the cell inflammatory factors.
In some embodiments, the enhancing skin antibacterial ability is up-regulation of expression of antimicrobial peptide-related genes S100A7, S100A8 and/or DEFB 4.
In some embodiments, the ameliorating hair loss comprises at least one of the following a) to f):
a) Up-regulating the expression of the hair follicle growth phase related gene beta-catenin and/or LEF 1;
b) Up-regulating the expression of hair follicle cell growth factor related genes VEGF, IGF-1 and FGF-2;
c) Up-regulating the expression of the hair follicle cell cycle regulatory factor related gene c-Myc, PP2A and/or Cyclin D1;
d) Up-regulating expression of a gene HSP27 related to inhibition of hair follicle apoptosis;
e) Up-regulating expression of hair follicle extracellular matrix-related genes CSPG4 and/or HSPG 2;
f) And the expression of the hair follicle resting phase related gene BMP4 is reduced.
In some embodiments, the product is a food product, a pharmaceutical product, or a cosmetic product.
Another object of the present invention is to provide a product for improving skin conditions, which is prepared from a material comprising bifidobacterium longum as described above.
In some embodiments, the bifidobacterium longum in the product comprises one or both of (1) or (2) as follows:
(1) A population and/or inoculum of bifidobacterium longum of claim 1;
(2) A culture, exosome, lysate and/or extract of Bifidobacterium longum of claim 1.
The invention discloses Bifidobacterium longum ProfMIC-231 with the preservation number of CCTCC NO: M20221386. Experiments show that ProfMIC-231 has the functions of resisting aging, improving the cell antibacterial ability and improving alopecia, and can be used for preparing foods, medicines, cosmetics and the like.
Biological preservation Instructions
Bifidobacterium longum (Bifidobacterium longum) ProfMIC-231 was deposited in China center for type culture Collection (CCTCC, address: wuhan university, wuhan's Wuchang region eight-way No. 299) at 9/13/2022, and the preservation number is CCTCC NO: M20221386.
Detailed Description
The invention provides bifidobacterium longum and application thereof. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
Bifidobacterium longum strain ProfMIC-231 of the present invention is derived from feces of an eight-year-old healthy girl and identified as Bifidobacterium longum (Bifidobacterium longum) by 16 SrDNA. The strain is gram-positive and rod-shaped under a microscope; the bacterial colony grows on a BBL flat plate, can form a round bacterial colony with a smooth and opaque surface, is white and has a regular edge; the strain grows uniformly and turbulently in a BBL liquid culture medium, and the strain is white precipitate after being placed for a long time, and the optimal growth temperature is 37 ℃.
Bifidobacterium longum (Bifidobacterium longum) ProfMIC-231, deposited in the collection: china center for type culture Collection, address: in the Wuhan university school of Wuhan 299 in the Wuchang area of Wuhan city, hubei province, the preservation date is as follows: 13 months 9 in 2022, the preservation number is CCTCC NO: M20221386.
Further, the present invention provides bifidobacterium longum ProfMIC-231 in a use or product according to the invention in a form that is live or dead or that has been subjected to sterilization, or in the form of a fermentation lysate and/or extract, or in the form of a bacterial product or in the form of a fermentation filtrate or in the form of a derivative, preferably selected from: metabolites, metabolic biological products, exosomes, prebiotics, cell walls and components thereof, exopolysaccharides, and compounds containing immunogenic components, preferably selected from: fermentation filtrate, fermentation lysate.
In vitro cell experiments show that the bifidobacterium longum ProfMIC-231 has the functions of up-regulating expression of an I-type collagen alpha chain gene COL1A1, a serine palmitoyltransferase gene SPTSSA, a tissue metalloproteinase inhibitor I gene TIMP1, a signal transduction protein gene SMAD3 and a microtubule-associated protein 1 light chain 3 beta gene LC3B related to a cell autophagy-associated gene, wherein the relative expression multiple of the genes is 1.12-2.04; has the function of down-regulating and degrading matrix metalloproteinase gene MMP1 related to extracellular matrix and tumor necrosis factor alpha gene TNF-alpha expression related to cell inflammatory factors, and the relative expression multiple of the genes is 0.44-0.75.
In vitro cell experiments show that the bifidobacterium longum ProfMIC-231 has the functions of up-regulating the expression of a collagen membrane protein 13 alpha chain gene COL13A1 related to HFF extracellular matrix, a serine palmitoyltransferase gene SPTSSA, an immunoregulatory factor related beta-endorphin receptor gene MOR, an antioxidant-related phosphatase with deletion of chromosome 10 and a tensin homolog PTEN, and inhibiting the expression of a B-lymphocytoma-2 gene BCL-2 related to apoptosis, and the relative expression multiple of the genes is 1.10-3.51 times; has the function of reducing the expression of matrix metalloproteinase family gene MMP related to degrading extracellular matrix, BCL2-Associated X protein gene BAX related to apoptosis, cysteine proteinase family gene Caspase and interleukin 6 gene IL-6 related to cell inflammatory factors, and the relative expression multiple of the genes is 0.33 to 0.83
In vitro cell experiments show that the bifidobacterium longum ProfMIC-231 has the function of up-regulating the expression of antibacterial peptide related genes S100A7, S100A8 and DEFB4, and the relative expression multiple of the genes is 1.20-1.91.
In vitro cell experiments show that the bifidobacterium longum ProfMIC-231 disclosed by the invention has the functions of up-regulating beta-catenin gene beta-catenin related to the hair follicle growth phase, lymphocyte binding enhancement factor 1LEF1, vascular endothelial cell growth factor VEGF related to the hair follicle cell growth factor, insulin-like growth factor 1IGF-1, fibroblast growth factor 2FGF-2, chondroitin sulfate proteoglycan gene CSPG related to hair follicle extracellular matrix, heparin sulfate proteoglycan gene HSPG, myc family protein gene c-Myc related to hair follicle cell cycle regulatory factor, cyclin D1, protein phosphatase 2A and protein apoptosis inhibition related heat shock protein 27 gene HSP27 expression, and the relative expression multiple of the genes is 1.10-2.04. The bone morphogenetic protein 4BMP4 related to the hair follicle development resting period is down regulated, and the relative expression multiple of the gene is 0.47-0.73.
The test materials adopted by the invention are all common commercial products and can be purchased commercially, and the invention is further explained by combining the following embodiments:
example 1
Isolation of ProfMIC-231:
samples were taken from eight year old healthy girls. Properly treating the sample, uniformly mixing the treated sample in normal saline by oscillation, taking the supernatant, streaking the supernatant on a BBL solid plate, carrying out anaerobic culture at the constant temperature of 37 ℃ for 48 hours, and then selecting a white bacterial colony for repeated inoculation and screening until a uniform single bacterial colony is obtained, wherein the single bacterial colony is named as ProfMIC-231.
Gram staining microscopy: the bacterial strain ProfMIC-231 is gram-positive and rod-shaped under a microscope; growing on a BBL flat plate to form white round microcolonies with smooth, mellow and non-transparent surfaces and regular edges; the bacteria grow uniformly and turbulently in BBL liquid culture medium, and the white bacteria precipitate after long-term placement.
Example 2
Nucleic acid identification of ProfMIC-231:
1. 16S rDNA gene sequence analysis:
picking single colony to be placed in MRS liquid culture medium, carrying out anaerobic culture at 37 ℃ overnight, then rotating at 12000 ℃ for 1min to collect thalli, and operating according to the steps of the DNA extraction kit. The primers adopt bacterial universal primers 27F and 1492R, a PCR amplification system is a 50 mu L system, and the pre-denaturation is carried out for 5min at 95 ℃;94 ℃ 15s,57 ℃ 15s,72 ℃ 40s,35 cycles; extension at 72 ℃ for 10min.
2. Results
The homology comparison (BLASTN) of the PCR product sequencing results with the published standard sequences in GenBank indicates that the ProfMIC-231 strain is Bifidobacterium longum (Bifidobacterium longum).
Example 3
ProfMIC-231 regulates photoaging HaCaT extracellular matrix/autophagy/degradation extracellular matrix/inflammatory factor-related gene expression experiments:
1. preparation of ProfMIC-231 fermentation filtrate and fermentation lysate:
selecting a single bifidobacterium longum ProfMIC-231 colony in a BBL liquid culture medium, carrying out anaerobic static culture in an incubator at 37 ℃ for 16-18 h, detecting by an enzyme labeling instrument, diluting by PBS and adjusting OD 600 Deactivating at 121 deg.C for 30min under high pressure, centrifuging at 12000 rpm for 2min, and filtering with 0.22 μm filter membrane to obtain fermentation filtrate. After centrifugation, the separated and precipitated thalli are washed twice by PBS and then added with liquid nitrogen for grinding and crushing, crushed bacterial mud is collected and resuspended to the volume during centrifugation by PBS, and then crushed for 20 minutes by ultrasonic wave, and inactivated at the temperature of 121 ℃ for 30 minutes under high pressure, thus obtaining the fermentation lysate.
2. HaCaT cell preparation and ultraviolet ray damage
HaCaT cells were digested and then digested at 0.5 ml/well (2X 10 contained therein) 5 Cells) were inoculated into 24-well plates and cultured overnight at 37 ℃ in a 5% carbon dioxide incubator. Total dose of 2J/cm was applied to cells in the wells 2 Ultraviolet UVB radiation damage.
3. ProfMIC-231 addition
5% (V/V) of the fermentation filtrate and 10% (V/V) of the fermentation lysate were added to the stimulated HaCaT cells (control group replaced the volume of PBS for the fermentation filtrate/fermentation lysate, respectively). Each group 3 was incubated overnight at 37 ℃.
4. qPCR method for detecting relative expression multiple of extracellular matrix/cell autophagy/extracellular matrix degradation/inflammatory factor related genes
Removing the culture medium of the cells, adding lysis solution, extracting total RNA of the cells, detecting the concentration and purity of the RNA, performing reverse transcription to obtain cDNA, detecting extracellular matrix related genes COL1A1, SPTSSA, TIMP1 and SMAD3 by using GAPDH as an internal reference gene and adopting real-time qPCR, and performing cell self-detectionThe expression of the phagocytosis-related gene LC3B, the degradation extracellular matrix-related gene MMP1 and the inflammatory factor-related gene TNF-alpha. Relative expression multiple F =1 of control group gene, using 2 -ΔΔCT The F value of each sample was calculated.
The formula: f =2 -ΔΔCT Wherein:
△CT experiment of ＝CT Experiment of -CT Internal reference (experiment) ；
△CT Control ＝CT Control -CT Internal reference (contrast) ；
△△CT＝△CT Experiment of -△CT Control 。
The fermentation filtrate up-regulates the extracellular matrix related gene TIMP1; down-regulating and degrading extracellular matrix related gene MMP1. The results are shown in the following table:
the fermentation lysate upregulates extracellular matrix genes COL1A1, SPTSSA, TIMP1 and SMAD3, autophagy-related genes LC3B; down-regulating the inflammatory factor-related gene TNF-alpha. The results are shown in the following table:
the result shows that the addition of ProfMIC-231 has the anti-aging effects of promoting the synthesis of HaCaT extracellular matrix, promoting the autophagy of cells, reducing the degradation of the extracellular matrix and reducing inflammatory factors.
Example 4
ProfMIC-231 assay for modulating oxidative damage to HFF extracellular matrix/immunomodulatory factor/antioxidant/apoptosis inhibiting/extracellular matrix degrading/apoptosis/inflammatory factor-related gene expression:
1. preparation of ProfMIC-231 fermentation filtrate and fermentation lysate:
the preparation method refers to example 3.
2. HFF cell preparation and H2O 2-induced oxidative damage
The HFF cells cultured with DMEM were digested at 0.5 ml/well (2X 10 contained therein) 5 Cells) were inoculated into 24-well plates and cultured overnight at 37 ℃ in a 5% carbon dioxide incubator. H was added to each well to a final concentration of 200. Mu.M 2 O 2 Stimulating, and standing at 37 ℃ for 1h.
ProfMIC-231 addition
The fermentation filtrate 5% (V/V), fermentation lysate 10% (V/V) were added to the stimulated HFF cells (control group replaced equal volume of PBS for fermentation filtrate/fermentation lysate, respectively). Each group 3 was incubated overnight at 37 ℃.
4. qPCR method for detecting relative expression multiple of extracellular matrix/immunoregulatory factor/antioxidation/apoptosis inhibition/extracellular matrix degradation/apoptosis/inflammatory factor related genes
Removing a culture medium from the cells, adding a lysis solution, extracting total RNA of the cells, detecting the concentration and purity of the RNA, performing reverse transcription to obtain cDNA, detecting COL13A1 and SPTSSA related genes of an extracellular matrix, MOR related genes of an immune regulatory factor, PTEN related genes and BCL-2 related genes of apoptosis inhibition by using GAPDH as an internal reference gene and adopting real-time qPCR; and degrading the expression of MMP family of extracellular matrix related genes, BAX and Caspase family genes of apoptosis related genes and IL-6 of inflammatory factor related genes. Relative expression multiple F =1 of control group gene, using 2 -ΔΔCT The F value of each sample was calculated.
The fermentation filtrate up-regulates extracellular matrix related gene COL13A1, immunoregulation factor related gene MOR, and inhibits apoptosis related gene BCL-2; down-regulating and degrading extracellular matrix related genes MMP family, apoptosis related genes BAX and Caspase family and inflammatory factor related genes IL-6. The results are shown in the following table:
the fermentation lysate up-regulates extracellular matrix related gene SPTSSA, antioxidant related gene PTEN, apoptosis-inhibiting gene related gene BCL-2; down-regulating apoptosis related genes BAX and Caspase-3. The results are shown in the following table:
the results show that the addition of ProfMIC-231 has the anti-aging effects of promoting the synthesis of HFF extracellular matrix, increasing cellular immune regulatory factors, increasing oxidation resistance, reducing apoptosis, reducing extracellular matrix degradation and reducing inflammatory factors.
Example 5
ProfMIC-231 up-regulated HaCaT antibacterial peptide related gene expression experiment:
1. preparation of ProfMIC-231 fermentation lysate:
the preparation method refers to example 3.
2. HaCaT cell preparation
HaCaT cells were digested and then dispensed at 0.5 ml/well (2X 10 contents) 5 Cells) were seeded into 24-well plates and cultured overnight at 37 ℃ in a 5% carbon dioxide incubator.
3. ProfMIC-231 addition and LPS stimulation
The fermentation lysate (10% (V/V) was added to HaCaT cells cultured overnight (control group replaced with an equal volume of PBS), and after 2h, 0.5ml of LPS solution with a concentration of 0.2. Mu.g/ml was added to induce cell inflammation, and the cells were cultured in a 5% carbon dioxide incubator at 37 ℃ for 20h.
4. qPCR method for detecting relative expression multiple of antibacterial peptide related gene
Removing the culture medium of the cells, adding a lysis solution, extracting total RNA of the cells, detecting the concentration and purity of the RNA, performing reverse transcription to obtain cDNA, and detecting the expression of S100A7, S100A8 and DEFB4 genes by using a real-time qPCR (quantitative polymerase chain reaction) by using GAPDH as an internal reference gene. Control with an equal volume of PBS-treated group (gene relative expression fold F = 1) using 2 -ΔΔCT The F value of each sample was calculated.
The inactivated thallus can up-regulate antibacterial peptide genes S100A7, S100A8 and DEFB4, and the results are shown in the following table:
the result shows that ProfMIC-231 has the function of promoting the expression of the antibacterial peptide gene.
Example 6
ProfMIC-231 regulates oxidative damage HDPCs hair follicle growth phase/hair follicle cell growth factor/hair follicle extracellular matrix/hair follicle cell cycle regulatory factor/hair follicle resting phase-related gene expression experiment:
1. preparation of ProfMIC-231 fermentation filtrate and fermentation lysate:
the preparation method refers to example 3.
2. HDPCs Hair follicle cell preparation and H 2 O 2 Inducing oxidative damage
The hair follicle cells of HDPCs cultured in fibroblast culture medium were digested and then digested at a rate of 2 ml/well (3X 10 content) 5 Cells) were inoculated into 6-well plates and cultured overnight at 37 ℃ in a 5% carbon dioxide incubator. H was added to each well to a final concentration of 200. Mu.M 2 O 2 Stimulating, and standing at 37 ℃ for 1h.
ProfMIC-231 addition
The fermentation filtrate 5% (V/V) and the fermentation lysate 10% (V/V) were added to the stimulated HDPCs cells (control group replaced equal volume of PBS for fermentation filtrate/fermentation lysate, respectively). Each group 3 was incubated overnight at 37 ℃.
4. qPCR method for detecting relative expression multiple of related genes of hair follicle growth phase/hair follicle cell growth factor/hair follicle extracellular matrix/hair follicle cell cycle regulatory factor/hair follicle resting phase
Removing a culture medium from the cells, adding a lysis solution, extracting total RNA of the cells, detecting the concentration and purity of the RNA, performing reverse transcription to obtain cDNA, detecting related genes beta-catenin and LEF1 in a hair follicle growth phase, related genes VEGF, IGF-1 and FGF-2 of hair follicle cell growth factors, related genes CSPG4 and HSPG2 of hair follicle extracellular matrix, related genes HSP27 for inhibiting hair follicle apoptosis, related genes c-Myc, cyclin D1 and PP2A of hair follicle cell cycle regulation factors by using GAPDH as an internal reference gene and adopting real-time qPCR (quantitative polymerase chain reaction); and of BMP4, a gene involved in telogen phase of hair folliclesAnd (4) expressing. Relative expression multiple F =1 of control group gene, using 2 -ΔΔCT F value was calculated for each sample.
The fermentation filtrate up-regulates related genes beta-catenin and LEF1 in the hair follicle growth phase, related genes VEGF, IGF-1 and FGF-2 of hair follicle cell growth factor, related genes CSPG4 and HSPG2 of hair follicle extracellular matrix, related genes HSP27 for inhibiting hair follicle cell apoptosis, related genes c-Myc, cyclin D1 and PP2A of hair follicle cell cycle regulatory factor; down-regulating a hair follicle resting phase related gene BMP4, and the results are shown in the following table:
the fermentation lysate up-regulates hair follicle growth phase related gene LEF1, hair follicle cell growth factor related genes VEGF, FGF-2, hair follicle cell apoptosis related gene HSP27 and hair follicle cell cycle regulatory factor related gene c-Myc; down-regulating a hair follicle resting phase related gene BMP4, and the results are shown in the following table:
the result shows that the addition of ProfMIC-231 has the effects of promoting the hair follicle growth phase of HDPCs, hair follicle cell growth factors, hair follicle extracellular matrix, inhibiting hair follicle cell apoptosis and related gene expression of hair follicle cell cycle regulatory factors; has the function of reducing the expression of related genes of the resting stage of hair follicles and improving alopecia.
Example 7
Preparation example of bifidobacterium longum ProfMIC-231 fermentation filtrate granules:
preparation of ProfMIC-231 fermentation filtrate granules, the formula is shown in the following table:
| raw materials | Mass ratio (%) |
| ProfMIC-231 baking powder | 50 |
| Recombinant collagen powder | 30 |
| Juicy peach powder | 18.8 |
| Vitamin C | 0.8 |
| Silicon dioxide | 0.4 |
The preparation process comprises the following steps: and (3) spray-drying the ProfMIC-231 fermentation filtrate prepared in the example 3 to prepare ProfMIC-231 fermentation powder, adding recombinant collagen powder, vitamin C, juicy peach fruit powder and silicon dioxide, and uniformly stirring and dispersing to obtain the medicinal granules. The obtained instant granules of ProfMIC-231 are soaked in water, have sour and sweet taste, good palatability and convenient carrying, and can be used as oral beverage for skin care.
Example 8
Example of bifidobacterium longum ProfMIC-231 fermentation lysate emulsion preparation:
preparation of emulsion of ProfMIC-231 inactivated thallus, and the formula is shown in the following table
| Starting materials | Mass ratio (%) |
| ProfMIC-231 fermentation lysate | 50.00 |
| Cyclopentasiloxane | 15.00 |
| Glycerol | 12.00 |
| Vaseline | 2.00 |
| 1,3 butanediol | 1.50 |
| Tween-80 | 0.75 |
| Vitamin E acetate | 0.20 |
| Chinese gum | 0.20 |
| Carbopol Ultrez 10 | 0.10 |
| Triethanolamine | Proper amount of |
| Deionized water | To 100 |
The preparation process comprises the following steps: mixing cyclopentasiloxane, vaseline, tween-80, vitamin E acetate, and glycerol, heating to 75 deg.C to obtain phase A; then taking the ProfMIC-231 fermentation lysate prepared in example 3, adding xanthan gum, carbopol Ultrez 10, 1,3-butanediol and deionized water, mixing and dispersing uniformly, and heating to 75 ℃ to be used as a phase B; slowly adding the preheated phase A into the preheated phase B, homogenizing completely, adding triethanolamine to adjust pH to 6-7, stirring, and cooling to 35 deg.C. The emulsion of the ProfMIC-231 fermentation lysate is lubricated and comfortable.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.
Claims (10)
1. Bifidobacterium longum (Bifidobacterium longum) is Bifidobacterium longum ProfMIC-231, which has been deposited in China center for type culture Collection at 9/13/2022 with the preservation number of CCTCC NO: M20221386.
2. Use of bifidobacterium longum according to claim 1 for the preparation of a product for improving skin conditions.
3. The use of claim 2, wherein the improvement in skin condition is at least one of anti-aging, increasing cellular antibacterial ability, improving hair loss.
4. The use according to claim 3, wherein the anti-aging is up-regulation of the expression of extracellular matrix-related genes; the extracellular matrix-related gene is at least one of COL1A1, COL13A1, SPTSSA, TIMP1 and/or SMAD 3;
the anti-aging is the expression of up-regulated apoptosis-inhibiting related gene BCL-2;
the anti-aging is the expression of up-regulated cell autophagy related gene LC3B;
the anti-aging is the expression of up-regulated immune regulation factor related gene MOR;
the anti-aging is the expression of up-regulated anti-oxidation related gene PTEN.
5. The use according to claim 3, wherein the anti-aging is down-regulation of expression of extracellular matrix-associated genes; the extracellular matrix-degrading related gene is at least one of the MMP families;
the anti-aging is the expression of down-regulation apoptosis related genes; the apoptosis-related gene is expression of at least one of BAX and/or Caspase families;
the anti-aging is the down-regulation of the expression of the genes IL-6 and/or TNF-alpha related to the cell inflammatory factors.
6. The use of claim 3, wherein the improvement of skin antibacterial ability is an up-regulation of the expression of antimicrobial peptide-related genes S100A7, S100A8 and/or DEFB 4.
7. The use according to claim 3, wherein said improvement in hair loss comprises at least one of the following a) to f):
a) Up-regulating the expression of the hair follicle growth phase related gene beta-catenin and/or LEF 1;
b) Up-regulating the expression of hair follicle cell growth factor related genes VEGF, IGF-1 and FGF-2;
c) Up-regulating the expression of the hair follicle cell cycle regulatory factor related gene c-Myc, PP2A and/or Cyclin D1;
d) Up-regulating expression of a gene HSP27 related to inhibition of hair follicle apoptosis;
e) Up-regulating expression of hair follicle extracellular matrix-related genes CSPG4 and/or HSPG 2;
f) And the expression of the hair follicle resting phase related gene BMP4 is reduced.
8. Use according to any one of claims 1 to 7, wherein the product is a food product, a pharmaceutical product or a cosmetic product.
9. A product for improving skin condition, prepared from a material comprising bifidobacterium longum as claimed in claim 1.
10. The product according to claim 9, wherein the bifidobacterium longum in the product comprises one or both of the following (1) or (2):
(1) A population and/or a microbial agent of bifidobacterium longum according to claim 1;
(2) A culture, exosome, lysate and/or extract of Bifidobacterium longum of claim 1.
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