CN111235058A - Bifidobacterium breve with effects of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging, and its preparation method and application - Google Patents
Bifidobacterium breve with effects of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging, and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a Bifidobacterium breve with the functions of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging and a preparation method and application thereof, and relates to the technical field of microorganisms, wherein the Bifidobacterium breve with the functions of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging is named as Bifidobacterium breve, and the preservation number is as follows: CGMCC No. 12915. The test result shows that the bifidobacterium breve has the function of repairing skin injury and inflammation caused by ultraviolet rays and can improve the skin photoaging phenomenon caused by long-term repeated irradiation of the ultraviolet rays.
Description
Technical Field
The invention relates to the technical field of microorganisms, and particularly relates to bifidobacterium breve with effects of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging, and a preparation method and application thereof.
Background
The skin is the first physiological barrier for the body to contact with the outside, and the long-term solar ultraviolet radiation can cause the symptoms of skin weakness, wrinkling, roughness, dryness, laxity, hyperpigmentation and the like, namely photoaging. Ultraviolet rays are classified into UVA, UVB, UVC according to wavelength, wherein UVC is absorbed and dispersed by the atmospheric ozone layer and does not reach the earth's surface. UVA has strong penetration capacity to skin, can affect dermis and subcutaneous tissue areas to cause cell damage, such as fibroblasts, vascular endothelial cells, Langerhans cells and the like, and changes the structure of dermis to cause skin relaxation and wrinkling; UVB has high energy but relatively weak penetrating power, mainly affects skin keratinocytes and melanocytes, and can thicken horny layer, proliferate epidermis and aggravate inflammatory reaction.
The skin type I and type III collagen is the most main protein component constituting the skin dermal extracellular matrix, has the functions of keeping skin moisture, supporting the normal structure of the skin and maintaining the skin toughness, is mainly distributed in the dermal layer and is synthesized and secreted only by fibroblasts. Current research shows that cells are induced to express Matrix Metalloproteinases (MMPs) after UV radiation, and degradation of collagen and elastin is catalyzed, wherein MMP-1 (interstitial collagenase) and MMP-3 (stromelysin) are main acting proteinases. High concentrations of ROS (reactive oxygen species) generated in the skin after UV irradiation, including superoxide anion, singlet oxygen, hydrogen peroxide, hydroxyl radical, etc., normal skin has a relatively perfect antioxidant defense system, and enzymatic antioxidant substances include: glutathione reductase, glutathione peroxidase, superoxide dismutase, catalase and the like, if ROS cannot be removed in time, damage to nucleic acid, lipid and protein in tissues can be caused, and related cell signal transduction and gene expression are influenced, so that skin photoaging is caused. With the continuous increase of environmental pollution, the ozone layer is increasingly damaged, the ultraviolet rays radiated to the surface of the earth are gradually increased, the skin is more and more damaged by the ultraviolet rays, and the physical and psychological health of people is affected.
At present, chemical exfoliation, photoelectric repair (intense pulse light, lattice laser, near infrared light and the like) and injection molding (botulinum toxin, hyaluronic acid injection and autologous fat filling) are mainly adopted clinically to relieve the problem of skin photoaging. Chemical exfoliation and injection molding can cause symptoms such as swelling, itching and the like of skin, and have risks, and photoelectric repair has small side effects but is expensive, so that the wide application is limited. In recent years, probiotics are recognized by the medical and scientific circles to be beneficial to human health and have no side effect, so that the probiotics capable of repairing skin ultraviolet injury, relieving inflammation and preventing skin photoaging are needed to be provided.
Disclosure of Invention
The invention aims to solve the technical problem of providing bifidobacterium breve with the effects of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging, and a preparation method and application thereof, and solves the problems of side effect or high price existing in the existing problem of relieving skin photoaging.
In order to solve the technical problems, the technical scheme of the invention is as follows:
bifidobacterium breve with the functions of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging is named as Bifidobacterium breve with the preservation number: CGMCC No. 12915.
A preparation method of the bifidobacterium breve with the effects of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging comprises the following steps:
(1) collecting a fecal sample, dissolving with sterile water, diluting in a gradient manner, and adding into a sterilized flat plate;
(2) adding a hydrogen peroxide solution into an MRS culture medium at the temperature of 50-50 ℃ to ensure that the initial hydrogen peroxide concentration in the culture medium is 0.5mmol/L to obtain a sample culture medium;
(3) pouring the sample culture medium into the flat plate in the step (1) and uniformly mixing, selecting a strain with good growth vigor of bacterial colonies on the flat plate, performing gene sequence comparison, and determining the strain type, thereby screening the lactobacillus with hydrogen peroxide tolerance;
(4) the selected lactobacillus with hydrogen peroxide tolerance has antioxidant performance, namely clearance rate of DPPH free radical, ABTS free radical and hydroxyl free radical, as re-screening index, and then the strain with higher antioxidant activity is selected, and the required bifidobacterium breve is obtained by separation.
Preferably, the MRS medium in step (2) is: the following raw materials in parts by weight: 10.0 parts of peptone, 8.0 parts of beef extract, 4.0 parts of yeast extract, 20.0 parts of lactose, 2.0 parts of dipotassium hydrogen phosphate, 5.0 parts of sodium acetate, 0.2 part of magnesium sulfate, 0.05 part of manganese sulfate, 1.0 part of L-cysteine hydrochloride, 2.0 parts of triammonium citrate, 801.0 parts of tween and 20.0 parts of agar, adjusting the pH to 6.0-6.5, adding distilled water to 1000mL, and sterilizing at 125 ℃ of 120-.
Application of the bifidobacterium breve with the effects of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging is disclosed, and the bifidobacterium breve is added into cosmetics, skin care products or medicaments for repairing skin ultraviolet injury and preventing skin photoaging.
Preferably, the bifidobacterium breve is added into lotion, emulsion, cream, mask, essence, foundation solution, concealer, sunscreen cream, oral liquid, granules, tablets, pills, powder, capsules or dropping pills for repairing ultraviolet damage of skin and preventing photoaging of skin.
Preferably, the cosmetic, skin care product or pharmaceutical product can be added with auxiliary materials.
Preferably, the auxiliary material is at least one of grease, surfactant, polyalcohol, humectant, defoamer, thickener, preservative, antioxidant, pH regulator, lubricant, emulsifier or adhesive.
By adopting the technical scheme, the test result shows that the bifidobacterium breve has the effect of repairing skin injury and inflammation caused by ultraviolet rays and can improve the skin photoaging phenomenon caused by long-term repeated irradiation of the ultraviolet rays.
Drawings
FIG. 1 is a graph showing the results of measurements of the antioxidant capacity of different lactic acid bacteria;
FIG. 2 is a graph showing the results of the cytotoxicity test of Bifidobacterium breve;
FIG. 3 is a graph showing the result of ROS detection of Bifidobacterium breve acting on cells; wherein, FIG. 3-1 is the result of acting on human dermal fibroblasts; FIG. 3-2 is a graph of effects on keratinocytes;
FIG. 4 is a graph showing the results of detecting the Bifidobacterium breve using human dermal fibroblast matrix metalloproteinases MMP-1 and MMP-3;
FIG. 5 is a graph showing the results of the measurement of the degree of collagen synthesis and crosslinking of Bifidobacterium breve in human dermal fibroblasts; wherein, FIG. 5-1 is a diagram showing the results of detecting the collagen content of type I and type III; FIG. 5-2 is a graph showing the result of measurement of hydroxyproline content; FIG. 5-3 is a graph showing the results of measurement of the hyaluronic acid content;
FIG. 6 is a graph of the test results of Bifidobacterium breve on the effects of inflammatory reaction and lipid peroxidation damage of mice after ultraviolet irradiation, wherein FIG. 6-1 is a graph of the test results of IL-1 β, IL-6 and TNF- α contents, and FIG. 6-2 is a graph of the test results of MDA, SOD and CAT contents.
Detailed Description
The following further describes embodiments of the present invention with reference to the drawings. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Example 1
Collecting a fecal sample, dissolving with sterile water, diluting in a gradient manner, adding into a sterilized flat plate, adding a hydrogen peroxide solution into an MRS culture medium at about 55 ℃ to ensure that the initial hydrogen peroxide concentration in the culture medium is 0.5mmol/L, pouring the culture medium into the flat plate, and uniformly mixing, wherein the MRS culture medium has the formula: 10.0g of peptone, 8.0g of beef extract, 4.0g of yeast extract, 20.0g of lactose, 2.0g of dipotassium phosphate, 5.0g of sodium acetate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate, 1.0g of L-cysteine hydrochloride, 2.0g of triammonium citrate, 801.0g of Tween, 20.0g of agar, pH of between 6.0 and 6.5, adding distilled water to 1000mL, and sterilizing at 121 ℃ for 20 min. Selecting strains with better colony growth on a flat plate, carrying out gene sequence comparison, determining the strain category, thereby screening lactobacillus with hydrogen peroxide tolerance, taking the antioxidant performance, namely the clearance rate of DPPH free radicals, ABTS free radicals and hydroxyl free radicals as a re-screening index, and the experimental result is shown in figure 1, screening the strains with higher antioxidant activity, observing the separated strains under a microscope to be club-shaped, cone-shaped, short, thin, capable of fermenting lactose, glucose and the like to generate lactic acid and acetic acid, comparing the 16S rDNA sequence of the strain with a model strain, identifying the strain as Bifidobacterium breve, namely Bifidobacterium breve, and preserving the unit code: CGMCC-China general microbiological culture Collection center; address: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North; the detection result is as follows: survival; the preservation date is as follows: 2016, 08 months, 29 days; the preservation number is: CGMCC No. 12915.
Example 2
Detecting the influence of Bifidobacterium breve (BBr 60 for short) on the proliferation activity of human dermal fibroblasts and keratinocytes:
2.1 preparation of bacterial liquid
BBr60 to 10mL of MRS medium was inoculated from the frozen tube and cultured anaerobically at 37 ℃ for 20 hours. The next day, 200. mu.l of the culture broth was inoculated into 10mL of MRS medium (2% secondary activation), and incubated under anaerobic conditions at 37 ℃ for 20 hours. 2mL of the cultured bacterial solution was taken out and centrifuged (13000rmp, 1min), after two times of bacteria in PBS buffer, the supernatant was removed, 1mL of PBS was taken to suspend the pellet, after uniform mixing, OD600 was measured, the number of bacteria was adjusted to 1x1011cfu/mL, and water bath at 95 ℃ for 10min was performed for standby.
2.2 cell culture
Human dermal fibroblasts or keratinocytes were thawed in a 37 ℃ water bath, and cultured in DMEM medium containing 20% Fetal Bovine Serum (FBS), 100U/mL penicillin and 100U/mL streptomycin at 37 ℃ in a 5% CO2 incubator.
2.3 evaluation of cytotoxicity
Collecting human dermal fibroblast or keratinocyte in logarithmic growth phase, digesting with 0.25% trypsin, blowing into single cell suspension, and making into powder with a size of 1 × 104The concentration per well was transferred to serum-free medium. After the growth of the cells attached to the wall for 24 hours in a constant temperature incubator with 5 percent CO2 at 37 ℃, 100 mu L of the cells containing 1x10 are respectively added11cfu/mL、5x1010cfu/mL、1x1010cfu/mL、8x109cfu/mL、4x109cfu/mL、2x109cfu/mL、8x108cfu/mL、4x108cfu/mL Bifidobacterium breve, with drug-free medium as blank control (5 multiple wells per concentration group), incubated in CO2 incubator for 24h, then 20. mu.L MTT solution (5mg/mL) was added to each well, and after 4h of incubation, the culture medium in the wells was aspirated, 150. mu.L DMSO was added to each well, and shaking was performed for 5min to ensure complete mixing. The absorbance at 490nm was measured three times with an enzyme linked immunosorbent assay, the data were recorded, and the results were calculated as the average of the three times. Three groups of proper drug concentrations are selected for subsequent experiments according to the cell activity detection result.
As shown in fig. 2, after culturing human dermal fibroblasts or keratinocytes in DMEM containing BBr60 at different concentrations for 24 hours, the effect of BBr60 on the proliferation activity of human dermal fibroblasts and keratinocytes was analyzed by MTT assay. The results show that the concentration ofBBr60 can produce cytotoxicity and promote apoptosis, when the concentration of BBr60 is less than or equal to 8x109At cfu/mL, the activity of the cells gradually decreases with the increase of the concentration of BBr60, and when the concentration is more than 8x109The cell activity is obviously reduced (p is less than 0.05) when cfu/mL is adopted, and the cell survival rate is less than 80%. The BBr60 concentration is selected to be 8x109cfu/mL、2x109cfu/mL、8x108cfu/mL。
Example 3
Testing the repairing or preventing effect of BBr60 on human skin fibroblasts and keratinocytes:
3.1 bacterial liquid culture
BBr60 to 10ml of MRS medium was inoculated from the frozen tube and cultured anaerobically at 37 ℃ for 20 hours. The next day, 200. mu.l of the culture broth was inoculated into 10ml of MRS medium (2% secondary activation), and incubated under anaerobic conditions at 37 ℃ for 20 hours.
3.2 cell culture
Human dermal fibroblasts or keratinocytes were thawed in a 37 ℃ water bath, and cultured in DMEM medium containing 20% Fetal Bovine Serum (FBS), 100U/ml penicillin and 100U/ml streptomycin at 37 ℃ in a 5% CO2 incubator.
3.3 Experimental groups
The test method comprises blank group, model group and administration group. Blank group: the culture was carried out in untreated DMEM medium. Model group: the cultures were performed in untreated DMEM medium, fibroblasts were subjected to UVA irradiation, and keratinocytes were subjected to UVB irradiation. Administration group: DMEM medium containing different concentrations of BBr60 was administered for culture, fibroblasts were irradiated with UVA, and keratinocytes were irradiated with UVB. The cells of each group are provided with 5 multiple holes.
3.4 ultraviolet irradiation
Inoculating human dermal fibroblast or keratinocyte in logarithmic growth phase into a 6-well plate according to a certain quantity, culturing at 37 ℃ in a constant-temperature incubator with 5% CO2, discarding culture solution in a culture dish when the human dermal fibroblast or keratinocyte grows to be 80% -90% fused, washing with PBS for 2 times, covering the cells with a small amount of DMEM without fetal calf serum, placing in an ultraviolet lamp box, controlling lamps of UVA and UVB to be 15cm away from a table top, respectively irradiating the human dermal fibroblast or keratinocyte with UVA and UVB, wherein the average irradiation power is 12.7mW/cm, the single irradiation time is 780S, the dose is 9.9J/cm, irradiating for 1 time every day, continuously irradiating for 7 days, continuously culturing for 24 hours after irradiation by changing new DMEM culture medium, adding 10 muL of WST-8 staining solution in a dark place, incubating for 30min at 37 ℃, using an enzyme linked immunosorbent assay detector to absorb light at 450nm, mixing blank control with the WST-8 staining solution with the cell-free culture solution with the same amount, and calculating the cell survival rate.
The drug groups were pretreated for 4h by adding 100 μ L of BBr60 at various concentrations and washed 2 times with PBS. The model group and the drug group are placed in an ultraviolet lamp box, UVA and UVB lamp tubes are 15cm away from a table top, the control group is placed outside the lamp box in a similar environment, the culture solution is discarded after irradiation, the three groups of cells are replaced by DMEM (serum-free) culture medium, and the light damage index is detected after continuous culture for 24 hours.
Control group: no ultraviolet irradiation is carried out; UVA: UVA irradiation (model set); administration group: 8x109cfu/mL、2x109cfu/mL、8x108cfu/mL (different concentrations of BBr60 treated human dermal fibroblasts).
Control group: no ultraviolet irradiation is carried out; UVB: UVB irradiation (model set); administration group: 8x109cfu/mL、2x109cfu/mL、8x108cfu/mL (different concentrations of BBr60 treated keratinocytes).
3.5 index for detecting antioxidant damage
Human dermal fibroblasts or keratinocytes intracellular ROS levels were determined by DCFH-DA. After 24h of culture, each group of cells was washed with PBS 2 times, incubated with DCFH-DA for 30min, digested with 0.25% trypsin, centrifuged (1000rmp, 10min), washed with PBS 2 times, centrifuged and the cells collected. The cells were resuspended in PBS and observed with a fluorescence microscope. Fluorescence intensity was measured at an optimum excitation wavelength of 502nm and an optimum emission wavelength of 530 nm.
The results of the experiment are shown in fig. 3, and the intracellular ROS levels of human dermal fibroblasts and keratinocytes after exposure to UVA and UVB, respectively, were significantly increased compared to the control group. ROS levels were significantly reduced in fibroblasts and keratinocytes after BBr60 treatment compared to the model group, with the differences being statistically significant (p < 0.05).
3.6 testing the protective Effect of BBr60 on human dermal fibroblasts
3.6.1 detection of matrix Metalloproteinase Activity in human dermal fibroblasts
The activity of matrix metalloproteinases MMP-1 and MMP-3 in human dermal fibroblasts was detected by ELISA. After each group of cells was cultured for 24 hours, the supernatant was collected, quantified, and stored at-80 ℃. Adding anti-human MMP-1 and MMP-3 primary antibody and secondary antibody for incubation according to the instruction, and calculating the expression quantity of matrix metalloprotease MMP-1 and MMP-3 according to a standard curve equation.
As shown in FIG. 4, the expression levels of matrix metalloproteinases MMP-1 and MMP-3 in human dermal fibroblasts were significantly increased after UVA irradiation. However, when BBr60 is treated and then irradiated by ultraviolet, the expression of matrix metalloproteinases MMP-1 and MMP-3 in human dermal fibroblasts is reduced, when the BBr60 treatment concentration is 8x109cfu/mL、2x109The comparison difference between cfu/mL and the model group is significant, and the statistical significance is achieved (p)<0.05)。
3.6.2 detection of collagen Synthesis and Cross-linking degree of human dermal fibroblasts
The contents of type I collagen, type III collagen, hydroxyproline and hyaluronic acid secreted by human dermal fibroblasts are measured by an ELISA kit. Culturing each group of cells for 24h, collecting supernatant, freezing at-80 deg.C for testing, performing 0.4% trypan blue staining on each group of cells, and converting into 1 × 10 according to the number of cells in each well5The level secreted by individual cells. Strictly operating according to the specification, measuring the light absorption value at 450nm of the microplate reader, and calculating the contents of type I collagen, type III collagen, hydroxyproline and hyaluronic acid according to a standard curve equation.
As shown in FIG. 5, the amounts of type I, type III collagen, hydroxyproline and hyaluronic acid in human dermal fibroblasts significantly decreased after exposure to UVA (p) compared with the control group<0.05), but the content of I type collagen, III type collagen, hydroxyproline and hyaluronic acid in cells is improved when BBr60 is treated and then ultraviolet irradiation is carried out, and when the treatment concentration of BBr60 is 8x109cfu/mL、2x109The comparison with the model group is obvious when cfu/mL (p)<0.05)。
3.7 animal test for repairing action on photodamage
3.7.1 photo-aging animal model establishment
48 male ICR mice with the age of 9 weeks are adaptively raised in a well-ventilated and sanitary animal room for 1 week (the temperature is 22 +/-2) DEG C, the relative humidity is 40% -70%, the illumination is 150-300 lx, and the illumination time is 12 h/day) under the control that the weight is 25-30 g. Randomly grouping after 1 week (control group, not receiving ultraviolet radiation, only coating pure matrix (glycerol), model group, receiving ultraviolet radiation, administration group, adding BBr60 bacterial solution into glycerol, and controlling final concentration to 8X109cfu/mL、2x109cfu/mL、8x108cfu/mL). The mice were anesthetized with ether, exposing hairless skin 1.0cm x 1.0cm on the back, after which the mouse hair was trimmed appropriately according to growth. Respectively placing the model group and the administration group mice in a rectangular experimental box with the size of 3cm multiplied by 6cm, respectively coating the exposed skin with a pure matrix and a preparation containing BBr60, wherein each medicine is 0.1mL, and the coating thickness is 0.1 cm; pretreating for 15min, and sequentially irradiating with UVA and UVB (UVA dose of 1.55J/cm)218min, UVB dose of 0.95J/cm211min), the irradiation height is 30 cm; other parts of the mouse body are shielded by a shading plate. 1 time daily for 7 days.
3.7.2 detection of various biochemical indexes
After irradiation, a mouse is killed by 20% urethane anesthesia, wound tissue is separated and the operation is carried out, wherein partial tissue is taken and cut into pieces, the cut tissue is placed into a homogenate tube according to the proportion of 1g (tissue) to 9mL (normal saline), 10% tissue homogenate is prepared, cells are completely crushed, and related biochemical indexes are detected, (1) ELISA is used for detecting the content of IL-1 β, IL-6 and TNF- α in skin tissue, (1) the skin tissue sample is treated according to the corresponding ELISA kit instruction, the content of IL-1 β, IL-6 and TNF- α is respectively measured to determine the optical density value at the wavelength of 450nm, (2) the content of the skin tissue, MDA and CAT is measured by using the improved trace thiobarbituric acid (TBA) colorimetric method, the content of MDA, IL-1 β, IL-6 and TNF- α in each unit of protein is calculated according to the protein concentration measured by the BCA method, (2) the sample is treated by using the SOD and CAT detection kit instruction, the D value of 450nm and the standard SOD curve is drawn in the sample.
As shown in FIG. 6, the chronic medium-long wave UV irradiation can increase the content of inflammatory factors IL-1 β, IL-6 and TNF- α in the irradiated skin tissue of mice, while the BBr60 pretreatment of skin can improve the abnormal content of inflammatory factors in the skin tissue caused by UV irradiation when the treatment concentration is 8x109cfu/mL、2x109cfu/mL, statistical significance with respect to model group (p)<0.05). The BBr60 can relieve lipid peroxidation caused by ultraviolet irradiation and improve skin oxidative stress caused by ultraviolet irradiation.
In actual use, the bifidobacterium breve of the present invention may be added to cosmetics, skin care products or pharmaceuticals for repairing ultraviolet damage of skin and preventing photoaging of skin, for example: astringent, lotion, cream, facial mask, essence, foundation solution, concealer, sunscreen cream, oral liquid, granule, tablet, pill, powder, capsule or drop pill, etc. Adjuvants can also be added into cosmetics, skin care products or pharmaceuticals, such as: grease, surfactant, polyalcohol, humectant, defoamer, thickener, preservative, antioxidant, pH regulator, lubricant, emulsifier or adhesive, etc.
The embodiments of the present invention have been described in detail with reference to the accompanying drawings, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and the scope of protection is still within the scope of the invention.
Claims (7)
1. Bifidobacterium breve with the functions of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging is characterized in that: the Bifidobacterium breve is named as Bifidobacterium breve, and the preservation number is as follows: CGMCC No. 12915.
2. A method for preparing Bifidobacterium breve having effects of repairing UV damage, relieving inflammation, and preventing skin photoaging according to claim 1, comprising: the method comprises the following steps:
(1) collecting a fecal sample, dissolving with sterile water, diluting in a gradient manner, and adding into a sterilized flat plate;
(2) adding a hydrogen peroxide solution into an MRS culture medium at the temperature of 50-50 ℃ to ensure that the initial hydrogen peroxide concentration in the culture medium is 0.5mmol/L to obtain a sample culture medium;
(3) pouring the sample culture medium into the flat plate in the step (1) and uniformly mixing, selecting a strain with good growth vigor of bacterial colonies on the flat plate, performing gene sequence comparison, and determining the strain type, thereby screening the lactobacillus with hydrogen peroxide tolerance;
(4) the selected lactobacillus with hydrogen peroxide tolerance has antioxidant performance, namely clearance rate of DPPH free radical, ABTS free radical and hydroxyl free radical, as re-screening index, and then the strain with higher antioxidant activity is selected, and the required bifidobacterium breve is obtained by separation.
3. The method for preparing bifidobacterium breve having effects of repairing ultraviolet injury, relieving inflammation and preventing skin photoaging according to claim 2, wherein the method comprises the following steps: the MRS culture medium in the step (2) is as follows: the following raw materials in parts by weight: 10.0 parts of peptone, 8.0 parts of beef extract, 4.0 parts of yeast extract, 20.0 parts of lactose, 2.0 parts of dipotassium hydrogen phosphate, 5.0 parts of sodium acetate, 0.2 part of magnesium sulfate, 0.05 part of manganese sulfate, 1.0 part of L-cysteine hydrochloride, 2.0 parts of triammonium citrate, 801.0 parts of tween and 20.0 parts of agar, adjusting the pH to 6.0-6.5, adding distilled water to 1000mL, and sterilizing at 125 ℃ of 120-.
4. Use of bifidobacterium breve having effects of repairing uv damage, relieving inflammation, preventing skin photoaging according to claim 1, wherein: the Bifidobacterium breve is added into cosmetics, skin care products or medicaments for repairing skin ultraviolet injury and preventing skin photoaging.
5. The use of Bifidobacterium breve having effects of repairing UV damage, relieving inflammation, preventing skin photoaging according to claim 4, wherein: the bifidobacterium breve is added into lotion, emulsion, cream, mask, essence, foundation solution, concealer, sunscreen cream, oral liquid, granules, tablets, pills, powder, capsules or dropping pills for repairing ultraviolet injury of skin and preventing photoaging of skin.
6. The use of Bifidobacterium breve having effects of repairing UV damage, relieving inflammation, preventing skin photoaging according to claim 4, wherein: the cosmetic, skin care product or medicinal product can be added with adjuvants.
7. The use of Bifidobacterium breve having effects of repairing UV damage, relieving inflammation, preventing skin photoaging according to claim 6, wherein: the auxiliary materials are at least one of grease, surfactant, polyalcohol, humectant, defoamer, thickener, preservative, antioxidant, pH regulator, lubricant, emulsifier or adhesive.
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Address after: 215000 No. 1033, long Qiao Road, Wujiang economic and Technological Development Zone, Suzhou, Jiangsu Patentee after: Weikang probiotics (Suzhou) Co.,Ltd. Address before: 215000 No. 1033, long Qiao Road, Wujiang economic and Technological Development Zone, Suzhou, Jiangsu Patentee before: JIANGSU WECARE BIOTECHNOLOGY Co.,Ltd. |
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