Bifidobacterium breve with effects of repairing ultraviolet injury, relieving inflammation and preventing skin light aging, and preparation method and application thereof
A Bifidobacterium breve, skin photoaging technology, applied in the field of microbiology, can solve the problems of expensive, side effects, etc.
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Embodiment 1
[0027] Collect feces samples, dissolve them in sterile water, serially dilute them, add them to a sterilized plate, add hydrogen peroxide solution to the MRS medium at about 55°C, and make the initial concentration of hydrogen peroxide in the medium be 0.5mmol / L, pour the culture medium on the plate and mix well, the formula of MRS medium is: peptone 10.0g, beef extract 8.0g, yeast extract 4.0g, lactose 20.0g, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, sulfuric acid Magnesium 0.2g, manganese sulfate 0.05g, L-cysteine hydrochloride 1.0g, triammonium citrate 2.0g, Tween 801.0g, agar 20.0g, pH6.0~6.5, add distilled water to 1000mL, Sterilize at 121°C for 20 minutes. Select the strains with good colony growth on the plate, compare the gene sequences, and determine the strain type, so as to screen out the lactobacilli with hydrogen peroxide tolerance, and use the antioxidant properties to resist DPPH free radicals, ABTS free radicals and hydroxyl free radicals. T...
Embodiment 2
[0029] To detect the effect of Bifidobacterium breve (referred to as BBr60) on the proliferation activity of human dermal fibroblasts and keratinocytes:
[0030] 2.1 Bacteria solution preparation
[0031] Inoculate BBr60 to 10 mL of MRS medium from the frozen tube, and culture it statically under anaerobic conditions at 37°C for 20 h. On the next day, 200 μl of the culture solution was inoculated into 10 mL of MRS medium (2% secondary activation), and cultured statically at 37° C. under anaerobic conditions for 20 h. Take out 2ml of the cultured bacteria solution and centrifuge (13000rmp, 1min). After the PBS buffer has bacterium twice, remove the supernatant, take 1mL PBS to suspend the bacteria block, mix well, measure OD600, and adjust the number of bacteria to 1x1011cfu / mL, in a water bath at 95°C for 10 minutes, for later use.
[0032] 2.2 Cell culture
[0033] Human dermal fibroblasts or keratinocytes were revived in a 37°C water bath, placed in DMEM medium containin...
Embodiment 3
[0038] Detection of BBr60 repair or preventive effect on human skin fibroblasts and keratinocytes:
[0039] 3.1 Bacteria culture
[0040] Inoculate BBr60 to 10ml of MRS medium from the frozen tube, and culture it statically under anaerobic conditions at 37°C for 20h. On the next day, 200 μl of the culture solution was inoculated into 10 ml of MRS medium (2% secondary activation), and cultured statically at 37° C. under anaerobic conditions for 20 h.
[0041] 3.2 Cell culture
[0042] Human dermal fibroblasts or keratinocytes were revived in a water bath at 37°C, placed in DMEM medium containing 20% fetal bovine serum (FBS), 100U / ml penicillin and 100U / ml streptomycin, kept at 37°C, 5% CO2 cultured in an incubator.
[0043] 3.3 Experimental grouping
[0044] Divided into blank group, model group, drug treatment group. Blank group: cultured in untreated DMEM medium. Model group: cultured in untreated DMEM medium, fibroblasts were irradiated with UVA, and keratinocytes we...
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