A kind of Bifidobacterium bifidum capable of alleviating colorectal cancer and its application
A technology of Bifidobacterium bifidum and microbial strains, applied in the field of microbiology, can solve the problems of not being able to be used in food and unclear regulation of tumor-related signaling pathways, and achieve the level of relieving inflammation, good gastric acid resistance and bile salt resistance, The effect of broad application prospects
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Embodiment 1
[0069] Example 1: Bifidobacterium bifidum HNJ6 has good tolerance to simulated gastrointestinal fluid
[0070] Streak inoculation of cryopreserved Bifidobacterium bifidum HNJ6 in mMRS medium (MRS medium + 0.05% cysteine hydrochloride) solid medium, anaerobic culture at 37°C for 48h, and then mMRS culture After 2 to 3 times of subculture, take the culture solution of Bifidobacterium bifidum HNJ6, centrifuge at 8000×g for 5 minutes to collect the bacteria, and resuspend in (1:1) artificial simulated gastric juice of pH 2.5 (containing 1% pepsin, pH = 2.5 mMRS medium) mixed, and then anaerobic culture at 37 ° C, samples were taken at the beginning (0h), 0.5h, 1h and 2h, respectively, and the mMRS agar medium was poured and cultured for plate colony counting to determine the number of viable bacteria and calculate its survival rate. The survival rate is the ratio of the number of viable bacteria in the culture medium to the number of viable bacteria at 0h, expressed in %.
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Embodiment 2
[0076] Example 2: Regulation of Bifidobacterium bifidum HNJ6 on Notch1, Notch2, VEGFR2 and colon cancer-related signaling pathways in HT-29 cells
[0077] Use RPMI1640 cell culture medium without antibiotics to resuspend the bacteria sludge of Bifidobacterium bifidum HNJ6 (or control Bifidobacterium bifidum BB37, LGG, Escherichia coli) after washing twice with PBS, and adjust the cell density to about 2×10 8 CFU / mL; add the HNJ6 (or BB37, LGG, E.coli) suspension into the 6-well plate cultured with HT-29 according to the amount of 2 mL of bacterial suspension per well (the cell fusion degree reaches 95%), and the blank group Add 2 mL of antibiotic-free cell culture medium and place at 37 °C with 5% CO 2 Incubate in a cell culture incubator for 2 hours; wash the cells 3 times with PBS, add 1 mL TRIzol to each well (6-well plate), let stand at room temperature for 5 minutes, repeatedly pipette with an enzyme-free pipette tip, and then move to an enzyme-free 1.5 mL EP tube, and ...
Embodiment 3
[0081] Example 3: Bifidobacterium bifidum HNJ6 has no acute toxic side effects on mice
[0082] Bifidobacterium bifidum HNJ6 was resuspended in 2% (w / v) sucrose solution, and the cell density was 4.0×10 9 CFU / mL. Take 10 healthy male BALB / c mice with a body weight of about 25 g, give the suspension of this concentration orally once a day, observe for a week, and record the death and body weight.
[0083] The results of these tests are listed in Table 4. These results indicated that feeding concentrations of 4.0×10 9CFU / mL Bifidobacterium bifidum HNJ6 had no significant effect on mice, no significant change in body weight, and no death. The appearance of the mice had no obvious pathological symptoms.
[0084] Table 4 Changes in body weight and death conditions of mice
[0085] time (days) 1 2 3 4 5 6 7 weight (g) 24.7±1.8 25.1±1.2 25.4±1.3 25.5±1.5 25.7±1.3 25.8±1.1 26.0±1.7 Circumstances of death - - - - - - -
[0086] Note: -...
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