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

Active Publication Date: 2020-10-27
JIANGNAN UNIV YANGZHOU FOOD BIOTECH INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] CN104523761A discloses a genetically engineered strain of Lactococcus lactis capable of inducibly expressing interleukin 12, which can inhibit the growth of intestinal tumors, but this bacterium is not obtained from nature and can only be used for medicine and not for food
The above three patented strains are clear and have clear functions, but the animal model involved uses colon cancer cell lines subcutaneously bearing tumors, which is quite different from the tumor environment in the intestine, and the role in the regulation of tumor-related signaling pathways is not clear

Method used

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  • A kind of Bifidobacterium bifidum capable of alleviating colorectal cancer and its application
  • A kind of Bifidobacterium bifidum capable of alleviating colorectal cancer and its application
  • A kind of Bifidobacterium bifidum capable of alleviating colorectal cancer and its application

Examples

Experimental program
Comparison scheme
Effect test

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 %.

[0...

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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Abstract

The invention discloses a Bifidobacterium bifidum capable of significantly inhibiting the occurrence of colorectal cancer and its use, belonging to the technical field of microbes. Bifidobacterium bifidum HNJ6 provided by the present invention has a preservation number of GDMCC No.60255, can tolerate gastric acid and bile salts, and can significantly reduce the inflammatory response in the colorectal part of colorectal cancer model mice, by regulating colorectal tissue The expression of Notch1 and Notch2 signaling pathways and VEGFR2 molecules in the tumor can reduce the number of tumors in the nodal and rectal regions of model mice. In addition, Bifidobacterium bifidum HNJ6 can also improve the structure of intestinal flora and the level of short-chain fatty acids in the intestine. The bifidobacterium bifidum HNJ6 is used to prepare fermented food that inhibits the occurrence of colorectal cancer, and has very broad application prospects.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Bifidobacterium bifidum capable of significantly inhibiting the occurrence of colorectal cancer by regulating Notch1, Notch2 and VEGFR2 signaling pathways and its application. Background technique [0002] Colorectal cancer is a malignant gastrointestinal tumor. As the morbidity and mortality of colorectal cancer continue to rise in my country, its threat to the health of our people is gradually deepening. At present, the mainstream methods for the clinical treatment of colorectal cancer all have relatively large side effects, which greatly affect the quality of life of patients. As a safe edible food, probiotics have been gradually confirmed by researches to alleviate and treat various diseases, such as promoting digestion and absorption, enhancing immunity, preventing reproductive system infections, alleviating allergic reactions, preventing and inhibiting Tumors, pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L33/135A23L19/00A23L11/00A23C11/10A23C9/12A23C13/16A23C19/032C12R1/01A23L11/50A23L11/65
Inventor 陈卫王刚王尔胤赵建新张灏
Owner JIANGNAN UNIV YANGZHOU FOOD BIOTECH INST
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