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Bifidobacterium bifidum capable of relieving colorectal cancer and application of Bifidobacterium bifidum

A technology of Bifidobacterium bifidum and microbial strains, applied in the field of microbiology, can solve the problems of unclear regulation of tumor-related signaling pathways and can not be used in food, etc., achieve good gastric acid and bile salt resistance, relieve inflammation, The effect of improving integrity

Active Publication Date: 2018-01-26
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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  • Bifidobacterium bifidum capable of relieving colorectal cancer and application of Bifidobacterium bifidum
  • Bifidobacterium bifidum capable of relieving colorectal cancer and application of Bifidobacterium bifidum
  • Bifidobacterium bifidum capable of relieving colorectal cancer and application of Bifidobacterium bifidum

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)

[0086] Note: -: no death of mice

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Abstract

The invention discloses Bifidobacterium bifidum with a remarkable colorectal cancer inhibition function and an application of the Bifidobacterium bifidum and belongs to the technical field of microorganisms. The Bifidobacterium bifidum HNJ6 with the preservation number being CGMCC No. 60255 can tolerate gastric acid and cholate and remarkably relieve inflammatory response at a colorectal part of acolorectal cancer model mouse, and the quantity of tumors at the colorectal part of the model mouse is reduced by regulating Notch1 and Notch2 signaling pathways in colorectal tissue and expression of VEGFR2 (vascular endothelial growth factor receptor 2) molecules. Besides, the Bifidobacterium bifidum HNJ6 can improve intestinal flora structure and short-chain fatty acid level in the intestinaltract. The Bifidobacterium bifidum HNJ6 can be used for preparing fermented food capable of inhibiting colorectal cancer and has quite extensive application prospect.

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 use. 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, prevention...

Claims

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Application Information

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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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