Bifidobacterium bifidum NX-7 and application of NX-7 in preparation of medicine for treating ischemic diseases

A Bifidobacterium bifidum and NX-7 technology, applied in the field of microorganisms, can solve problems such as not necessarily suitable, less research and application of probiotics, and lack of scientific research evidence for the function of probiotics, so as to promote damage repair, regeneration and reconstruction , the effect of promoting the growth of zebrafish inferior intestinal vein

Active Publication Date: 2021-02-23
FOSHAN LANGSEE BIOTECHNOLOGY CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current research and application of probiotics in ischemic diseases is less
At the same time, the current international patent applications for probiotics are concentrated in the United States, Japan, and Russia, traditional research and development powerhouses, while my country lacks functional strains with independent intellectual property rights
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Method used

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  • Bifidobacterium bifidum NX-7 and application of NX-7 in preparation of medicine for treating ischemic diseases
  • Bifidobacterium bifidum NX-7 and application of NX-7 in preparation of medicine for treating ischemic diseases
  • Bifidobacterium bifidum NX-7 and application of NX-7 in preparation of medicine for treating ischemic diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Isolation, Identification and Preservation of Bifidobacterium bifidum NX-7

[0028] (1) Separation: Inoculate the feces of infants and young children into TPY solid medium, MRS solid medium, BHI solid medium, and BS solid medium after gradient dilution, culture anaerobically at 37°C for 48 hours, and pick the feces on the plate. Single colonies were streaked to obtain pure colonies. The pure colony on the plate was inoculated in BS liquid medium, cultured anaerobically at 37°C for 12-16h, added 20% glycerol, and stored in a -80°C refrigerator.

[0029] (2) Morphological identification of bacterial strains: Gram-stained strains were observed under a microscope, Gram-positive bacteria were purple, and Gram-negative bacteria were red.

[0030] (3) Molecular biological identification of the strain: Genomic DNA was extracted from the obtained strain, and the full-length 16S rDNA fragment was amplified by PCR using 16S rDNA universal primers 27F and 1492R, followed...

Embodiment 2

[0036] Preparation of embodiment 2 Bifidobacterium bifidum NX-7 fermentation supernatant (extracellular secretion), bacterium suspension (thalline)

[0037] Bifidobacterium bifidum NX-7 was activated and cultured and inoculated in BS liquid medium. After culturing at 37°C for 15 hours, the concentration of fermentation bacteria was adjusted to 1×10 6 CFU / mL, centrifuged at 4°C, 6000r / min for 10min to obtain the culture supernatant and cell pellet, the supernatant was filtered through a 0.22μm filter membrane to obtain the fermentation supernatant (extracellular secretion); the cell pellet was filtered through PBS twice After the first wash, the cells were resuspended with PBS, and the cell concentration was adjusted to 1×10 6 CFU / mL to obtain bacterial suspension (cell); fermentation supernatant (extracellular secretion) and bacterial suspension (bacterial body) were heated at 121°C for 15 minutes to make heat-inactivated fermentation supernatant (extracellular secretion ), b...

Embodiment 3

[0038] Example 3 Effect of Bifidobacterium bifidum NX-7 on the development of inferior intestinal vein in zebrafish

[0039] Select healthy zebrafish Tg(fli1:EGFP) developed to 2hpf (hours post fertilization) and place them in a 6-well cell culture plate, 15 cells / well, add PBS to the blank group, add non-inactivated or inactivated bacteria to the bacterial suspension group Add non-inactivated or inactivated fermentation supernatant to the fermentation supernatant group, 3 mL per well, change the solution every 24 hours, incubate at 28°C for 72 hours, anesthetize the zebrafish with tricaine, observe under a fluorescent microscope The growth of the inferior intestinal vein was taken and photographed, and then the number of budding of the inferior intestinal vein was counted. SPSS19.0 software was used to statistically process the data, and the experimental data were all used Data are represented by one-way analysis of variance. Comparing each experimental group with blank gr...

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Abstract

The invention discloses bifidobacterium bifidum NX-7 and application of NX-7 in preparation of a medicine for treating ischemic diseases, and belongs to the technical field of microorganisms. The preservation number of the bifidobacterium bifidum NX-7 disclosed by the invention is CGMCC (China General Microbiological Culture Collection Center) No. 20115. The inactivated and non-inactivated fermentation supernatant and bacterial suspension of the bifidobacterium bifidum NX-7 can significantly promote the growth of the zebra fish subintestinal veins in vivo, significantly repair the injury of the zebra fish subintestinal veins induced by a vascular endothelial cell growth factor receptor inhibitor (PTK787), significantly promote the repair of the injury of the zebra fish tail fin, has the potential of promoting microcirculation regeneration and reconstruction, and shows the effect of being applied to in-vivo treatment of ischemic diseases. The bifidobacterium bifidum NX-7 disclosed by the invention has a huge potential application prospect in the aspect of treating ischemic diseases.

Description

technical field [0001] The invention relates to the technical field of microbes, and more specifically relates to Bifidobacterium bifidum NX-7 and its application in the preparation of medicines for treating ischemic diseases. Background technique [0002] At present, the main treatment methods for ischemic cardiovascular disease and cerebrovascular disease are percutaneous coronary intervention, surgical bypass, and drug thrombolysis. There are certain limitations. Promote cell proliferation, angiogenesis and repair through drug therapy or other intervention methods, and finally achieve tissue regeneration and microcirculation reconstruction, providing a new idea for the treatment of cardiovascular and cerebrovascular diseases. Therefore, therapeutic angiogenesis has become one of the research hotspots in the treatment of various ischemic diseases. Local administration of a single pro-angiogenic growth factor (VECF, bFGF) can effectively promote the growth of blood vessel...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/745A61P9/00A61P9/10C12R1/01
CPCA61K35/745A61P9/00A61P9/10C12R2001/01C12N1/205
Inventor 张召郑康帝马燕赵英杰陈涛
Owner FOSHAN LANGSEE BIOTECHNOLOGY CO LTD
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