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Application of lactobacillus acidophilus LA-03 in preparation of anti-helicobacter-pylori drug

A technology against Helicobacter pylori and Lactobacillus acidophilus, applied in the field of microorganisms, can solve the problems of lack of functional strains, not necessarily suitable, and dependence on imports of probiotic strains, etc., to inhibit the growth of Helicobacter pylori, improve inflammatory response, and improve The effect of the inflammatory response

Pending Publication Date: 2022-01-28
广东一元兰欣生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the current international patent applications for probiotics are concentrated in the traditional research and development powerhouses of the United States, Japan, and Russia, while my country lacks functional strains with independent intellectual property rights
The probiotic strains used by domestic production enterprises have long relied on imports, and foreign strains may not be suitable for the physiological conditions of the gastrointestinal tract of Chinese residents
In addition, the function of probiotics lacks strong scientific research evidence, which seriously affects the promotion of probiotics and their products

Method used

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  • Application of lactobacillus acidophilus LA-03 in preparation of anti-helicobacter-pylori drug
  • Application of lactobacillus acidophilus LA-03 in preparation of anti-helicobacter-pylori drug
  • Application of lactobacillus acidophilus LA-03 in preparation of anti-helicobacter-pylori drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Lactobacillus acidophilus LA-03 isolation, identification and preservation

[0023] (1) Separation: after gradient dilution of the spicy cabbage fermentation broth, inoculate them in BHI solid medium, TPY solid medium, MRS solid medium and BDS solid medium respectively, culture anaerobically at 37°C for 48 hours, and pick the A single colony was streaked to obtain pure colonies. The pure colony on the plate was inoculated into MRS liquid medium, cultured anaerobically at 37°C for 12-16h, added 20% glycerol, and stored in a -80°C refrigerator.

[0024] (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.

[0025] (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

[0032] Preparation of embodiment 2 Lactobacillus acidophilus LA-03 fermentation supernatant (extracellular secretion), bacterium suspension (thalline)

[0033] Lactobacillus acidophilus LA-03 was activated and cultured and inoculated in MRS liquid medium. After culturing at 37°C for 15 hours, the concentration of fermentation bacteria was adjusted to 1.5×10 7 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 1.5×10 7 CFU / mL to obtain bacterial suspension (bacteria).

Embodiment 3

[0034] The inhibitory effect of embodiment 3 Lactobacillus acidophilus LA-03 on the growth of Helicobacter pylori

[0035] The concentration of 100μL is 1×10 8 CFU / mL of Helicobacter pylori 26695 (ATCC 700392), Helicobacter pylori 11637 (ATCC 43504), and Helicobacter pylori 11916 (ATCC 43526) were evenly spread on Columbia agar plates (plate diameter 90 mm), without antibiotics. Then put 3 Oxford cups on each Columbia agar plate, and add 120 μL of Lactobacillus acidophilus LA-03 suspension, 120 μL of fermentation supernatant, and 120 μL of PBS to the 3 Oxford cups respectively, and put the plate in microneed Incubate at 37°C for 72h under oxygen condition, and then measure the diameter of the inhibition zone. PBS was used as blank control group. The experiment was repeated three times.

[0036] SPSS 19.0 software was used to statistically process the data, and the experimental data were expressed as x±SEM data, and one-way analysis of variance was used. Compared with blank...

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Abstract

The invention discloses application of lactobacillus acidophilus LA-03 in preparation of an anti-helicobacter-pylori drug, and belongs to the technical field of microorganisms. The fermentation supernatant and bacterial suspension of the lactobacillus acidophilus LA-03 disclosed by the invention can significantly inhibit the growth of helicobacter pylori, significantly inhibit the adhesion of the helicobacter pylori to gastric epithelial cells AGS, and significantly inhibit the helicobacter pylori from inducing the gastric epithelial cells AGS to secrete inflammatory factors IL-8, and show good effects of inhibiting the growth of the helicobacter pylori and improving the inflammatory reaction caused by the helicobacter pylori. The lactobacillus acidophilus LA-03 disclosed by the invention has a huge potential application prospect in the aspects of inhibiting the growth of helicobacter pylori and improving inflammation caused by the helicobacter pylori.

Description

technical field [0001] The invention relates to the technical field of microorganisms, and more specifically relates to the application of Lactobacillus acidophilus LA-03 in the preparation of anti-helicobacter pylori drugs. Background technique [0002] Helicobacter pylori is a major pathogen responsible for a range of gastric diseases, including gastroduodenal inflammation, peptic ulcer and gastric cancer. H. pylori infection induces the production of pro-inflammatory cytokines and chemokines such as interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), leading to gastric inflammation characterized by infiltration of the gastric mucosa by plasma cells, lymphocytes, neutrophils, and monocytes. IL-8 secreted by gastric epithelial cells is a potent neutrophil activator and chemoattractant and plays a major role in Helicobacter pylori-induced mucosal inflammation. Increased levels of IL-8 have been reported in gastric fluid and ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P1/04A61P31/04C12R1/23
CPCA61K35/747A61P31/04A61P1/04Y02A50/30
Inventor 张召郑康帝刘彦陈涛
Owner 广东一元兰欣生物科技有限公司
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