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 produ

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

Example Embodiment

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

[0023] (1) Separation: After gradient dilution, the chili cabbage fermentation broth was inoculated into BHI solid medium, TPY solid medium, MRS solid medium and BDS solid medium, respectively, cultured at 37°C for 48h anaerobic, and picked out the plates. The single colonies were streaked to obtain pure colonies. The pure colonies on the plate were inoculated into MRS liquid medium, cultured anaerobic at 37°C for 12-16 hours, added with 20% glycerol, and stored in a -80°C refrigerator.

[0024] (2) Morphological identification of strains: The screened strains were Gram-stained and observed under a microscope. Gram-positive bacteria were purple and Gram-negative bacteria were red.

[0025] (3) Molecular biological identification of strains: genomic DNA was extracted from the obtained strains, and the full-length fragments of 16S rDNA were amplified by PCR technology using 16S rDNA...

Example Embodiment

[0032] Example 2 Preparation of Lactobacillus acidophilus LA-03 fermentation supernatant (extracellular secretion) and bacterial suspension (cell)

[0033] Lactobacillus acidophilus LA-03 was activated and cultured and then inoculated into MRS liquid medium. After culturing at 37°C for 15 hours, the concentration of fermented bacteria was adjusted to 1.5×10 7 CFU / mL, 4°C, 6000r / min centrifugation for 10min to obtain the culture supernatant and bacterial cell precipitate, the supernatant liquid was filtered through a 0.22μm filter to obtain the fermentation supernatant (extracellular secretion); After washing, the cells were resuspended in PBS, and the cell concentration was adjusted to 1.5×10 7 CFU / mL to obtain bacterial suspension (cell).

Example Embodiment

[0034] Example 3 Inhibitory effect of Lactobacillus acidophilus LA-03 on the growth of Helicobacter pylori

[0035] The concentration of 100 μL is 1×10 8 CFU / mL 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 Lactobacillus acidophilus LA-03 bacterial suspension, 120 μL fermentation supernatant, and 120 μL PBS to the 3 Oxford cups respectively. Incubate at 37 °C for 72 h under oxygen conditions, and then measure the diameter of the inhibition zone. PBS served as blank control group. The experiment was repeated three times.

[0036] SPSS 19.0 software was used for statistical processing of data, experimental data were expressed as x±SEM data, and one-way analysis of variance was used. Compared with blank control group: ***P<0.005.

[0037] The res...

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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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