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Application of L.p (lactobacillus paracasei) R3-10 in preparation of medicines for preventing or treating oral inflammatory diseases

A technology of oral inflammation and Lactobacillus, applied in the field of microbiology, can solve the problems of dependence on import of probiotic strains, lack of scientific research evidence for the function of probiotics, and lack of functional strains, etc., to achieve the goal of inhibiting aggregation, promoting clearance, and promoting damage repair Effect

Active Publication Date: 2020-11-10
广东南芯医疗科技有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current probiotic products on the domestic market are mainly aimed at intestinal problems, while there are few probiotic products for the oral cavity
At the same time, 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
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 oral cavity and 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 L.p (lactobacillus paracasei) R3-10 in preparation of medicines for preventing or treating oral inflammatory diseases
  • Application of L.p (lactobacillus paracasei) R3-10 in preparation of medicines for preventing or treating oral inflammatory diseases
  • Application of L.p (lactobacillus paracasei) R3-10 in preparation of medicines for preventing or treating oral inflammatory diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The isolation and identification of the precursor strain L.p R3 (Lactobacillus paracasei) of embodiment 1 L.p R3-10

[0037] (1) Sample source

[0038] Healthy babies aged 0-6 months were selected from the families of school staff as volunteers. Two weeks before sampling, a normal diet is required and there is no recent history of intestinal infection or antibiotic use. Take the morning stool on the day of sampling, and quickly use the intelligent microbial separation system of Nanjing Famet Company to separate the fecal bacteria after collection. After the separation was completed, the crude fecal bacteria liquid was collected quickly, added with cryopreservation protection solution, and then stored in a -80°C ultra-low temperature refrigerator for future use.

[0039] (2) Isolation, culture and identification of L.p R3

[0040] Take 1mL of the crude fecal extract liquid and add it to 9mL of normal saline, mix well and perform gradient dilution. Draw the dilution co...

Embodiment 2

[0043] Example 2 Induction and Identification of L.p R3-10

[0044] (1) Low nutrient gradient tolerance method induces L.p R3 to become L.p R3-10

[0045] Take out the frozen L.p R3 from the -80°C refrigerator and put it in a warm water bath at 37°C to melt it quickly. The thawed bacterial solution was poured into an anaerobic blood agar plate, and placed in an anaerobic condition at 37°C for static culture for 48 hours. Observe the growth of the colony on the plate and whether there is a hemolytic ring, observe the shape of the strain under a Gram staining microscope, and confirm that there is no pollution, transfer to the MRS agar plate, culture it under anaerobic conditions at 37°C for 24 hours, and pick a single cell on the plate. The colonies were inoculated in 6mL MRS liquid medium, and cultured and activated under anaerobic conditions at 37°C for 16-18h. The activated bacterial solution was inoculated into 100 mL of MRS broth with an inoculum amount of 3% (v / v), cultu...

Embodiment 3

[0055] Preparation of embodiment 3 Lactobacillus paracasei L.p R3-10 fermentation supernatant (extracellular secretion), bacterium suspension (thalline)

[0056] Lactobacillus paracasei L.p R3-10 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 2×10 8 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 2×10 8 CFU / mL to obtain bacterial suspension (bacteria). Fermentation supernatant (extracellular secretions), bacterial suspension (cells) were heated at 100°C for 20 minutes to prepare heat-inactivated fermentation supernatant (extracellular secreti...

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Abstract

The invention discloses an application of L.p R3-10 in preparation of medicines for preventing or treating oral inflammatory diseases, and belongs to the technical field of microorganisms. Inactivatedand non-inactivated fermentation supernatant and bacterial suspension of the L.p R3-10 disclosed by the invention have the effect of inhibiting LPS from stimulating human gingival fibroblasts to secrete cell inflammatory factors IL6, TNF-alpha and PGE2 in vitro; in an in-vivo inflammation model, neutrophils and macrophages can be remarkably inhibited from being gathered to the inflammation position of the tail fin of a zebra fish, and clearing of the neutrophils and the macrophages at the inflammation position of the tail fin of the zebra fish is remarkably promoted; and meanwhile, damage repair of the tail fin of the zebra fish can be remarkably promoted. The L.p R3-10 disclosed by the invention has a huge potential application prospect in the aspect of treating and / or preventing the oral inflammatory diseases.

Description

technical field [0001] The invention relates to the technical field of microorganisms, and more specifically relates to the application of a strain of Lactobacillus paracasei L.p R3-10 in the preparation of medicines for preventing or treating oral inflammatory diseases. Background technique [0002] Periodontal disease, mainly including gingivitis and periodontitis, is one of the common oral diseases and the leading cause of tooth loss in adults. Gingivitis is caused by the buildup of plaque on the tooth surface near the gum margin. In particular, the endotoxin lipopolysaccharide (LPS) produced by Gram-negative bacteria increases in content as plaque matures, triggering a nonspecific inflammatory immune response. This response is produced by gingival epithelial cells, fibroblasts, and leukocytes secreting proinflammatory factors (such as tumor necrosis factor-α [TNF-α] and interleukin-1 [IL-1]), chemokines (such as IL-8 ) and prostaglandins (such as prostaglandin E2 [PGE2...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/747A61P1/02A61P29/00A61K8/99A61Q11/00
CPCA61K8/99A61K35/747A61Q11/00A61P1/02A61P29/00
Inventor 张召曾今诚郑康帝
Owner 广东南芯医疗科技有限公司
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