Lactiplantibacillus plantarum HNU082 with antagonistic effect and application thereof

A technology of HNU082 and Lactobacillus plantarum, applied in the field of microorganisms, can solve the problems of destroying normal intestinal flora colonization resistance, intestinal flora damage, aggravating inflammation, etc., to enhance the intestinal and systemic immune response, and better antagonism Function, effect of increasing content

Active Publication Date: 2021-07-16
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, treatment with antibiotics has an impact on the gut microbiota of mice, as it non-selectively kills other bacteria, alters the gut microbiota structure, reduces overall microbial diversity, and leads to microbiota disturbance
At the same time, antibiotics disrupt the colonization resistance of the normal gut flora, making it easier for pathogens or potential pathogens to multiply
In addition, antibiotics can have a temporary or permanent effect on the metabolic capacity of gut microbes, which is irreversible damage to the gut microbiota
To specifically inhibit the proliferation of Fusobacterium nucleatum, the researchers used phage therapy, but this may exacerbate inflammation because phages increase epithelial cell permeability and alter intestinal immunity

Method used

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  • Lactiplantibacillus plantarum HNU082 with antagonistic effect and application thereof
  • Lactiplantibacillus plantarum HNU082 with antagonistic effect and application thereof
  • Lactiplantibacillus plantarum HNU082 with antagonistic effect and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Screening and Identification of Lactobacillus plantarum HNU082

[0034] Preliminary screening: from farmers’ markets in different regions of Hainan Province, China (including eastern regions: Wanning, Wenchang, Qionghai, Lingshui; western regions: Lingao, Dongfang, Changjiang, Baisha; middle regions: Qiongzhong, Wuzhishan) Collect 36 parts of fermented vegetable samples, accurately weigh 10g fermented vegetable juice and mix in 90mL sterile NaCl solution (0.85%, W / V), and prepare a gradient of 10 by 10-fold dilution method. -1 -10 -5 sample diluent, take 10 -3 、10 -4 、10 -5 100 μL of concentration gradient dilutions were spread on MRS agar medium respectively, anaerobically cultured at 37°C for 48 hours, and 29 strains for primary screening were obtained after repeated isolation and purification.

[0035] Re-screening: Test the acid resistance and bile salt resistance of the above 29 primary screened strains, and further screen out the strains with strong ...

Embodiment 2

[0051] Example 2 In vitro antagonism experiment of Lactobacillus plantarum HNU082 on Fusobacterium nucleatum

[0052] In order to study the antagonism of Lactobacillus plantarum HNU082 and Fusobacterium nucleatum in vitro, the above two strains were inoculated into BHIS liquid medium (MRS liquid medium), co-cultivated at 37 ° C, 150 rpm / min, and at different times Point (2h, 6h, 12h, 24h, 48h) to extract DNA, using real-time fluorescence quantitative polymerase chain reaction (quantitative real-time Polymerase Chain Reaction, qPCR) method to obtain the abundance of F. nucleatum and L. plantarum HNU082. The experiment was divided into 4 groups, and the specific information is shown in Table 3:

[0053] Table 3 Experimental Grouping Situation

[0054] project Grouping First group BHIS+10 7 CFU / mL L. plantarum HNU082+10 7 CFU / mL F.nucleatum

Second Group BHIS+10 7 CFU / mL L. plantarum HNU082+10 5 CFU / mL F.nucleatum

The third group MRS+10...

Embodiment 3

[0065] Example 3 Lactobacillus plantarum HNU082 antagonizes Fusobacterium nucleatum in vivo animal experiment

[0066] Twenty-four 4-week-old C57BL / 6J male mice were taken. After 14 days of normal feeding and adaptation, the mice were randomly divided into 3 groups. The specific groups are shown in Table 4. There were 8 mice in each group, and the daily gavage volume of each group was 300 μL; all mice were intervened for 6 weeks, and on the last day, the mice were euthanized after fasting for 24 hours, and then the blood was collected from the eyeball, and the colon and rectum were collected. operate.

[0067] Table 4 Grouping of experimental mice

[0068]

[0069] 3.1 Antagonism test of Lactobacillus plantarum HNU082 on Fusobacterium nucleatum in mice

[0070] Take the intestinal contents of the mice in each experimental group, part of which is used for the test of antagonism, and the other part is used for the test of short-chain fatty acids. Specifically, adopt the me...

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Abstract

The invention particularly discloses Lactiplantibacillus plantarum HNU082 with an antagonistic effect and application thereof. The strain is preserved in Guangdong Microbial Culture Collection Center (GDMCC) on March 9, 2021, the preservation number is GDMCC NO 61552: and the preservation address is the 5 floor, Laboratory Building of the Institute of Microbiology, #100, Middle Xianlie Road, Guangzhou City, China; and the taxonomic name is Lactiplantibacillus plantarum. According to the Lactiplantibacillus plantarum HNU082 disclosed by the invention, one strain of Lactiplantibacillus plantarum HNU082 is screened from a lot of samples; the strain has a relatively good antagonistic effect on Fusobacterium nucleatum in vitro or in vivo; furthermore, the Lactiplantibacillus plantarum HNU082 can be used for inducing the generation of anti-inflammatory cytokines and enhancing intestinal and systemic immune responses; the growth of pathogenic bacteria is inhibited through reducing the pH (potential of Hydrogen) value and the content of short-chain fatty acid is increased, so that the expression of inflammatory factors is reduced; and a feasible intervention strategy is provided for in-vivo antagonism of probiotics on the Fusobacterium nucleatum and a reliable basis is provided for preventing early-stage colorectal cancer.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to an antagonistic Lactobacillus plantarum HNU082 and its application. Background technique [0002] Colorectal cancer (CRC) is a common gastrointestinal malignancy and ranks second in the world's leading cause of cancer death. Especially in people under the age of 50, the incidence of colorectal cancer is on the rise. It is estimated that by 2030, the number of colorectal cancer patients will increase by 60%, with more than 2.2 million new cases and 1.1 million deaths. Many factors are related to the pathogenesis of CRC, such as diet, genetics and other factors; in addition to the above factors, a large number of studies have shown that CRC is closely related to some bacteria in the gastrointestinal tract, and more and more evidence shows that Fusobacterium nucleatum (Fusobacterium nucleatum) is A potential biomarker that is positively associated with the develo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/747A61P31/04A61P29/00A61P35/00C12R1/25
CPCA61K35/747A61P31/04A61P29/00A61P35/00Y02A50/30
Inventor 张家超汪媛媛黎嘉和霍冬雪张琳韩哲沈思源
Owner HAINAN UNIVERSITY
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