Applications of Bacteroides fragilis in preparation of inducer for inducing polarization of macrophages into M1 macrophages

A technology of Bacteroides fragilis and macrophages, applied in the application field of Bacteroides fragilis

Inactive Publication Date: 2018-01-26
GUANGZHOU ZHIYI PHARMA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, the immune regulation mechanism involved in Bacteroides fragilis is being explored by many research institutions, and the research results a...

Method used

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  • Applications of Bacteroides fragilis in preparation of inducer for inducing polarization of macrophages into M1 macrophages
  • Applications of Bacteroides fragilis in preparation of inducer for inducing polarization of macrophages into M1 macrophages
  • Applications of Bacteroides fragilis in preparation of inducer for inducing polarization of macrophages into M1 macrophages

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Isolation of Bacteroides fragilis and preparation of samples

[0036] (1) Enrichment culture of Bacteroides fragilis and preparation of live bacteria solution

[0037] Streak inoculation of strains on blood plate, 37 ℃, anaerobic culture for 48h. Observe the colony morphological characteristics, staining characteristics, size, club shape and distribution, etc.

[0038] Colony characteristics: Bacteroides fragilis ZY-312, after being cultured on a blood plate for 48 hours, is slightly convex, translucent, white, smooth, non-hemolytic, and the diameter of the colony is between 1-3mm. See figure 1 .

[0039] Morphology under the microscope: Bacteroides fragilis ZY-312 was examined by Gram staining. It is a Gram-negative bacterium with a typical rod shape, blunt rounded ends and dense staining. The uncolored part in the middle of the bacteria is like a vacuole. figure 2 .

[0040] Select a single colony and inoculate it in the improved tryptone broth for enri...

Embodiment 2

[0050] Effect of Example 2 Sample on Macrophage Polarization

[0051] The live Bacteroides fragilis solution, inactivated bacteria solution, lysate solution and culture supernatant prepared in Example 1 were used for experiments respectively. In this example, fluorescent quantitative real-time PCR is used to detect 6 factors, the 6 factors are M1 macrophage-related factors iNOs, IL-1β, TNF-α and IL-12, and M2 macrophage-related factors Factors Arg-1, IL-10. At the same time, the internal reference is set, and the internal reference is GAPDH. The 6 factors and internal reference were detected by fluorescent quantitative real-time PCR to determine the phenotype of macrophages. The specific experimental steps are as follows:

[0052] 1. Design primers

[0053] Primers were designed for the above seven factors and internal references. The details are shown in Table 2:

[0054] Table 2 Related factor primers and internal reference primers

[0055]

[0056]

[0057] 2. ...

Embodiment 3

[0106] Example 3 Effects of Different Infection Time on Macrophage Polarization

[0107] In order to detect the influence of different infection times of the above samples on the polarization of macrophages, this example uses the live Bacteroides fragilis liquid sample C, the culture supernatant sample F, the inactivated bacterial liquid sample I and the lysate prepared in Example 1 Solution samples L were tested separately. Referring to the method of Example 2, take a 24-well plate covered with BMDM in good condition, discard the cell culture supernatant, and add the above four samples according to grouping, 1 mL per well, and place at 37 ° C, 5% CO 2 Incubate in the incubator for 3, 6, and 12 hours, respectively.

[0108] Using the operation method of Example 2, the M1-type macrophage-related factors iNOs, IL-1β, TNF-α and IL-12, and the M2-type macrophage-related factors Arg-1 and IL-10 were detected respectively. The specific results are shown in Tables 11-12 and Figur...

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Abstract

The present invention relates to applications of Bacteroides fragilis in preparation of an inducer for inducing the polarization of macrophages into M1 macrophages. According to the present invention,the results of a large number of experiments prove that Bacteroides fragilis can well induce the polarization of macrophages into M1 macrophages, such that it is indicated that Bacteroides fragilis can regulate the type and the function of immune reactions; and the viable bacteria, the inactivated bacteria, the lysate or the culture supernatant of Bacteroides fragilis as the intestinal bacteria can be used for preparing the inducer for inducing the polarization of macrophages into M1 macrophages.

Description

technical field [0001] The invention relates to the technical field of application of Bacteroides fragilis, in particular to the application of Bacteroides fragilis in preparing an inducer for inducing macrophage polarization. Background technique [0002] Macrophages are important phagocytic and antigen-presenting cells in the body, and play an important role in the body's processing of exogenous pathogenic microorganisms and danger signals generated in the body. Macrophages develop primarily from bone marrow stem cells. Under the stimulation of multi-colony-stimulating factor (multiCSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF), bone marrow stem cells develop into mononuclear blasts, which further differentiate into promonocytes and enter the bloodstream. Here differentiate into mature monocytes. Monocytes cross the vascular endothelium, migrate to different tissues, and differentiate into tissue-specific macrophages. Because macrophages are a hetero...

Claims

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Application Information

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IPC IPC(8): A61K35/741A61P43/00A23L33/135
Inventor 智发朝白杨王晔刘洋洋邓慧敏
Owner GUANGZHOU ZHIYI PHARMA INC
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