Arabinoxylan-bifidobacterium longum synbiotics and application of arabinoxylan-bifidobacterium longum synbiotics in regulation of arachidic acid metabolism
By preparing Arabidopsis-Bifibacterium longan synbiotics, using sodium alginate-chitosan nanoparticle embedding technology, the problem of unclear influence mechanisms of prebiotics and probiotics in regulating arachid-like metabolism was solved, effectively regulated intestinal flora and inflammatory response, and had significant disease improvement effects.
Patent Information
- Application Number
- CN202510612746.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-13
- Publication Date
- 2025-08-12
AI Technical Summary
The mechanism of the influence of prebiotics and probiotics on arachidoid metabolism in the prior art is unclear, especially in the study of synbiotics (probiotics + prebiotics), and its effect in regulating intestinal flora and inflammatory response is limited.
Arabidianxyllan-Bifitan longan synbiotic was prepared by optimizing raw material extraction, strain fermentation and microcapsule embedding techniques, and sodium alginate-chitosan nanoparticles were used as the embedding carrier to improve their tolerance and bioavailability in the gastrointestinal tract.
It significantly regulates the intestinal flora, increases the concentration of short-chain fatty acids (SCFAs), reduces the level of inflammatory factors, and improves chronic inflammation-related diseases, and has wide application prospects.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of microbial preparations, and in particular relates to an arabinoxylan-Bifidobacterium longum synbiotic and an application thereof in regulating eicosanoid metabolism. Background Art
[0002] Eicosanoids are a class of bioactive lipids produced by the metabolism of polyunsaturated fatty acids such as arachidonic acid (AA) and eicosapentaenoic acid (EPA). They include prostaglandins (PGs), thromboxanes (TXs), leukotrienes (LTs), and lipoxins (LXs), and have important influences on inflammatory responses, immune regulation, and vascular homeostasis. Eicosanoid metabolic pathways include three main pathways: COX, LOX, and CYP. Gut microbes can influence the expression of genes such as COX-2 and affect the synthesis of eicosanoids. For example, they can regulate the COX-2 pathway and affect the synthesis of PGE2. The mechanisms by which prebiotics and probiotics affect eicosanoid metabolism are not yet fully understood in existing research, and research and application of synbiotics (probiotics + prebiotics) are even rarer. Summary of the Invention
[0003] The purpose of the present invention is to provide an arabinoxylan-Bifidobacterium longum synbiotic, which improves the stability and functional activity of the synbiotic by optimizing raw material extraction, strain fermentation and microcapsule embedding technology.
[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:
[0005] The invention provides an arabinoxylan-Bifidobacterium longum synbiotic. The preparation method of the arabinoxylan-Bifidobacterium longum synbiotic comprises the following steps: mixing Bifidobacterium longum and arabinoxylan, adding a mixed solution of sodium alginate and chitosan for embedding, and freeze-drying to obtain the arabinoxylan-Bifidobacterium longum synbiotic.
[0006] Preferably, the mass ratio of the Bifidobacterium longum to arabinoxylan is 1:3-5.
[0007] Preferably, the number of viable Bifidobacterium longum is ≥1×10 10 CFU / g, and the purity of the arabinoxylan is ≥85%.
[0008] Preferably, the volume ratio of the mixture of Bifidobacterium longum and arabinoxylan to the mixed solution of sodium alginate and chitosan is 1-2:1-2.
[0009] Preferably, in the mixed solution of sodium alginate and chitosan, the concentration of sodium alginate is 1-3% w / v, and the concentration of chitosan is 0.5-2% w / v.
[0010] Preferably, the embedding conditions are: 20-30° C., 600-800 rpm, stirring for 55-65 min, and an embedding rate ≥85%.
[0011] Preferably, after freeze-drying, the water content of the arabinoxylan-Bifidobacterium longum synbiotic is ≤5%.
[0012] The present invention also provides the use of the arabinoxylan-Bifidobacterium longum synbiotic in the preparation of products for improving intestinal flora, regulating eicosanoid metabolism or reducing inflammatory responses.
[0013] The present invention utilizes Bifidobacterium longum as a probiotic, arabinoxylan as a prebiotic, and sodium alginate-chitosan nanoparticles as an encapsulation carrier. Nanoencapsulation technology significantly improves the gastrointestinal tolerance of the synbiotic and enhances its bioavailability. The combined use of arabinoxylan and Bifidobacterium longum has a better effect on regulating SCFA and eicosanoid metabolism than using either prebiotic or probiotic alone. This synbiotic can influence inflammatory factors through the "intestinal flora-SCFAs-eicosanoid metabolism axis," and has broad application prospects for chronic inflammatory diseases. BRIEF DESCRIPTION OF THE DRAWINGS
[0014] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.
[0015] Figure 1 is a comparison chart of serum COX-2, PGE2, LTB4 and LXA4 levels;
[0016] Figure 2 This is a comparison of inflammatory factor levels in mouse colon tissue;
[0017] Figure 3 is the concentration of EET and DHET in mouse serum;
[0018] Figure 4 is the DiHETE concentration in mouse serum;
[0019] Figure 5 Comparison chart of SCFAs levels in mice and relative expression levels of genes related to eicosanoid metabolism. DETAILED DESCRIPTION
[0020] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0021] Example 1
[0022] (1) Extraction of arabinoxylan:
[0023] Wheat bran was dried at 100°C and then crushed to pass through a 40-mesh sieve. It was then placed in a 0.25 mol / L NaOH solution (solid-liquid ratio 1:15, g / ml) and treated at 65°C for 30 min under ultrasonic power of 500 W, microwave power of 800 W, and filtration. The pH of the extract was adjusted to 4.5, and 95% ethanol solution was added for precipitation for 24 h (the supernatant was mixed with 95% ethanol in a ratio of 1:2 v / v). The arabinoxylan with a purity of ≥85% was obtained by freeze-drying.
[0024] (2) Bifidobacterium longum (CICC 6186) fermentation:
[0025] Activation of strains: Bifidobacterium longum was inoculated into MRS medium (2 mL of Bifidobacterium longum glycerol stock solution was inoculated into every 100 mL of MRS medium, and the initial bacterial concentration in the glycerol stock solution was about 1×10 8 CFU / mL), cultured anaerobically at 37°C for 24 h;
[0026] Optimize fermentation: Inoculate 10% of the activated bacterial solution by volume into a medium containing arabinoxylan (based on MRS desugared basal medium supplemented with 1.5% (w / v) arabinoxylan) and ferment anaerobically at 37°C for 18 hours with arabinoxylan as the sole carbon source until the viable cell count reaches 1×10 11 CFU / g.
[0027] (3) Compounding and embedding:
[0028] Adjust the number of viable Bifidobacterium longum to 1×10 10 CFU / g, and then mixed with the arabinoxylan prepared in step (1) at a mass ratio of 1:4; then slowly mixed with a mixed solution of sodium alginate (2% w / v) and chitosan (1% w / v) at a volume ratio of 1:1, and formed nanoparticles through ionic crosslinking, the reaction system was stirred at 700 rpm for 60 minutes at room temperature (25°C) to make the embedding rate ≥85%, and freeze-dried (pre-freezing at -80°C for 6 hours, freeze-drying temperature at -60°C, vacuum degree 20 Pa, freeze-drying time 48 hours) to obtain arabinoxylan-Bifidobacterium longum synbiotic powder with a water content of ≤5%.
[0029] Example 2
[0030] Effects of arabinoxylan-Bifidobacterium longum synbiotics on eicosanoid metabolism in obese mice
[0031] 1. Experimental Grouping
[0032] Forty C57BL / 6 male mice were randomly divided into two groups:
[0033] High-fat diet control group (HFD): fed with 45% high-fat feed;
[0034] HFD+synbiotic group: 50 mg / kg synbiotics were administered orally daily on the basis of HFD for 4 weeks.
[0035] 2. Experimental Design
[0036] After 28 days of oral gavage, the mice were tail-clipped and blood was collected. The levels of COX-2, PGE2, LTB4, and LXA4 in serum were detected using ELISA kits ( Figure 1 ), inflammatory factor levels in mouse colon tissue ( Figure 2 ) and the concentrations of EET, DHET, and DiHETE ( Figure 3 and Figure 4 ), the total amount of SCFAs was detected by gas chromatography, and the expression levels of related genes were determined by RT-qPCR and Western Blot ( Figure 5 ).
[0037] 3. Results and Analysis
[0038] In the HFD+synbiotic group, PGE2 levels were 50% lower than in the HFD control group. Serum levels of inflammatory factors such as PGE2 and TNF-α decreased, while levels of eicosanoid metabolites such as DHETs and DiHETEs were altered. Intestinal concentrations of SCFAs, including butyrate, were elevated. Total SCFA levels increased by 48.7% compared to the HFD group. Expression of eicosanoid metabolism genes, including COX-2 and sEH, was decreased, and the Firmicutes / Bacteroidetes ratio was reduced. This suggests that this synbiotic can influence inflammatory factors through the "gut flora-SCFAs-eicosanoid metabolism axis," and has broad application prospects for chronic inflammatory diseases.
[0039] Comparative Example 1
[0040] Effects of arabinoxylan or Bifidobacterium longum alone on eicosanoid metabolism in obese mice
[0041] 1. Experimental Design (Using the obese mouse model of Example 2, the detection method is the same as Example 2) Two additional groups of mice were set up:
[0042] HFD+arabinoxylan group (HFD+AX): 50 mg / kg arabinoxylan was administered orally daily;
[0043] HFD+Bifidobacterium longum group (HFD+BL): 1×10 10 CFU / mL Bifidobacterium longum 50mL / kg.
[0044] 2. Results Analysis
[0045] HFD+AX group: total SCFAs increased by 19.5% (lower than the synbiotic group); however, PGE2 and LTB4 decreased to a lesser extent (only 10-15%).
[0046] HFD+BL group: There was no significant increase in SCFAs; there was no significant change in eicosanoid metabolism (PGE2 decreased by 7.3%).
[0047] It can be seen that the combined use of arabinoxylan + Bifidobacterium longum has a better effect on regulating SCFAs and eicosanoid metabolism than the use of prebiotics or probiotics alone.
[0048] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.
Claims
1. An arabinoxylan-Bifidobacterium longum synbiotic, characterized in that The preparation method comprises the following steps: mixing pre-fermented bifidobacterium longum and arabinoxylan, adding a mixed solution of sodium alginate and chitosan for embedding, and freeze-drying to obtain the arabinoxylan-bifidobacterium longum synbiotic.
2. The arabinoxylan-Bifidobacterium longum synbiotic according to claim 1, characterized in that The mass ratio of the Bifidobacterium longum to arabinoxylan is 1:3-5.
3. The arabinoxylan-Bifidobacterium longum synbiotic according to claim 2, characterized in that The number of viable Bifidobacterium longum is ≥1×10 10 CFU / g, and the purity of the arabinoxylan is ≥85%.
4. The arabinoxylan-Bifidobacterium longum synbiotic according to claim 3, characterized in that The method for pre-fermentation of Bifidobacterium longum comprises: activating Bifidobacterium longum and then inoculating it into an arabinoxylan culture medium and fermenting it for 15 to 20 hours; The arabinoxylan culture medium is based on an MRS desugared basal culture medium supplemented with 1-2% (w / v) arabinoxylan.
5. The arabinoxylan-Bifidobacterium longum synbiotic according to claim 4, characterized in that The volume ratio of the mixture of Bifidobacterium longum and arabinoxylan to the mixed solution of sodium alginate and chitosan is 1-2:1-2.
6. The arabinoxylan-Bifidobacterium longum synbiotic according to claim 5, characterized in that In the mixed solution of sodium alginate and chitosan, the concentration of sodium alginate is 1-3% w / v, and the concentration of chitosan is 0.5-2% w / v.
7. The arabinoxylan-Bifidobacterium longum synbiotic according to claim 6, characterized in that The embedding conditions are: 20-30° C., 600-800 rpm, stirring for 55-65 minutes, and an embedding rate of ≥85%.
8. The arabinoxylan-Bifidobacterium longum synbiotic according to claim 7, characterized in that After freeze-drying, the moisture content of the arabinoxylan-Bifidobacterium longum synbiotic is ≤5%.
9. Use of the arabinoxylan-Bifidobacterium longum synbiotic according to claim 8 in the preparation of a product for improving intestinal flora, regulating the eicosanoid metabolic axis or reducing inflammatory response.