YLGB-1496 and prebiotic composition for relieving constipation

Abstract

The invention provides a YLGB-1496 and prebiotic composition capable of relieving constipation and a preparation method of the YLGB-1496 and prebiotic composition. Specifically, the invention provides an application of a composition containing Bifidobacterium longum subsp.infantis and prebiotics in preparation of a composition helpful for preventing and / or improving constipation, and the Bifidobacterium longum subsp.infantis is the Bifidobacterium longum subsp.infantis with the preservation number of CCTCC (China Center For Type Culture Collection) NO: M2011122. The bifidobacterium longum subsp. Infantis can be combined with galactooligosaccharide to serve as an active component beneficial to prevention and / or improvement of constipation, and generation of intestinal gases and short-chain organic acids of people with constipation is regulated and controlled.

Description

Technical Field

[0001] The present invention relates to a new application of Bifidobacterium longum subsp. infantis YLGB-1496 (preservation number CCTCC NO: M2011122), and specifically, the present invention relates to an application of Bifidobacterium longum subsp. infantis YLGB-1496 and a prebiotic in combination for regulating intestinal gas and short-chain organic acid production in constipated people. Background Art

[0002] Constipation is a common intestinal health abnormality that affects 10% to 20% of the global population. It threatens intestinal health by reducing bowel movements and prolonging bowel movements, which in turn leads to the proliferation of harmful bacteria. In addition, constipation increases the risk of cardiovascular diseases, including atherosclerosis and arrhythmias, which adversely affects the daily lives of these patients.

[0003] Currently, the common method to solve or relieve constipation is to take prescription laxatives, especially osmotic or secretory laxatives. However, these methods can cause various side effects, including dizziness, dehydration and / or coronary artery constriction in subjects. Therefore, more than 50% of subjects are not completely satisfied with this method.

[0004] Studies have reported that probiotics colonize in the intestines, regulate intestinal flora, and relieve constipation. However, the effect of probiotics on constipation relief is strain-specific.

[0005] CN114634884A discloses Bifidobacterium longum subsp. infantis YLGB-1496, whose deposit number is CCTCC NO: M2011122. It is a probiotic. Studies have shown that YLGB-1496 has good effects of improving resistance to intestinal bacterial infection and intestinal immunity.

[0006] Upon investigation, there is no report in the prior art on whether the Bifidobacterium longum subspecies infantis YLGB-1496 strain can solve constipation. Summary of the invention

[0007] One object of the present invention is to provide a new application of Bifidobacterium longum subsp. infantis YLGB-1496.

[0008] Bifidobacterium longum subsp. infantis YLGB-1496 strain was deposited in China Center for Type Culture Collection (CCTCC) on April 10, 2011, with a deposit number of CCTCC NO: M2011122. Bifidobacterium longum subsp. infantis YLGB-1496 strain is a biological material published in patent documents CN114634884A, CN113841898A, etc.

[0009] The inventors of this case discovered through experiments in their research that Bifidobacterium longum subsp. infantis (i.e., Bifidobacterium longum subsp. infantis YLGB-1496) with a preservation number of CCTCC NO: M2011122 is beneficial for improving constipation, especially when used in combination with galacto-oligosaccharides (GOS), it can synergistically regulate the production of intestinal gas and short-chain organic acids in people with constipation, increase the intestinal acetic acid content and reduce the intestinal isobutyric acid and / or isovaleric acid content without significantly affecting the total intestinal acid production; it can reduce the total intestinal gas production and reduce the intestinal CO2, H2S and / or CH4 production.

[0010] Thus, on the one hand, the present invention provides a use of a composition comprising Bifidobacterium longum subsp. infantis and a prebiotic in the preparation of a composition that helps prevent and / or improve constipation, wherein the Bifidobacterium longum subsp. infantis is the Bifidobacterium longum subsp. infantis with a deposit number of CCTCC NO: M2011122. That is, the present invention provides a use of Bifidobacterium longum subsp. infantis YLGB-1496.

[0011] According to a specific embodiment of the present invention, in the present invention, the prebiotic is galacto-oligosaccharide. The Bifidobacterium longum subsp. infantis YLGB-1496 (Bifidobacterium longum subsp. infantis with a deposit number of CCTCC NO: M2011122) of the present invention is combined with galacto-oligosaccharide as an active component that helps prevent and / or improve constipation.

[0012] The “improvement” mentioned in the present invention includes alleviation of relevant symptoms.

[0013] According to a specific embodiment of the present invention, the method of helping to prevent and / or improve constipation includes regulating the intestinal gas and / or short-chain organic acid production in constipated people.

[0014] According to a specific embodiment of the present invention, the method of helping to prevent and / or improve constipation comprises:

[0015] Increase intestinal acetic acid content;

[0016] Reduce intestinal isobutyric acid and / or isovaleric acid levels;

[0017] Reduce total intestinal gas production;

[0018] Reduce intestinal CO2, H2S and / or CH4 production.

[0019] According to a specific embodiment of the present invention, in the application of Bifidobacterium longum subspecies infantis YLGB-1496 of the present invention, the composition includes a food composition, a feed composition or a pharmaceutical composition.

[0020] According to a specific embodiment of the present invention, in the application of Bifidobacterium longum subspecies infantis YLGB-1496 of the present invention, the Bifidobacterium longum subspecies infantis can be used to prepare the composition in the form of a solid or liquid bacterial preparation.

[0021] According to a specific embodiment of the present invention, in the application of Bifidobacterium longum subspecies infantis YLGB-1496 of the present invention, the Bifidobacterium longum subspecies infantis can be used to prepare the composition in the form of live bacteria or inactivated bacteria.

[0022] According to a specific embodiment of the present invention, in the application of Bifidobacterium longum infantis subspecies YLGB-1496 of the present invention, the application amount of Bifidobacterium longum infantis subspecies is 1.0×10 3 CFU / day - 1.0 × 10 12 CFU / day, preferably 1.0×10 7 CFU / day - 1.0 × 10 11 CFU / day.

[0023] On the other hand, the present invention also provides a composition for preventing and / or improving constipation, the composition comprising Bifidobacterium longum subsp. infantis with a deposit number of CCTCC NO: M2011122 and a prebiotic.

[0024] According to a specific embodiment of the present invention, the prebiotic is galacto-oligosaccharide.

[0025] According to a specific embodiment of the present invention, in the composition, the ratio of galacto-oligosaccharide to Bifidobacterium longum subsp. infantis is: 0.1-1.0 mg: 5×10 8 CFU.

[0026] On the other hand, the present invention further provides a food, the raw material composition of which includes the Bifidobacterium longum subspecies infantis with the preservation number CCTCC NO: M2011122 of the present invention and a composition of prebiotics.

[0027] According to a specific embodiment of the present invention, the composition of the present invention may also include conventional material components in the field. For example, for pharmaceutical compositions, an appropriate amount of auxiliary materials may be included, and the auxiliary materials may be excipients, diluents, fillers and / or absorption enhancers, etc. For food compositions, the longum infantis subspecies of the present invention may be produced according to the food containing longum infantis subspecies of Bifidobacterium in the prior art, and the oligosaccharide may be added within the scope of the amount allowed by laws and regulations. The composition may be in different forms according to the needs of the recipient. For example, powders, tablets, granulations, microcapsules and / or liquid preparations, etc.

[0028] According to some specific embodiments of the present invention, in the present invention, the food can be a fermented dairy product, cheese, a milk-containing beverage, a solid beverage, a compressed candy, a milk tablet, an ice cream or milk powder.

[0029] In summary, the present invention provides a new application of the combination of Bifidobacterium longum subspecies infantis YLGB-1496 and galacto-oligosaccharide GOS for preparing a composition that helps prevent and / or improve constipation. The present invention proves through experiments that the combination of YLGB-1496 and GOS can synergistically regulate the production of intestinal gas and short-chain organic acids in constipated people, and can increase the intestinal acetic acid content and reduce the intestinal isobutyric acid and / or isovaleric acid content without significantly affecting the total intestinal acid production; it can reduce the total intestinal gas production and reduce intestinal CO 2、 The present invention is of great significance for exploring the mechanism of the combination of probiotics and prebiotics in relieving constipation. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1A The percentage of acetic acid produced in each experimental group in Example 1 is shown.

[0031] Figure 1B The percentage of isobutyric acid produced in each experimental group in Example 1 is shown.

[0032] Figure 1C The percentage of isovaleric acid produced in each experimental group in Example 1 is shown.

[0033] Figure 1D The total acid production of each experimental group in Example 1 is shown.

[0034] Figure 2A Shows the total gas volume produced by each experimental group in Example 2.

[0035] Figure 2B Shows the CO2 gas production ratio of each experimental group in Example 2.

[0036] Figure 2C Shows the CH4 gas production ratio of each experimental group in Example 2.

[0037] Figure 2DShows the H2S gas production ratio of each experimental group in Example 2.

[0038] In each figure, *P<0.05, **P<0.01, ***P<0.001. DETAILED DESCRIPTION

[0039] In order to have a clearer understanding of the technical features, purposes and beneficial effects of the present invention, the technical scheme of the present invention is described in detail below in conjunction with specific embodiments and drawings. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. In the embodiments, each raw reagent material can be obtained commercially, and the experimental method without specifying specific conditions is a conventional method and conventional conditions well known in the art, or according to the conditions recommended by the instrument manufacturer.

[0040] Fecal sample collection and in vitro fermentation

[0041] A total of 10 patients with constipation were included in this study (Table 1). The inclusion criteria were as follows: patients who met the Rome IV diagnostic criteria for constipation (16 cases); aged 20-50 years; Wexner scale requirements (score of 30 points, the score was positively correlated with the severity of constipation (17 points)), and the number of bowel movements was 1-3 times / week. Exclusion criteria: use of any probiotic products within two weeks before the start of the study; patients with gastrointestinal, nervous system, cardiovascular, endocrine, renal and / or other chronic diseases that may affect intestinal motility; patients with chronic diarrhea; patients who underwent gastric banding or gastric bypass surgery and used conventional laxatives, as well as pregnant or lactating women. The design of this study was approved by the Ethics Committee of Hangzhou Normal University (approval number: 20190061). All participants in this study participated voluntarily and signed informed consent.

[0042] The collected specimens were placed in sterile stool collection tubes, transported to the laboratory at low temperature, and stored at -80°C for future use. The samples of each participant were divided into four groups for in vitro fermentation experiments. 0.8±0.02 g of the sample was placed in a HALO-F100 stool processor (Suzhou Hailu Biotechnology, Jiangsu, China) to obtain a 10% stool suspension. Subsequently, 0.5 mL of the clarified suspension was injected into a fermentation bottle containing YCFA medium. The samples were divided into a blank control group (no addition), YLGB-1496 (5×10 8 cfu / mL), GOS (concentration 0.4‰), synbiotic group (YLGB-1496 concentration 5×10 8 cfu / mL, GOS concentration 0.4‰) for a total of 4 groups. All samples were tested after fermentation at 37℃ for 24h.

[0043] Table 1 Basic information of subjects

[0044]

[0045] Example 1. Detection of short-chain organic acids

[0046] The level of short-chain fatty acids (SCFAs) in the fermentation broth of each group was determined by gas phase method. Briefly, 0.5 mL of 10% fecal suspension was mixed with 0.1 mL of 25% metaphosphoric acid at -20°C for 24 hours. The mixture was then thawed, centrifuged at 4°C and 3000g for 20 min, and filtered with a 0.22 μm membrane for later use. The content of short-chain fatty acids was determined by a gas chromatograph (GC, Shimadzu, GC-2010 Plus, Kyoto, Japan) equipped with a DB-FFAP column. The determination method includes total acid, acetic acid, isobutyric acid, and isovaleric acid content (purity [propionic acid, acetic acid, butyric acid standards]> 99%, chromatographic grade, purchased from Ehrensorfer Standard Company, Germany), and the internal standard is 2-ethylbutyric acid.

[0047] The results of the measurement can be found in Figures 1A to 1D . The experimental results showed that compared with the YCFA group, the acetic acid content in the GOS group was significantly higher than that in the YCFA group (P<0.05), and that in the YLGB-1496+GOS group (synbiotic group) was higher than that in the YCFA group (P<0.01). The isovaleric acid content in the GOS group was significantly reduced (P<0.05), and the isobutyric acid and isovaleric acid contents in the YLGB-1496+GOS group (synbiotic group) were further significantly reduced (P<0.01). There was no significant difference in the total acid content between the YLGB-1496, GOS, and YLGB-1496+GOS groups (synbiotic group) and the YCFA group (P>0.05).

[0048] Example 2. Gas Detection

[0049] The concentrations of H2, CO2, CH4, and H2S and the total gas release of in vitro fermentation were measured using a gas analyzer (HL-QT01, Hailu Biotechnology, Hunan, China). The fermented gas entered the gas analyzer through a sampling tube, where the gas content was measured by a gas sensor. Finally, the measurement results were analyzed using the “Multi-Gas Analyzer.exe” software.

[0050] After 24 hours of in vitro fermentation, the gas production of each group was measured. FIG. 2A to FIG. 2D. Compared with the YCFA group, the total gas content in the GOS group and the YLGB-1496+GOS group (synbiotic group) was significantly reduced (P<0.05), and the total gas production in the YLGB-1496+GOS group (synbiotic group) was significantly reduced compared with the individual YLGB-1496 and GOS. The GOS group and the YLGB-1496+GOS group significantly reduced the production of CO2 and H2S (P<0.001). The CH4 content in the YLGB-1496 group decreased compared with that in the YCFA group, but there was no significant difference. The CH4 production in the YLGB-1496+GOS group (synbiotic group) was significantly reduced compared with that in the YCFA group (P<0.05). The combination of YLGB-1496 and GOS had a synergistic effect on the total gas production and CH4 production compared with the individual YLGB-1496 and GOS.

Claims

1. Use of a composition comprising Bifidobacterium longum subsp. infantis and a prebiotic in the preparation of a composition that helps prevent and / or improve constipation, wherein: The Bifidobacterium longum subsp. infantis is the Bifidobacterium longum subsp. infantis with a deposit number of CCTCC NO: M2011122.

2. The use according to claim 1, wherein: The prebiotic is galacto-oligosaccharide, and the Bifidobacterium longum subspecies infantis with the deposit number CCTCC NO: M2011122 is combined with galacto-oligosaccharide as an active component that helps prevent and / or improve constipation.

3. The use according to claim 1 or 2, wherein: The method for helping to prevent and / or improve constipation includes regulating the intestinal gas and / or short-chain organic acid production in constipated people.

4. The use according to claim 3, wherein: Some of the things that may help prevent and / or improve constipation include: Increase intestinal acetic acid content; Reduce intestinal isobutyric acid and / or isovaleric acid levels; Reduce overall intestinal gas production; and / or Reduce intestinal CO2, H2S and / or CH4 production.

5. The use according to claim 1 or 2, wherein: The composition includes a food composition, a feed composition or a pharmaceutical composition.

6. The use according to claim 1 or 2, wherein: The Bifidobacterium longum subspecies infantis is used to prepare the composition in the form of a solid or liquid bacterial preparation; or The Bifidobacterium longum subspecies infantis is used to prepare the composition in the form of live bacteria or inactivated bacteria.

7. The use according to any one of claims 1 to 6, wherein: The application amount of Bifidobacterium longum subspecies infantis is 1.0×10 3 CFU / day - 1.0 × 10 12 CFU / day, preferably 1.0×10 7 CFU / day - 1.0 × 10 11 CFU / day.

8. The use according to any one of claims 2 to 7, wherein: In the composition, the dosage ratio of galacto-oligosaccharide to Bifidobacterium longum subspecies infantis is: 0.1-1.0 mg: 5×10 8 CFU.

9. A composition for preventing and / or improving constipation, comprising Bifidobacterium longum subsp. infantis with a deposit number of CCTCC NO: M2011122 and a prebiotic; Preferably, the prebiotic is galacto-oligosaccharide; More preferably, the ratio of galacto-oligosaccharide to Bifidobacterium longum subsp. infantis is: 0.1-1.0 mg: 5×10 8 CFU.

10. A food, the raw material composition of which includes the composition according to claim 9; Preferably, the food is a fermented dairy product, cheese, a milk-containing beverage, a solid beverage, a compressed candy, a milk tablet, an ice cream or milk powder.

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