Use of a polysaccharide composition in the manufacture of a product for improving gut homeostasis

Abstract

The application discloses application of a polysaccharide composition in preparation of a product for improving intestinal homeostasis, and the polysaccharide composition comprises an active ingredient combination of oat beta-glucan and 2'-fucosyllactose and a food or pharmaceutically acceptable carrier or adjuvant. The application can simultaneously improve aging or inflammation-induced intestinal homeostasis damage from multiple dimensions of intestinal barrier repair, inflammation relief, oxidative stress reduction and intestinal flora regulation through mechanism complementation and synergistic effect of oat beta-glucan and 2'-fucosyllactose, and the effect is significantly better than that of a single active ingredient. The active ingredients are all of natural origin, high in safety, suitable for long-term taking, and can be developed into functional food, health products or medicines, and have a wide application prospect.

Description

Technical Field

[0001] This invention relates to the fields of food and biomedicine, and more specifically to the application of a polysaccharide composition in the preparation of products that improve intestinal homeostasis. Background Technology

[0002] Gut homeostasis is a core foundation for maintaining normal physiological functions. It is jointly maintained by the intestinal epithelial barrier, gut microbiota homeostasis, and the balance of intestinal immune inflammation. Among these, the integrity of the intestinal epithelial barrier is a structural prerequisite for the normal functioning of gut homeostasis. The aging process induces a series of physiological degenerative changes in the gut, manifested as damage to the tight junction structure of the intestinal epithelium, abnormally increased barrier permeability, increased levels of circulating endotoxins, upregulation of local pro-inflammatory factors (such as TNF-α and IL-6) and a chronic low-grade inflammatory state, increased oxidative stress, accelerated epithelial cell apoptosis, gut microbiota dysbiosis, decreased abundance of probiotics, and insufficient production of short-chain fatty acids. These pathological changes interact, further accelerating the imbalance of gut homeostasis, leading not only to decreased digestive and absorptive functions but also increasing the risk of metabolic syndrome, autoimmune diseases, and even tumors, severely impacting quality of life.

[0003] Active polysaccharides have become a research hotspot in the field of intestinal health due to their natural origin, high safety, and diverse biological activities. Among them, oat β-glucan, extracted from oat bran, has activities such as regulating blood lipids, immune regulation, and promoting the proliferation of probiotics. Existing studies have confirmed that it can mildly regulate the composition of intestinal flora. However, the mechanism of action of oat β-glucan alone is singular, and its effect on improving the multidimensional intestinal homeostasis imbalance caused by aging or inflammation, such as intestinal barrier damage, chronic inflammation, and oxidative stress, is limited. It cannot effectively improve existing intestinal pathological damage. Therefore, there is an urgent need to develop a safe, efficient, and long-term usable polysaccharide composition that can improve aging or inflammation-induced intestinal homeostasis damage from multiple dimensions. Summary of the Invention

[0004] The purpose of this invention is to solve the above-mentioned technical problems and provide an application of a polysaccharide composition in the preparation of products that improve intestinal homeostasis. This composition can effectively improve multidimensional intestinal homeostasis damage induced by aging or inflammation, and achieve multidimensional improvement in intestinal barrier repair, inflammation relief, oxidative stress reduction and intestinal flora regulation.

[0005] The above-mentioned objective of the present invention is achieved through the following technical solution:

[0006] The use of a polysaccharide composition in the preparation of a product for improving intestinal homeostasis, the polysaccharide composition comprising a combination of active ingredients and a food- or pharmaceutically acceptable carrier or excipient, the combination of active ingredients being oat β-glucan (BG) and 2'-fucosylated lactose (2'-FL).

[0007] This invention combines oat β-glucan and 2'-fucosylated lactose to improve aging- or inflammation-induced intestinal homeostasis damage. The two components exert a synergistic effect through different mechanisms of action: oat β-glucan acts as a prebiotic to promote the proliferation of beneficial intestinal bacteria, while 2'-fucosylated lactose directly regulates intestinal immunity, inhibits inflammatory responses, and promotes epithelial barrier repair. When used together, oat β-glucan improves the intestinal microenvironment, providing a favorable physiological basis for 2'-fucosylated lactose to exert its immunomodulatory and barrier-repairing effects. Furthermore, the repaired intestinal barrier by 2'-fucosylated lactose better preserves the intestinal flora and its metabolites. Therefore, this combination simultaneously improves intestinal homeostasis damage from multiple dimensions, including improving flora structure, repairing barrier integrity, alleviating chronic inflammation, and reducing oxidative stress, with significantly better results than either single active ingredient.

[0008] Furthermore, the mass ratio of oat β-glucan to 2'-fucosylated lactose is 1:3 to 3:1, preferably 1:1. By adjusting the mass ratio of oat β-glucan to 2'-fucosylated lactose, the preventive and ameliorative effects of the polysaccharide composition on aging- or inflammation-induced intestinal homeostasis damage can be further enhanced.

[0009] Furthermore, each unit dose is defined as a single administration per kilogram of body weight, and each unit dose of the polysaccharide composition contains 50-200 mg of oat β-glucan and 50-200 mg of 2'-fucosylated lactose.

[0010] Preferably, a single dose per kilogram of body weight is considered as one unit dose, and each unit dose of the polysaccharide composition contains 100-150 mg of oat β-glucan and 100-150 mg of 2'-fucosylated lactose.

[0011] Furthermore, the polysaccharide composition is a combination formulation comprising separately packaged first and second formulations; the active ingredient of the first formulation is oat β-glucan, and the active ingredient of the second formulation is 2'-fucosylated lactose. The combination formulation can prevent, alleviate, and / or treat aging or inflammatory bowel disease.

[0012] Furthermore, the dosage form of the polysaccharide composition is selected from pills, powders, capsules, tablets, granules, films, or liquids.

[0013] Furthermore, the product for improving intestinal homeostasis is a product that improves intestinal homeostasis by having at least one of the following functions:

[0014] (1) Improves intestinal barrier permeability damage;

[0015] (2) Relieves intestinal inflammation;

[0016] (3) Reduce intestinal oxidative stress levels;

[0017] (4) Regulate the structure of intestinal flora and increase the abundance of probiotics in the gut;

[0018] (5) Promotes the metabolism and production of short-chain fatty acids.

[0019] Furthermore, the intestinal barrier permeability impairment, intestinal inflammatory response, or intestinal oxidative stress are induced by aging or inflammation.

[0020] Furthermore, the product for improving gut homeostasis is a functional food, health product, or medicine.

[0021] Furthermore, the food- or pharmaceutically acceptable carrier or excipient is selected from one or more of excipients, diluents, binders, disintegrants, lubricants, flow aids, surfactants, buffers, preservatives, flavoring agents, and coloring agents.

[0022] Furthermore, the food- or pharmaceutically acceptable carrier or excipient may also include dispersants, suspending agents, granulation agents, fillers, liquid mediators, isotonic agents, thickeners, emulsifiers, solvents, etc.

[0023] Furthermore, the excipient is selected from one or more of starch, lactose, and dextrin.

[0024] Furthermore, the diluent is selected from one or more of calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, and corn starch.

[0025] The above-described technical solution of the present invention has the following beneficial effects:

[0026] The polysaccharide composition provided by this invention can complement and synergistically enhance the effects of oat β-glucan and 2'-fucosylated lactose, thereby improving intestinal barrier permeability damage, alleviating inflammation, reducing oxidative stress levels, improving intestinal flora structure, and increasing probiotic abundance. It can effectively improve intestinal homeostasis and promote health. Both active ingredients used are widely used natural sources, with high safety, no toxic side effects, and are suitable for long-term use. They can be directly developed into functional foods, health products, or pharmaceuticals, with broad application prospects. Attached Figure Description

[0027] Figure 1 Figure 1 shows the changes in body weight of aging model mice after intervention in the CON group, BG group, 2'-FL group, and BG+2'-FL group.

[0028] Figure 2 The images show the histopathological changes of the jejunum and ileum of aging model mice after intervention in the CON group, BG group, 2'-FL group, and BG+2'-FL group, stained with H&E.

[0029] Figure 3 The figure shows the results of detecting the mRNA expression levels of intestinal inflammatory factors and tight junction proteins in the jejunum and ileum of aging model mice after intervention in the CON group, BG group, 2'-FL group and BG+2'-FL group; where (A) is ZO-1, (B) is Occludin, (C) is Claudin, (D) is IL-6, (E) is IL-1β and (F) is TNF-α.

[0030] Figure 4 Figure 1 shows the results of intestinal oxidative stress level detection in aging model mice after intervention in the CON group, BG group, 2'-FL group and BG+2'-FL group; where (A) is SOD activity in the jejunum, (B) is MDA level in the jejunum, (C) is GSH content in the jejunum, (D) is SOD activity in the ileum, (E) is MDA level in the ileum and (F) is GSH content in the ileum.

[0031] Figure 5 The diagram shows the changes in gut microbiota structure in aging model mice after intervention in the CON group, BG group, 2'-FL group and BG+2'-FL group; where (A) is the Chao1 index analysis diagram, (B) is the Simpson index analysis diagram, (C) is the Shannon index analysis diagram, and (D) is the ACE index analysis diagram.

[0032] Figure 6 The relative abundance of gut microbiota at the phylum level in aging model mice after intervention in the CON group, BG group, 2'-FL group, and BG+2'-FL group.

[0033] Figure 7 The relative abundance of gut microbiota at the genus level in aging model mice after intervention in the CON group, BG group, 2'-FL group, and BG+2'-FL group. Detailed Implementation

[0034] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0035] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0036] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, and the materials and reagents used are commercially available.

[0037] The BG used in the following examples was purchased from Shaanxi Baichuan Biotechnology Co., Ltd., with a purity >99% and a relative molecular weight of 92.47 kDa; 2'-FL was purchased from Xi'an Youkangfulai Biotechnology Co., Ltd., with a purity of 95%.

[0038] Example 1

[0039] A polysaccharide composition, wherein the polysaccharide composition is a combined formulation comprising a first formulation and a second formulation, which are separately packaged; the active ingredient of the first formulation is BG, and the active ingredient of the second formulation is 2'-FL; the mass ratio of BG to 2'-FL is 1:1.

[0040] Example 2

[0041] A polysaccharide composition comprising an active ingredient combination and microcrystalline cellulose, wherein the active ingredient combination is BG and 2'-FL, and the polysaccharide composition is prepared by the following method:

[0042] (1) Weigh 50 g each of dried BG and 2'-FL, put them into a three-dimensional mixer and mix for 15 min to obtain premixed active powder;

[0043] (2) Add 20 g of microcrystalline cellulose to the above premixed active powder and continue mixing for 20 min until the mixture is uniform to obtain the total mixed powder;

[0044] (3) The total mixed powder is dry granulated, passed through a 20-40 mesh sieve for granulation, and packaged to obtain a polysaccharide composition.

[0045] Test Example 1

[0046] Forty 18-month-old male C57BL / 6J aging model mice were randomly divided into a normal control group (CON), an oat β-glucan group (BG), a 2'-fucosylated lactose group (2'-FL), and an oat β-glucan and 2'-fucosylated lactose combined group (Example 1, BG+2'-FL, mass ratio 1:1), with 10 mice in each group. After one week of acclimatization, the CON group was administered 200 mg of saline per mouse via gavage daily; the BG group was administered 200 mg BG / kg / day of oat β-glucan solution (saline solution); the 2'-FL group was administered 200 mg 2'-FL / kg / day of 2'-fucosylated lactose solution (saline solution); and the BG+2'-FL group was administered a combination of 100 mg BG / kg / day of oat β-glucan solution (saline solution) and 100 mg 2'-FL / kg / day of 2'-fucosylated lactose solution (saline solution). This intervention lasted for 70 consecutive days. Mouse weight, fecal morphology, and anal condition were recorded daily during the intervention. After the intervention, mice were sacrificed, and the following indicators were measured:

[0047] (1) Monitor mouse weight changes by weighing and recording the weight of each group of mice at a fixed time each week. The test results are as follows: Figure 1 As shown, there was no statistically significant difference in body weight among the groups of mice during the intervention period. However, the aging model mice showed a progressive trend of weight loss after the intervention, suggesting that this trend is related to age-related metabolic function decline and muscle loss.

[0048] (2) After the intervention, intestinal tissue samples were collected. Mice were anesthetized and blood and intestinal tissues and organs of each segment were collected. 0.5 cm of jejunum and ileum were cut off, fixed with 4% paraformaldehyde solution, and stored at 4 ℃ for intestinal tissue pathological analysis. The remaining intestinal tissue was rapidly quenched with liquid nitrogen and stored at -80 ℃ for later use. Formalin-fixed jejunum and ileum specimens were taken, embedded in paraffin, and sectioned. Hematoxylin-eosin (H&E) staining was performed, and the intestinal mucosal structure of the sections was observed and the height of the intestinal villi was measured using an optical microscope.

[0049] Test results are as follows Figure 2 As shown, the jejunum and ileum of the CON group mice exhibited pathological features such as uneven villi, villi separation from the lamina propria, dilation of the lamina propria interstitial space, and occasional small lymphocyte aggregation. The villi length of the jejunum and ileum in the BG group and the 2'-FL group was significantly increased (p<0.05), with the increase being more significant in the BG+2'-FL group (p<0.01). At the same time, the disordered structure of epithelial cells and lamina propria and the infiltration of inflammatory cells were also significantly improved.

[0050] (3) The expression levels of intestinal inflammatory factors and tight junction proteins mRNA were detected. The test method was to extract total RNA from mouse jejunum and ileum tissues and use reverse transcription polymerase chain reaction (RT-PCR) technology to detect the mRNA expression levels of three tight junction proteins, tight junction protein 1 (ZO-1), occludin, and claudin, as well as three pro-inflammatory factors, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β), at the transcriptional level.

[0051] Test results are as follows Figure 3 As shown, compared with the CON group, the BG+2'-FL group significantly increased the mRNA expression levels of ZO-1, Occludin, and Claudin in the jejunum and ileum, while significantly decreasing the mRNA expression levels of IL-6, TNF-α, and IL-1β in the above three intestinal segments. This indicates that the BG+2'-FL composition can effectively enhance the expression of intestinal tight junction proteins in aging model mice and alleviate the aging-induced chronic intestinal inflammation.

[0052] (4) Intestinal oxidative stress level was detected. The test method was to take fresh jejunum and ileum tissues, weigh them accurately, mix them with pre-cooled physiological saline at a ratio of 1:9 (g / mL), and homogenize them at low temperature using a high-throughput tissue homogenizer. The homogenate was centrifuged at 4℃ and 2500 rpm for 10 min, and the supernatant was collected. Superoxide dismutase (SOD) activity, glutathione peroxidase (GSH) content, and malondialdehyde (MDA) level were detected using commercial kits.

[0053] Test results are as follows Figure 4 As shown, compared with the CON group, the SOD activity and GSH content in the jejunum of the BG+2'-FL group were significantly increased, and the MDA content was significantly decreased; the SOD activity in the ileum was significantly increased, the MDA level was significantly decreased, and the GSH content was significantly increased. This indicates that the BG+2'-FL composition can enhance the endogenous antioxidant capacity of the intestine, effectively scavenge reactive oxygen free radicals, and reduce the damage of oxidative stress to intestinal epithelial cells.

[0054] (5) 16S rRNA amplicon sequencing analysis of gut microbiota was performed. The test method was to collect mouse fecal samples using metabolic cages in the last week before the end of the intervention and freeze them at -80 ℃ for later use. Total microbial DNA was extracted from the frozen mouse fecal samples, and the V3-V4 region of the 16S rRNA gene was amplified using primers 341F (5'-CCTACGGGGNGGCWGCAG-3') and 806R (5'-GGACTACHVGGGGTATCTAAT-3'). After purification, the amplicon was ligated with sequencing adapters to construct a sequencing library. High-throughput sequencing was performed using the Illumina platform, and the sequencing data were analyzed using bioinformatics.

[0055] Test results are as follows Figures 5-7 As shown, compared with the CON group, there were no significant differences in the microbial abundance index (Chao1 and ACE) and species diversity index (Shannon and Simpson) of the gut microbiota among the intervention groups, indicating that polysaccharide intervention did not significantly alter the species richness and evenness of the gut microbiota in aging model mice. However, the trends among the groups suggest that polysaccharide intervention did have an impact on the gut microbiota of aging model mice. Figure 6 At the phylum level, the abundance of Bacteroidetes was significantly increased in the BG+2'-FL group (p<0.001), while the abundance of Firmicutes was significantly decreased (p<0.001). Bacteroidetes and other gut microbes can efficiently utilize carbon sources such as complex plant polysaccharides and dietary fiber. Figure 7 At the genus level, the BG+2'-FL group significantly upregulated multiple bacterial species that contribute to delaying aging, including *Prevotellamassilia*, *Parabacteroides*, *Collinsella*, *Faecalibacterium*, and *Hallella*. *Prevotellamassilia* primarily participates in carbohydrate metabolism, generating precursors of SCFAs to provide energy for the gut and, to some extent, improve age-related intestinal metabolic imbalances. *Parabacteroides* possesses anti-inflammatory regulatory functions; its metabolites can reduce the release of pro-inflammatory cytokines, enhance intestinal barrier integrity, and alleviate age-related inflammatory responses. *Faecalibacterium* is a key butyrate producer in the gut, providing energy for intestinal epithelial cells and modulating the immune system to exert anti-inflammatory effects. The upregulation of these bacterial species can synergistically promote intestinal homeostasis through mechanisms such as short-chain fatty acid synthesis, immune function regulation, and intestinal barrier protection.

[0056] The above results indicate that the polysaccharide composition of the present invention, containing oat β-glucan and 2'-fucosylated lactose, can effectively improve age-related intestinal degenerative changes, improve intestinal homeostasis damage through multi-dimensional synergistic effects, and all active ingredients used are of natural origin, have high safety, are suitable for long-term use, and have extremely high development value.

[0057] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art should understand that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively describe all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. The application of a polysaccharide composition in the preparation of products that improve intestinal homeostasis, characterized in that, The polysaccharide composition comprises a combination of active ingredients and a food- or pharmaceutically acceptable carrier or excipient, wherein the combination of active ingredients is oat β-glucan and 2'-fucosylated lactose.

2. The application according to claim 1, characterized in that, The mass ratio of oat β-glucan to 2'-fucosylated lactose is 1:3 to 3:

1.

3. The application according to claim 1, characterized in that, A single dose is administered per kilogram of body weight, and each unit dose of the polysaccharide composition contains 50-200 mg of oat β-glucan and 50-200 mg of 2'-fucosylated lactose.

4. The application according to claim 1, characterized in that, The polysaccharide composition is a combined formulation comprising a first formulation and a second formulation packaged separately; the active ingredient of the first formulation is oat β-glucan, and the active ingredient of the second formulation is 2'-fucosylated lactose.

5. The application according to claim 4, characterized in that, The dosage form of the polysaccharide composition is selected from pills, powders, capsules, tablets, granules, films, or liquids.

6. The application according to claim 1, characterized in that, The product that improves gut homeostasis is one that has at least one of the following functions: (1) Improves intestinal barrier permeability damage; (2) Relieves intestinal inflammation; (3) Reduce intestinal oxidative stress levels; (4) Regulate the structure of intestinal flora and increase the abundance of probiotics in the gut; (5) Promotes the metabolism and production of short-chain fatty acids.

7. The application according to claim 6, characterized in that, The intestinal barrier permeability impairment, intestinal inflammatory response, or intestinal oxidative stress are induced by aging or inflammation.

8. The application according to claim 1, characterized in that, The products mentioned for improving gut homeostasis are functional foods, health supplements, or pharmaceuticals.

9. The application according to claim 1, characterized in that, The food- or pharmaceutically acceptable carrier or excipient is selected from one or more of the following: excipients, diluents, binders, disintegrants, lubricants, flow aids, surfactants, buffers, preservatives, flavoring agents, and coloring agents.

10. The application according to claim 9, characterized in that, The excipient is selected from one or more of starch, lactose, and dextrin; the diluent is selected from one or more of calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, and corn starch.

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