A complex prebiotic composition and its use in improving cognitive impairment in diabetes

Abstract

The application provides a composite prebiotic composition and application thereof in improving diabetic cognitive impairment, and the composite prebiotic composition comprises components in the following mass fractions: 15-25 parts of functional fructan, 0.03-0.04 parts of polysaccharide and 0.10-0.14 parts of polyphenol; wherein the functional fructan comprises long-chain inulin with an average polymerization degree DP of 23 or more and short-chain inulin with an average polymerization degree DP of 3-5 in a mass ratio of (1-4):1. The composite prebiotic composition of the application can significantly improve intestinal flora diversity, specifically enrich beneficial flora and up-regulate serum beta-MCA and GLP-1 levels after intervention for 3-6 weeks.

Description

Technical Field

[0001] This invention belongs to the field of functional food, health product and pharmaceutical technology, and in particular relates to a compound prebiotic composition and its application in improving cognitive impairment in diabetes. Background Technology

[0002] Diabetic cognitive impairment (DE) is a significant central nervous system complication of diabetes, characterized by cognitive decline, memory loss, and reduced learning ability, and can progress to dementia in severe cases. DE shares similar neurodegenerative features with Alzheimer's disease (AD), both involving hippocampal β-amyloid (Aβ) deposition and tau protein hyperphosphorylation. With over 550 million people living with diabetes globally, approximately one-quarter of whom are in China, the cognitive impairment caused by DE has become a serious public health problem.

[0003] The gut microbiota plays a crucial regulatory role in central nervous system metabolism and inflammation through the microbiota-gut-brain axis. Studies have shown that the diversity and stability of the gut microbiota are significantly reduced in diabetic patients. Dysbiosis promotes increased lipopolysaccharide (LPS) levels and blood-brain barrier (BBB) ​​damage, thereby accelerating Aβ deposition and Tau protein phosphorylation, exacerbating cognitive impairment. Therefore, intervening in the gut microbiota to improve diabetic dementia (DE) has important scientific basis and clinical translational value.

[0004] Glucagon-like peptide-1 (GLP-1) receptor agonists have significant neuroprotective effects: related studies report that GLP-1 receptor agonists can reduce the incidence of dementia in diabetic patients by more than 50%, and exert protective effects through multiple targets such as inhibiting inflammation, reducing oxidative stress, and reducing Aβ and Tau pathology. β-Muricholic Acid (β-MCA), a secondary bile acid, is a metabolic product of gut microbiota and can activate intestinal L cells and central GLP-1 receptors, exerting neuroregulatory effects.

[0005] Functional fructans are a class of water-soluble dietary fibers composed of fructose molecules linked by β-2,1 (inulin, oligofructose) or β-2,6 (Levan fructan) glycosidic bonds. Because the human body lacks the enzymes to break down these glycosidic bonds, they cannot be directly digested and absorbed. However, they are fermented and utilized by probiotics in the colon, exhibiting significant prebiotic properties. Inulin (C6H) is a particularly valuable component. 11 O5(C6H 10 O5) n OH) is a naturally sourced fructan-based dietary fiber that selectively promotes the growth of Bifidobacteria (Bifidobacteria). Bifidobacterium ), Lactobacillus ( Lactobacillus Bacteroides ( BacteroidesThe prebiotic effect of promoting the proliferation of beneficial bacteria such as Trichophyton ( ) can also be achieved by promoting the growth of beneficial bacteria in the Trichophytonceae family ( ). Lachnospiraceae The enrichment of key gut microbiota such as inulin drives the synthesis of secondary bile acids such as β-MCA. However, most commercially available products are simple blends of inulin alone or with large amounts of fructooligosaccharides. There are currently no compound prebiotic products or related patents that specifically target the gut microbiota-β-MCA-GLP-1 receptor axis and are designed to improve dementia (DE). Therefore, this invention provides a compound prebiotic composition with clearly defined ingredients and a scientifically formulated ratio, using high-purity inulin as the core ingredient and blended with trace amounts of immunopolysaccharides and plant polyphenols.

[0006] The shortcomings of existing technologies are mainly reflected in: (1) the lack of specific prebiotic intervention programs for cognitive impairment (DE); (2) existing prebiotic products mainly target blood glucose regulation, neglecting the role of the gut-brain axis in cognitive protection; and (3) the lack of systematic regulation of the gut microbiota-bile acid-GLP-1 pathway. To address these shortcomings, this invention provides a compound prebiotic composition with inulin as the core, synergistically regulating gut microbiota and GLP-1 signaling, and its application. Summary of the Invention

[0007] In view of this, the present invention aims to overcome the deficiencies in the prior art and proposes a compound prebiotic composition and its application in improving diabetic cognitive impairment. The compound prebiotic composition of the present invention improves diabetic cognitive impairment by remodeling the gut microbiota structure, driving β-MCA enrichment, activating intestinal L cells and central GLP-1 receptors, inhibiting hippocampal microglia neuroinflammation, and reducing Aβ deposition and Tau protein phosphorylation.

[0008] To achieve the above objectives, the technical solution of the present invention is implemented as follows: In a first aspect, the present invention provides a compound prebiotic composition, the compound prebiotic composition comprising the following components in parts by weight: The product contains 15-25 parts of functional fructan, 0.03-0.04 parts of polysaccharide, and 0.10-0.14 parts of polyphenol; wherein the functional fructan comprises long-chain inulin with an average degree of polymerization (DP) ≥ 23 and short-chain inulin with an average degree of polymerization (DP) of 3-5, in a mass ratio of (1-4):1.

[0009] Preferably, both the long-chain inulin and the short-chain inulin are derived from chicory, and the inulin content in the compound prebiotic composition is 70%-90% by mass.

[0010] Preferably, the functional fructan further includes fructooligosaccharides, wherein the mass fraction of the fructooligosaccharides is less than 10% of the total mass of the complex prebiotic composition.

[0011] Preferably, the polysaccharide is yeast-glucan.

[0012] Preferably, the polyphenol is luteolin.

[0013] Preferably, the compound prebiotic composition further comprises RS2 resistant starch as an auxiliary dietary fiber, wherein the mass fraction of RS2 resistant starch is 5-10 parts.

[0014] Preferably, the compound prebiotic composition further comprises B vitamins as an excipient for supporting nerve metabolism, wherein the B vitamins are selected from one or more of vitamin B1, vitamin B6, and vitamin B12, and the mass fraction of the B vitamins is 0.05 parts.

[0015] Preferably, the compound prebiotic composition comprises the following components in parts by weight: The composition includes 22.5 parts inulin, 0.035 parts yeast β-glucan, 0.12 parts luteolin, 2.5 parts fructooligosaccharides, 2.45 parts RS2 resistant starch, and 0.05 parts B vitamins. The inulin is composed of long-chain inulin with an average degree of polymerization (DP) ≥ 23 and short-chain inulin with an average degree of polymerization (DP) of 3-5, in a mass ratio of 3:1.

[0016] Secondly, the present invention provides the application of this complex prebiotic composition in the preparation of products that target and regulate the gut microbiota-bile acid-GLP-1 receptor signaling axis.

[0017] Thirdly, the present invention provides the use of this compound prebiotic composition in the preparation of health products, functional foods, dietary supplements or special medical purpose foods for improving cognitive impairment in diabetes.

[0018] Compared with the prior art, the present invention has the following advantages: (1) The compound prebiotic composition of the present invention uses long-chain and short-chain inulin derived from chicory roots as the core, which can be synergistically combined with polysaccharides and polyphenols to exert multiple neuroprotective effects from the intestine to the central nervous system through multi-target synergistic action. It targets and regulates the intestinal flora-β-mouse bile acid (β-MCA)-GLP-1 receptor signaling axis. The inulin drives the increase of secondary bile acid β-MCA synthesis by selectively enriching key bacteria such as Trichophyton, thereby activating intestinal L cells and hippocampal central GLP-1 receptors, inhibiting microglial neuroinflammation (M1→M2 polarization), reducing Aβ deposition and Tau protein hyperphosphorylation in the hippocampus, and improving diabetic cognitive impairment. Animal experiments show that the compound prebiotic composition of the present invention can significantly improve intestinal flora diversity, specifically enrich beneficial bacteria, and upregulate serum β-MCA and GLP-1 levels after 3-6 weeks of intervention.

[0019] (2) This invention provides a new strategy for non-pharmacological intervention of diabetic cognitive impairment, which has the characteristics of high safety, strong targeting and sustained effect. Attached Figure Description

[0020] Figure 1 The results of the DE model mice were used to evaluate their cognitive abilities in the water maze test (Figure A shows the results of the water maze test for the four groups of mice; the shorter the path, the stronger the cognitive ability; Figure B shows the number of times the mice entered the target quadrant in the water maze; the higher the number of times, the stronger the cognitive ability). Figure 2 Immunohistochemical results of Aβ protein, total Tau protein, and phosphorylated Tau protein in the hippocampus of four groups of mice; Figure 3 A comparison chart (A) of gut microbiota α diversity (Shannon index) and β diversity (Bray-Curtis PCoA) analysis chart (B) for mice in each group before intervention (week 0), week 3 and week 6 of intervention. Figure 4 A heatmap of serum metabolomics (A) and a comparison of GLP-1 levels (B) between the compound prebiotic intervention group and the DE model group. Figure 5 ELISA detection of GLP-1 levels in intestinal L cells treated with different β-MCA (A), detection of neuroinflammatory factors in neurons and microglia (B), and qPCR detection of polarization markers (C) were performed. In the figure, Ctrl is the blank control, HG is the high glucose treatment group, GLP1 is the HG group + GLP-1 preparation, Butyrate is the HG group + sodium butyrate treatment of GLUTag cell supernatant, and TβMCA is the HG group + taurine β-cholic acid treatment of GLUTag cell supernatant. Figure 6 This is a schematic diagram illustrating the mechanism of action of the present invention. Detailed Implementation

[0021] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0022] In this document, 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 application pertains.

[0023] In this document, when values ​​are described as ranges, it should be understood that such disclosure includes disclosure of all possible subranges within that range, as well as the specific numerical values ​​that fall within that range, regardless of whether the specific numerical value or specific subrange is explicitly specified.

[0024] In this article, the terms "multiple" or "more than" are used unless otherwise specified, referring to a quantity greater than or equal to 2. For example, "one or more" means one or more types.

[0025] In this document, the terms "preferred" and "more preferred" are used only to describe implementation methods or embodiments with better effects, and should be understood as not constituting a limitation on the scope of protection of this invention.

[0026] In this document, terms such as "further" are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of this invention.

[0027] In this article, the term "and / or" describes an association between objects, indicating that three relationships can exist. For example, A and / or B means: A or B, or A and B.

[0028] In this document, the term "about" means a specified value of + / - 10%, preferably + / - 5%, and more preferably + / - 1%.

[0029] In this article, the terms “include,” “including,” “have,” “contain,” etc., are all open-ended terms, meaning that they include but are not limited to.

[0030] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar to or equivalent to those described herein may be used in the implementation or testing of this invention.

[0031] The present invention will be described in detail below with reference to the embodiments.

[0032] Example 1: The compound prebiotic composition of the present invention is mainly composed of inulin. The active ingredients are weighed according to the recommended daily intake ratio. The composition contains multiple ingredients as listed in Table 1: Table 1 Composition Table of the Composition

[0033] The preparation method of the above-mentioned compound prebiotic composition includes the following steps: The preparation method of the compound prebiotic composition of the present invention includes the following steps:

[0034] (1) Weigh each raw material component precisely according to the set ratio: 22.500 parts of chicory inulin (the ratio of long-chain inulin with DP≥23 to short-chain inulin with an average degree of polymerization of DP3-5 is 4:1, i.e., 18 parts and 4.5 parts respectively), 0.035 parts of yeast β-glucan, 0.120 parts of luteolin, 2.450 parts of RS2 type resistant starch and 0.050 parts of vitamin B complex; (2) Mix each component thoroughly and evenly in a mixing device for no less than 20 minutes to obtain a mixed powder; (3) To facilitate preservation, the mixture is further formulated into a hard capsule dosage form. Conventional pharmaceutical grade excipients are added to the mixed powder: 50,000 parts of pharmaceutical grade filler microcrystalline cellulose and 2,000 parts of lubricant magnesium stearate are mixed, and then standard No. 0 hard capsule shells are selected for precision filling, granulation, and packaging. (4) Ensure that the filling amount of each capsule is stably controlled at about 0.500 parts, and after passing the quality inspection, seal the package and store it in a cool and dry place to obtain compound prebiotic capsules.

[0035] The recommended dosage of the composition described in this invention is 15 capsules daily, taken in multiple doses with meals. Based on the formulation process, each capsule prepared in this embodiment contains, on average, the following active ingredients: approximately 0.1476 parts inulin, approximately 0.0164 parts resistant starch and B vitamins complex, approximately 0.00078 parts luteolin, and approximately 0.00023 parts yeast β-glucan. The continuous intervention period is not less than 4 weeks, preferably more than 6 weeks, to achieve the optimal effects of gut microbiota remodeling and cognitive improvement.

[0036] Example 2: Application of compound prebiotic intervention in improving cognitive impairment in diabetes

[0037] DE model mouse construction: SAMP8 rapidly aging mice were selected, and a diabetic cognitive impairment (DE) mouse model was established by inducing a high-fat diet (HFD) combined with streptozotocin (STZ). The experiment was divided into the following groups: Negative control group (R1): SAMP8 normal control R1 mice, fed normally without any intervention; Positive control group (P8): SAMP8 rapidly aging mice, fed normally without any intervention; DE model group (DE group): no prebiotic intervention was given after modeling; Compound prebiotic group (Inulin group): DE model mice were treated with the composition of this invention. Intervention time points: baseline (week 0), week 3, and week 6. Fecal samples were collected from each group at each time point for 16S rRNA gene sequencing, and blood samples were collected simultaneously to detect metabolic indicators such as GLP-1 and β-MCA.

[0038] Morris water maze test to assess cognitive abilities in DE model mice: The cognitive abilities of four groups of mice were assessed using the Morris water maze behavioral test. Intervention with compound prebiotics significantly improved spatial learning and memory abilities and working memory accuracy in DE model mice. Figure 1 A), the latency period for escaping the water maze was significantly shortened in the compound prebiotic intervention group (P < 0.01). Figure 1 B).

[0039] Detection of Aβ and Tau protein phosphorylation in the hippocampus: Immunohistochemical detection of Aβ deposition and Tau phosphorylation in brain tissue pathology. Figure 2The results showed that, compared with the DE group, the expression of Aβ protein in the CA3 region of the hippocampus and the cerebral cortex was significantly reduced after the compound prebiotic intervention (P<0.01), and the expression of Tau protein and Tau pS404 protein was significantly reduced (P<0.01), further verifying the improvement of water maze cognition.

[0040] Gut microbiota diversity detection: such as Figure 3 As shown, compared with the negative control group, the α-diversity of gut microbiota was significantly increased in the compound prebiotic group at weeks 3 and 6. The improvement in Shannon index in the compound prebiotic group was significantly better than that in the positive control group (P<0.01). β-diversity analysis showed that after 6 weeks of intervention, the microbiota structure of the compound prebiotic group was significantly separated from that of the R1, P8 control groups and the DE group (PERMANOVA, P=0.001, R²=0.49), indicating that the product of this invention has unique microbiota remodeling characteristics.

[0041] Compound prebiotic intervention upregulates serum β-MCA and GLP-1 levels: such as Figure 4 As shown, the serum metabolome of the compound prebiotic intervention group and the DE model group showed that β-MCA was significantly increased after compound prebiotic intervention; serum GLP-1 level was also significantly increased, indicating that compound prebiotic intervention can exert a neuroprotective effect through GLP-1.

[0042] Example 3: Compound prebiotic intervention upregulates serum β-MCA and GLP-1 levels

[0043] To further elucidate the mechanism by which compound prebiotics increase β-MCA and thereby regulate the secretion of GLP-1 by intestinal L cells, and how GLP-1 reduces neuronal inflammation and improves cognition, such as... Figure 5 As shown, mouse GLUTag cells were treated with β-MCA using a compound prebiotic. ELISA analysis revealed that GLP-1 levels increased with increasing β-MCA concentration. Furthermore, GLP-1 produced by β-MCA-treated intestinal L cells was collected and used in experiments on neurons and microglia. In the high-glucose group, the expression of the inflammatory factors IL1B, IL6, and TNFβ in hippocampal microglia decreased with the M1 marker CD86 and increased with the M2 marker CD206, suggesting a shift in the inflammatory microenvironment towards anti-inflammatory activity.

[0044] The above results confirm that the compound prebiotic composition of the present invention can effectively inhibit neuroinflammation through the gut microbiota-β-MCA-GLP-1 receptor axis, reduce Aβ deposition and Tau protein phosphorylation associated with diabetic cognitive impairment, and simultaneously remodel the gut microbiota and enrich... LachnospiraceaeKey gut microbiota drive β-MCA synthesis and promote GLP-1 secretion, forming a complete signaling axis from gut microbiota to β-MCA to GLP-1, thereby exerting a cognitive protective mechanism. Figure 6 ).

[0045] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A compound prebiotic composition, characterized in that: The compound prebiotic composition comprises the following components in parts by weight: The product contains 15-25 parts of functional fructan, 0.03-0.04 parts of polysaccharide, and 0.10-0.14 parts of polyphenol; wherein the functional fructan comprises long-chain inulin with an average degree of polymerization (DP) ≥ 23 and short-chain inulin with an average degree of polymerization (DP) of 3-5, in a mass ratio of (1-4):

1.

2. The compound prebiotic composition according to claim 1, characterized in that: Both the long-chain inulin and the short-chain inulin are derived from chicory, and the inulin content in the compound prebiotic composition is 70%-90% by mass.

3. The compound prebiotic composition according to claim 1, characterized in that: The functional fructan also includes fructooligosaccharides, wherein the mass fraction of the fructooligosaccharides is less than 10% of the total mass of the complex prebiotic composition.

4. The compound prebiotic composition according to claim 1, characterized in that: The polysaccharide is yeast-glucan.

5. The compound prebiotic composition according to claim 1, characterized in that: The polyphenol is luteolin.

6. The compound prebiotic composition according to claim 1, characterized in that: The compound prebiotic composition also contains RS2 resistant starch as an auxiliary dietary fiber, wherein the mass fraction of RS2 resistant starch is 5-10 parts.

7. The compound prebiotic composition according to claim 1, characterized in that: The compound prebiotic composition also contains B vitamins as an excipient to support nerve metabolism, wherein the B vitamins are selected from one or more of vitamin B1, vitamin B6, and vitamin B12, and the mass fraction of the B vitamins is 0.05-parts.

8. The compound prebiotic composition according to claim 1, characterized in that: The compound prebiotic composition comprises the following components in parts by weight: The composition includes 22.5 parts inulin, 0.035 parts yeast β-glucan, 0.12 parts luteolin, 2.5 parts fructooligosaccharides, 2.45 parts RS2 resistant starch, and 0.05 parts B vitamins. The inulin is composed of long-chain inulin with an average degree of polymerization (DP) ≥ 23 and short-chain inulin with an average degree of polymerization (DP) of 3-5, in a mass ratio of 3:

1.

9. The use of the compound prebiotic composition according to any one of claims 1-8 in the preparation of products that target and regulate the gut microbiota-bile acid-GLP-1 receptor signaling axis.

10. The use of the compound prebiotic composition according to any one of claims 1-8 in the preparation of health products, functional foods, dietary supplements or special medical purpose foods for improving cognitive impairment in diabetes.

Images