A composition for modulating neural excitability and uses thereof
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-06-03
- Publication Date
- 2026-08-11
AI Technical Summary
然而，现有技术中关于天然来源活性成分及合成生物原料在调节神经兴奋性方面的应用，多局限于单一成分的提取与利用，缺乏对多种原料协同作用的系统性研究与揭示
本发明提供的调节神经兴奋性的组合物包括牛乳外泌体、母乳低聚糖和功能蛋白。母乳低聚糖可作用于肠道受体并介导神经内分泌通路，调控神经递质的合成与转运；功能蛋白通过螯合金属离子维持神经元胞内离子稳态，以稳定细胞膜电位，调控突触传递效率；牛乳外泌体可通过递送功能性核酸、蛋白质等活性物质调控肠-脑轴信号传导，从而调节神经元离子通道与神经递质释放。因此本发明通过三种核心组分的复配，实现了协同增效，经肠-脑轴及神经-内分泌网络共同发挥调节作用，维持神经兴奋状态处于正常生理区间。
Smart Images
Figure SMS_1 
Figure SMS_2 
Figure SMS_3
Abstract
Description
Technical Field
[0001] This invention relates to the field of food technology, and more particularly to a composition for regulating nerve excitability and its application. Background Technology
[0002] The nervous system is one of the most complex functional systems in the human body. The central nervous system (CNS) is responsible for integrating and regulating sensory, motor, and higher cognitive functions throughout the body. In recent years, with the accelerated pace of modern society, increased work pressure, and the aging population, various nervous system dysfunctions caused by imbalances in neural excitability have become increasingly common, such as anxiety, insomnia, memory loss, attention deficit, and neurodegenerative diseases. Most of these diseases are closely related to the disruption of central nervous system homeostasis, manifesting as excessive neuronal excitation or insufficient inhibition, leading to a series of pathological changes such as decreased synaptic plasticity, neurotransmitter metabolic disorders, and enhanced neuroinflammatory responses.
[0003] Currently, clinical treatments for abnormal neurological excitability mainly include intervention with chemically synthesized drugs, such as benzodiazepines, antiepileptic drugs, and antidepressants. Although these drugs have certain efficacy in the short term, they generally have high risks of dependence, significant side effects (such as drowsiness, dizziness, and cognitive impairment), and a high relapse rate after discontinuation. Furthermore, existing treatments often focus on relieving individual symptoms and lack mechanisms for regulating the overall homeostasis of the central nervous system, making it difficult to fundamentally improve neurological function.
[0004] Meanwhile, naturally derived active ingredients and synthetic biological raw materials are used in health foods or foods for special medical purposes (foods for special medical purposes) and are increasingly being applied to the field of nervous system health. However, existing technologies regarding the application of naturally derived active ingredients and synthetic biological raw materials in regulating nerve excitability are mostly limited to the extraction and utilization of single components, lacking systematic research and elucidation of the synergistic effects of multiple raw materials. Therefore, there is an urgent need in this field to provide a composition that is safe in its raw material sources, convenient to consume, and capable of synergistically regulating nerve excitability and promoting the restoration of central nervous system homeostasis, in order to compensate for the shortcomings of existing drugs and traditional foods for special medical purposes in maintaining nervous system health. Summary of the Invention
[0005] To address the aforementioned problems, this invention provides a composition for regulating nerve excitability and its application. Through the design of the composition, nerve excitability is effectively regulated, maintaining the homeostatic balance of the central nervous system.
[0006] To achieve the above-mentioned objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a composition for regulating nerve excitability, comprising bovine milk exosomes, human milk oligosaccharides, and functional proteins.
[0007] The gut-brain axis (GBA) is a bidirectional, multi-pathway information exchange system between the gut and the central nervous system. The gut is the largest endocrine organ in the human body, secreting various hormones that act on the brain via blood circulation or indirectly transmit signals by activating vagal nerve afferent fibers. These hormones mainly include serotonin (5-HT), glucagon-like peptide-1 (GLP-1), cholecystokinin (CCK), ghrelin, and leptin. 5-HT and GLP-1, as key gut-derived signaling molecules, synergistically participate in the regulation of central nervous system excitability homeostasis through the gut-brain axis. The concentration difference between the central nervous system and peripheral tissues can significantly regulate neuronal excitability and the body's physiological homeostasis.
[0008] Compared with the prior art, the composition for regulating nerve excitability provided by the present invention regulates the transmission of gut-derived signals to the central nervous system by regulating the expression of tryptophan hydroxylase 1 gene (TPH1) and GLP-1 in intestinal epithelial Caco-2 cells, thereby regulating nerve excitability, maintaining the homeostasis of the central nervous system, and reducing agitation.
[0009] Preferably, the composition comprises the following components in parts by weight: 5 to 50 parts bovine milk exosomes, 50 to 150 parts human milk oligosaccharides, and 10 to 30 parts functional protein.
[0010] Preferably, the human milk oligosaccharide includes at least two of 2'-fucosylated lactose (2'-FL), 3'-sialylated lactose (3'-SL), lactose-N-tetrasaccharide (LNT), or 6'-sialylated lactose (6'-SL).
[0011] More preferably, the human milk oligosaccharide comprises 2'-fucosylated lactose and 3'-sialylated lactose.
[0012] More preferably, the mass ratio of 2'-fucosylated lactose to 3'-sialylated lactose in the human milk oligosaccharide is (9~18):1.
[0013] More preferably, the mass ratio of 2'-fucosylated lactose to 3'-sialylated lactose in the human milk oligosaccharide is (9~15):1.
[0014] Preferably, the functional protein includes at least one of casein phosphopeptide or osteopontin.
[0015] In this invention, if the functional protein includes casein phosphopeptide and osteopontin, there is no specific limitation on the ratio of casein phosphopeptide and osteopontin; any ratio is acceptable.
[0016] More preferably, the composition comprises the following components in parts by weight: 5 to 50 parts of bovine milk exosomes, 47 to 135 parts of 2'-fucosylated lactose, 3 to 15 parts of 3'-sialylated lactose, and 10 to 25 parts of casein phosphopeptide.
[0017] More preferably, the composition comprises the following components in parts by weight: 10 to 40 parts of bovine milk exosomes, 60 to 120 parts of 2'-fucosylated lactose, 6 to 15 parts of 3'-sialylated lactose, and 10 to 20 parts of casein phosphopeptide.
[0018] More preferably, the composition comprises the following components in parts by weight: 5 to 50 parts of bovine milk exosomes, 47 to 135 parts of 2'-fucosylated lactose, 3 to 15 parts of 3'-sialylated lactose, and 15 to 30 parts of osteopontin.
[0019] More preferably, the composition comprises the following components in parts by weight: 10 to 40 parts of bovine milk exosomes, 70 to 110 parts of 2'-fucosylated lactose, 5 to 12 parts of 3'-sialylated lactose, and 15 to 25 parts of osteopontin.
[0020] Secondly, the present invention provides a method for preparing the composition for regulating nerve excitability, comprising the following steps: Weigh each component according to the designed ratio, mix bovine milk exosomes, human milk oligosaccharides and functional proteins evenly to obtain a composition for regulating nerve excitability.
[0021] Thirdly, the present invention provides a product comprising the aforementioned composition for regulating nerve excitability.
[0022] Preferably, the product includes foods for special medical purposes.
[0023] The composition for regulating nerve excitability provided by the present invention can be used to prepare special medical purpose formula foods, and its form can be liquid preparation, powder, capsule, etc.
[0024] Preferably, the product is a dairy product.
[0025] More preferably, the dairy product includes at least one of liquid milk, flavored milk, yogurt, cheese, flavored milk powder, milk powder, or dairy beverage.
[0026] The present invention has the following beneficial effects: The composition for regulating neuronal excitability provided by this invention comprises bovine milk exosomes, human milk oligosaccharides, and functional proteins. Human milk oligosaccharides act on intestinal receptors and mediate neuroendocrine pathways, regulating the synthesis and transport of neurotransmitters. Functional proteins maintain intracellular ion homeostasis in neurons by chelating metal ions, thereby stabilizing cell membrane potential and regulating synaptic transmission efficiency. Bovine milk exosomes can regulate gut-brain axis signal transduction by delivering functional nucleic acids, proteins, and other active substances, thereby regulating neuronal ion channels and neurotransmitter release. Therefore, this invention achieves synergistic effects through the combination of these three core components, working together via the gut-brain axis and neuroendocrine network to maintain neuronal excitability within the normal physiological range. Detailed Implementation
[0027] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.
[0028] Unless otherwise specified, all materials used in the embodiments of this invention can be obtained through commercial channels or prepared by conventional methods in the art.
[0029] Example 1 This embodiment provides a composition for regulating nerve excitability, comprising the following components in parts by weight: 10 parts bovine milk exosomes, 120 parts human milk oligosaccharides, and 10 parts osteopontin. The human milk oligosaccharides are composed of 2'-FL and 3'-SL in a mass ratio of 14:1, i.e., 112 parts 2'-FL and 8 parts 3'-SL.
[0030] The preparation method of the above-mentioned composition for regulating nerve excitability includes the following steps: Weigh each component according to the designed proportions, mix them evenly, and you will get the composition for regulating nerve excitability.
[0031] Example 2 This embodiment provides a composition for regulating nerve excitability, comprising the following components in parts by weight: 25 parts bovine milk exosomes, 100 parts human milk oligosaccharides, and 15 parts osteopontin. The human milk oligosaccharides are composed of 2'-FL and 3'-SL in a mass ratio of 9:1, i.e., 90 parts 2'-FL and 10 parts 3'-SL.
[0032] The preparation method of the above-mentioned composition for regulating nerve excitability is the same as that in Example 1, and will not be repeated here.
[0033] Example 3 This embodiment provides a composition for regulating nerve excitability, comprising the following components in parts by weight: 40 parts bovine milk exosomes, 80 parts human milk oligosaccharides, and 25 parts casein phosphopeptides. The human milk oligosaccharides are composed of 2'-FL and 3'-SL in a mass ratio of 12.3:1, i.e., 74 parts 2'-FL and 6 parts 3'-SL.
[0034] The preparation method of the above-mentioned composition for regulating nerve excitability is the same as that in Example 1, and will not be repeated here.
[0035] Example 4 This embodiment provides a composition for regulating nerve excitability, similar to Example 2, except that 2'-FL is replaced with an equal mass of lactose-N-tetrasaccharide (LNT). Specifically, the composition comprises the following components in parts by mass: 25 parts bovine milk exosomes, 100 parts human milk oligosaccharides, and 15 parts osteopontin. The human milk oligosaccharides consist of LNT and 3'-SL in a mass ratio of 9:1, i.e., 90 parts LNT and 10 parts 3'-SL.
[0036] The preparation method of the above-mentioned composition for regulating nerve excitability is the same as that in Example 1, and will not be repeated here.
[0037] Example 5 This embodiment provides a composition for regulating nerve excitability, similar to Example 2, except that 3'-SL is replaced with an equal mass of 6'-SL. Specifically, the composition comprises the following components in parts by mass: 25 parts bovine milk exosomes, 100 parts human milk oligosaccharides, and 15 parts osteopontin. The human milk oligosaccharides are composed of 2'-FL and 6'-SL in a mass ratio of 9:1, i.e., 90 parts 2'-FL and 10 parts 6'-SL.
[0038] The preparation method of the above-mentioned composition for regulating nerve excitability is the same as that in Example 1, and will not be repeated here.
[0039] Comparative Example 1 This comparative example provides a composition similar to that of Example 2, except that it does not contain functional proteins and the human milk oligosaccharide is 2'-FL (without 3'-SL). Specifically, the composition comprises the following components in parts by weight: 25 parts bovine milk exosomes and 100 parts 2'-FL.
[0040] The preparation method of the above composition is the same as that in Example 1 (without the addition of osteopontin), and will not be repeated here.
[0041] Comparative Example 2 This comparative example provides a composition similar to that of Example 2, except that the bovine exosomes are replaced with an equal mass of bovine milk. Specifically, the composition comprises the following components in parts by weight: 25 parts bovine milk, 100 parts human milk oligosaccharides, and 15 parts osteopontin. The human milk oligosaccharides consist of 2'-FL and 3'-SL in a mass ratio of 9:1, i.e., 90 parts 2'-FL and 10 parts 3'-SL.
[0042] The preparation method of the above composition is the same as that in Example 1 (by replacing bovine milk exosomes with bovine milk), and will not be repeated here.
[0043] Comparative Example 3 This comparative example provides a composition similar to that of Example 2, except that the human milk oligosaccharides are replaced with an equal mass of galactooligosaccharides (GOS). Specifically, the composition comprises the following components in parts by mass: 25 parts bovine exosomes, 100 parts GOS, and 15 parts osteopontin.
[0044] The preparation method of the above composition is the same as that in Example 1 (with human milk oligosaccharides replaced by GOS), and will not be repeated here.
[0045] Cellular experimental verification 1. Cell experiments and grouping Caco-2 cells were activated in DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin, and cultured at 37°C with 5% CO2, with the medium changed every other day. Cells with a confluence greater than 85% in the logarithmic growth phase were passaged, washed twice with PBS, digested with 0.25% trypsin for 2 min, and then digested with complete culture medium to stop the digestion. Cells were pipetted until they detached from the flask wall, transferred to centrifuge tubes, centrifuged at 1500 rpm for 5 min, and the supernatant was removed. Cells were then passaged at a ratio of 1:2. Cells in the logarithmic growth phase were used for experiments.
[0046] 2. The composition regulates the expression of TPH1 and GLP-1 genes. Caco-2 cells were divided into 1×10 6 Cells were seeded per well in cell culture plates and cultured for 12 h until adherence. The culture medium was removed, and the cells were washed twice with sterile PBS buffer. 2 mL of a mixture prepared with DMEM (1% w / v) was added to each experimental group, while the control group received an equal volume of DMEM medium. Cells were cultured at 37°C and 5% CO2 for 24 h. After culture, the supernatant was discarded, and the cells were washed twice with sterile PBS buffer. Cells were scraped and collected, and total RNA was extracted according to the reagent instructions. The concentration and purity were determined. After extraction, the RNA was reverse transcribed into cDNA, and the expression levels of TPH1 and GLP-1 were determined using qPCR.
[0047] The experimental groups are shown in Table 1. After adding each composition to DMEM medium, the concentration of the composition in each experimental group was 1% w / v. Note: The composition in each experimental group can be a single component, such as experimental group 1 and experimental group 3.
[0048] Table 1. Cell Experiment Group Design
[0049] Elevated central 5-HT concentrations can mediate neuronal hyperpolarization by activating 5-HT1A receptors, inhibiting the release of excitatory neurotransmitters such as glutamate, and enhancing the activity of the GABA (gamma-aminobutyric acid, an inhibitory neurotransmitter) inhibitory pathway, thereby reducing central neuronal excitability, manifested as a more stable mood and reduced anxiety and impulsive behavior. However, excessive accumulation of central 5-HT can lead to excessive inhibition of neuronal excitability, resulting in phenotypes such as apathy, slowed thinking, decreased activity, and drowsiness and fatigue. TPH1 in intestinal tissue is a key rate-limiting enzyme in 5-HT synthesis; its upregulation can increase the synthesis and secretion of peripheral 5-HT. GLP-1 receptors are widely distributed in the central nervous system; in brain regions such as the amygdala and hippocampus, GLP-1 signaling can enhance the release and efficacy of GABA, leading to a more sedative state in neural networks. Intestinal secretions of 5-HT and GLP-1 can mediate bidirectional communication along the gut-brain axis via the vagus nerve pathway and circulatory pathway, further mitigating excessive central nervous system excitation and inhibiting excessive secretion of stress hormones. These factors play a crucial physiological role in maintaining central nervous system homeostasis and regulating early neural development. The gene expression levels of TPH1 and GLP-1 are shown in Table 2.
[0050] Table 2 Gene expression levels of TPH1 and GLP-1
[0051] Note: Each experimental group was compared with the control group. *, **, and *** indicate significant differences. P < 0.05 was marked as *, P < 0.01 as **, and P < 0.001 as ***.
[0052] Table 2 shows that the composition provided by this invention is superior to compositions containing only a single component and compositions containing any two components. Compared with experimental groups 12-14, experimental groups 8 showed increased expression levels of related genes, indicating that the composition of bovine exosomes, human milk oligosaccharides, and functional proteins has a regulatory effect on nerve excitation compared to bovine milk and GOS. When any two components of bovine exosomes, human milk oligosaccharides, and osteopontin were combined (experimental groups 4-6), the gene expression levels of TPH1 and GLP-1 increased significantly (P < 0.05). When the three components were combined synergistically (experimental groups 7-9), the effects of different ratios of the compositions on the gene expression levels of TPH1 and GLP-1 varied. When the mass ratio of bovine exosomes, human milk oligosaccharides, and functional proteins was 25:100:15, the gene expression levels of TPH1 and GLP-1 both reached their maximum values (P < 0.001), suggesting the existence of a dose-dependent optimal ratio.
[0053] Animal experiments verified 1. Laboratory animals Zebrafish were housed in aquarium water at 28°C, and their husbandry and management met the requirements of the International Committee for Assessment and Certification of Laboratory Animal Husbandry and Fertility (CIAH). Water quality requirements: 200 mg of readily soluble sea salt was added per 1 L of reverse osmosis water; conductivity was 450 μS / cm–550 μS / cm; pH was 6.5–8.5; hardness was 50 mg / L–100 mg / L CaCO3. Wild-type AB strain zebrafish, 5 days after fertilization (5dpf, at which point their organs were mature), were randomly selected and placed in cell culture plates. Each well contained 30 zebrafish, and each well contained 3 mL of feeding medium.
[0054] 2. Grouping and Model Establishment of Zebrafish The zebrafish were randomly divided into 16 groups, including a blank control group, a model control group, and 14 sample treatment groups, as shown in Table 3. The blank control group received no treatment. Both the model control group and the sample treatment groups ingested 140 μg / mL of penetrazol (PTZ) to induce agitation. The sample treatment groups ingested PTZ along with the various compositions. All compositions were thoroughly mixed and dissolved in water at a concentration of 1% w / v.
[0055] Table 3 Animal Experiment Group Design
[0056] 3. Effects of sample treatment groups on zebrafish behavior After treatment at 28℃ for 1 day, 10 zebrafish were randomly selected from each group and transferred to 96-well plates, one zebrafish per well, with a feeding volume of 200 μL per well. Behavioral analysis was then performed on the zebrafish to evaluate the effects of different compositions on their behavior. The results are shown in Table 4.
[0057] Table 4. Zebrafish behavioral data
[0058] As shown in Table 4, compared with the blank control group, the model control group after sample treatment successfully established the agitation model in zebrafish, with the highest number of awakening rounds and total awakening time. After treatment with different compositions, sample treatment group 8 showed the greatest reduction in the number of awakening rounds (16.66 times) and the highest reduction in total awakening time (1027.02 s), indicating that the specific ratio of bovine milk exosomes, human milk oligosaccharides, and functional proteins has excellent effects in reducing nerve excitability and maintaining central nervous system homeostasis.
[0059] In summary, the composition for regulating nerve excitability provided by this invention exhibits a super-additive synergistic effect among bovine milk exosomes, human milk oligosaccharides, and functional proteins. By regulating the expression of TPH1 and GLP-1 in intestinal epithelial Caco-2 cells, it modulates the transmission of gut-derived signals to the central nervous system, thereby regulating nerve excitability, maintaining homeostasis of the central nervous system, and reducing agitation.
[0060] 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 or improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A composition for modulating neural excitability, characterized by: It includes bovine milk exosomes, human milk oligosaccharides, and functional proteins.
2. The composition for regulating nerve excitability according to claim 1, wherein: The composition comprises the following components in parts by weight: 5 to 50 parts bovine milk exosomes, 50 to 150 parts human milk oligosaccharides, and 10 to 30 parts functional protein.
3. The composition for regulating neural excitability according to claim 1, wherein: The human milk oligosaccharides include 2'-fucosylated lactose and 3'-sialylated lactose.
4. The composition for regulating nerve excitability according to claim 3, wherein: The mass ratio of 2'-fucosylated lactose to 3'-sialylated lactose in the human milk oligosaccharide is (9~18):
1.
5. The composition for regulating nerve excitability as described in claim 1, characterized in that: The functional protein includes at least one of casein phosphopeptide or osteopontin.
6. The composition for regulating nerve excitability according to any one of claims 1 to 5, wherein: The composition comprises the following components in parts by weight: 5 to 50 parts bovine milk exosomes, 47 to 135 parts 2'-fucosylated lactose, 3 to 15 parts 3'-sialylated lactose, and 10 to 25 parts casein phosphopeptide.
7. The composition for regulating nerve excitability according to any one of claims 1 to 5, wherein: The composition comprises the following components in parts by weight: 5 to 50 parts bovine milk exosomes, 47 to 135 parts 2'-fucosylated lactose, 3 to 15 parts 3'-sialylated lactose, and 15 to 30 parts osteopontin.
8. A product characterized by: The composition comprising any one of claims 1 to 7 for regulating nerve excitability.