A composition containing yak milk fat globule membrane capable of improving cognitive function decline and application thereof

Abstract

The present application belongs to the technical field of pharmaceutical compositions, and particularly relates to a composition containing yak milk fat globule membrane and capable of improving cognitive function decline and application thereof. The composition takes yak milk fat globule membrane as a core active ingredient, and takes taurine, selenium-rich yeast and yam extract as auxiliary active ingredients. Through synergistic effect of the components, the composition can target inhibit acetylcholinesterase activity, relieve oxidative stress damage of brain nerve cells, reduce the level of pro-inflammatory factors in the brain, and then significantly improve cognitive function. The composition takes into account functionality and nutrition, and shows broad application prospects in the development of products related to improving learning and memory ability, delaying cognitive function decline and improving cognitive impairment.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical composition technology, and specifically relates to a composition containing yak milk fat globule membrane that can improve cognitive decline and its application. Background Technology

[0002] Cognitive function is a core function of the human central nervous system, encompassing key activities such as learning and memory, and spatial recognition, and is the foundation for normal life and social interaction. Influenced by multiple factors including age and lifestyle, cognitive decline mainly manifests as reduced learning and memory, and spatial cognitive impairment, not only lowering the individual's quality of life but also placing a heavy burden on families and society. Its pathogenesis is complex, corely involving decreased synaptic plasticity and cholinergic neuronal disorders. Among these, excessive activation of acetylcholinesterase (AChE), neuroinflammatory cascade reactions, and oxidative damage to brain neurons are key factors mediating cognitive decline; these three factors synergistically exacerbate neuronal damage and accelerate functional decline.

[0003] Currently, interventions for cognitive decline mainly include drug therapy, behavioral intervention, and functional food conditioning. While commonly used drugs can alleviate symptoms, they have limitations such as side effects and drug resistance; behavioral interventions are slow to take effect and vary greatly from person to person. In contrast, functional foods have become a research hotspot due to their safety, long-term suitability, and multi-target regulation advantages, with the development of highly effective and safe natural active compound formulas being the core direction.

[0004] Milk fat globule membrane (MFGM) is rich in active lipids and has neuroprotective functions. Among them, yak milk MFGM has a unique lipid composition due to the high-altitude environment, and its cognitive protection potential is superior to that of cow milk MFGM.

[0005] Existing technologies mostly focus on the study of single active ingredients, and have not yet scientifically combined MFGM with other components, and lack systematic research on its synergistic mechanism and its application in improving cognitive decline.

[0006] Based on this, the present invention uses yak milk MFGM as the core and combines it with three natural active ingredients (taurine, selenium-enriched yeast, and yam extract) to construct a composition. It improves cognitive function decline through multi-pathway regulation, provides new raw materials and technical solutions for the development of novel cognitive intervention products, and has important industrialization value. Summary of the Invention

[0007] To address the shortcomings of existing technologies, this invention aims to provide a composition containing yak milk fat globule membrane that can improve cognitive decline and its applications. The composition uses yak milk fat globule membrane as the core active ingredient and taurine, selenium-enriched yeast, and yam extract as auxiliary active ingredients. Through the synergistic effect of these components, the composition can target and inhibit acetylcholinesterase activity, alleviate oxidative stress damage to brain nerve cells, and reduce the level of pro-inflammatory factors in the brain, thereby significantly improving cognitive function. This composition combines functionality and nutritional value, demonstrating broad application prospects in improving cognitive function.

[0008] To achieve the above objectives, in a first aspect, the present invention provides a composition comprising yak milk fat globule membrane, taurine, selenium-enriched yeast, and yam extract, wherein the mass ratio of the yak milk fat globule membrane, taurine, selenium-enriched yeast, and yam extract is 15–35:2–8:3–10:0.5–2. For example, 17:2:2.5:0.5, 28:4:5:1, and 35:8:10:2.

[0009] Furthermore, the mass ratio of the yak milk fat globule membrane, taurine, selenium-enriched yeast, and yam extract is 15–35: 3–5: 4–6: 0.5–1.5.

[0010] Furthermore, the mass ratio of the yak milk fat globule membrane, taurine, selenium-enriched yeast, and yam extract is 15–35:4:5:1.

[0011] Furthermore, the mass ratio of the yak milk fat globule membrane, taurine, selenium-enriched yeast, and yam extract is 28:4:5:1.

[0012] In a second aspect, the present invention provides a product comprising the composition described in the first aspect.

[0013] Furthermore, the product is a health food or a medicine; Furthermore, the product also includes excipients acceptable for use in health foods or pharmaceuticals.

[0014] Thirdly, the present invention provides the use of the composition described in the first aspect in the preparation of products capable of improving cognitive function; Furthermore, the product is a pharmaceutical product.

[0015] Furthermore, the application is in the preparation of products with acetylcholinesterase inhibitory function.

[0016] Furthermore, the application is in the preparation of products that enhance learning and memory abilities.

[0017] Furthermore, the application includes at least one of the following: (1) Application in the preparation of products that can improve object recognition capabilities; (2) Application in the preparation of products that can enhance exploration capabilities; (3) Application in the preparation of products that can enhance the body's anti-inflammatory capabilities; (4) Application in the preparation of products that can improve the body’s antioxidant capacity and / or inhibit lipid peroxidation.

[0018] In specific embodiments of the present invention, the improvements in object recognition ability, exploration ability, anti-inflammatory capacity, antioxidant capacity, and / or inhibition of lipid peroxidation are demonstrated in the context of cognitive decline.

[0019] Furthermore, the enhancement of the body's anti-inflammatory capacity includes reducing the level of pro-inflammatory factors in the brain; preferably, the pro-inflammatory factors in the brain include IL-6 or COX-2 in the brain.

[0020] In a specific embodiment of the present invention, abnormal levels of pro-inflammatory factors in the brain are caused by a decline in cognitive function.

[0021] Furthermore, the enhancement of the body's antioxidant capacity and / or inhibition of lipid peroxidation includes improving the level of antioxidant factors in brain tissue; preferably, the antioxidant factors include GSH or MDA.

[0022] In a specific embodiment of the present invention, abnormal antioxidant capacity and abnormal lipid peroxidation levels are caused by a decline in cognitive function.

[0023] Fourthly, the present invention provides the use of the composition described in the first aspect in the preparation of products capable of assisting in improving memory; Furthermore, the product is a health food or a medicine.

[0024] Compared with the prior art, the present invention has the following advantages: (1) This invention demonstrates that the combined intervention of yak milk fat globule membrane, taurine, selenium-enriched yeast and yam extract can help improve cognitive function, and these four components have a significant synergistic effect. (2) The composition of the present invention can improve the decline of cognitive function, has a synergistic effect in inhibiting acetylcholinesterase and improving learning and memory ability, and has the effects of improving object recognition ability, improving exploration ability, improving the body's anti-inflammatory ability, improving the body's antioxidant ability and / or inhibiting lipid peroxidation ability. Attached Figure Description

[0025] Figure 1The contents of glycerophospholipids and sphingomyelins in yak milk MFGM and bovine milk MFGM were analyzed, where A represents the contents of glycerophospholipids and sphingomyelins in bovine MFGM; and B represents the contents of glycerophospholipids and sphingomyelins in yak MFGM.

[0026] Figure 2 The contents of glycerophospholipids and sphingomyelin in yak milk MFGM and cow milk MFGM were analyzed, where A is the content of phosphatidylcholine (PC); B is the content of phosphatidic acid (PA); C is the content of phosphatidylinositol (PI); D is the content of phosphatidylserine (PS); E is the content of phosphatidylethanolamine (PE); and F is the content of sphingomyelin (SM).

[0027] Figure 3 Lipid profiles of MFGM in yak milk and MFGM in cow milk.

[0028] Figure 4 Differential lipid analysis was performed on the lipidomes of yak milk MFGM and bovine milk MFGM. In this study, A was principal component analysis (PCA) and B was partial least squares analysis (OPLS-DA).

[0029] Figure 5 Differences in lipid variation trends between yak milk MFGM and cow milk MFGM.

[0030] Figure 6 This study analyzed the enrichment and upregulated lipid function of differentially regulated lipids in yak milk MFGM using the KEGG pathway. A represents the KEGG analysis of differentially regulated lipids in yak and cow milk MFGM, and B represents the KEGG enrichment map of upregulated lipids in yak milk MFGM.

[0031] Figure 7 The inhibitory activity of MFGM from yak milk against AChE was investigated.

[0032] Figure 8 The effects of each component on the Y-maze test in mice with D-galactose-induced cognitive decline.

[0033] Figure 9 The effects of each component on the novel object recognition test in mice with D-galactose-induced cognitive decline.

[0034] Figure 10 The effects of each component on the water maze test in mice with D-galactose-induced cognitive decline were shown; where A represents the number of times the target area was traversed and B represents the time spent in the target quadrant.

[0035] Figure 11 The effects of each component on brain inflammation in mice with D-galactose-induced cognitive decline were investigated; where A represents IL-6 content and B represents COX-2 content.

[0036] Figure 12The effects of each component on oxidative damage in the brain of mice with D-galactose-induced cognitive decline were investigated; where A represents GSH content and B represents MDA content. Detailed Implementation

[0037] The following detailed embodiments further illustrate the concept and technical effects of the present invention to fully understand its purpose, features, and effects. Unless otherwise specified, all methods described are conventional methods. Unless otherwise specified, all materials are available from publicly available commercial sources. The illustrative embodiments and descriptions of the present invention are used to explain the invention and do not constitute an undue limitation thereof. It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0038] Example 1: Preparation of milk fat globule membrane 1. Raw material preparation: Select healthy adult yaks from Aba Prefecture, Sichuan Province and healthy dairy cows from Beijing Sanshi Dairy Farm Co., Ltd., and collect 3000 g of raw milk from each during the lactation period for later use. During the collection process, keep the temperature of the raw milk below 10 ℃ to avoid microbial growth and lipid oxidation.

[0039] 2. Centrifugation to separate cream: Centrifuge 3000 g of raw milk at 3000×g and 4 ℃ for 15 min. After centrifugation, collect the supernatant cream, seal it, and refrigerate it at 4 ℃ for 24 h. Add 0.1M phosphate buffer (pH=6.8) to the yak milk fat, mix thoroughly, and wash to remove residual milk protein. Separate and collect the fat phase, and transfer it to 4 ℃ for crystallization.

[0040] 3. Whisk and filter to obtain MFGM: Take out the chilled butter and pour it into the mixing bowl of a high-speed mixer. Pour the mixture into a glass funnel lined with three layers of sterilized gauze. Collect the light yellow aqueous phase obtained by filtration, which is yak MFGM and dairy cow MFGM (hereinafter referred to as cow MFGM). Then place it in a refrigerator at 4 ℃ for temporary storage.

[0041] Example 2: Lipidome characteristics of milk fat globule membrane An ACQUITY UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm, Waters, USA) was used; column temperature 55℃; injection volume 5 μL; mobile phase A was acetonitrile-water (60:40, v / v, containing 0.1% formic acid and 10 mM NH4COOH), and mobile phase B was acetonitrile-isopropanol (1:1, v / v, containing 0.1% formic acid and 10 mM NH4COOH); gradient elution program: 0 min 0% B, 1.5 min 0% B, 5 min 55% B, 10 min 60% B, 13 min 70% B, 15 min 90% B, 16 min 100% B, 18 min 100% B, 18.1 min 0% B, 20 min 0% B; flow rate 0.35 mL / min.

[0042] Mass spectrometry conditions: Electrospray ionization (ESI) source, positive and negative ion scanning mode; ion source voltage: +5500V (positive ion) / -4500V (negative ion); source temperature 400℃; sheath gas (GS1) 50psi, auxiliary gas (GS2) 55psi, curtain gas 40psi; collision gas (CAD) at medium intensity; quantitative analysis was performed using Schedule-MRM mode, controlled by Analyst 1.7 software.

[0043] Glycerol phospholipids and sphingomyelin lipids have been shown to have cognitive-improving activities. For example... Figure 1 The figures show the content of glycerophospholipids and sphingomyelin in yak MFGM and bovine MFGM. The content of glycerophospholipids in yak milk fat globule membrane was significantly higher than that in bovine milk fat globule membrane. The contents of phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidic acid (PA), phosphatidylinositol (PI), and phosphatidylethanolamine (PE) among the glycerophospholipids all showed significant differences. p < 0.05, Figure 2 AE); at the same time, the content of sphingomyelin (SM) in yak milk fat globule membrane is also significantly higher than that in bovine milk fat globule membrane, possessing a unique lipid composition advantage (AE); p < 0.05, Figure 2 F).

[0044] From the perspective of mechanism of action, the above five types of lipids can jointly promote the improvement of cognitive functions such as learning and memory through the synergistic effect of structural support and functional regulation: PC, as a precursor to the synthesis of the neurotransmitter acetylcholine, can directly participate in signal transmission in the central nervous system; PS can optimize the efficiency of signal transmission between nerve cells by regulating the activity of signaling molecules on the neuronal membrane; PA is a core precursor of glycerophospholipids, participating in membrane structural stability and nerve signal transmission, and its polyunsaturated fatty acid chains (such as molecules containing EPA and DHA) are the key material basis for cognitive improvement; PI, as an important precursor of signaling molecules, is deeply involved in the regulation of multiple signaling pathways within nerve cells, affecting nerve cell proliferation, differentiation, and synaptic plasticity; PE can maintain the homeostasis of nerve membrane structure and the efficiency of synaptic signal transmission, and alleviate nerve damage through anti-inflammatory and antioxidant effects; SM is a core component of the myelin sheath, which can ensure the integrity of the myelin sheath structure and maintain the efficient transmission of nerve impulses. In summary, compared with bovine milk fat globule membrane, yak milk fat globule membrane, with its unique and high-content characteristic lipid composition, shows more prominent potential application value in cognitive function protection and nerve function regulation.

[0045] Figure 3 The figure shows the difference in lipid expression between yak milk fat globule membrane and bovine milk fat globule membrane. Samples 1, 2, and 3 in the figure are parallel samples. The lipid profiles of the two groups of samples exhibit obvious grouping and clustering characteristics. In the yak MFGM group (1-3), most lipids (such as phosphatidylcholine (PC), some ceramides (Cer), phosphatidylserine (PS), sphingomyelin (SM), etc.) are highly expressed, which are lipids related to cognitive improvement. In contrast, in the bovine MFGM group (1-3), most lipids are mainly expressed at low levels, with only a few lipids showing relatively high expression. Combined with lipid function analysis, it can be seen that the lipids highly expressed in yak milk fat globule membrane, such as phosphatidylcholine (PC), phosphatidylserine (PS), sphingomyelin (SM), and ceramide (Cer), are precisely the core components related to cognitive improvement. The above results suggest that the lipid composition of yak milk fat globule membrane has a potential advantage over bovine milk fat globule membrane in terms of cognitive protection.

[0046] Example 3: Screening of differential lipids in yak milk fat globule membrane Principal component analysis (PCA) was performed on the MFGM of yaks and cattle to preliminarily understand the overall metabolic differences between the two groups and the magnitude of variability within each group. Figure 4 As shown in Figure A, significant clustering exists within the lipidomes of yak MFGM and bovine MFGM, and they can be clearly separated along the first principal component (PC1) axis, indicating a significant difference in lipid composition between the two groups. Lipidome data were analyzed using the OPLS-DA model, and score plots were generated for each group. Figure 4 (B) further demonstrates the differences between yak MFGM and cow MFGM.

[0047] Figure 5 The study demonstrates the trends in lipid changes between the two groups and the statistical significance of the differences. Among the differentially expressed lipids in yak MFGM and bovine MFGM, a total of 483 lipids were upregulated in yak milk, while 291 lipids were downregulated in yak milk.

[0048] The lipid molecules unique to yak milk fat globule membranes are mainly glycerophospholipids. As shown in Table 1, among the lipid molecules uniquely expressed in yak milk fat globule membranes (MFGMs), those belonging to the PS subclass are the most abundant, including PS (16:0–19:0); followed by the PC subclass, including PC (18:1–22:6); PA subclass, including PA (18:1–22:6); PI subclass, including PI (18:2–16:1); PG subclass, including PG (17:0–18:0); and PE subclass, including PE (22:6–18:1). These lipid molecules are present in higher amounts in yak MFGMs than other specific lipid molecules, and are important auxiliary active lipids for improving cognitive function in yak MFGMs.

[0049] Table 1. Lipid molecules unique to yak milk fat globule membrane

[0050] Example 4: Differential lipid screening and cognitive pathway enrichment Differential lipid fractions were imported into the MetaboAnalyst database for pathway enrichment analysis, and the results are as follows: Figure 6 As shown in Figures AB, KEGG analysis of differentially expressed lipids in MFGM from bovine and yak milk revealed their primary involvement in neural regulation, lipid metabolism, and disease and inflammatory signaling pathways. The neural regulation involved primarily includes retrograde endocannabinoid signaling, the nerve growth factor (NGF) signaling pathway, serotonin synapses, and glutamatergic synaptic regulation. Specifically, Cer was the main lipid involved in the NGF signaling pathway, while PGE2 was the main lipid involved in the serotonergic synaptic pathway. PC and PE were the main lipids involved in retrograde endocannabinoid signaling, with PC (18:1–22:6) and PE (22:6–18:1) being unique to yak MFGM and participating in the retrograde endocannabinoid signaling pathway. Lipid metabolism involved in cognitive improvement mainly included glycerophospholipid and sphingolipid metabolism, while inflammatory signaling pathways involved primarily included the MAPK signaling pathway and the NF-kappa B signaling pathway.

[0051] In summary, based on lipidomics data and mechanistic analysis, the abundance of brain-targeting lipids in yak milk fat globule membranes can be translated into a clear advantage in cognitive pathway regulation. For example, the contents of PC, PA, PI, PS, PE, and SM in yak milk fat globule membranes are 2.78 times, 5.13 times, and 3.6 times, and 5.99 times, 5.67 times, and 2.71 times, respectively, compared to bovine MFGM. The overall cognitive pathway regulatory effect far exceeds that of bovine MFGM.

[0052] Example 5: Effect of yak MFGM on AChE inhibitory activity Acetylcholinesterase (AChE) is responsible for degrading the neurotransmitter acetylcholine. Inhibiting its activity can increase the level of acetylcholine in the synaptic cleft, enhance cholinergic neurotransmission, and thus improve cognitive functions such as learning and memory. Acetylcholinesterase (AChE) inhibitory activity was detected on 96-well microplates. Experimental group (S), experimental background group (SB), control group (C), and blank group (B) were set up, with three parallel wells in each group. The specific procedures were as follows: For the experimental group, 100 μL of pH 8.0 PBS buffer, 50 μL of yak MFGM and bovine MFGM solution, and 20 μL of 1.0 U / mL AChE were added sequentially. After mixing thoroughly, the mixture was incubated at 37 ℃ for 20 min. Then, 20 μL of 0.075 M ACTI and 20 μL of 0.01 M DTNB were added sequentially, and the mixture was incubated at 37 ℃ in the dark for 20 min before the absorbance was measured at 412 nm. For the experimental background group, 20 μL of pH 8.0 PBS buffer was used instead of the 20 μL 1 U / mL AChE solution in the experimental group, with the remaining procedures being the same as the experimental group, to eliminate the influence of the binding of the experimental sample and the chromogenic substance on the absorbance. For the control group, 50 μL of sterile water was used instead of the 50 μL of sterile water in the experimental group. The control group used 50 μL of different yak MFGM and bovine MFGM solutions instead of the experimental group, and 20 μL of pH 8.0 PBS buffer instead of the experimental group, with the remaining procedures being the same as the experimental group, to eliminate the influence of substrate and chromogenic substances on absorbance.

[0053] The AChE activity inhibition rate was calculated using the following formula: AChE activity inhibition rate (%) = [1-(OD S -OD SB ) / (OD C -OD B )]*100% OD S : Absorbance of the experimental group OD SB : Absorbance of the experimental background group ODC : Absorbance of the control group OD B Absorbance of the blank group The results are as follows Figure 7 As shown, MFGM intervention can reduce AChE inhibitory activity, and the inhibitory activity of yak MFGM on AChE is significantly higher than that of bovine MFGM. p < 0.05).

[0054] Example 6: Raw material screening for the composition Using the method described in Example 5, the acetylcholinesterase (AChE) inhibitory activity of each candidate raw material was determined to complete the raw material screening. Yak MFGM, taurine (purchased from Qianjiang Yong'an Pharmaceutical Co., Ltd.), arabinoxylan (purchased from Shanghai Yaji Biotechnology Co., Ltd.), lutein (purchased from Chenguang Biotech Group Co., Ltd.), selenium-enriched yeast (purchased from Angel Yeast Co., Ltd.), yam extract (purchased from Tianyu Biotechnology Co., Ltd.), and fructooligosaccharides (purchased from Guangdong Mingtong Biotechnology Co., Ltd.) were selected for AChE inhibitory activity testing. The inhibition rate of AChE activity of the above raw materials was determined according to the method described in Example 5. The results are shown in Table 2, and all of the above substances showed a certain AChE inhibitory effect. Considering the biological activity, production cost, and sensory characteristics of each raw material, yak MFGM, taurine, selenium-enriched yeast, and yam extract were finally determined as the raw materials for the preparation of the subsequent composition.

[0055] Table 2. Inhibitory activity of different active substances against AChE

[0056] According to the proportions shown in Table 3, three or four of the following were mixed in different proportions to obtain solid composition powders, and the AChE inhibitory activity of each composition was determined.

[0057] Table 3 Formulation ratio settings for each group of compositions

[0058] Note: - indicates that the substance is not added. The same mass of each group of compositions was weighed and the inhibitory activity of different compositions on AChE was determined. The results are shown in Table 2. Among them, composition II (yak MFGM: taurine: selenium-enriched yeast: yam extract = 28: 4: 5: 1) had the highest inhibitory activity on AChE, which was 52.41% ± 0.86%.

[0059] Table 4. Inhibition rate of the composition against acetylcholinesteride

[0060] Example 7 Effects of the composition on mice with cognitive decline Eight-week-old male SPF-grade C57BL / 6J mice were housed in an environment with a temperature of 20±2℃, relative humidity of 65±5%, and a 12-hour light-dark cycle, ensuring good ventilation. Eight mice were per cage, fed mouse feed and given free access to water. The mice were housed for one week prior to the experiment to acclimatize. Mice were randomly divided into a negative control group, a model group, a yak MFGM group, a taurine group, a selenium-enriched yeast group, a yam extract group, a composition group (fed with composition II prepared in Example 6), and a positive control group. The feeding methods for each group were as follows: Negative control group: 0.1 mL / 10 g BW sterile saline was administered by gavage, and 0.1 mL / 10 g BW sterile saline was injected by injection; Model group: 0.1 mL / 10 g BW sterile saline was administered by gavage, followed by an injection of 300 mg / kg BW D-galactose; Yak MFGM group: 150 mg / kg BW yak MFGM was administered by gavage, followed by injection of 300 mg / kg BW D-galactose; Taurine group: 150 mg / kg BW taurine was administered by gavage, followed by injection of 300 mg / kg BW D-galactose; Selenium-enriched yeast group: 150 mg / kg BW selenium-enriched yeast was administered by gavage, followed by an injection of 300 mg / kg BW D-galactose; Yam extract group: 150 mg / kg BW yam extract was administered by gavage, followed by injection of 300 mg / kg BW D-galactose; Composition group: 150 mg / kg BW Composition II by gavage, followed by injection of 300 mg / kg BW D-galactose; Positive control group: administered 1 mg / kg BW donepezil by gavage and injected 300 mg / kg BW D-galactose.

[0061] Except for the negative control group, the injection volume of D-galactose solution per unit body weight of mice in each group was consistent with that in the negative control group. The mice were administered the drug via gavage daily, followed by intraperitoneal injection 4 hours later, for 8 consecutive weeks. After the intervention, behavioral tests were performed, followed by anesthesia and euthanasia by cervical dislocation, and organ and blood samples were collected.

[0062] 1. The regulatory effect of the composition on the behavior of mice with cognitive decline. 1.1 Y-maze experiment The spontaneous alternation rate in the Y-maze experiment is used to evaluate the cognitive function of mice. A higher spontaneous alternation rate generally indicates better working memory and spatial cognition. The Y-maze consists of three identical arms, each measuring 34 cm × 5 cm × 15 cm (length × width × height), with an angle of 120° between the arms. Mice are placed in the center of the Y-maze and allowed to explore freely for 8 minutes. The total number of times mice enter an arm and the number of consecutive entries into three different arms are recorded using an analysis system, and the spontaneous alternation rate is calculated according to the formula.

[0063] Spontaneous alternation rate = (Number of consecutive entries into three different arms) / (Total number of arm entries - 2) × 100% Depend on Figure 8 It can be seen that the spontaneous alternation rate of the Y-maze in the model group mice was 37.73%±1.13%, which was significantly lower than that in the negative control group ( p < 0.05. Compared with the model group, all interventions significantly increased the spontaneous alternation rate of the mouse Y-maze. Among the single-component comparisons, the yak MFGM group was superior to the taurine group, the selenium-enriched yeast group, and the yam extract group. Meanwhile, there was no significant difference in levels between the combined treatment group and the negative control group (…). p (>0.05); Compared with the single-drug interventions of the yak MFGM group, taurine group, selenium-enriched yeast group, and yam extract group, the spontaneous alternation rate of the combined group was significantly increased, indicating that the synergistic effect of each component in the composition is better than the effect of individual interventions of each component at the same dose. The above results indicate that the composition can significantly improve the spontaneous alternation behavior in the Y-maze of mice with cognitive decline and improve the cognitive function of the mice.

[0064] 1.2 New Object Recognition Experiment A novel object recognition experiment was used to evaluate the object discrimination ability of mice in a free-roaming state. During the first day of adaptation, mice were allowed to explore freely for 5 minutes. 24 hours after adaptation, training began. On the second day of training, two identical objects were placed at one-quarter of the height of a box. The mouse was placed into the box from the center of the side wall, equidistant from the two identical objects, and allowed to explore freely for 7 minutes. 24 hours after training, testing was conducted on the third day. A new object of different color and shape was used to replace one of the old objects. The positions of the two objects were the same as in the second day of training. The mouse was placed into the box from the center of the side wall, equidistant from the two identical objects, and allowed to explore freely for 7 minutes. After each experiment, the box and the recognized object were thoroughly disinfected with 75% ethanol. The exploration time of the mice for the new and old objects was manually recorded, and the preference index for the new object was calculated using a formula.

[0065] Preference Index = Time spent exploring new objects / (Time spent exploring new objects + Time spent exploring old objects) Depend on Figure 9As can be seen, compared with the negative control group, the novel object recognition index of mice in the model group was significantly reduced, indicating a significant decline in their short-term and long-term memory abilities, and the model was successfully established. Compared with the model group, all intervention groups significantly improved the novel object recognition index of mice, with the combined treatment group and the positive control group showing the most significant improvement, and there was no significant difference between the two groups. p The concentration of the compound was >0.05%, indicating that the composition was comparable to the positive control in improving the novel object recognition index in mice. Compared with the composition group, the novel object recognition index of the yak MFGM group, taurine group, selenium-enriched yeast group, and yam extract group was lower, indicating that the intervention effect of this composition was superior to that of each individual component at the same dose. The above results show that the composition can effectively improve the novel object recognition index in mice with declining cognitive function and improve cognitive decline; yak MFGM, taurine, selenium-enriched yeast, and yam extract have a significant synergistic effect in improving object recognition and exploration ability and delaying cognitive decline.

[0066] 1.3 Water Maze Experiment The Morris water maze was used to assess the spatial learning and memory abilities of mice, particularly those dependent on hippocampal function. The maze consisted of a circular pool (120 cm in diameter, 45 cm in height) and a hidden platform (10 cm in diameter, 20 cm in height), with the water temperature maintained at 20 ± 2 °C. White pigment was added to the water, and software was used to track and identify the mice. The Morris water maze consisted of four quadrants. The hidden platform was placed 1 cm below the water surface in the third quadrant as the mouse's escape target, and this quadrant was recorded as the target quadrant.

[0067] The first four days were the training phase. Mice freely explored the water maze with escape platforms for 90 seconds. Each time, mice were placed into the water pool facing the wall from different quadrants. The time it took for the mice to find the platform—the latency period (escape late negative control y)—was recorded using an image acquisition system. For mice that did not find the platform within 90 seconds, the latency period was recorded as 90 seconds, and the mice were guided to the platform and stayed there for 15 seconds. Day 5 was the localization and navigation experiment: mice freely explored the water maze with escape platforms for 60 seconds. The time it took for the mice to reach the platform was recorded using an image acquisition system, and the escape latency period was recorded. For mice that did not find the platform, the escape latency period was recorded as 60 seconds, and the mice were guided to the platform and allowed to stay there for 15 seconds. Day 6 was the spatial exploration experiment. The platforms were removed, and mice freely explored the water maze without escape platforms for 60 seconds. The swimming trajectory of the mice was tracked and timed using an image acquisition system. The number of times the mice crossed the original platform location within 60 seconds and the time spent in the target quadrant were recorded.

[0068] In the spatial exploration experiment on day 6 of the water maze, the number of times mice traversed the target area and the time spent in the target quadrant were as follows: Figure 10As shown in AB, the number of times the model group mice crossed the target area was 2.625 ± 0.52, significantly lower than that of the negative control group ( p <0.05; compared with the model group, the number of crossings in the yak MFGM group and the combined group was significantly increased ( p < 0.05), and there was no significant difference between the composition group and the negative control group and the positive control group ( p > 0.05). The time spent in the target quadrant by the model group mice was 23.99 ± 0.48 s, which was significantly shorter than that of the negative control group ( p < 0.05); Compared with the model group, the time spent in the target quadrant was significantly prolonged in both the yak MFGM group and the combined group of mice ( p The concentration of the compound was < 0.05, and there was no significant difference between the composition group and the negative control group, indicating that the composition can effectively restore the spatial exploration ability of mice.

[0069] 2. Effects of the composition on inflammation in mice with cognitive decline IL-6, a key pro-inflammatory cytokine, can exacerbate neuroinflammatory responses and impair synaptic plasticity and normal neuronal function through abnormally high expression. COX-2 mediates the synthesis of inflammatory mediators, and its sustained high expression can further amplify the neuroinflammatory cascade. Both IL-6 and COX-2 jointly participate in the occurrence and development of cognitive impairment. Figure 11 As shown in AB, the levels of IL-6 and COX-2 in the brain tissue of mice in the model group were significantly higher than those in the negative control group. Compared with the model group, the levels of IL-6 and COX-2 in the brains of mice in each intervention group were significantly decreased. p < 0.05). Compared with the yak MFGM group, taurine group, selenium-enriched yeast group, and yam extract group, the combined group showed a more significant decrease in IL-6 and COX-2 levels, suggesting that the anti-inflammatory effect of this combination is superior to that of each individual component at the same dosage. The above results confirm that yak MFGM, taurine, selenium-enriched yeast, and yam extract have a synergistic effect in improving the inflammatory state of the brain in mice with cognitive decline. The combination of these four components can effectively reduce the level of pro-inflammatory factors in the brain of mice with cognitive decline and enhance the body's anti-inflammatory capacity.

[0070] 3. Effects of the composition on oxidative damage in mice with cognitive decline Reduced glutathione (GSH) is a key non-enzymatic antioxidant in the body that can directly scavenge reactive oxygen species and maintain the body's redox homeostasis. A decrease in its content will exacerbate oxidative stress in the brain and aggravate neuronal damage. Malondialdehyde (MDA) is the end product of lipid peroxidation. An increase in its level can be used as a marker of aggravated oxidative damage. It will damage cell membrane integrity and synaptic function. Both GSH and MDA are involved in the occurrence and development of cognitive impairment.

[0071] like Figure 12As shown in AB, compared with the negative control group, GSH activity was significantly decreased and MDA content was significantly increased in the brain tissue of the model group mice. p < 0.05%. Compared with the model group, the yak MFGM group, taurine group, selenium-enriched yeast group, yam extract group, and combined group all significantly increased GSH activity in the brain and decreased MDA content ( p < 0.05). Compared with the single-component intervention groups, the combined group had higher GSH levels and lower MDA levels, indicating that the antioxidant effect of the combined group was significantly better than that of the single components at the same dosage. The above results indicate that yak MFGM, taurine, selenium-enriched yeast, and yam extract have a synergistic effect in reducing oxidative damage in mice with cognitive decline. The combination of these four ingredients can effectively enhance the body's antioxidant capacity, inhibit lipid peroxidation, and significantly improve the antioxidant level in the brain of mice with cognitive decline.

[0072] The embodiments described above are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

Claims

1. A composition, characterized in that, The composition comprises yak milk fat globule membrane, taurine, selenium-enriched yeast, and yam extract, wherein the mass ratio of yak milk fat globule membrane, taurine, selenium-enriched yeast, and yam extract is 15-35:2-8:3-10:0.5-2.

2. The composition according to claim 1, characterized in that, The mass ratio of the yak milk fat globule membrane, taurine, selenium-enriched yeast, and yam extract is 15–35: 3–5: 4–6: 0.5–1.

5.

3. A health food product, characterized in that, The health food product includes the composition according to claim 1 or 2.

4. A medicine, characterized in that, The medicine comprises the composition according to claim 1 or 2.

5. The use of the composition according to claim 1 or 2 in the preparation of products capable of improving cognitive function, characterized in that, The product in question is a pharmaceutical product.

6. The application according to claim 5, characterized in that, The application is in the preparation of products with acetylcholinesterase inhibitory function.

7. The application according to claim 5, characterized in that, The application includes at least one of the following: (1) Application in the preparation of products that can improve object recognition capabilities; (2) Application in the preparation of products that can enhance exploration capabilities; (3) Application in the preparation of products that can enhance the body's anti-inflammatory capabilities; (4) Application in the preparation of products that can improve the body’s antioxidant capacity and / or inhibit lipid peroxidation.

8. The application according to claim 7, characterized in that, Enhancing the body's anti-inflammatory capacity includes reducing the level of pro-inflammatory factors in the brain.

9. The application according to claim 7, characterized in that, Enhancing the body's antioxidant capacity and / or inhibiting lipid peroxidation includes improving the level of antioxidant factors in brain tissue.

10. The use of the composition according to claim 1 or 2 in the preparation of a product capable of assisting in improving memory, characterized in that, The product is either a health food or a medicine.

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