A composition capable of improving cognition and its application in the prevention and treatment of Alzheimer's disease

Abstract

The present invention discloses a composition capable of improving cognition and its application in the prevention and treatment of Alzheimer's disease. The present invention relates to the field of biomedicine, and provides a composition for improving cognition, which is composed of a neuron cell membrane protectant (omega-3 fatty acids and / or phospholipid compounds) and a nerve repair factor (D vitamins and / or B vitamins), or is composed of the neuron cell membrane protectant, the nerve repair factor and an enhanced protectant (taurine and / or carotenoids), or is composed of the nerve repair factor. The present invention has confirmed through in vivo and in vitro experiments that the composition provided by the present invention can have a significant therapeutic effect on the improvement of cognitive ability; at the same time, it also has a certain preventive and therapeutic effect on Alzheimer's disease.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a composition capable of improving cognition and application thereof in the prevention and treatment of Alzheimer's disease. Background Art

[0002] Alzheimer's disease (AD) and related dementias have had a huge impact on global public health and socioeconomics. According to the 2018 estimate of the Alzheimer's Disease International, the number of people with dementia worldwide is about 50 million, and it is expected to double to about 100 million by 2050. AD is the most common neurodegenerative disease in the elderly, characterized by cognitive decline, Aβ deposition and neurofibrillary tangles, and other pathological features include progressive neuronal and synaptic degeneration / loss, neuroinflammation and oxidative damage.

[0003] AD is a complex clinical syndrome characterized by progressive decline in multiple cognitive domains, including memory, language, executive function, visuospatial ability, personality, and behavior. This decline in cognitive function ultimately leads to the inability of patients to independently complete instrumental and / or basic activities of daily life.

[0004] AD is a progressive process. The pathological changes of AD may begin without obvious clinical symptoms, then progress to the stage of mild cognitive impairment, and eventually lead to obvious dementia symptoms.

[0005] Currently, there are limited drug treatment options for AD patients, which mainly include two categories: (1) Cholinesterase inhibitors (such as donepezil, rivastigmine and galantamine): suitable for patients with mild, moderate or severe AD and patients with Parkinson's disease-related dementia. (2) N-methyl-D-aspartate (NMDA) receptor antagonists (such as memantine): as non-competitive NMDA receptor antagonists and dopamine agonists, they are mainly used to treat moderate to severe AD patients, especially those who show difficulties with attention and vigilance. However, these drugs have limited efficacy and may cause a series of side effects, so new drugs for the prevention and treatment of AD are expected to emerge. Summary of the invention

[0006] The purpose of the present invention is to provide a composition capable of improving cognition and its application in the prevention and treatment of Alzheimer's disease.

[0007] In a first aspect, the present invention claims a composition for improving cognition.

[0008] The composition claimed in the present invention also has the following functions: treating and / or preventing the neurodegenerative disease Alzheimer's disease; improving the symptoms of neurodegenerative diseases; and reducing the toxicity of β-amyloid protein to nerve cells. The neurodegenerative disease is particularly a cognitive degenerative disease, such as Alzheimer's disease.

[0009] The composition is composition A or composition B or composition C;

[0010] The composition A is composed of a neuron cell membrane protective agent and a nerve repair factor;

[0011] The composition B is composed of the neuron cell membrane protective agent, the nerve repair factor and the enhanced protective agent;

[0012] The composition C consists of the nerve repair factor;

[0013] The neuron cell membrane protective agent is composed of all or part of the following: omega-3 fatty acids and phospholipid compounds;

[0014] The omega-3 fatty acid is docosahexaenoic acid (DHA) and / or eicosapentaenoic acid (EPA); the phospholipid compound is phosphatidylserine, phosphatidylcholine and / or phosphatidylethanolamine; and the composition can significantly improve the stability and integrity of neuronal cell membranes.

[0015] The nerve repair factor is composed of all or part of the following: D vitamins and B vitamins;

[0016] The D vitamins are vitamin D2 and / or vitamin D3; the B vitamins are selected from vitamin B12, vitamin B6, vitamin B1 and / or vitamin B9; and the composition can promote the growth and repair of neuronal cells.

[0017] The enhanced protective agent consists of all or part of the following: taurine and carotenoids; wherein the taurine can be selectively replaced partially or completely by a taurine derivative (such as -acetyltaurine); the carotenoids are astaxanthin, β-carotene and / or lutein; and the composition can enhance the antioxidant capacity and energy metabolism of neuronal cells.

[0018] Furthermore, the neuronal cell membrane protective agent may be any of the following:

[0019] (a1) is composed of the following mass parts of the omega-3 fatty acid and the phospholipid compound (corresponding to animal experiments):

[0020] The omega-3 fatty acid (such as DHA): 1-10 parts by mass (such as 3.3 parts by mass);

[0021] The phospholipid compound (such as phosphatidylserine): 1 part by mass;

[0022] (a2) is composed of the following molar proportions of the omega-3 fatty acid and the phospholipid compound (corresponding to cell experiments):

[0023] The omega-3 fatty acid (such as DHA): 0.5-2 molar parts (such as 1 molar part);

[0024] The phospholipid compound (such as phosphatidylserine): 1 molar part.

[0025] In one embodiment of the present invention, the neuron cell membrane protective agent is composed of DHA and phosphatidylserine in a mass ratio of 3.3:1 (corresponding to animal experiments). In another embodiment of the present invention, the neuron cell membrane protective agent is composed of DHA and phosphatidylserine in a molar ratio of 1:1 (corresponding to cell experiments).

[0026] Furthermore, the neural repair factor is any one of the following:

[0027] (b1) is composed of the following amounts by weight of vitamin D, vitamin B12, vitamin B6, vitamin B1 and vitamin B9:

[0028] Vitamin D (such as vitamin D3): 0.1 parts by mass;

[0029] Vitamin B12: 0.05-1 parts by mass (e.g. 0.1 parts by mass);

[0030] Vitamin B6: 5-50 parts by mass (e.g., 10 parts by mass);

[0031] Vitamin B1: 10-100 parts by mass (e.g., 20 parts by mass);

[0032] Vitamin B9: 0.4-4 parts by mass (e.g. 0.8 parts by mass);

[0033] (b2) is composed of the following molar proportions of vitamin D, vitamin B12, vitamin B6, vitamin B1 and vitamin B9:

[0034] Vitamin D (as vitamin D3): 1 mol;

[0035] Vitamin B12: 0.5-2 mol parts (e.g. 1 mol part);

[0036] Vitamin B6: 0.5-2 mol parts (e.g. 1 mol part);

[0037] Vitamin B1: 0.5-2 mol parts (e.g. 1 mol part);

[0038] Vitamin B9: 0.5-2 molar parts (e.g. 1 molar part).

[0039] In one embodiment of the present invention, the nerve repair factor is composed of vitamin D3, vitamin B12, vitamin B6, vitamin B1 and vitamin B9 in a mass ratio of 0.1:0.1:10:20:0.8 (corresponding to animal experiments). In another embodiment of the present invention, the nerve repair factor is composed of vitamin D3, vitamin B12, vitamin B6, vitamin B1 and vitamin B9 in a molar ratio of 1:1:1:1:1 (corresponding to cell experiments).

[0040] Furthermore, the reinforcing protective agent is any one of the following:

[0041] (c1) is composed of the following parts by weight of taurine and the carotenoid:

[0042] Taurine: 1-10 parts by mass (e.g., 5 parts by mass);

[0043] The carotenoid (such as astaxanthin): 1 part by mass;

[0044] (c2) is composed of the following molar proportions of taurine and the carotenoid:

[0045] Taurine: 0.5-2 mol parts (e.g. 1 mol part);

[0046] The carotenoid (such as astaxanthin): 1 mol part;

[0047] Wherein, the taurine can be selectively replaced partially or completely by a taurine derivative (such as -acetyltaurine).

[0048] In one embodiment of the present invention, the enhanced protective agent is composed of taurine and astaxanthin in a mass ratio of 5:1 (corresponding to animal experiments). In another embodiment of the present invention, the enhanced protective agent is composed of taurine and astaxanthin in a molar ratio of 1:1 (corresponding to cell experiments).

[0049] Furthermore, the composition is any of the following:

[0050] (d1) is composed of the following parts by weight of the omega-3 fatty acids, the phospholipid compounds, vitamin D, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine and the carotenoids:

[0051] The omega-3 fatty acid (such as DHA): 8000-24000 parts by mass (such as 16000 parts by mass);

[0052] The phospholipid compound (such as phosphatidylserine): 2400-7200 parts by mass (such as 4800 parts by mass);

[0053] Vitamin D (such as vitamin D3): 1 part by mass;

[0054] Vitamin B12: 0.5-2 parts by mass (e.g. 1 part by mass);

[0055] Vitamin B6: 50-150 parts by mass (e.g. 100 parts by mass);

[0056] Vitamin B1: 100-300 parts by mass (e.g. 200 parts by mass);

[0057] Vitamin B9: 4-12 parts by mass (e.g. 8 parts by mass);

[0058] Taurine: 2500-7500 parts by mass (e.g., 5000 parts by mass);

[0059] The carotenoid (such as astaxanthin): 500-1500 parts by mass (such as 1000 parts by mass);

[0060] (d2) is composed of the following parts by weight of the omega-3 fatty acids, the phospholipid compounds, vitamin D, vitamin B12, vitamin B6, vitamin B1 and vitamin B9:

[0061] The omega-3 fatty acid (such as DHA): 8000-24000 parts by mass (such as 16000 parts by mass);

[0062] The phospholipid compound (such as phosphatidylserine): 2400-7200 parts by mass (such as 4800 parts by mass);

[0063] Vitamin D (such as vitamin D3): 1 part by mass;

[0064] Vitamin B12: 0.5-2 parts by mass (e.g. 1 part by mass);

[0065] Vitamin B6: 50-150 parts by mass (e.g. 100 parts by mass);

[0066] Vitamin B1: 100-300 parts by mass (e.g. 200 parts by mass);

[0067] Vitamin B9: 4-12 parts by mass (e.g. 8 parts by mass);

[0068] (d3) is composed of the following amounts by weight of vitamin D, vitamin B12, vitamin B6, vitamin B1 and vitamin B9:

[0069] Vitamin D (such as vitamin D3): 1 part by mass;

[0070] Vitamin B12: 0.5-2 parts by mass (e.g. 1 part by mass);

[0071] Vitamin B6: 50-150 parts by mass (e.g. 100 parts by mass);

[0072] Vitamin B1: 100-300 parts by mass (e.g. 200 parts by mass);

[0073] Vitamin B9: 4-12 parts by mass (e.g. 8 parts by mass);

[0074] (d4) is composed of the following molar proportions of the omega-3 fatty acids, the phospholipid compounds, vitamin D, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine and the carotenoids:

[0075] The omega-3 fatty acid (such as DHA): 0.5-2 molar parts (such as 1 molar part);

[0076] The phospholipid compound (such as phosphatidylserine): 0.5-2 molar parts (such as 1 molar part);

[0077] Vitamin D (as vitamin D3): 1 mol;

[0078] Vitamin B12: 0.5-2 mol parts (e.g. 1 mol part);

[0079] Vitamin B6: 0.5-2 mol parts (e.g. 1 mol part);

[0080] Vitamin B1: 0.5-2 mol parts (e.g. 1 mol part);

[0081] Vitamin B9: 0.5-2 mol parts (e.g. 1 mol part);

[0082] Taurine: 0.5-2 mol parts (e.g. 1 mol part);

[0083] The carotenoid (such as astaxanthin): 0.5-2 molar parts (such as 1 molar part);

[0084] (d5) is composed of the following molar proportions of the omega-3 fatty acids, the phospholipid compounds, vitamin D, vitamin B12, vitamin B6, vitamin B1 and vitamin B9:

[0085] The omega-3 fatty acid (such as DHA): 0.5-2 molar parts (such as 1 molar part);

[0086] The phospholipid compound (such as phosphatidylserine): 0.5-2 molar parts (such as 1 molar part);

[0087] Vitamin D (as vitamin D3): 1 mol;

[0088] Vitamin B12: 0.5-2 mol parts (e.g. 1 mol part);

[0089] Vitamin B6: 0.5-2 mol parts (e.g. 1 mol part);

[0090] Vitamin B1: 0.5-2 mol parts (e.g. 1 mol part);

[0091] Vitamin B9: 0.5-2 mol parts (e.g. 1 mol part);

[0092] (D6) is composed of the following molar proportions of vitamin D, vitamin B12, vitamin B6, vitamin B1 and vitamin B9:

[0093] Vitamin D (as vitamin D3): 1 mol;

[0094] Vitamin B12: 0.5-2 mol parts (e.g. 1 mol part);

[0095] Vitamin B6: 0.5-2 mol parts (e.g. 1 mol part);

[0096] Vitamin B1: 0.5-2 mol parts (e.g. 1 mol part);

[0097] Vitamin B9: 0.5-2 mol parts (e.g. 1 mol part);

[0098] In (d1) and (d4), the taurine may be selectively replaced partially or completely by a taurine derivative (such as acetyltaurine).

[0099] In one embodiment of the present invention, the composition is composed of DHA, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine and astaxanthin in a mass ratio of 16000:4800:1:1:100:200:8:5000:1000 (corresponding to animal experiments Vit+DP+TA, i.e., a combination of a nerve repair factor, a neuron cell membrane protective agent and an enhanced protective agent). In another embodiment of the present invention, the composition is composed of DHA, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1 and vitamin B9 in a mass ratio of 16000:4800:1:1:100:200:8 (corresponding to animal experiments Vit+DP, i.e., a combination of a nerve repair factor and a neuron cell membrane protective agent). In another embodiment of the present invention, the composition is composed of vitamin D3, vitamin B12, vitamin B6, vitamin B1 and vitamin B9 in a mass ratio of 1:1:100:200:8 (corresponding to animal experiment Vit, i.e., nerve repair factor group). In another embodiment of the present invention, the composition is composed of DHA, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine and astaxanthin in a molar ratio of 1:1:1:1:1:1:1:1:1 (corresponding to cell experiment Vit+DP+TA). In another embodiment of the present invention, the composition is composed of DHA, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1 and vitamin B9 in a molar ratio of 1:1:1:1:1:1:1 (corresponding to cell experiment Vit+DP). In another embodiment of the present invention, the composition consists of vitamin D3, vitamin B12, vitamin B6, vitamin B1 and vitamin B9 in a molar ratio of 1:1:1:1:1 (corresponding to cell experiment Vit).

[0100] In a second aspect, the present invention claims the use of the composition described in the first aspect above in improving cognition or preparing a product for improving cognition.

[0101] Among them, the improved cognition may include spatial learning and memory ability, new object recognition ability, etc.

[0102] In a third aspect, the present invention claims the use of the composition described in the first aspect above in any of the following:

[0103] (A1) Preparation of products for the treatment and / or prevention of neurodegenerative diseases; or, treatment and / or prevention of neurodegenerative diseases;

[0104] (A2) Preparing a product for improving symptoms of a neurodegenerative disease; or, improving symptoms of a neurodegenerative disease.

[0105] Wherein, the neurodegenerative disease may be a cognitive degenerative disease, such as Alzheimer's disease.

[0106] In a fourth aspect, the present invention claims protection for the use of the composition described in the first aspect above in reducing the toxicity of neurotoxic proteins to nerve cells or in preparing a product capable of reducing the toxicity of neurotoxic proteins to nerve cells.

[0107] Wherein, the neurotoxic protein includes but is not limited to β-amyloid protein, tau protein, TDP-43 protein and α-synuclein, etc. In one embodiment of the present invention, the neurotoxic protein is β-amyloid protein.

[0108] The neural cells include, but are not limited to, primary neural cells, SH-SY5Y cells, and neural cells induced by iPSC pluripotent stem cells, etc. In one embodiment of the present invention, the neural cells are SH-SY5Y cells.

[0109] In a fifth aspect, the present invention claims protection for a product having any of the following functions:

[0110] (B1) Improve cognition;

[0111] (B2) treatment and / or prevention of neurodegenerative diseases;

[0112] (B3) Improve symptoms of neurodegenerative diseases;

[0113] (B4) Reduce the toxic effects of neurotoxic proteins on nerve cells.

[0114] In (B2) and (B3), the neurodegenerative disease may be a cognitive degenerative disease, such as Alzheimer's disease.

[0115] In (B4), the neurotoxic proteins include but are not limited to β-amyloid protein, tau protein, TDP-43 protein and α-synuclein, etc. The neural cells include but are not limited to primary neuronal cells, SH-SY5Y cells and neuronal cells induced by iPSC pluripotent stem cells, etc.

[0116] The product claimed in the present invention comprises the composition described in the first aspect above.

[0117] In the above related aspects, the product may be a pharmaceutical product.

[0118] In a sixth aspect, the present invention further claims a method for improving cognition, comprising the following steps: administering an effective dose of the composition described in the first aspect above to a subject.

[0119] In a seventh aspect, the present invention further claims a method for treating and / or preventing neurodegenerative diseases, comprising the following steps: administering an effective dose of the composition described in the first aspect above to a subject.

[0120] In an eighth aspect, the present invention further claims a method for improving symptoms of a neurodegenerative disease, comprising the following steps: administering an effective dose of the composition described in the first aspect above to a subject.

[0121] In a ninth aspect, the present invention further claims a method for reducing the toxicity of neurotoxic proteins to nerve cells, comprising the following steps: treating nerve cells damaged or about to be damaged by neurotoxic proteins with an effective dose of the composition described in the first aspect above.

[0122] In the above-mentioned related aspects, the subject is a patient with a neurodegenerative disease. The neurodegenerative disease may be a cognitive degenerative disease, such as Alzheimer's disease. The neurotoxic protein includes but is not limited to β-amyloid protein, tau protein and α-synuclein, etc. The neural cells include but are not limited to primary neuronal cells, SH-SY5Y cells and neuronal cells induced by iPSC pluripotent stem cells, etc.

[0123] In the above-mentioned related aspects, the improvement of Alzheimer's disease symptoms can be specifically reflected in behavioral improvement and / or pathological improvement. Among them, the behavioral improvement can be improved cognition. The pathological improvement can be as follows: reducing Aβ deposition and microglial activation in the dentate gyrus (DG) region of the hippocampus, reducing APP and phosphorylated APP deposition in the DG region, reducing the levels of APP and Tau proteins, and restoring the secretion of Reelin in the dentate gyrus of the hippocampus.

[0124] The present invention has confirmed through in vitro and in vivo experiments that the composition provided by the present invention can significantly improve the pathological conditions of the Alzheimer's disease model. In in vitro experiments, the composition of the present invention can reduce the toxic effects of Aβ in SH-SY5Y cells. In in vivo experiments, the cognitive ability and pathological sections of 5xFAD mice showed that the composition of the present invention can significantly improve the cognitive ability of 5xFAD mice. Overall, the composition of the present invention can have a significant therapeutic effect on the improvement of cognitive ability; at the same time, it also has a certain preventive and therapeutic effect on Alzheimer's disease.

[0125] In addition, the composition of the composition provided by the present invention has a milder mechanism of action, fewer side effects, and is suitable for long-term use. It may be beneficial to overall health, not only for AD symptoms, but also for improving systemic organ dysfunction caused by aging. It can reduce the risk of combined medication, especially for elderly patients who need to take multiple medications. BRIEF DESCRIPTION OF THE DRAWINGS

[0126] Figure 1 The combination of neural repair factor and neuronal cell membrane protector (Vit+DP) can reduce the toxic effect of Aβ in SH-SY5Y cells.

[0127] Figure 2 The combination of taurine and astaxanthin as a neuroprotectant (TA) can alleviate the toxic effects of Aβ in SH-SY5Y cells.

[0128] Figure 3 The combination of neural repair factors, neuronal cell membrane protectants and enhanced protectants (Vit+DP+TA) can alleviate the toxic effects of Aβ in SH-SY5Y cells.

[0129] Figure 4 The Y-maze test was used to evaluate the effects of Vit+DP+TA treatment on spatial memory and cognitive function. A is a representative heat map image of the Y-maze test; B is the number of entries into the novel arm; C is the spontaneous alternation rate.

[0130] Figure 5 The effect of Vit+DP+TA treatment on cognitive function was evaluated for the novel object recognition test. A is a representative heat map image of the novel object recognition (NOR) test. Red circles represent novel objects, while black circles represent familiar objects; B is the percentage of time spent exploring novel objects; data are expressed as mean ± standard error (SEM).

[0131] Figure 6 Morris water maze test was used to evaluate the effect of Vit+DP+TA treatment on cognitive function. A is the representative swimming trajectory of mice in the learning stage or memory stage; B is the escape latency and swimming path in the learning stage; C is the escape latency in the memory stage; and D is the number of times the platform was crossed in the memory stage.

[0132] Figure 7 Treatment with neurorepair factor (Vit) reduces Aβ deposition and microglial activation in the dentate gyrus of the hippocampus.

[0133] Figure 8 Treatment with neurorepair factor (Vit) reduced the deposition of APP and phosphorylated APP in the dentate gyrus of the hippocampus.

[0134] Fig. 9 Treatment with the neurorepair factor (Vit) reduced the levels of pathological phosphorylated APP and Tau proteins in 5xFAD mice.

[0135] Fig.10 Treatment with neurorepair factor (Vit) increases the expression of brain protective reelin. DETAILED DESCRIPTION

[0136] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0137] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0138] The composition provided by the present invention includes all or part of the following components: DHA, taurine, phosphatidylserine, astaxanthin, vitamin B12, vitamin D3, vitamin B1, vitamin B6, and vitamin B9. Among them, DHA and phosphatidylserine are neuron cell membrane protectors (DP for short); vitamin D3, vitamin B12, vitamin B6, vitamin B1, and vitamin B9 are nerve repair factors (Vit for short); taurine and astaxanthin are enhanced protectors (TA for short).

[0139] The specific classification and functions of these compositions are shown in Table 1:

[0140]

[0141] The following examples will test the effectiveness of the composition provided by the present invention in improving cognition and preventing and treating Alzheimer's disease (AD) from two dimensions: in vitro experiments (cell experiments) and in vivo experiments (mouse experiments).

[0142] Example 1: In vitro effectiveness evaluation of the composition of the present invention

[0143] The cell line used in the in vitro cell experiment of the present invention is specifically the SH-SY5Y cell line. As the preferred cell model for AD research, SH-SY5Y cells have neuronal characteristics and can differentiate, providing an ideal platform for AD research. Its immortalized properties ensure experimental stability and repeatability. SH-SY5Y cells are highly sensitive to Aβ toxicity and are a powerful tool for studying the key pathological mechanisms of AD. As a human cell, it is closer to human neurons than other animal cells and may more accurately reflect certain pathological processes of AD. The SH-SY5Y cell line used in this embodiment is a product of BeNa Culture Collection (Shanghai, China).

[0144] 1. Combination of nerve repair factor and neuron cell membrane protector (Vit+DP)

[0145] SH-SY5Y cells were seeded in 96-well plates containing complete medium and cultured at 37°C for 24 hours. They were then treated in the following groups for 24 hours each:

[0146] Blank control group: SH-SY5Y cells only;

[0147] Positive control group (Aβ treatment): SH-SY5Y cells + 1 μM Aβ-42 (Sigma-Aldrich # A9810, the same below);

[0148] Neural repair factor group (Aβ+Vit treatment): SH-SY5Y cells + 1μM Aβ-42 + 10μM neural repair factor;

[0149] Neuronal cell membrane protective agent group (Aβ+DP treatment): SH-SY5Y cells + 1μM Aβ-42 + 10μM neuronal cell membrane protective agent;

[0150] Neural repair factor + neuronal cell membrane protective agent group (Aβ+Vit+DP treatment): SH-SY5Y cells + 1μM Aβ-42 + 10μM neural repair factor + 10μM neuronal cell membrane protective agent.

[0151] Note: In each group, "1μM Aβ-42" means that the final concentration of Aβ-42 in the SH-SY5Y cell culture system is 1μM; "10μM neural repair factor" means that the final concentrations of vitamin D3, vitamin B12, vitamin B6, vitamin B1 and vitamin B9 in the SH-SY5Y cell culture system are all 10μM; "10μM neuronal cell membrane protective agent" means that the final concentrations of DHA and phosphatidylserine in the SH-SY5Y cell culture system are all 10μM.

[0152] After treatment, SH-SY5Y cells in each group were incubated with MTT (final concentration of 0.5 mg / mL in the incubation system) for 4 h, and then incubated with 10% SDS solution at 37 °C for another 15 min. The absorbance was measured at 560 nm using a microplate reader (Spectra Max M2, Molecular Devices, San Jose, CA, USA). This was done using the MTT Cell Proliferation Assay Kit (MTT Cell Proliferation and Cytotoxicity Assay Kit, Wobo Bio).

[0153] Calculate the relative cell viability: cell survival rate (%) = (OD560 value of treatment group - OD560 value of blank control) / (OD560 value of control group - OD560 value of blank control) × 100%;

[0154] In the formula, the "treatment group OD560 value" is the 560nm absorbance of the cell sample after experimental treatment; the "blank control OD560 value" is the 560nm absorbance of the well containing only culture medium and reagents but no cells; and the "control group OD560 value" is the 560nm absorbance of untreated normal cells.

[0155] Each group was repeated 5 times, and the results were given as mean ± standard deviation.

[0156] The results are as follows Figure 1 As shown in the figure. It can be seen from the figure that there are differences in cell survival rates under different treatment conditions. The cell survival rate of the blank control group (SH-SY5Y cells only) is the highest, close to 100%; the cell survival rate of the positive control group (Aβ treatment) is significantly reduced to about 60%, which is extremely significant compared with the blank control group (****p<0.0001); the cell survival rates of the two groups of the neural repair factor group (Aβ+Vit treatment) and the neuronal cell membrane protective agent group (Aβ+DP) are slightly increased, but the difference is not significant compared with the Aβ treatment group; the neural repair factor + neuronal cell membrane protective agent group (Aβ+Vit+DP treatment) shows the best therapeutic effect. The cell survival rate of this group is significantly higher than that of the Aβ treatment group alone, close to 80%, and it is the only drug treatment group that reaches statistical significance compared with the Aβ treatment group (****p<0.0001). This result shows that the combined use of neural repair factor (Vit) and neuronal cell membrane protective agent (DP) has a synergistic effect in protecting cells from the toxic effects of Aβ.

[0157] 2. Enhanced Protective Agent Combination (TA)

[0158] SH-SY5Y cells were seeded in 96-well plates containing complete medium and cultured at 37°C for 24 hours. They were then treated in the following groups for 24 hours each:

[0159] Blank control group: SH-SY5Y cells only;

[0160] Positive control group (Aβ-treated): SH-SY5Y cells + 1 μM Aβ-42;

[0161] Taur group (Aβ+Taur treatment): SH-SY5Y cells + 1μM Aβ-42 + 10μM taurine;

[0162] Astaxanthin (Asta) group (Aβ+Asta treatment): SH-SY5Y cells + 1μM Aβ-42 + 10μM astaxanthin;

[0163] Taur + astaxanthin group (Aβ + Taur + Asta treatment): SH-SY5Y cells + 1μM Aβ-42 + 10μM taur + 10μM astaxanthin;

[0164] Note: In each group, "1μM Aβ-42" means that the final concentration of Aβ-42 in the SH-SY5Y cell culture system is 1μM; "10μM taurine" means that the final concentration of taurine in the SH-SY5Y cell culture system is 10μM; "10μM astaxanthin" means that the final concentration of astaxanthin in the SH-SY5Y cell culture system is 10μM.

[0165] After treatment, SH-SY5Y cells in each group were incubated with MTT (final concentration of 0.5 mg / mL in the incubation system) for 4 h, and then incubated with 10% SDS solution at 37 °C for another 15 min. The absorbance was measured at 560 nm using a microplate reader (Spectra Max M2, Molecular Devices, San Jose, CA, USA), and the cell viability was calculated. For details, see the previous text. Each group was set up with 5 replicates, and the results are given as mean ± standard deviation.

[0166] The results are as follows Figure 2 As shown. As can be seen from the figure, there are differences in cell survival rates under different treatment conditions. The cell survival rate of the blank control group (SH-SY5Y cells only) is the highest, close to 100%; the cell survival rate of the positive control group (Aβ treatment) is the lowest, about 60%, indicating that Aβ has a significant toxic effect on cells (****p<0.0001); the cell survival rate of the taurine group (Aβ+Taur treatment) is slightly improved, but the effect is not obvious; the cell survival rate of the astaxanthin group (Aβ+Asta treatment) is higher than that of the Aβ treatment alone, reaching about 70%; the cell survival rate of the taurine+astaxanthin group (Aβ+Taur+Asta treatment) is further improved, reaching about 75-80%, and is significantly different from the Aβ treatment group (****p<0.0001). This shows that the combination of the two types of substances is more effective than using them alone.

[0167] Astaxanthin or taurine alone has a certain effect on counteracting Aβ toxicity; the combination of the two substances is more effective than either alone.

[0168] 3. Combination of nerve repair factor, neuron cell membrane protectant and enhanced protectant (Vit+DP+TA)

[0169] SH-SY5Y cells were seeded in 96-well plates containing complete medium and cultured at 37°C for 24 hours. They were then treated in the following groups for 24 hours each.

[0170] Blank control group: SH-SY5Y cells only;

[0171] Positive control group (Aβ-treated): SH-SY5Y cells + 1 μM Aβ-42;

[0172] Neural repair factor + neuronal cell membrane protective agent group (Aβ + Vit + DP treatment): SH-SY5Y cells + 1μM Aβ-42 + 10μM neural repair factor + 10μM neuronal cell membrane protective agent;

[0173] Enhanced protectant group (Aβ+TA treatment): SH-SY5Y cells + 1μM Aβ-42 + 10μM taurine + 10μM astaxanthin;

[0174] Neural repair factor + neuronal cell membrane protective agent + enhanced protective agent group (Aβ+Vit+DP+TA treatment): SH-SY5Y cells + 1μM Aβ-42 + 10μM neural repair factor + 10μM neuronal cell membrane protective agent + 10μM taurine + 10μM astaxanthin.

[0175] Note: In each group, "1μM Aβ-42" means that the final concentration of Aβ-42 in the SH-SY5Y cell culture system is 1μM; "10μM neural repair factor" means that the final concentrations of vitamin D3, vitamin B12, vitamin B6, vitamin B1 and vitamin B9 in the SH-SY5Y cell culture system are all 10μM; "10μM neuronal cell membrane protective agent" means that the final concentrations of DHA and phosphatidylserine in the SH-SY5Y cell culture system are both 10μM; "10μM taurine" means that the final concentration of taurine in the SH-SY5Y cell culture system is 10μM; "10μM astaxanthin" means that the final concentration of astaxanthin in the SH-SY5Y cell culture system is 10μM.

[0176] After treatment, SH-SY5Y cells in each group were incubated with MTT (final concentration of 0.5 mg / mL in the incubation system) for 4 h, and then incubated with 10% SDS solution at 37 °C for another 15 min. The absorbance was measured at 560 nm using a microplate reader (Spectra Max M2, Molecular Devices, San Jose, CA, USA), and the cell viability was calculated. For details, see the previous text. Each group was set up with 5 replicates, and the results are given as mean ± standard deviation.

[0177] The results are as follows Figure 3As shown in the figure, it can be seen that there are differences in cell survival rates under different treatment conditions: the blank control group (SH-SY5Y cells only) has the highest cell survival rate, close to 100%; the cell survival rate of the positive control group (Aβ treatment) is significantly reduced to about 60%, which is extremely significantly different from the blank control group (****p<0.0001); the cell survival rates of the neural repair factor + neuronal cell membrane protective agent group (Aβ+Vit+DP treatment) and the enhanced protective agent group (Aβ+TA treatment) are improved; the neural repair factor + neuronal cell membrane protective agent + enhanced protective agent group (Aβ+Vit+DP+TA treatment) shows the best therapeutic effect, and the cell survival rate of this group is significantly higher than that of the Aβ treatment group alone, close to 90%, and the difference is the smallest compared with the blank control group. This group shows an extremely significant difference compared with the Aβ treatment group (****p<0.0001).

[0178] It is particularly noteworthy that the mean cell survival rate of the Aβ-treated group was 62.6%, the mean cell survival rate of the Aβ+Vit+DP-treated group was 71.0%, the mean cell survival rate of the Aβ+TA-treated group was 77.8%, and the mean cell survival rate of the Aβ+Vit+DP+TA-treated group was 89.2%. According to calculations, compared with the Aβ-treated group, the cell survival rate of the Aβ+Vit+DP-treated group increased by 8.4%, the cell survival rate of the Aβ+TA-treated group increased by 15.2%, and the cell survival rate of the Aβ+Vit+DP+TA-treated group increased by as much as 26.6%, which exceeded the sum of the increases of the Aβ+Vit+DP-treated group and the Aβ+TA-treated group. This confirms that the combined use of TA and Vit+DP can produce significant synergistic effects, providing new ideas for the treatment strategy of neurodegenerative diseases.

[0179] Example 2: In vivo effectiveness evaluation of the composition of the present invention

[0180] The in vivo mouse experiments of the present invention used 5xFAD disease model mice (Cat# 034848-JAX, RRID:MMRRC_034848-JAX). This model mouse carries five familial Alzheimer's disease (AD) mutations. 5xFAD mice carry the K670N / M67L (Sweden), I716V (Florida) and V717I (London) mutations of human APP (695) in the brain, as well as the M146L and L286V mutations of the human PSEN1 gene. 5xFAD mice are a transgenic model that rapidly develops pathological features of Alzheimer's disease. These mice begin to develop amyloid plaques at 2 months of age and show significant neuronal loss at 4-5 months of age. Their cognitive behavioral abnormalities usually begin to gradually appear at 2-3 months of age, become obvious at 4-5 months of age, and continue to worsen thereafter. The 5xFAD model is widely used to study the pathological mechanisms of AD, test potential therapeutic strategies, and investigate neuroinflammatory and neurodegenerative processes.

[0181] 1. Behavioral Experiment

[0182] A total of 14 5xFAD mice and 7 wild-type littermates (C57BL / 6 mice, all female) were used in this animal experiment. All mice were 5 months old. The 5xFAD mice were randomly divided into positive administration (N=7), positive control group (N=7) and wild-type mouse group (N=7). The positive administration group was treated with nerve repair factor + neuronal cell membrane protectant + enhanced protectant composition (Vit+DP+TA) by gavage every day (see Table 1 for the specific amount of each substance in the composition); the positive control group was treated with pure solvent (1% Tween 80 + 1% methylcellulose + 98% water, mass ratio) by gavage; the duration of administration was 3 months. Then animal behavior experiments were conducted.

[0183] For animal behavior experiments, Y-maze-novel arm, Y-maze-spontaneous alternation, novel object recognition, and Morris water maze were conducted to verify changes in their cognitive abilities.

[0184] 1. Y-maze-novel arm and spontaneous alternation

[0185] The Y-maze experiment was used to evaluate the short-term spatial cognition and memory ability of mice. The Y-maze apparatus consisted of three arms of equal length (30 cm long, 8 cm wide, and 15 cm high) that were 120° apart.

[0186] Before the novel arm exploration experiment of the Y-maze experiment, adjust the light intensity of the experimental field to about 100 Lux, and move the mice into the experimental field to adapt for at least 30 minutes. Use a baffle to block one arm of the Y-maze as the novel arm (Novel), then place the mouse with its back to the maze in the starting arm (Start), let it explore freely for 5 minutes, and return it to the breeding cage after the training. After an interval of 15 minutes, remove the baffle to open the novel arm, then place the mouse with its back to the maze in the starting arm (Start), let it explore freely for 5 minutes. When the mouse's limbs completely enter a certain arm, it is judged that the mouse has entered the arm, and manually record the number of times the mouse enters the novel arm and the total number of times it enters all arms. After the experiment, clean the open field with 75% ethanol to remove the smell left by the mouse.

[0187] Before the spontaneous alternation experiment of the Y-maze test, adjust the light intensity of the experimental field to about 100 Lux, and move the mice into the experimental field to adapt for at least 30 minutes. During the experiment, place the mice in one arm with their backs facing the maze and let them explore freely for 8 minutes. The mice are judged to have entered an arm only when their four limbs have completely entered an arm, and the order in which the mice enter the arm is recorded. After the experiment, clean the open field with 75% ethanol to remove the smell left by the mice.

[0188] The results are as follows Figure 4 As shown, it includes three parts: heat map, number of entries into the novel arm, and spontaneous alternation rate. The heat map shows that the wild-type mouse group has uniform activity in the three arms, the positive control group (pure solvent) has reduced activity in the novel arm, and the positive drug group (Vit+DP+TA treatment) has recovered activity in the novel arm close to that of the wild-type mouse group. In terms of the number of entries into the novel arm and the spontaneous alternation rate, the positive control group (pure solvent) was significantly lower than the wild-type mouse group, showing a decline in cognitive function; while the Vit+DP+TA treatment group showed significant improvement. These results clearly show that Vit+DP+TA treatment has a significant effect and can effectively improve the spatial memory and working memory of mice.

[0189] 2. New object recognition

[0190] The new object recognition experiment was conducted in an open field box of 40cm×40cm×40cm to evaluate the exploration and cognitive ability of mice. The experiment was conducted over three days: the mice were allowed to move freely for 5 minutes during the first day of the adaptation period; the mice were allowed to explore two identical objects for 5 minutes during the second day of the training period; the mice were allowed to explore two identical objects for 5 minutes during the third day of the test period, and one object was replaced with a new object after 15 minutes and then explored for another 5 minutes. The exploration time was analyzed using Tracking Master V3.1.62 software, and the cognitive index was calculated as new object exploration time / (new object exploration time + old object exploration time)×100%. The experimental environment illumination was maintained at about 100Lux, and the open field was cleaned with 75% ethanol after each experiment.

[0191] The results are as follows Figure 5 The results of the novel object recognition (NOR) test are shown in Figure 2, including a heat map and a recognition index. From the heat map, the wild-type mice group and the positive drug administration group (Vit+DP+TA treatment) were more active around the novel object, while the positive control group (pure solvent) paid less attention to the novel object. The recognition index graph showed that the recognition ability of the mice in the positive control group (pure solvent) was significantly lower than that of the wild-type mice group, while the mice in the Vit+DP+TA treatment group recovered a considerable degree of recognition ability.

[0192] These results clearly show that Vit+DP+TA treatment has a significant effect and can effectively improve the object recognition ability and cognitive function of mice.

[0193] 3. Morris water maze

[0194] The Morris water maze experiment evaluates the learning, memory and spatial cognitive abilities of mice. A circular pool with a diameter of 1.2m and a height of 0.5m is used, divided into four quadrants, one of which is a hidden platform with a diameter of 5cm. The experiment is divided into two stages: learning period (platform hiding period) and memory period (no platform period). Four experiments were conducted every day during the learning period, each for 60 seconds, and the time and path of the mice to find the hidden platform were recorded; the memory period was conducted 24 hours after the end of the learning period, the platform was removed, and the mice were allowed to swim freely for 60 seconds, and the swimming path, the number of times the original platform was crossed, and the time spent in the original platform quadrant was recorded. The light in the experimental environment was maintained at about 100Lux, and food-grade titanium dioxide was added to the pool to make the water milky white. The mice adapted to the experimental environment for at least 30 minutes. These data were used to analyze the spatial learning and memory abilities of mice.

[0195] The results are as follows Figure 6 As shown in the figure, compared with the positive control group (pure solvent) mice, in the platform hiding period, the time (escape latency) and moving distance of the positive drug administration group (Vit+DP+TA treatment) mice to find the hidden platform significantly decreased as the number of learning days increased, indicating that Vit+DP+TA treatment can significantly improve the learning and memory function of mice. In the non-platform period, the escape latency of mice in the Vit+DP+TA treatment group was shorter than that of mice in the positive control group (pure solvent), and their retention time in the platform quadrant increased, and the number of times they crossed the platform position increased significantly, indicating that the memory function of mice was significantly improved.

[0196] 2. Pathological testing

[0197] This animal experiment used a total of 6 5xFAD mice and 3 wild-type littermates (C57BL / 6 mice, all female), all 5 months old: 5xFAD mice were randomly divided into positive treatment (N=3), positive control group (N=3) and wild-type mouse group (N=3). The positive administration group was treated with nerve repair factor (Vit) by gavage every day (see Table 1 for the specific amount of each substance in the composition); the positive control group was treated with pure solvent (1% Tween 80 + 1% methylcellulose + 98% water, mass ratio) by gavage control; the administration period was 3 months. Then animal pathology experiments were carried out.

[0198] 1. Neurorepair factor treatment reduces Aβ deposition and microglial activation in the dentate gyrus (DG) region of the hippocampus

[0199] The experiment used 3 μm thick paraffin sections of mouse spinal cord and analyzed by immunofluorescence staining. The experimental groups included wild-type control, positive control group and positive drug administration group, with 3 mice in each group. The tissue was fixed with 4% paraformaldehyde for 24 hours, dehydrated, transparent, and embedded in wax. The sections were dewaxed with xylene, rehydrated with gradient ethanol, and antigen repaired with 0.01M citric acid buffer (pH 6.0) (95°C, 20 minutes).

[0200] Immunofluorescence staining used Iba1 primary antibody and Aβ (6E10) antibody, and incubated at 4°C overnight. The next day, Alexa Fluor488-labeled secondary antibody was used for incubation at room temperature for 2 hours, and DAPI (1:1000) was used to stain the nucleus for 10 minutes. Images were collected using a Zeiss LSM 880 confocal microscope with a 20× objective lens. ImageJ software was used to calculate the cell area of ​​each sample.

[0201] like Figure 7As shown, the Aβ deposition and microglial activation in the DG region of wild-type mice (WT), positive control group (pure solvent) mice, and positive drug administration group (i.e., 5xFAD mice treated with neurorepair factors) were compared. On the left are Aβ (6E10 antibody labeled, green), microglial marker Iba1 (red), DAPI staining (blue), and the merged images of the three. It can be seen that in WT mice, there are almost no Aβ plaques and microglial activation; in the positive control group (pure solvent) mice, obvious Aβ plaques (green) and microglial activation (red) were observed; after treatment with neurorepair factors, Aβ deposition and microglial activation in the positive drug administration group mice were significantly reduced. The bar graph on the right quantifies the Aβ plaque area and Iba1-positive microglial area in the DG region, respectively. Compared with the WT group, the Aβ plaque area of ​​the positive control group (pure solvent) mice increased significantly (****p<0.0001), and the Iba1-positive area increased significantly (****p<0.0001). After treatment with neurorepair factors, the Aβ plaque area and Iba1-positive area of ​​mice in the positive administration group were significantly reduced (*p<0.05 or **p<0.01). Scale bar: 50μm.

[0202] 2. Neurorepair factor treatment reduces APP and phosphorylated APP deposition in the dentate gyrus (DG) region of the hippocampus

[0203] The experiment used 3 μm thick paraffin sections of mouse spinal cord and analyzed by immunofluorescence staining. The experimental groups included wild-type control, positive control group and positive drug administration group, with 3 mice in each group. The tissue was fixed with 4% paraformaldehyde for 24 hours, dehydrated, transparent, and embedded in wax. The sections were dewaxed with xylene, rehydrated with gradient ethanol, and antigen repaired with 0.01M citric acid buffer (pH 6.0) (95°C, 20 minutes).

[0204] Immunofluorescence staining used phosphorylated-APP (p-APP) primary antibody and APP primary antibody, incubated at 4°C overnight. The next day, Alexa Fluor 488-labeled secondary antibody was used for incubation at room temperature for 2 hours, and DAPI (1:1000) was used to stain the nucleus for 10 minutes. Images were collected using a Zeiss LSM880 confocal microscope with a 20× objective lens. ImageJ software was used to calculate the cell area of ​​each sample.

[0205] like Figure 8As shown, the expression of APP and p-APP (Thr668 site) in the dentate gyrus (DG) region of the hippocampus of wild-type mice (WT), positive control group (pure solvent) mice, and positive drug administration group (i.e., 5xFAD mice treated with neurorepair factors). On the left are APP (green), p-APP (Thr668, red), DAPI staining (blue), and the merged image of the three. In the WT group, the expression of APP and p-APP was low; in the positive control group (pure solvent) mice, APP (green) and p-APP (red) increased significantly in the DG region, indicating the accumulation of pathological APP and p-APP; after treatment with neurorepair factors, the expression of APP and p-APP in the positive drug administration group mice was significantly reduced. The bar graph on the right quantifies the p-APP (Thr668) positive area in the DG region. Compared with the WT group, the p-APP in the positive control group (pure solvent) mice increased significantly (***p<0.001), and the p-APP area in the positive drug administration group mice was significantly reduced after treatment with neurorepair factors (**p<0.01). Scale bar: 50 μm.

[0206] 3. Neurorepair factor treatment reduces the levels of APP and Tau proteins

[0207] Western blot was used to detect the expression and phosphorylation levels of APP and Tau proteins in mouse brain tissues. First, protein samples were extracted from the brain tissues of wild-type control, positive control and positive drug-treated mice. After protein quantification using the BCA method, equal amounts of samples were separated by electrophoresis on SDS-PAGE gels and then transferred to PVDF membranes. After blocking with 5% skim milk powder, anti-p-APP (Thr668), APP, p-Tau (AT180) (Thr231), Tau5 and GAPDH (internal reference) antibodies were added and incubated overnight at 4°C, and HRP-labeled secondary antibodies were added the next day for incubation at room temperature for 1-2 hours.

[0208] After chemiluminescence development using ECL substrate, the grayscale of the bands was analyzed using Image J software to calculate the relative expression levels of the target protein and GAPDH.

[0209] The results are as follows Fig. 9As shown, the expression analysis of p-APP (Thr668), APP, p-Tau (AT180) and Tau5 proteins in wild-type mice (WT), positive control group (pure solvent) mice and positive drug administration group (i.e. 5xFAD mice treated with neural repair factors) is shown in Western blot, and GAPDH is used as an internal reference. It can be seen that: compared with WT mice, the expression levels of p-APP (Thr668) and p-Tau (AT180) proteins in the positive control group (pure solvent) mice increased significantly; after treatment with neural repair factors, the expression levels of p-APP (Thr668) and p-Tau (AT180) in the positive drug administration group mice decreased. Quantitative analysis bar graphs show the ratios of p-APP / APP and p-Tau / Tau5. Compared with the WT group, the p-APP / APP ratio of mice in the positive control group (pure solvent) was significantly increased (**p<0.01), while this ratio was significantly decreased after treatment with neural repair factors (*p<0.05); the p-Tau / Tau5 ratio of mice in the positive control group (pure solvent) was significantly increased (*p<0.05), while this ratio was significantly decreased after treatment with neural repair factors (*p<0.05).

[0210] 4. Neural repair factor treatment improves the secretion of Reelin in the dentate gyrus of the hippocampus of mice

[0211] The experiment used 3 μm thick paraffin sections of mouse spinal cord and analyzed by immunofluorescence staining. The experimental groups included wild-type control, positive control group and positive drug administration group, with 3 mice in each group. The tissue was fixed with 4% paraformaldehyde for 24 hours, dehydrated, transparent, and embedded in wax. The sections were dewaxed with xylene, rehydrated with gradient ethanol, and antigen repaired with 0.01M citric acid buffer (pH 6.0) (95°C, 20 minutes).

[0212] Immunofluorescence staining used Reelin primary antibody and incubated overnight at 4°C. The next day, Alexa Fluor 488-labeled secondary antibody was used for incubation at room temperature for 2 hours, and DAPI (1:1000) was used to stain the nucleus for 10 minutes. Images were collected using a Zeiss LSM 880 confocal microscope with a 20× objective lens. ImageJ software was used to calculate the cell area of ​​each sample.

[0213] The results are as follows Fig.10As shown, the figure shows the expression of Reelin protein in the dentate gyrus (DG) region of the hippocampus of wild-type mice (WT), positive control group (pure solvent) mice, and positive drug administration group (i.e., 5xFAD mice treated with neural repair factors). The images from left to right are Reelin (green), DAPI (blue), and the merged image of the three. In the WT group of mice, Reelin was abundantly expressed in the DG region; in the positive control group (pure solvent) mice, Reelin expression was significantly reduced; after treatment with neural repair factors, Reelin expression in the positive drug administration group mice was significantly restored. The bar graph on the right shows the quantitative analysis of the Reelin-positive area in the DG region. Compared with the WT group, the expression of Reelin in the positive control group (pure solvent) mice was significantly reduced (*p<0.05), and after treatment with neural repair factors, the expression of Reelin was significantly increased (*p<0.05). Scale bar: 50μm.

[0214] The present invention has been described in detail above. For those skilled in the art, without departing from the purpose and scope of the present invention, and without the need to carry out unnecessary experimental conditions, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. Although the present invention provides specific embodiments, it should be understood that the present invention can be further improved. In a word, according to the principles of the present invention, the application is intended to include any changes, uses or improvements to the present invention, including departure from the disclosed scope in the application, and changes made with conventional techniques known in the art.

Claims

1. A composition for improving cognition, characterized in that: The composition is any of the following: (d1) Docosahexaenoic acid, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine and astaxanthin in the following amounts by weight: composition: Docosahexaenoic acid: 8000-24000 parts by mass; Phosphatidylserine: 2400-7200 parts by mass; Vitamin D3: 1 part by mass; Vitamin B12: 0.5-2 parts by weight; Vitamin B6: 50-150 parts by weight; Vitamin B1: 100-300 parts by mass; Vitamin B9: 4-12 parts by weight; Taurine: 2500-7500 parts by mass; Astaxanthin: 500-1500 parts by mass; (d2) is composed of the following molar proportions of docosahexaenoic acid, phosphatidylserine, vitamin D3, vitamin B12, vitamin B6, vitamin B1, vitamin B9, taurine and astaxanthin composition: Docosahexaenoic acid: 0.5-2 mol parts; Phosphatidylserine: 0.5-2 mol parts; Vitamin D3: 1 mol; Vitamin B12: 0.5-2 mol; Vitamin B6: 0.5-2 mol; Vitamin B1: 0.5-2 mol; Vitamin B9: 0.5-2 mol; Taurine: 0.5-2 mol parts; Astaxanthin: 0.5-2 molar parts.

2. The composition according to claim 1, characterized in that: In (d1), the mass parts of each substance are as follows: Docosahexaenoic acid: 16000 parts by mass; Phosphatidylserine: 4800 parts by mass; Vitamin D3: 1 part by mass; Vitamin B12: 1 part by mass; Vitamin B6: 100 parts by mass; Vitamin B1: 200 parts by mass; Vitamin B9: 8 parts by weight; Taurine: 5000 parts by mass; Astaxanthin: 1000 parts by mass.

3. The composition according to claim 1, characterized in that: In said (d2), the molar fractions of each substance are as follows: Docosahexaenoic acid: 1 mol part; Phosphatidylserine: 1 mol; Vitamin D3: 1 mol; Vitamin B12: 1 mol; Vitamin B6: 1 mol; Vitamin B1: 1 mol; Vitamin B9: 1 mol; Taurine: 0.5-2 mol parts; Astaxanthin: 1 mol.

4. Use of the composition according to any one of claims 1 to 3 in the preparation of a product for improving cognition.

5. Use of the composition according to any one of claims 1 to 3 in any of the following: (A1) Preparation of products for the treatment and / or prevention of neurodegenerative diseases; (A2) Preparing a product for ameliorating symptoms of a neurodegenerative disease.

6. Use of the composition according to any one of claims 1 to 3 in the preparation of a product capable of reducing the toxicity of neurotoxic proteins to nerve cells.

7. The use according to claim 6, characterized in that: The neural cells are primary neural cells, SH-SY5Y cells or neural cells induced by iPSC pluripotent stem cells.

8. A product comprising the composition according to any one of claims 1 to 3; The product has any of the following functions: (B1) Improve cognition; (B2) treatment and / or prevention of neurodegenerative diseases; (B3) Improve symptoms of neurodegenerative diseases; (B4) Reduce the toxic effects of neurotoxic proteins on nerve cells.

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