Composition for assisting in improving memory and application thereof

Abstract

The invention discloses a composition for assisting in improving memory and application of the composition, and particularly relates to the technical field of health care products. The composition comprises broccoli leaf polypeptide, pleurotus nebrodensis polypeptide, gastrodia elata protein peptide and N-acetylneuraminic acid in a mass ratio of 1: (0.5-1.5): (2-3): (0.1-0.3). According to the invention, the four components cooperate to promote secretion of acetylcholine and reduce the content of acetylcholin esterase, so that the composition has the efficacy of assisting in improving memory and cognitive ability, and can be applied to health care products.

Description

Technical Field

[0001] This invention relates to the field of health product technology, specifically to a composition that helps improve memory and its application. Background Technology

[0002] In the field of functional products that help improve memory, peptides are widely studied and applied due to their small molecular weight and high bioactivity. However, existing compositions suffer from the following significant bottlenecks: The synergistic mechanism is unclear: Existing compositions often use single or simply mixed peptide components, lacking scientific compatibility, resulting in insufficient synergistic effect between active ingredients, making it difficult to fully exert the effect of assisting in improving memory.

[0003] Poor stability: Some peptide components are susceptible to environmental factors (such as temperature and pH), which can lead to reduced bioactivity and affect product shelf life and efficacy stability.

[0004] Low cost-effectiveness: The complexity of some compositions or the high cost of raw materials limit their large-scale application in the health supplement field.

[0005] Based on this, the present invention scientifically combines specific polypeptide components (broccoli leaf polypeptide, white lingzhi polypeptide, and gastrodia protein peptide) with N-acetylneuraminic acid to construct a composition with a clear synergistic effect mechanism, and verifies its regulatory effect on acetylcholine secretion and acetylcholinesterase content, thereby overcoming the defects of the prior art and providing a highly efficient and stable composition to help improve memory. Summary of the Invention

[0006] In view of the shortcomings of the existing technology, the present invention provides a composition for improving memory and its application, which has a significant effect on improving memory.

[0007] In a first aspect, the present invention provides a composition for assisting in improving memory, the composition comprising the following raw materials: broccoli leaf polypeptide, white lingzhi polypeptide, gastrodia protein peptide, and N-acetylneuraminic acid, in a mass ratio of 1:0.5-1.5:2-3:0.1-0.3.

[0008] Preferably, the mass ratio of broccoli leaf peptide, white lingzhi peptide, gastrodia protein peptide, and N-acetylneuraminic acid in the composition is 1:0.5-1:2-2.5:0.1-0.2.

[0009] More preferably, the mass ratio of broccoli leaf peptide, white lingzhi peptide, gastrodia protein peptide, and N-acetylneuraminic acid in the composition is 1:1:2.5:0.2.

[0010] Secondly, the present invention provides the use of the composition described in the first aspect for improving memory in the preparation of health products having the effect of improving memory.

[0011] Thirdly, the present invention provides a health product with the effect of assisting in improving memory, the health product comprising 20-30 wt% of the composition for assisting in improving memory described in the first aspect.

[0012] Preferably, the health product further includes at least one of sweetener, thickener, nicotinamide, and milk powder.

[0013] Preferably, the sweetener is at least one selected from white sugar, brown sugar, black sugar, glucose, fructose, cyclamate, acesulfame potassium, sucralose, sorbitol, maltitol, xylitol, mannitol, and erythritol.

[0014] Preferably, the thickener is at least one of xanthan gum, tamarind gum, resistant dextrin, carrageenan, and gelatin.

[0015] More preferably, the thickener is tamarind polysaccharide gum, resistant dextrin, and carrageenan, with a mass ratio of 1:2-4:0.5-1.5.

[0016] Preferably, the milk powder is whole milk powder or skim milk powder; Preferably, the health product with the function of assisting in improving memory comprises the following components: a composition, a sweetener, a thickener, niacinamide, and milk powder, and its preparation method includes the following steps: Mix the composition, sweetener, thickener, nicotinamide, and milk powder evenly, sterilize by irradiation, and store in a sealed container.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows: The composition provided by this invention has significant effects in improving memory. Specifically, the composition of this invention can moderately increase the level of acetylcholine (ACh) and decrease the content of acetylcholinesterase in mice with cognitive impairment. The moderate increase in acetylcholine level can enhance communication between neurons, and the decrease in acetylcholinesterase content can increase the retention time of acetylcholine in the synaptic cleft. This makes the transmission of neural signals more persistent and helps to strengthen the memory and learning process, which is crucial for the formation of short-term memory and the consolidation of long-term memory. Detailed Implementation

[0018] To better understand the present invention, the present invention will be further described below with reference to specific embodiments. The terminology used in the compositions or test examples is for describing specific specific embodiments and does not constitute a limitation on the scope of protection of the present invention.

[0019] For the following compositions or test examples, unless specific conditions are specified, the test methods are generally performed under standard conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0020] The sources of some of the raw materials involved in this invention are as follows: Broccoli leaf peptides: purchased from Lanzhou Waterles Biotechnology Co., Ltd.

[0021] White lingzhi mushroom polypeptide: purchased from Shaanxi Sinote Biotechnology Co., Ltd.

[0022] Gastrodia elata protein peptides: purchased from Fufeng Sinote Biotechnology Co., Ltd.

[0023] Acetylneuraminic acid: purchased from Xi'an Haotian Biotechnology Co., Ltd.

[0024] In a specific embodiment of the present invention, the total mass fraction of the composition for assisting in improving memory is 100 parts, and the raw materials and their mass fractions are as follows: Composition 1: It is composed of broccoli leaf peptides, white lingzhi peptides, gastrodia protein peptides, and N-acetylneuraminic acid in a mass ratio of 1:1:2.5:0.2.

[0025] Composition 2: It is composed of broccoli leaf polypeptide, white lingzhi polypeptide, gastrodia protein peptide and N-acetylneuraminic acid in a mass ratio of 1:0.5:2:0.1.

[0026] Composition 3: It is composed of broccoli leaf polypeptide, white lingzhi polypeptide, gastrodia protein peptide and N-acetylneuraminic acid in a mass ratio of 1:1.5:3:0.3.

[0027] Composition ①: It is composed of white lingzhi peptide, gastrodia protein peptide and N-acetylneuraminic acid in a mass ratio of 1:2.5:0.2.

[0028] Composition ②: It is composed of broccoli leaf polypeptide, gastrodia protein peptide and N-acetylneuraminic acid in a mass ratio of 1:2.5:0.2.

[0029] Composition ③: It is composed of broccoli leaf polypeptide, white ginseng polypeptide and N-acetylneuraminic acid in a mass ratio of 1:1:0.2.

[0030] Composition ④: It is composed of broccoli leaf peptides, white lingzhi peptides, and gastrodia protein peptides in a mass ratio of 1:1:2.5.

[0031] Composition ⑤: It is composed of broccoli leaf polypeptide, white lingzhi polypeptide, gastrodia protein peptide and N-acetylneuraminic acid in a mass ratio of 1:2:1:0.5.

[0032] The preparation method of the above-mentioned composition for improving memory includes the following steps: mixing each raw material evenly according to the formula amount, and sealing and storing it in a light-proof container.

[0033] The health supplement with the function of assisting in improving memory in this invention comprises the following components by weight percentage: Health Supplement 1: Composition: 25 wt% Sweetener: 0.4 wt%; Thickener: 2wt% Nicotinamide: 0.5 wt% Milk powder: supplemented to 100 wt%; Wherein, the composition is composition 1; the sweetener is sucralose; the thickener is composed of tamarind polysaccharide gum, resistant dextrin, and carrageenan in a mass ratio of 1:3:1; and the milk powder is whole milk powder. Preparation method of health product 1: Mix the composition, sweetener, thickener, nicotinamide and milk powder evenly, sterilize by irradiation, and store in a sealed container.

[0034] Health Supplement 2: Composition: 20wt% Sweetener: 0.4 wt%; Thickener: 2wt% Nicotinamide: 0.5 wt% Milk powder: supplemented to 100 wt%; Wherein, the composition is composition 1; the sweetener is sucralose; the thickener is composed of tamarind polysaccharide gum, resistant dextrin, and carrageenan in a mass ratio of 1:3:1; and the milk powder is whole milk powder. The preparation method of health product 2 is the same as that of health product 1.

[0035] Health Supplement 3: Composition: 30 wt% Sweetener: 0.4 wt%; Thickener: 2wt% Nicotinamide: 0.5 wt% Milk powder: supplemented to 100 wt%; Wherein, the composition is composition 1; the sweetener is sucralose; the thickener is composed of tamarind polysaccharide gum, resistant dextrin, and carrageenan in a mass ratio of 1:3:1; and the milk powder is whole milk powder. The preparation method of health supplement 3 is the same as that of health supplement 1.

[0036] Health Supplement 4: Composition: 25 wt% Sweetener: 0.4 wt%; Thickener: 2wt% Nicotinamide: 0.5 wt% Milk powder: supplemented to 100 wt%; Wherein, the composition is composition 2; the sweetener is sucralose; the thickener is composed of tamarind polysaccharide gum, resistant dextrin, and carrageenan in a mass ratio of 1:3:1; and the milk powder is whole milk powder. The preparation method of health supplement 4 is the same as that of health supplement 1.

[0037] Health Supplement 5: Composition: 25 wt% Sweetener: 10 wt% Thickener: 2wt% Nicotinamide: 0.5 wt% Whole milk powder: supplemented to 100 wt%; Wherein, the composition is composition 3; the sweetener is white granulated sugar; the thickener is composed of tamarind polysaccharide gum, resistant dextrin and carrageenan in a mass ratio of 1:3:1; The preparation method of health supplement 5 is the same as that of health supplement 1.

[0038] Blank health supplements: Unlike health supplement 1, this product uses an equal amount of whole milk powder to replace the composition, while the remaining ingredients and their mass percentages are the same as those in health supplement 1. Preparation method of blank health products: Mix sweetener, thickener, nicotinamide and milk powder evenly, sterilize by irradiation, and store in a sealed container.

[0039] Test Example 1: Behavioral Evaluation 1.1. Test sample solution and its preparation method: The test samples were compositions 1-3 and compositions ①-⑤; The test sample solution is prepared by using physiological saline to prepare a 1 mg / mL test sample solution, which is prepared as needed.

[0040] 1.2. Animal handling and grouping Seven-week-old male C57BL / 6 mice (SPF grade) were housed in an environment with a temperature of 22±2℃, humidity of 55±2%, and a 12h:12h light-dark cycle, with free access to food and water. After one week of acclimatization, the mice were randomly divided into a control group, a model group, and an experimental group, for a total of 10 groups, with 20 mice in each group. The 20 mice in each group were then randomly and equally divided into two groups, A and B, to participate in two behavioral evaluation tests.

[0041] The control group received intraperitoneal injections of saline solution followed by gavage. The model group received daily intraperitoneal injections of D-galactose at a dose of 800 mg / kg body weight, followed by gavage of saline solution. The experimental group received daily intraperitoneal injections of D-galactose at a dose of 800 mg / kg body weight, followed by gavage of the experimental sample solution. The intraperitoneal injection and gavage volume was 0.2 mL per mouse.

[0042] Twelve weeks later, a series of behavioral tests were conducted on each group. After the behavioral tests were completed, the mice were decapitated and the hippocampus was collected, immediately frozen in liquid nitrogen, and stored at -80°C.

[0043] 1.3. Behavioral Evaluation of Nesting Experiments Mice were placed individually in a clean cage containing 32 square pieces of shredded paper (5cm x 5cm) evenly distributed within the cage. The mice were allowed free movement in the cage for 12 hours before evaluation. Nest scores were then statistically analyzed according to the following criteria (out of 4): No nest found: 1 point; No obvious nest, nest paper exceeds 1 / 3 of the cage space: 2 points; Obvious nest building, nest paper occupies 1 / 4-1 / 3 of the cage space: 3 points; Bowl-shaped nest can be seen or nest paper is less than 1 / 4 of the cage space: 4 points.

[0044] The average score of each group of mice was taken (retaining one decimal place), as shown in Table 1 below.

[0045] 1.4. Behavioral Evaluation of the Dark Avoidance Test The darkness avoidance test, also known as the passive avoidance test, is a test chamber designed to utilize the dark-seeking and light-avoidance behavior of laboratory mice. The chamber is half dark and half bright, connected by a small door in the middle. The bottom of the dark chamber is lined with an electrified copper grid; animals entering the dark chamber will receive an electric shock.

[0046] The darkness avoidance experiment lasted for 3 days. The training experiment started at 9:00 am on the first day, and then the mice were tested 3 times at 3:00 pm on the same day, 9:00 am on the second day, and 9:00 am on the third day. The number of electric shocks to the mice's feet within 5 minutes was recorded. The average number of electric shocks for each group of mice was taken (retaining 1 decimal place), as shown in Table 1 below.

[0047] Table 1 Behavioral evaluation results Grouping Nest building score Number of electric shocks control group 3.1 8.3 Model group 1.2 24.4 Composition 1 2.9 9.7 Composition 2 2.8 9.9 Composition 3 2.7 10.4 Composition ① 2.2 16.3 Composition ② 1.8 17.7 Composition ③ 1.9 17.9 Composition ④ 1.6 19.8 Composition ⑤ 2.5 14.1 Results Analysis: In the nesting behavior test, the nesting score of the model group was significantly lower than that of the experimental sample group and the control group. However, the nesting scores of the experimental sample groups (compositions 1-3) were not significantly different from those of the control group, indicating that the composition of this invention can effectively improve cognitive impairment in mice. Comparing the nesting scores of composition 1, compositions ①-④, and the model group, it can be seen that although composition 1 and compositions ①-④ both have a certain effect on improving cognitive impairment in mice, the improvement effect of compositions ①-④ is significantly worse than that of composition 1. This indicates that the broccoli leaf peptide, white lingzhi peptide, gastrodia protein peptide, and N-acetylneuraminic acid used in this invention have a synergistic effect on improving cognitive impairment in mice. Comparing the scores of composition 1 and composition ⑤, it can be seen that the composition prepared using the mass ratio of broccoli leaf peptide, white lingzhi peptide, gastrodia protein peptide, and N-acetylneuraminic acid of this invention has a more significant effect on improving cognitive impairment in mice.

[0048] The avoidance test in mice is highly sensitive to memory processes, especially memory retrieval ability. In the avoidance test, the number of foot shocks in the experimental sample group was significantly lower than that in the model group, indicating that the composition effectively enhances the memory retrieval ability of mice. Comparing the number of shocks in Composition 1, Compositions ①-④, and the model group, it can be seen that although Composition 1 and Compositions ①-④ all have a certain effect on enhancing the memory retrieval ability of mice, the enhancement effect of Composition 1 is more significant than that of Compositions ①-④. This indicates that the broccoli leaf peptide, Pleurotus eryngii peptide, Gastrodia elata protein peptide, and N-acetylneuraminic acid used in this invention have a synergistic effect on enhancing the memory retrieval ability of mice. Comparing the number of shocks in Composition 1 and Composition ⑤, it can be seen that the composition prepared using the mass ratio of broccoli leaf peptide, Pleurotus eryngii peptide, Gastrodia elata protein peptide, and N-acetylneuraminic acid specified in this invention has a more significant effect on enhancing the memory retrieval ability of mice.

[0049] Test Example 2: Physicochemical Test Acetylcholine (ACh) is a neurotransmitter that is essential for brain and nervous system function, including memory and cognitive functions, emotional control and muscle control.

[0050] Equal masses of hippocampal bodies from the aforementioned experimental mice were placed in centrifuge tubes, and 0.5 mL of pre-chilled PBS buffer was added. The tissues were homogenized using a tissue homogenizer (frequency: 500 Hz, time: 300 s, temperature: -30℃), centrifuged (speed: 8000 rpm, time: 10 min, temperature: 4℃), and the supernatant was collected. The levels of acetylcholine (ACh) and acetylcholinesterase (AChE) were detected using an ELISA kit. The results are expressed as mean ± standard deviation, as shown in Table 2 below.

[0051] Table 2 Effects of the composition on the levels of acetylcholine and acetylcholinesterase in the mouse hippocampus Group ACh (μg / mL) AchE (μg / mL) control group 427.01±10.60* 159.86±11.75* Model group 277.21±8.18* 274.18±10.53* Composition 1 349.07±15.05 179.69±13.57 Composition 2 337.47±11.95 180.47±12.04 Composition 3 332.41±10.46 185.79±9.51 Composition ① 297.03±10.82* 236.26±9.23* Composition ② 288.05±10.78* 243.33±6.88* Composition ③ 291.25±11.07* 238.56±7.29* Composition ④ 280.73±8.52* 246.59±6.12* Composition ⑤ 309.04±7.23* 222.03±6.25* Note: "*" indicates p < 0.05 compared to composition 1. Comparing the results of composition 1 with those of compositions ①-⑤ in Table 2, it can be seen that the compositions provided by the present invention can significantly increase the content of acetylcholine in the hippocampus of mice and reduce the content of acetylcholinesterase, which is consistent with the behavioral test results.

[0052] Test Example 3: Sensory Characteristics Test samples: health supplements 1-5 and blank health supplements.

[0053] Sample preparation method: Take the health product, mix it with water, stir well, and cool it to room temperature. The mass ratio of health product to water is 1:3. The water is drinking water with a temperature of 90-100℃.

[0054] Sensory evaluators with professional training were invited to conduct sensory evaluations of the color, appearance, taste, dispersibility, texture, and mouthfeel of the health products after brewing. The evaluation results are shown in Table 3.

[0055] Table 3. Sensory characteristics evaluation of health supplements before and after brewing.

[0056] As shown in Table 3, the health product prepared by this invention has a uniform texture, no lumps, no off-odors, good dispersibility after being brewed with water, uniform brewing, and a rich milky aroma.

[0057] The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and these variations still fall within the protection scope of the present invention.

Claims

1. A composition for assisting in improving memory, characterized by comprising: The composition comprises the following raw materials: broccoli leaf polypeptide, white mushroom polypeptide, gastrodin protein peptide, and N-acetylneuraminic acid, and the mass ratio is 1:0.5-1.5:2-3:0.1-0.

3.

2. The composition of claim 1, wherein, The mass ratio of the broccoli leaf polypeptide, white mushroom polypeptide, gastrodin protein peptide, and N-acetylneuraminic acid in the composition is 1:0.5-1:2-2.5:0.1-0.

2.

3. The composition of claim 2, wherein, The mass ratio of the broccoli leaf polypeptide, white mushroom polypeptide, gastrodin protein peptide, and N-acetylneuraminic acid in the composition is 1:1:2.5:0.

2.

4. Use of the composition according to any one of claims 1-3 in the preparation of health products with the effect of assisting in improving memory.

5. A health care product having an effect of assisting improvement of memory, characterized by comprising the extract according to any one of claims 1 to 4. The health product comprises 20-30 wt% of the composition for assisting in improving memory according to any one of claims 1-3.

6. The health care product according to claim 5, wherein The health product further comprises at least one of a sweetener, a thickening agent, nicotinamide, and milk powder.

7. The health care product according to claim 6, wherein The milk powder is full-fat milk powder or skimmed milk powder.

8. The health care product according to claim 6, wherein The sweetener is at least one of white granulated sugar, brown sugar, black sugar, glucose, fructose, sweetener, acesulfame, chloro sucrose, sorbitol, maltitol, xylitol, mannitol, and erythritol.

9. The health care product according to claim 6, wherein The thickening agent is at least one of xanthan gum, tamarind polysaccharide gum, resistant dextrin, carrageenan, and gelatin.

10. The health care product according to claim 9, wherein The thickening agent is tamarind polysaccharide gum, resistant dextrin, and carrageenan, and the mass ratio is 1:2-4:0.5-1.5.

Images