Food formula for improving learning and memory ability and preparation method thereof

Abstract

The invention discloses a food formula for improving learning and memory ability and a preparation method thereof. The food formula comprises the following components in parts by mass: 1.6 parts of broccoli seed water extract; 2 parts of konjak; 2 parts of Chinese yam; and 0.000052% of vitamin A-0. 00052% 2 to 0.0002 part of vitamin B; and 0.00003 of folic acid. All the components are ground into composite powder which is used for brewing meal replacement. The learning and memory ability under the AD symptom can be healthily and sustainably prevented and improved.

Description

Technical Field

[0001] This invention relates to food, specifically to food formulations and preparation methods for improving learning and memory abilities. Background Technology

[0002] Alzheimer's disease (AD) is a neurodegenerative disease that commonly affects people over the age of 60. It is characterized by changes in memory, calculation, thinking, language, orientation, emotion, and personality, as well as a decline in social activity. It can ultimately lead to death and its incidence rate is second only to cardiovascular disease and cancer.

[0003] Currently, the main method for studying Alzheimer's disease (AD) is the establishment of animal models. Using scopolamine to create an AD model with impaired cholinergic function is one of the most representative animal models available.

[0004] Using functional foods and supplements to alleviate Alzheimer's disease (AD) is a healthy and sustainable treatment approach. Summary of the Invention

[0005] In view of the shortcomings of the existing technology, the purpose of this invention is to provide a food formula and preparation method for improving learning and memory abilities, which can prevent and improve learning and memory abilities under AD symptoms.

[0006] To achieve the above objectives, the present invention provides the following technical solution: A food formula for improving learning and memory abilities, comprising the following ingredients in the indicated mass ratios. Broccoli seed water extract -- 1.6; Konjac -- 2; Yam -- 2; Vitamin A -- 0.000052; Vitamin B2 -- 0.0002; Folic acid -- 0.00003.

[0007] The above-mentioned proportions of ingredients can improve learning and memory function under AD symptoms.

[0008] As a further improvement, the following components by mass ratio are also included: Xiaomi -- 2.7; Oats -- 2.7; Corn -- 2.7; Soybeans -- 2; Peas -- 2; Job's tears -- 2; Buckwheat - 2; Carrots -- 1.3 Hickory nuts -- 0.7; Sunflower seeds--0.7; Black sesame seeds -- 0.7g; Calcium -- 0.052; Vitamin -- 0.0000007.

[0009] In addition, the above ingredients can be added to adjust the nutritional ratio, making it suitable for use as a meal replacement and achieving healthy and sustained symptom improvement.

[0010] As a further improvement of the present invention, all ingredients are ground into powder to form a composite powder for use in preparing meal replacements.

[0011] A method for preparing a food formula to improve learning and memory is also provided, comprising obtaining the formula as described above and grinding and mixing it to obtain a powder. After grinding and mixing, it can be taken as a meal replacement, with each spoonful mixed with water to achieve healthy and sustainable prevention and improvement.

[0012] As a further improvement to this invention, 26g of powder is individually packaged. This makes it convenient to prepare as a meal replacement and allows for measured dosage.

[0013] The beneficial effects of this invention are that it can prevent and improve learning, memory and recognition abilities under AD symptoms to a certain extent, and it exhibits a certain dose-response relationship. It also has a significant neuroprotective effect on localization learning and memory abilities. Attached Figure Description

[0014] Figure 1 This is a flowchart of the experimental process of the present invention; Figure 2 This invention relates to a novel object recognition experiment in mice. Figure 3 The swimming trajectories of the control group and model group after modeling in this invention are shown (A is the control group, B is the model group). Figure 4 The escape latency of mice in each group was compared according to the present invention (*P<0.05 compared with the control group; &P<0.05 compared with the positive control group; #P<0.05 compared with the model group). Figure 5 The average training time for mice in each group was compared according to the present invention (*P<0.05 compared with the control group; &P<0.05 compared with the positive control group; #P<0.05 compared with the model group). Figure 6 The average time to find the target in each group of mice in this invention was compared (*P<0.05 compared with the control group; &P<0.05 compared with the positive control group; #P<0.05 compared with the model group). Figure 7Comparison of the average target distance for each group of mice in the present invention (*P < 0.05 compared with the control group; &P < 0.05 compared with the positive control group; #P < 0.05 compared with the model group); Figure 8 Comparison of the residence time within quadrants for each group of mice in the present invention (*P < 0.05 compared with the control group; &P < 0.05 compared with the positive control group; #P < 0.05 compared with the model group); Figure 9 Comparison of the moving distance within the target quadrant for each group of mice in the present invention (*P < 0.05 compared with the control group; &P < 0.05 compared with the positive control group; #P < 0.05 compared with the model group); Figure 10 Comparison of the number of times of crossing the platform for each group of mice in the present invention (*P < 0.05 compared with the control group; &P < 0.05 compared with the positive control group; #P < 0.05 compared with the model group); Figure 11 Comparison of the novelty object DI levels for each group of mice in the present invention (*P < 0.05 compared with the control group; &P < 0.05 compared with the positive control group; #P < 0.05 compared with the model group); Detailed implementation manners

[0015] The present invention will be further described in detail below with reference to the embodiments given in the drawings.

[0016] 1 Materials and methods 1.1 Experimental animals SPF-grade male Kunming mice, 6 - 8 months old, were purchased from Tongxiang Branch of Zhejiang Vital River Co., Ltd. (SCXK (Zhe) 2019 - 0001). The mice were housed in the SPF-grade animal room of the Animal Experiment Center of Wenzhou Medical University. The animal feeding completely complied with the regulations of the Animal Experiment Management Committee of the experiment center. The day and night were alternated for 12 hours, the temperature was 22 ± 2 °C, and the humidity was 55 ± 5%. The mouse feed was purchased from Medison and was SPF-grade 60Co irradiated and sterilized. The mouse drinking water was strictly treated by high-pressure sterilization.

[0017] 1.2 Experimental materials Water maze experiment instrument (Reword Life Science Co., Ltd.); Novel object recognition experiment instrument (Reword Life Science Co., Ltd.); Donepezil hydrochloride; Sodium carboxymethylcellulose (Shanghai Aladdin Reagent Co., Ltd.); Scopolamine (Shanghai Aladdin Reagent Co., Ltd.); Formula food was prepared according to the aforementioned mass ratio: water extract of broccoli seeds (the active ingredient sulforaphane is about 30%), konjac, yam, vitamin A, vitamin B2, folic acid, millet, oats, corn, soybeans, peas, coix seeds, buckwheat, carrots, pecans, sunflower kernels, black sesame seeds, vitamin D, calcium.

[0018] 1.3 Experimental Methods (Refer to...) Figure 1 (As shown) 1.3.1 Intervention and Model Establishment Seventy-five male Kunming mice were randomly divided into four groups: a blank control group (Group C), a model group (Group S), and a positive control group (Group Y), with 12 AD mice in each group, and 13 mice in each of the intervention groups (low, medium, and high concentrations). The concentrations of LMH in the intervention group were set at 0.1161 g / kg, 0.2322 g / kg, and 0.4643 g / kg, according to the formula food of the pre-experimental intervention group. After 7 days of oral administration of sodium carboxymethyl cellulose to Groups C, Y, and S, Group S was given donepezil hydrochloride (dissolved in sodium carboxymethyl cellulose) by oral administration for 3 days, followed by intraperitoneal injection of scopolamine (dissolved in physiological saline) to establish the model for 8 days.

[0019] 1.3.2 Water Maze Experiment For the next 6 days, after 30 minutes of gavage, mice were injected intraperitoneally and then subjected to behavioral experiments after 30 minutes (10 mice were selected from each group for the water maze experiment in the first 4 days, and 60 mice were subjected to the novel object recognition experiment in the last 2 days).

[0020] One day before the experiment, we moved the mice to the behavioral testing room to facilitate their early familiarization and adaptation to their surroundings. During the experiment, we observed the mice's activity and water intake daily. Before the experiment began, water was added to a predetermined height of approximately 35 cm and heated to about 20°C. A circular platform with a diameter of approximately 9 cm was then placed in the center of the first quadrant. We observed the mice's behavioral changes daily and recorded their drinking time and feeding. Note that each mouse needed a 1-hour interval before each of their two water insertions.

[0021] A camera is installed directly above the pool, and the camera, which adjusts the pool surface, obscures the view of the pool. The computer system sets the swimming time to 60 seconds; using a mouse experiment, the red circle is adjusted to precisely define the area of ​​the pool and platform, and tap water is injected into the pool. To ensure that the mice's judgment is not disturbed by the surrounding environment, the lighting is adjusted to avoid water reflection while avoiding hanging objects on the walls, and the room must be kept completely silent.

[0022] (1) Adaptive training The circular platform in the first quadrant was adjusted to be about 1.0 cm above the water surface. The mice were then observed to adapt for 60 seconds. Next, the mice were placed into the water in the first, second, third, and fourth quadrants in sequence (with their heads facing the pool wall) and allowed to search in the water for 60 seconds. If the mice failed to find the platform, they were guided to the platform with a long stick and stayed there for 30 seconds.

[0023] (2) Positioning and navigation experiment In this experiment, water was added to submerge the circular platform, making the platform approximately 1.0 cm below the water surface. Each mouse was sequentially dropped into the Morris water maze, facing the pool wall, from quadrants one through four. The entry process was recorded until the mouse found the underwater platform and stood for 5 seconds; this time was termed the escape latency. The mouse's path was also recorded. Adaptability to environmental changes was evaluated by comparing the dwell time and path length in each area. If a mouse did not find the platform within 60 seconds, it was gently guided to the platform using a long stick, stood for 30 seconds, and its escape latency was recorded as 60 seconds.

[0024] (3) Space exploration experiments In spatial exploration experiments, the circular platform needs to be disassembled; however, the platform's location still needs to be marked with a circle in the computer recording. To address this issue, this experiment designed multiple models to simulate the effects of platforms of different shapes and sizes on mouse behavior. Next, mice in each group were systematically released into the water, approaching the pool wall from the second, third, and fourth quadrants. The swimming routes, the path and time spent in the first quadrant, and the number of times they passed the platform location were recorded over 60 seconds.

[0025] 1.3.3 Novel Object Recognition Experiment Before each experiment, the objects were washed with a 75% ethanol solution to eliminate any odor. Mice explored the objects within a distance of 2 cm or less by touching or sniffing. Climbing or chewing did not meet the criteria for exploration.

[0026] The recognition index (DI = time spent exploring novel objects / time spent exploring novel + familiar objects). When the recognition index of a new object is higher than the random level (50%), it represents the recognition memory of the new object. As the recognition index increases, the recognition memory of the new object becomes stronger, and vice versa.

[0027] (1) Familiarization stage In the absence of test subjects, mice were placed alone in a container and allowed to explore for 5 minutes to allow them to acclimatize to their surroundings. (2) Identification stage Two identical objects were placed symmetrically on opposite sides. The mouse was allowed to explore the box for 5 minutes during the training process. Two stopwatches were used to record the time the mouse spent exploring the two objects. The timing was stopped after the mouse had been exploring the two objects for 20 seconds. The total time of the training phase was also recorded.

[0028] After approximately one hour of training, a familiar object was replaced by a completely new object. Two stopwatches were then used to record the time required to explore the familiar object. Timing was stopped when the total time spent exploring both objects reached 25 seconds, or when the total time during the testing phase reached 5 minutes, and the total training time was recorded. The experimental scenario is illustrated below. Figure 2 As shown.

[0029] Weigh the patient approximately every 4 days and adjust the dosage accordingly. Weigh the feed during this period and calculate the feed intake.

[0030] 1.4 Statistical Methods Data analysis was performed using SPSS 26.0 software. Data are expressed as mean ± standard error. One-way ANOVA was used to compare data between groups. If the variances were homogeneous, the LSD test was used; if the variances were unequal, Dunnett's T3 test was used, or an independent samples t-test was directly applied. A two-sided p-value < 0.05 was considered statistically significant. Origin 2018 was used for graphing.

[0031] 2 Results 2.1 Behavioral comparison between control group and model group mice like Figure 3 As shown, comparing the swimming trajectories of mice in the control group and the model group after modeling revealed that the swimming trajectories of the model group were scattered, random, and aimless. Figure 3 B; The control group showed a purposeful swimming preference and a linear swimming pattern, see Figure 3 A indicates that the spatial memory and orientation ability of the AD model mice were significantly reduced compared with the negative control group, indicating that the model was successfully established.

[0032] 2.2 Effects of formulated diet on learning and memory abilities in AD mice The escape latency in groups L (P<0.05) and M (P<0.05) was significantly longer than that in the positive control group. Although the mean latency in group H was longer than that in the positive control group, the difference between group H and the positive control group was not statistically significant (P>0.05). The escape latency of LMH was significantly longer than that in the control group (P<0.05) and significantly shorter than that in the model group (P<0.05). See [link to relevant documentation]. Figure 4 .

[0033] The training time in the intervention group was significantly longer than that in the control group compared to the LMH group (P<0.05). Although the mean training time in the LMH group was longer than that in the positive control group, the difference was not statistically significant (P>0.05). Compared to the model group, the training time in the intervention group was significantly shorter than that in the model group (P<0.05). Figure 5 .

[0034] The average time to find the target in group L was significantly longer than that in the control group (P<0.05), while the mean average time to find the target in group MH was not significantly different from that in the blank control group (P>0.05). Figure 6 The mean values ​​of the LM group were all higher than those of the positive control group, while the mean value of the H group was shorter than that of the positive control group. However, there was no statistically significant difference between the LMH and positive control groups (P>0.05). The mean values ​​of the intervention groups were all lower than those of the model group, and the average time to find the target in the H group was significantly shorter than that in the model group (P<0.05). Although the mean value of the LM group was also shorter than that in the model group, the difference was not statistically significant (P>0.05). See [link to relevant documentation]. Figure 6 .

[0035] The mean target finding distance in the LMH group was not significantly different from that in the control group and the positive control group (P>0.05), but it was significantly shorter than that in the model group (P<0.05). Figure 7 .

[0036] There was no significant difference in quadrant dwell time between the intervention group and the positive control group and the blank control group (P>0.05). However, compared with the model group, the mean quadrant dwell time in the intervention group was longer, and the M and H groups were significantly longer than the model group (P<0.05). But there was no significant difference between the model group and the L group (P>0.05). Figure 8 .

[0037] The mean quadrant movement distances in groups L and M were significantly shorter than those in the blank control group (P<0.05). While the distances in group H were also shorter, there was no significant difference (P>0.05). Compared to the positive control group, the distances in groups MH and L were all seemingly shorter, but not significantly so (P>0.05). However, the distances in group L were significantly shorter than those in the positive control group (P<0.05). The mean quadrant movement distances in groups LMH were longer than those in the model group. Groups M and H were significantly longer than those in the model group (P<0.05), while there was no significant difference in group L (P>0.05). See [link to relevant documentation]. Figure 9 .

[0038] Compared with the control group, the LMH group had fewer mean crossings, but the difference was not significant (P>0.05). The L group had fewer crossings compared with the positive control group, while the MH group had a higher mean, but again there was no significant difference (P>0.05). The LM group had a higher number of crossings than the model group, but the difference was not significant (P>0.05). The H group had significantly more crossings than the model group (P<0.05). See Figure 10 .

[0039] 2.3 Effects of formulated diets on novelty object recognition in AD mice The identification index (DI) in group L was significantly lower than that in the control group (P<0.05). Although the mean DI in group MH was lower than that in the control group, the difference was not significant (P>0.05). Similarly, the DI in group L was significantly lower than that in the positive control (P<0.05), while the mean DI in group MH was lower but not significantly different from that in the positive control group (P>0.05). Compared to the model group, except for the low-concentration group (P>0.05), the DI in group MH was significantly higher than that in group model (P<0.05). (See...) Figure 11 .

[0040] In summary, the experimental results show that, compared with the model group, the formula food containing sulforaphane (broccoli seed water extract) to a certain extent prevented and improved the learning, memory and recognition behaviors of mice, and showed a certain dose-response relationship.

[0041] By chemically damaging AD mice with scopolamine, cholinergic AD mice were induced. The successful modeling was confirmed by comparing the movement trajectories of the control and model groups in a water maze. The experiment showed that the diet containing sulforaphane (broccoli seed water extract) effectively improved the escape latency of the mice compared to the model group, a result similar to previous studies. Furthermore, compared to the model group, the intervention group showed significant improvements in average training time and average distance to the target. The bar chart revealed that the intervention group had higher time spent in the target quadrant, longer distance traveled within the target quadrant, and more platform crossings than the model group, while the average time to find the target was lower. Except for the low-dose intervention group, the other indicators showed no significant difference between the intervention group and the positive control group. The escape latency, average time to find the target, time spent in the target quadrant, longer distance traveled within the target quadrant, and more platform crossings all showed a dose-response relationship in the intervention group; that is, the higher the concentration of the diet, the more significant the difference compared to the model group. Experimental results showed that the formulated food containing sulforaphane had a significant neuroprotective effect on the localization learning and memory ability of AD mice, and its preventive and therapeutic effects gradually increased with the increase of the concentration of the formulated food (including broccoli seed water extract and multivitamins).

[0042] The above description is merely a preferred embodiment of the present invention. The scope of protection of the present invention is not limited to the above embodiments. All technical solutions falling within the scope of the present invention's concept are within the scope of protection of the present invention. It should be noted that for those skilled in the art, any improvements and modifications made without departing from the principles of the present invention should also be considered within the scope of protection of the present invention.

Claims

1. A food formula for improving learning and memory abilities, characterized in that, Including the following components in the mass ratio Broccoli seed water extract -- 1.6; Konjac -- 2; Yam -- 2; Vitamin A -- 0.000052; Vitamin B2 -- 0.0002; Folic acid -- 0.00003.

2. The food formula for improving learning and memory ability according to claim 1, characterized in that, It also includes the following components in the following mass ratios: Xiaomi -- 2.7; Oats -- 2.7; Corn -- 2.7; Soybeans -- 2; Peas -- 2; Job's tears -- 2; Buckwheat - 2; Carrots -- 1.3 Hickory nuts -- 0.7; Sunflower seeds--0.7; Black sesame seeds -- 0.7g; Calcium -- 0.052; Vitamin -- 0.0000007.

3. The food formula for improving learning and memory ability according to claim 2, characterized in that, All ingredients are ground into a compound powder for use in preparing meal replacements.

4. A method for preparing a food formula that improves learning and memory abilities, characterized in that, Obtain the formula as described in claim 2 and grind and mix it to obtain a powder.

5. The method for preparing the food formula for improving learning and memory ability according to claim 4, characterized in that, Weigh out 26g of powder and package it individually.

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