Application of pecan oil in resisting Alzheimer's disease, reducing blood fat, softening blood vessels and promoting skeletal development

Abstract

The invention relates to the technical field of food, in particular to application of pecan oil in resisting Alzheimer's disease, reducing blood fat, softening blood vessels and promoting skeletal development. By establishing different zebrafish animal models and utilizing fluorescent staining observation, it is found that the hickory nut oil has the effects of improving memory, resisting Alzheimer's disease, reducing blood fat, resisting atherosclerosis, softening blood vessels, reducing blood pressure, increasing bone mineral density and promoting skeletal development, and development of the hickory nut oil, especially wild hickory nut oil, can be promoted.

Description

Technical Field

[0001] The invention relates to the field of food technology, in particular to application of pecan oil in resisting Alzheimer's disease, lowering blood lipids, softening blood vessels and promoting bone development. Background Art

[0002] Walnut oil is a common edible oil with a high content of unsaturated fatty acids. Containing over 80% unsaturated fatty acids, including oleic, linoleic, palmitic, and linolenic acids, walnut oil serves as an important source of unsaturated fatty acids for the human body. Walnut oil also contains active ingredients that the human body cannot synthesize naturally, such as tocopherols, phytosterols, and squalene. Pecan oil is produced by pressing pecans, but its efficacy is less well-researched and unclear. Summary of the Invention

[0003] In order to solve the above problems, the present invention discovered that pecan oil has the effects of moistening the intestines and promoting bowel movements, resisting aging and enhancing immunity, thereby promoting the development of pecan oil.

[0004] In order to achieve the above object, the present invention provides the following technical solutions:

[0005] The present invention provides the use of pecan oil in preparing a product having one or more functions of improving memory, resisting Alzheimer's disease, lowering blood lipids, resisting atherosclerosis, softening blood vessels, lowering blood pressure, increasing bone density and promoting bone development.

[0006] Preferably, the pecan oil is wild pecan oil.

[0007] Preferably, the wild pecan oil contains fatty acids and vitamin E; the fatty acids include palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid and linolenic acid.

[0008] Preferably, the fatty acid is composed of the following components in percentage by mass: palmitic acid 2.4%, palmitoleic acid 0.2%, stearic acid 0.5%, oleic acid 17.0%, linoleic acid 69.7% and linolenic acid 10.2%.

[0009] Preferably, the content of vitamin E in the wild pecan oil is 56.7 mg / 100 g.

[0010] Preferably, the preparation method of the wild pecan oil comprises: crushing wild pecan kernels and then squeezing to obtain the wild pecan oil.

[0011] Preferably, the particle size of the wild pecan kernels after crushing is ≤1 cm.

[0012] Preferably, the raw materials for preparing the wild pecan oil include wild pecans from Yahe Village, Yahe Township, Huanren Manchu Autonomous County, Benxi City, Liaoning Province.

[0013] Preferably, the lipid-lowering comprises lowering triglyceride levels.

[0014] Preferably, the anti-atherosclerosis comprises lowering cholesterol levels.

[0015] Beneficial effects:

[0016] The present invention provides the use of pecan oil in preparing products that have one or more of the following effects: improving memory, combating Alzheimer's disease, lowering blood lipids, combating atherosclerosis, softening blood vessels, lowering blood pressure, increasing bone density, and promoting skeletal development. By establishing different zebrafish animal models, the present invention evaluated the memory-improving and Alzheimer's disease-fighting effects of pecan oil using fluorescence staining, a zebrafish behavioral trajectory tracking system, and analysis of zebrafish swimming distance. The findings indicate that pecan oil has efficacy in combating Alzheimer's disease, maintaining vascular health, increasing bone density, and promoting skeletal development. This research may promote the development of pecan oil, particularly wild pecan oil, and provide technical support for the development of safe and effective bone health intervention strategies. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments are briefly introduced below.

[0018] Figure 1 Results of the effects of different treatments on the movement distance of zebrafish;

[0019] Figure 2 This is a trajectory diagram showing the effects of different treatments on zebrafish behavior;

[0020] Figure 3 The triglyceride test results of zebrafish in different treatment groups in Example 4;

[0021] Figure 4 The cholesterol test results of zebrafish in different treatment groups in Example 4;

[0022] Figure 5 The results of nitric oxide content testing of zebrafish in different treatment groups in Example 5 are as follows;

[0023] Figure 6 The results of blood vessel diameter detection of zebrafish in different treatment groups in Example 6;

[0024] Figure 7 Fluorescence images of zebrafish blood vessels in different treatment groups in Example 6;

[0025] Figure 8 The average bone fluorescence intensity test results of zebrafish in different treatment groups in Example 7;

[0026] Figure 9This is the fluorescence image of zebrafish in different treatment groups in Example 7;

[0027] Figure 10 The average number of vertebrae in zebrafish treated in different groups in Example 8 is shown in Figure 8.

[0028] Figure 11 This is the fluorescence image of zebrafish in different treatment groups in Example 8;

[0029] Among them, * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001. DETAILED DESCRIPTION

[0030] The present invention provides the use of pecan oil in preparing a product having one or more functions of improving memory, resisting Alzheimer's disease, lowering blood lipids, resisting atherosclerosis, softening blood vessels, lowering blood pressure, increasing bone density and promoting bone development.

[0031] In one embodiment, the pecan oil is wild pecan oil. In one embodiment, the wild pecan oil contains fatty acids and vitamin E; the fatty acids include palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. In one embodiment, the fatty acids are composed of the following components in percentage by weight: 2.4% palmitic acid, 0.2% palmitoleic acid, 0.5% stearic acid, 17.0% oleic acid, 69.7% linoleic acid, and 10.2% linolenic acid. In one embodiment, the vitamin E content of the wild pecan oil is 56.7 mg / 100 g.

[0032] In one embodiment, the method for preparing wild pecan oil comprises: crushing wild pecan kernels and then pressing them to obtain wild pecan oil. In one embodiment, the particle size of the crushed wild pecan kernels is ≤1 cm. In another embodiment, the crushed wild pecan kernels are dried at 50°C for 24 hours, ground into powder after drying, and mechanically pressed in an oil press. In another embodiment, the raw materials for preparing the wild pecan oil include wild pecans from Yahe Village, Yahe Township, Huanren Manchu Autonomous County, Benxi City, Liaoning Province.

[0033] As an embodiment, the lipid-lowering includes lowering triglyceride levels.

[0034] In one embodiment, the anti-atherosclerosis includes lowering cholesterol levels.

[0035] The pecan oil provided by the present invention is a natural vegetable oil extracted from pecans and is rich in various bioactive ingredients such as monounsaturated fatty acids, polyphenol compounds (such as ellagic acid) and vitamin E. The present invention establishes different zebrafish animal models, uses fluorescent staining for observation, uses a zebrafish behavior trajectory tracking system, and analyzes the swimming distance of zebrafish to evaluate the efficacy of pecan oil in improving memory and preventing Alzheimer's disease. The present invention discovers the efficacy of pecan oil in preventing Alzheimer's disease, maintaining vascular health, increasing bone density, and promoting bone development. This can promote the development of pecan oil, especially wild pecan oil, and provide technical support for the development of safe and effective bone health intervention strategies.

[0036] To further illustrate the present invention, the application of the pecan oil provided by the present invention in combating Alzheimer's disease, lowering blood lipids, softening blood vessels and promoting bone development is described in detail below with reference to the examples and drawings, but they should not be construed as limiting the scope of protection of the present invention.

[0037] Example 1

[0038] Wild pecan oil is prepared by the following method: shelling wild pecans and crushing them into small pieces less than 1 cm in size, drying them at 50°C for 24 hours, grinding them into powder after drying, wrapping the powder in a filter bag, and placing it in an oil press for mechanical pressing to obtain wild pecan oil; the wild pecans were purchased from Yahe Village, Yahe Township, Huanren Manchu Autonomous County, Benxi City, Liaoning Province.

[0039] The fatty acid composition of the wild pecan oil was determined using the method of GB / T 22327-2008 "Walnut Oil", and the results were as follows: palmitic acid (C16:0) 2.4%, palmitoleic acid (C16:1) 0.2%, stearic acid (C18:0) 0.5%, oleic acid (C18:1) 17.0%, linoleic acid (C18:2) 69.7% and linolenic acid (C18:3) 10.2%.

[0040] The vitamin E content in the wild pecan oil was detected by the method of GB / T 5009.82-2003 and was 56.7 mg / 100 g.

[0041] Example 2

[0042] Feifan Standard Technology Services (Suzhou) Co., Ltd. was commissioned to test the anti-Alzheimer's disease efficacy of the wild pecan oil in Example 1, including the following steps:

[0043] Testing Principle: Zebrafish share a high degree of morphological, physiological, and genetic similarity with humans, and their nervous systems are similar to those of mammals. Excessive intake of aluminum chloride can lead to abnormal animal behavior, disrupt cholinergic nerve function, and increase amyloid production, ultimately leading to memory loss and reduced learning ability. The anti-Alzheimer's efficacy is evaluated by analyzing the swimming distance of zebrafish using a behavioral analyzer.

[0044] Experimental Materials: Wild-type AB strain zebrafish were used, with 30 per group housed in individual culture tanks. Adult fish were reared under strictly controlled conditions: water temperature was maintained at 28 ± 0.5°C, and the light-dark cycle was 14 hours light and 10 hours dark. Water quality parameters were as follows: pH 7.0-7.5, conductivity 450-500 μS / cm, total ammonia nitrogen <0.02 mg / L, and dissolved oxygen 5-8 mg / L. Live Artemia nauplii were fed twice daily prior to the experiment to ensure optimal growth and physiological conditions.

[0045] The tested zebrafish were divided into four groups, namely blank control group, model control group, positive control group and sample treatment group. Among them, the blank control group did not receive any treatment, while the model control group, sample treatment group and positive control group all ingested an equal amount of aluminum chloride hexahydrate (aluminum chloride hexahydrate was ingested into the zebrafish by dissolving it in the water used for fish farming). The positive control group ingested aluminum chloride hexahydrate and donepezil hydrochloride at the same time, while the sample treatment group ingested aluminum chloride hexahydrate and wild pecan oil solution (volume concentration was 2%). After a period of time, the behavior of the zebrafish was analyzed, and the total movement distance of the zebrafish was finally presented as the result. See the results. Figures 1-2 and Table 1.

[0046] Table 1 Movement distance of zebrafish in different groups (mm)

[0047] Blank control group Model control group Positive control group Sample processing group 819.2±378.6 147.5±85.6 534.7±257.5 568.2±228.2

[0048] This example systematically evaluated the effect of pecan oil on the improvement of neurobehavioral function in the zebrafish model of Alzheimer's disease induced by aluminum chloride through zebrafish movement distance analysis. The results showed that the movement distance of zebrafish in the model control group (147.5 ± 85.6 mm) was significantly lower than that in the blank control group (819.2 ± 378.6 mm), indicating that aluminum chloride hexahydrate successfully induced neurobehavioral dysfunction in zebrafish. This result is consistent with the results of previous studies. Aluminum chloride hexahydrate simulates the pathological characteristics of Alzheimer's disease by inducing oxidative stress, neuroinflammation and neuronal damage. The significant reduction in movement distance further verifies the reliability of the model and provides a basis for subsequent drug intervention experiments.

[0049] The zebrafish in the positive control group (treated with donepezil hydrochloride) moved significantly more distance (534.7±257.5 mm) than the model control group, indicating that donepezil hydrochloride can effectively improve aluminum chloride-induced neurobehavioral dysfunction. Donepezil, an acetylcholinesterase inhibitor, improves cognitive function by increasing acetylcholine levels in the synaptic cleft and enhancing neuronal signaling. The results of the positive control group validated the effectiveness of the experimental method and provide a reference for the efficacy of wild pecan oil.

[0050] The movement distance of zebrafish in the sample treatment group (568.2±228.2mm) was significantly improved compared with the model control group, and there was no statistically significant difference with the positive control group, indicating that the wild pecan oil provided by the present invention has a significant improvement effect on neurobehavioral function, and the neuroprotective effect is equivalent to that of donepezil.

[0051] This embodiment uses video trajectory tracking technology to visualize the movement pattern of zebrafish. The trajectory diagram shows the movement trajectory of zebrafish within a certain period of time and their preferences for visiting or staying. Figure 2 As shown, the model control group zebrafish showed a significant reduction in activity range compared to the blank control group, and the exploration desire and motor ability decreased. These behavioral characteristics are highly similar to the cognitive impairment and spatial exploration ability of Alzheimer's patients, indicating that aluminum chloride successfully induced the neurobehavioral dysfunction of zebrafish. The increase in limbic behavior may be due to the anxiety-like behavior of zebrafish under neurotoxicity. This "wall-tending behavior" is a typical anxiety-like performance. Studies have confirmed that it is significantly correlated with memory function impairment, further verifying the reliability of the model. The zebrafish movement trajectory and central area activity of the positive control group (donepezil treatment) were significantly restored, and the zebrafish movement trajectory and central area activity of the sample treatment group were significantly restored, and close to the level of the positive control group. This shows that the wild pecan oil provided by the present invention can effectively improve the neurobehavioral dysfunction induced by aluminum chloride, restore the exploration desire and motor ability of zebrafish, and show that the neuroprotective effect of the wild pecan oil provided by the present invention is comparable to donepezil. It can be seen that the wild pecan oil provided by the present invention, as a natural plant extract, has potential clinical application value, especially in the early prevention and intervention of Alzheimer's disease.

[0052] Example 3

[0053] Feifan Standard Technology Service (Suzhou) Co., Ltd. was commissioned to test the memory-improving effect of the wild pecan oil in Example 1, comprising the following steps:

[0054] Testing Principle: Zebrafish share a high degree of morphological, physiological, and genetic similarity with humans, and their nervous systems are similar to those of mammals. Excessive intake of aluminum chloride can lead to abnormal animal behavior, disrupt cholinergic nerve function, and increase amyloid production, ultimately leading to memory loss and reduced learning ability. Memory-enhancing efficacy is assessed by analyzing the distance zebrafish swim using a behavioral analyzer.

[0055] The test materials and methods are the same as in Example 2. Figures 1-2 and Table 1.

[0056] This example systematically evaluated the effect of wild pecan oil on the improvement of aluminum chloride-induced memory dysfunction by zebrafish movement distance analysis. The results showed that zebrafish in the model control group showed significant motor impairment and abnormal exploratory behavior, while pecan oil treatment effectively reversed these behavioral defects, with an effect comparable to that of the positive control drug donepezil ( Figure 1 These behavioral changes provide important evidence for the role of pecan oil in improving memory function.

[0057] Compared to the blank control group (819.2±378.6mm), the movement distance of zebrafish in the model control group was significantly reduced to 147.5±85.6mm, a phenomenon closely related to spatial memory impairment. In mammalian studies, hippocampal-dependent spatial memory impairment is often manifested as reduced exploratory behavior. As a vertebrate model, zebrafish have conserved neural mechanisms underlying their locomotor exploration and memory functions. Compared to the model control group, zebrafish in the sample-treated group showed significant improvement in movement distance (568.2±228.2mm), and the degree of improvement was not statistically different from that in the positive control group (534.7±257.5cm). This result suggests that pecan oil may improve memory by regulating the cholinergic system. Donepezil, a cholinesterase inhibitor, has a widely recognized mechanism of action for improving memory, and pecan oil can achieve similar effects, suggesting that it may act through similar pathways.

[0058] The trajectory map shows the movement trajectory of zebrafish in a certain period of time and their preference for visiting or staying ( Figure 2 Trajectory analysis showed that zebrafish in the blank control group exhibited a typical spatial exploration pattern. Their movement trajectories were evenly distributed, frequently crossed the central area, and exhibited continuous exploration of novel areas. This complex movement pattern reflects normal working memory and spatial memory function. In contrast, the movement trajectories of zebrafish in the model control group showed obvious characteristics of memory impairment: the range of movement was significantly reduced, the trajectory was mainly confined to the peripheral area, and a simple, repetitive circular movement pattern was presented. This "edge chemotaxis behavior" is a typical manifestation of memory dysfunction, suggesting that zebrafish have lost their normal environmental cognition and spatial positioning abilities.

[0059] The trajectory characteristics of zebrafish in the positive control group showed significant improvement: the range of movement expanded, the frequency of exploration in the central region increased, and the complexity of the trajectory increased significantly. Notably, the trajectory patterns of the pecan oil-treated group displayed similar improvements to those of the positive drug group: the movement trajectory was redistributed throughout the test area, exploration behavior in the central region was restored, and the trajectory exhibited normal random exploration characteristics. This recovery in movement patterns was particularly evident in three aspects: first, the area covered by the trajectory was significantly expanded, indicating improved spatial memory; second, the frequency of crossing the central region increased, reflecting the recovery of exploratory behavior; and finally, the randomness of the movement path increased, suggesting improved cognitive flexibility.

[0060] Through in-depth analysis of trajectory characteristics, the researchers discovered that zebrafish treated with wild pecan oil exhibited unique behavioral improvements. Compared to the positive drug group, their trajectory changes exhibited two characteristic differences: a higher frequency of turning points, indicating enhanced information processing; and a more systematic exploration path, indicating a more efficient spatial memory strategy. These characteristic movement pattern changes provide intuitive behavioral evidence for the memory-enhancing effects of wild pecan oil.

[0061] Example 4

[0062] Feifan Standard Technology Service (Suzhou) Co., Ltd. was commissioned to test the blood lipid lowering and anti-atherosclerosis efficacy of the wild pecan oil in Example 1, including the following steps:

[0063] Testing Principle: The zebrafish genome shares a high degree of homology with that of humans, and many genes associated with human diseases have corresponding homologous genes in zebrafish. The ingredients in a high-sugar, high-fat diet can simulate the excessive sugar and fat intake found in the human diet. This diet can lead to disrupted fat metabolism in the body and elevated levels of lipids such as triglycerides in the blood, thus inducing a condition in zebrafish similar to hyperlipidemia in humans. If the test sample can effectively lower triglyceride levels in zebrafish, it indicates that the sample has lipid-lowering properties. Atorvastatin calcium tablets are positive. Triglycerides (TG) and total cholesterol (TC) are also associated with atherosclerosis. Excessive levels of TG and TC may lead to disrupted lipid metabolism, thereby increasing the risk of atherosclerosis. Zebrafish are highly similar to humans. High-sugar, high-fat, high-fat diets are used to create hyperlipidemia in zebrafish. The anti-atherosclerotic efficacy is evaluated by testing whether blood lipids are reduced after adding the sample.

[0064] The test materials are the same as those in Example 2.

[0065] The zebrafish were divided into four groups: a blank control (NC), a high-fat diet control (HFD), a positive control (PC), and a wild pecan oil-treated (WPO) group. The NC group received no treatment, while the HFD, PC, and WPO groups all consumed an equal amount of a high-sugar, high-fat diet. The PC and WPO groups, while consuming the high-sugar, high-fat diet, also received atorvastatin calcium tablets and a test sample solution (5% by volume). The high-sugar, high-fat diet was prepared by dissolving 10g of egg yolk powder and 4g of glucose, homogenizing the resulting mixture on a magnetic stirrer, and then drying it in a freeze dryer overnight to produce the high-sugar, high-fat diet. The high-sugar, high-fat diet was dissolved in the fish culture water and administered to the zebrafish. After a period of incubation, blood lipid levels were measured in the zebrafish from the different groups. Five juvenile fish were randomly selected from each group for testing of various indicators. Triglyceride levels were measured using a lipid analyzer (PFS-30A, Guilin Unitech Electronics Group Co., Ltd.). Total cholesterol levels were measured using a cholesterol assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Each group had one well of zebrafish, with 30 zebrafish per well.

[0066] See the results Figures 3-4 and Tables 2-3.

[0067] Table 2 Triglyceride values (mmol / L) of zebrafish in different treatment groups

[0068] Blank control group Model control group Positive control group Sample processing group 1.05±0.09 2.81±0.05 0.85±0.04 1.08±0.02

[0069] Table 3 Cholesterol values (mmol / L) of zebrafish in different treatment groups

[0070] Blank control group Model control group Positive control group Sample processing group 1.88±0.11 2.15±0.04 1.92±0.11 1.86±0.17

[0071] The results showed that the triglyceride value of the model control group increased significantly compared with the blank control group, indicating that the modeling was successful. The triglyceride value of the sample treatment group decreased significantly compared with the model control group, indicating that the sample has the effect of lowering blood lipids ( Figure 3 and Table 2 ).

[0072] Compared with the blank control group, the cholesterol content of zebrafish in the model control group increased significantly, indicating that the modeling was successful. Compared with the model control group, the cholesterol content in the sample treatment group decreased significantly, indicating that the sample has anti-atherosclerosis effect ( Figure 4 and Table 3 ).

[0073] Example 5

[0074] Feifan Standard Technology Service (Suzhou) Co., Ltd. was commissioned to test the blood vessel softening effect of the wild pecan oil in Example 1, including the following steps:

[0075] Testing Principle: Traditional Chinese Medicine (TCM) theory holds that cardiovascular and cerebrovascular diseases are caused by insufficient nitric oxide (NO). Nitric oxide has important physiological functions, including softening blood vessels—increasing vascular elasticity, relaxing and softening blood vessels, lowering blood pressure, inhibiting smooth muscle cell proliferation and platelet adhesion, participating in immune responses, and killing tumor cells and microorganisms. It has important clinical significance in the pathogenesis of many cardiovascular diseases, including hypertension, myocardial ischemia, and stroke, as well as in the development and progression of autoimmune diseases, degenerative diseases, and inflammation. Nitric oxide dysfunction is a key factor in the development and progression of cardiovascular and cerebrovascular diseases.

[0076] The test materials are the same as those in Example 2.

[0077] The zebrafish were divided into two groups, namely the normal control group (NC) and the wild pecan oil treatment group (WPO). The NC group did not receive any treatment, and the wild pecan oil to be tested was added to the sample treatment group (treatment concentration was 500 μg / mL). After soaking for a period of time, five juvenile fish were randomly selected from each group. The zebrafish were crushed and the supernatant was taken, and ELISA was used to detect the nitric oxide content of zebrafish using an enzyme-linked immunosorbent assay (ELISA) kit. The data analysis results of the two groups were compared. Each group had one well of zebrafish, and each well had 30 zebrafish. The results are shown in Figure 5 .

[0078] The results showed that the nitric oxide content of zebrafish in the sample-treated group (23.21±0.81pg / mL) was significantly increased compared with the blank control group (21.25±0.38pg / mL), indicating that the sample has the effect of softening blood vessels and increasing vascular elasticity.

[0079] Example 6

[0080] Feifan Standard Technology Service (Suzhou) Co., Ltd. was commissioned to test the blood pressure lowering effect of the wild pecan oil in Example 1, including the following steps:

[0081] Testing Principle: Zebrafish genes share 87% similarity with human genes, and their blood systems share many similarities. Hypertension is primarily related to arterial diameter; smaller arterial diameters correlate with higher blood pressure. Measuring zebrafish artery diameters is used to assess a sample's ability to lower blood pressure.

[0082] Test materials: transgenic vascular fluorescent zebrafish, reared in the same manner as in Example 2.

[0083] The zebrafish were divided into two groups: a blank control group and a sample-treated group. The blank control group received no treatment, while the sample-treated group was treated with a solution of wild pecan oil (5% by volume). After a period of incubation, the zebrafish in both groups were observed and photographed using a fluorescence microscope. The diameter of the blood vessels was calculated using Image J 1.48 software, and the data analysis results of the two groups were compared. Figures 6-7 .

[0084] The results showed that the blood vessel diameter in the sample-treated group (26.59±5.92 pixels) was significantly increased compared with the blank control group (20.80±3.83 pixels), indicating that the sample has an auxiliary blood pressure-lowering effect.

[0085] Example 7

[0086] Feifan Standard Technology Service (Suzhou) Co., Ltd. was commissioned to test the bone density-increasing efficacy of the wild pecan oil in Example 1, including the following steps:

[0087] Detection Principle: Skeletal research using zebrafish as a model has become a hot topic in recent years, as bony fish and humans share a high degree of homology in genes and signaling pathways involved in skeletal development. Furthermore, compared to other animal models, zebrafish are smaller, making them suitable for high-throughput chemical screening, and their transparent juvenile bodies make skeletal development easier to observe. Bone density is largely determined by the calcium content in the bones. When fluorescent dyes specifically bind to calcium in the bones, the fluorescence intensity is positively correlated with bone density. Stronger fluorescence indicates higher bone density, greater calcium content in the bones, and therefore better skeletal development. Fluorescence intensity measured using a fluorescence microscope after zebrafish bone staining can objectively determine whether the sample is effective in increasing bone density.

[0088] The test materials are the same as those in Example 2.

[0089] The zebrafish were divided into a blank control group and a sample treatment group. The test substance stock solution was diluted with buffered water in an isocratic manner to a gradient solution of appropriate concentration, with a dilution factor of not less than 10 times, and set aside. The blank control group was not treated in any way, and the sample treatment group was added with a 5% volume concentration of organic wild walnut oil to be tested. After a period of culture, the zebrafish bones were stained with calcein, and the zebrafish bone fluorescence intensity data was collected using a fluorescence microscope. The results are shown in Figures 8-9 .

[0090] The results showed that the bone fluorescence intensity of the sample-treated group (89.05±15.89) was significantly increased compared with the blank control group (58.61±6.96), indicating that the sample has the effect of increasing bone density.

[0091] Example 8

[0092] Feifan Standard Technology Service (Suzhou) Co., Ltd. was commissioned to test the wild pecan oil in Example 1 for its efficacy in promoting bone development, comprising the following steps:

[0093] Testing Principle: During the early stages of development, the skeletal system of zebrafish larvae develops in a highly organized and genetically regulated process. By around seven days old, the vertebrae have begun to develop a specific morphology and structure, and this developmental process shares similar molecular mechanisms and signaling pathways with mammals, including humans. If a sample promotes an increase in the number of vertebrae in zebrafish larvae, this indicates that the sample has activated signaling pathways that promote bone formation or provided substances that promote skeletal cell proliferation and differentiation during these early stages of development, thus confirming its efficacy in promoting skeletal development.

[0094] The test materials, grouping and treatment methods are the same as in Example 7. Figures 10-11 .

[0095] The results showed that the average number of vertebrae in the sample-treated group (5.89±2.51) was significantly increased compared with the blank control group (2.89±0.87), indicating that the sample has the effect of promoting bone development.

[0096] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. The use of pecan oil in the preparation of products having one or more effects of improving memory, resisting Alzheimer's disease, lowering blood lipids, resisting atherosclerosis, softening blood vessels, lowering blood pressure, increasing bone density and promoting bone development.

2. The use according to claim 1, characterized in that The pecan oil is wild pecan oil.

3. The use according to claim 2, characterized in that The wild pecan oil contains fatty acids and vitamin E; the fatty acids include palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid and linolenic acid.

4. The use according to claim 3, characterized in that The fatty acid consists of the following components in percentage by mass: 2.4% of palmitic acid, 0.2% of palmitoleic acid, 0.5% of stearic acid, 17.0% of oleic acid, 69.7% of linoleic acid and 10.2% of linolenic acid.

5. The use according to claim 3, characterized in that The content of vitamin E in the wild pecan oil is 56.7 mg / 100 g.

6. The use according to claim 2 or 3, characterized in that The preparation method of the wild pecan oil comprises the following steps: crushing wild pecan kernels and then squeezing to obtain the wild pecan oil.

7. The use according to claim 6, characterized in that The particle size of the crushed wild pecan kernels is ≤1 cm.

8. The use according to claim 2 or 3, characterized in that The raw materials for preparing the wild pecan oil include wild pecans from Yahe Village, Yahe Township, Huanren Manchu Autonomous County, Benxi City, Liaoning Province.

9. The use according to claim 1, characterized in that The lipid-lowering includes lowering triglyceride levels.

10. The use according to claim 1, characterized in that The anti-atherosclerotic effect includes lowering cholesterol levels.

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