Nano-emulsion for preventing Alzheimer disease and preparation method thereof

Abstract

The invention belongs to the technical field of nano-emulsions, and particularly relates to a nano-emulsion for preventing Alzheimer's disease and a preparation method of the nano-emulsion. The nano-emulsion is prepared from the following raw materials in parts by weight: 11 to 22 parts of oil, 1.11 to 3.50 parts of an emulsifier, 1 to 3 parts of a processing aid, 0.31 to 0.86 part of a functional component, 0.01 to 0.03 part of an antioxidant and 70 to 90 parts of water, wherein the oil comprises acer truncatum seed oil and xanthoceras sorbifolia bunge oil. The acer truncatum seed oil and the xanthoceras sorbifolia bunge oil are reasonably matched and have a synergistic effect with functional ingredients, and it is found through a Morris water maze experiment that the learning ability of mice can be improved; mouse experiments show that the SOD activity of brain tissues, liver tissues and serum of mice with Alzheimer's disease can be improved, the activity of MDA (malondialdehyde) can be reduced, the ChAT activity of the brain tissues of the mice can be improved, and the activity of AChE can be reduced, so that the symptoms of the Alzheimer's disease can be prevented / relieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of nanoemulsions, and in particular relates to a nanoemulsion for preventing Alzheimer's disease and a preparation method thereof. Background Art

[0002] Alzheimer's disease (AD) is a progressive neurodegenerative disorder that primarily occurs in the elderly or pre-elderly. The disease is characterized by progressive cognitive impairment and behavioral impairment, which includes impairments in memory, thinking, analytical judgment, visual-spatial perception, self-care, and emotional control, all without any conscious effort.

[0003] Currently, the main hypotheses to explain the pathogenic mechanism of this disease include the cholinergic hypothesis, the β-amyloid protein hypothesis, the Tau protein hypothesis, the metal ion disorder hypothesis, etc., and a large number of recent studies have shown that lipid metabolism may be involved in the occurrence of AD.

[0004] Acer truncatum seed oil contains over 90% unsaturated fatty acids and is rich in neuraminic acid, which accounts for over 5%. As a long-chain monounsaturated fatty acid, neuraminic acid is the only substance that can repair and clear damaged nerve fibers in the brain's neural pathways and promote nerve cell regeneration, enhancing memory and improving brain function.

[0005] Traditional oil dosage forms have problems such as poor mouthfeel, low bioavailability, and poor stability, and are therefore often improved. For example, Chinese patent CN106901373A discloses a kind of Acer truncatum seed oil nano-microcapsule powder for middle-aged and elderly health food and its preparation method, the microcapsule powder comprises Acer truncatum seed oil 40g, octenylsuccinic acid arabic gum ester 5g, octenylsuccinic acid starch 1.5-15g, mono- and di-fatty acid glycerides 1.2g, diacetyl tartaric acid mono- and di-anhydride ester 0.2g, oligofructose syrup 35-55g, and distilled water 200-300g. In this patent, Acer truncatum seed oil is prepared into microcapsule powder, which is more conducive to the absorption of Acer truncatum seed oil by middle-aged and elderly people. However, in this patent, Acer truncatum seed oil is only used as raw material, and the efficacy of its single oil is relatively limited, and efficacy still needs to be improved. Summary of the Invention

[0006] To this end, the present invention provides a nanoemulsion for preventing Alzheimer's disease. This nanoemulsion is prepared by rationally compounding Acer truncatum seed oil and Xanthoceras sorbifolia oil and adding functional ingredients. The nanoemulsion exhibits Alzheimer's disease prevention effects. The present invention also provides a method for preparing the nanoemulsion, ensuring stable product quality, safety, and ease of industrial production.

[0007] In order to solve the above technical problems, the technical solution of the present invention is:

[0008] In one aspect, the present invention provides a nanoemulsion for preventing Alzheimer's disease, comprising the following ingredients, by weight: 11-22 parts of an oil, 1.11-3.50 parts of an emulsifier, 1-3 parts of a processing aid, 0.31-0.86 parts of a functional ingredient, 0.01-0.03 parts of an antioxidant, and 70-90 parts of water. The oils include Acer truncatum seed oil and Xanthoceras sorbifolia oil.

[0009] Preferably, the weight ratio of the Acer truncatum seed oil to the Xanthoceras sorbifolia oil is (6-12):(5-10).

[0010] Further preferably, the weight ratio of the Acer truncatum seed oil to the Xanthoceras sorbifolia oil is (0.5-2):1.

[0011] More preferably, the weight ratio of the Acer truncatum seed oil to the Xanthoceras sorbifolia oil is 1.125:1.

[0012] Preferably, the content of nervonic acid in the Acer truncatum seed oil is greater than 5%; further preferably, the content of nervonic acid in the Acer truncatum seed oil is ≥ 6%.

[0013] Preferably, the content of nervonic acid in the Xanthoceras sorbifolia oil is greater than 3%; further preferably, the content of nervonic acid in the Xanthoceras sorbifolia oil is ≥3.2%.

[0014] Preferably, the functional ingredients include one or more of sage extract, walnut peptide, fermented jujube kernel powder, and N-acetylneuraminic acid.

[0015] Further preferably, the functional ingredients include sage extract, walnut peptide, and fermented jujube kernel powder; the weight ratio of sage extract, walnut peptide, and fermented jujube kernel powder in the functional ingredients is (0.01-0.06): (0.1-0.3): (0.2-0.5).

[0016] More preferably, the weight ratio of the sage extract, walnut peptide and fermented jujube kernel powder is 1:(2-15):(8-20).

[0017] More preferably, the weight ratio of the sage extract, walnut peptide and fermented jujube kernel powder is 1:5:10.

[0018] Preferably, the sage extract contains rosmarinic acid, and the content of rosmarinic acid is not less than 2.5%.

[0019] Preferably, the walnut peptide is prepared from walnuts as raw materials through squeezing, enzymatic hydrolysis, enzyme inactivation, filtration and concentration.

[0020] Preferably, the fermented jujube seed powder contains total saponins, and the total saponin content is not less than 0.1%.

[0021] Preferably, the emulsifier comprises Tween 80, mono- and di-glycerol fatty acid esters and phospholipids;

[0022] The weight ratio of the Tween 80, mono- and diglycerol fatty acid esters and phospholipids is (0.01-0.05): (0.1-0.45): (1.0-3.0).

[0023] More preferably, the weight ratio of Tween 80, mono- and diglycerol fatty acid esters, and phospholipids is 0.035:0.25:2.0.

[0024] Preferably, the processing aid includes one or more of glycerol and ethanol; preferably, the processing aid of the present invention is glycerol.

[0025] Preferably, the antioxidant is vitamin E.

[0026] Preferably, the present invention provides a nanoemulsion for preventing Alzheimer's disease, comprising the following ingredients, by weight: 9 parts of Acer truncatum seed oil, 8 parts of Xanthoceras sorbifolia oil, 0.035 parts of Tween 80, 0.25 parts of mono- and diglycerol fatty acid esters, 2.0 parts of phospholipids, 0.035 parts of sage extract, 0.175 parts of walnut peptide, 0.35 parts of fermented jujube seed powder, 2 parts of glycerol, 0.02 parts of vitamin E, and 78.135 parts of water.

[0027] In another aspect, the present invention provides a method for preparing a nanoemulsion for preventing Alzheimer's disease, comprising the following steps:

[0028] (1) Mixing oil, emulsifier, processing aid and water, heating and stirring to fully dissolve the components, and obtaining a crude emulsion A; (2) Cooling the crude emulsion A to room temperature, adding an antioxidant and stirring to uniformly mix to obtain a crude emulsion B; (3) Shearing the crude emulsion B at room temperature, adding functional ingredients during the shearing process and adjusting the pH to 6-7, to obtain an emulsion C; (4) After high-pressure homogenization, cooling the emulsion C to room temperature to obtain a nanoemulsion.

[0029] Preferably, the heating and stirring in step (1) is carried out at a temperature of 60-75° C. for 15-30 min; more preferably, the temperature is 72° C. for 20 min.

[0030] Preferably, the stirring time in step (2) is 10-20 min; further preferably, the stirring time is 15 min.

[0031] Preferably, the shearing time in step (3) is 15-25 min, and the rotation speed is 6500-8000 rpm; more preferably, the shearing time is 20 min, and the rotation speed is 7000 rpm.

[0032] Preferably, in step (3), phosphate buffer is used to adjust the pH.

[0033] Further preferably, the preparation method of the phosphate buffer is: take 8.34g of potassium dihydrogen phosphate and 0.87g of dipotassium hydrogen phosphate, add water to dissolve them into 1000ml, and obtain the obtained solution.

[0034] Preferably, the homogenization pressure in step (4) is 200-400 bar, and the homogenization time is 20-80 min; more preferably, the homogenization pressure is 350 bar, and the homogenization time is 50 min.

[0035] Application of the above nanoemulsion in preparing products for preventing / improving Alzheimer's disease.

[0036] Compared with the prior art, the present invention has the following advantages:

[0037] 1. The nanoemulsion for preventing Alzheimer's disease provided by the present invention uses Acer truncatum seed oil and Xanthoceras sorbifolia oil as main raw materials, and adds functional ingredients sage extract, walnut peptide and fermented jujube seed powder. The above components work synergistically, as verified by Morris water maze experiments, and are found to improve the learning ability of mice. Experiments on mice have found that the nanoemulsion can increase the SOD activity of the brain tissue, liver tissue and serum of Alzheimer's mice, reduce the activity of MDA, and increase the ChAT activity of the mouse brain tissue, and reduce the activity of AChE, thereby preventing / improving the symptoms of Alzheimer's disease.

[0038] 2. The nanoemulsions provided by the present invention are made from natural plant oils and fats, making them safe and non-toxic. Nanoemulsions have the advantages of small particle size, high stability, high bioavailability, improved solubility and bioavailability, and can effectively improve the absorption and utilization of functional ingredients.

[0039] 3. The present invention provides a method for preparing a nanoemulsion for preventing Alzheimer's disease, which has a simple process and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 The state diagram of the nanoemulsion prepared in Examples 1-5

[0041] Figure 2 The state diagram of the nanoemulsion prepared in Comparative Examples 1-6

[0042] Figure 3 The state diagram of the nanoemulsion prepared in Comparative Example 7 DETAILED DESCRIPTION

[0043] The following non-limiting examples are provided to enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way. The following are merely illustrative of the scope of the present invention, and those skilled in the art may make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.

[0044] When numerical ranges are given in the examples, it should be understood that, unless otherwise specified herein, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the invention belongs.

[0045] The present invention is further described below by way of specific examples. Unless otherwise specified, the various chemical reagents used in the examples of the present invention were obtained through conventional commercial channels. The products of different manufacturers did not significantly affect the effects.

[0046] Acer truncatum seed oil: purchased from Heze Zhonghe Jianyuan Biotechnology Co., Ltd., with a neuraminic acid content of ≥6%; Xanthoceras sorbifolia oil: purchased from Heze Zhonghe Jianyuan Biotechnology Co., Ltd., with a neuraminic acid content of 3.2%; pine nut oil, purchased from Jiangxi Baolin Natural Flavor Co., Ltd., is obtained by cold pressing mature dried pine nut kernels, product number BL-21J; mono- and diglycerol fatty acid esters: food grade, purchased from Guangzhou Kaiwen Food Development Co., Ltd., product model: KVDMG9503A; Tween 80: food grade, purchased from Guangdong Runhua Chemical Co., Ltd., CAS number: 9005-65-6; phospholipids: 1000P, Shanghai Jiayi Biotechnology Co., Ltd.; sucrose fatty acid ester: S-90 (HLB value 9), Zhejiang Hetang Technology Co., Ltd.; sage extract: purchased from Guangzhou Kanglai Food Additive Co., Ltd., rosmarinic acid ≥2.5%; walnut peptide: food grade, purchased from Shandong Xinxiong Biotechnology Co., Ltd., product model WP002; fermented jujube kernel powder: purchased from Jiangsu Tianfu Biotechnology Co., Ltd., total saponin content ≥0.1%; glycerol: GLY-995FAG, purchased from Fengyi Oil Technology Co., Ltd.; vitamin E: purchased from Zhejiang Medicine Co., Ltd., CAS number: 59-02-09.

[0047] The preparation method of phosphate buffer is as follows: take 8.34g of potassium dihydrogen phosphate and 0.87g of dipotassium hydrogen phosphate, add water to dissolve them into 1000ml, and obtain the obtained solution.

[0048] Example 1

[0049] This embodiment provides a nanoemulsion for preventing Alzheimer's disease, which includes, by weight, 9 parts of Acer truncatum seed oil, 8 parts of Xanthoceras sorbifolia oil, 0.035 parts of Tween 80, 0.25 parts of mono- and diglycerol fatty acid esters, 2.0 parts of phospholipids, 0.035 parts of sage extract, 0.175 parts of walnut peptide, 0.35 parts of fermented jujube seed powder, 2 parts of glycerol, 0.02 parts of vitamin E, and 78.135 parts of water.

[0050] Its preparation method is:

[0051] (1) Weighing ingredients: Weigh all materials according to the proportion and set aside; (2) Mix Acer truncatum seed oil, Xanthoceras sorbifolia oil, Tween 80, mono- and diglycerol fatty acid esters, phospholipids, glycerol and water, heat and stir at 72°C for 20 min to fully dissolve the ingredients, and obtain a crude emulsion A; (3) Cool the crude emulsion A to room temperature, add vitamin E and stir for 15 min to mix evenly to obtain a crude emulsion B; (4) Shear the crude emulsion B at room temperature at a speed of 7000 rpm for 20 min, add sage extract, walnut peptide and fermented jujube kernel powder during the shearing process, and add phosphate buffer to adjust the pH to 6.4 to obtain an emulsion C; (5) High-pressure homogenize the emulsion C at a homogenization pressure of 350 bar and a homogenization time of 50 min, and then cool it to room temperature to obtain a nanoemulsion.

[0052] Example 2

[0053] This embodiment provides a nanoemulsion for preventing Alzheimer's disease, which includes, by weight, 6 parts of Acer truncatum seed oil, 9 parts of Xanthoceras sorbifolia oil, 0.01 parts of Tween 80, 0.10 parts of mono- and diglycerides of fatty acids, 1.5 parts of phospholipids, 0.01 parts of sage extract, 0.15 parts of walnut peptides, 0.20 parts of fermented jujube seed powder, 1 part of glycerol, 0.01 parts of vitamin E, and 82.02 parts of water.

[0054] Its preparation method is:

[0055] (1) Weighing ingredients: Weigh all materials according to the proportion and set aside; (2) Mix Acer truncatum seed oil, Xanthoceras sorbifolia oil, Tween 80, mono- and diglycerol fatty acid esters, phospholipids, glycerol and water, heat and stir at 60°C for 30 minutes to fully dissolve the ingredients, and obtain a crude emulsion A; (3) Cool the crude emulsion A to room temperature, add vitamin E and stir for 10 minutes to mix evenly to obtain a crude emulsion B; (4) Shear the crude emulsion B at room temperature at a speed of 6500 rpm for 25 minutes, add sage extract, walnut peptide and fermented jujube kernel powder during the shearing process, and add phosphate buffer to adjust the pH to 6.4 to obtain an emulsion C; (5) High-pressure homogenize the emulsion C at a homogenization pressure of 400 bar and a homogenization time of 20 minutes, and then cool it to room temperature to obtain a nanoemulsion.

[0056] Example 3

[0057] This embodiment provides a nanoemulsion for preventing Alzheimer's disease, which includes, by weight, 8 parts of Acer truncatum seed oil, 6 parts of Xanthoceras sorbifolia oil, 0.03 parts of Tween 80, 0.2 parts of mono- and diglycerol fatty acid esters, 1.0 parts of phospholipids, 0.03 parts of sage extract, 0.25 parts of walnut peptide, 0.30 parts of fermented jujube seed powder, 1.5 parts of glycerol, 0.015 parts of vitamin E, and 82.675 parts of water.

[0058] Its preparation method is:

[0059] (1) Weighing ingredients: Weigh all materials according to the proportion and set aside; (2) Mix Acer truncatum seed oil, Xanthoceras sorbifolia oil, Tween 80, mono- and diglycerol fatty acid esters, phospholipids, glycerol and water, heat and stir at 70°C for 15 minutes to fully dissolve the ingredients, and obtain a crude emulsion A; (3) Cool the crude emulsion A to room temperature, add vitamin E and stir for 20 minutes to mix evenly to obtain a crude emulsion B; (4) Shear the crude emulsion B at room temperature at a speed of 7500 rpm for 20 minutes, add sage extract, walnut peptide and fermented jujube kernel powder during the shearing process, and add phosphate buffer to adjust the pH to 6.4 to obtain an emulsion C; (5) High-pressure homogenize the emulsion C at a homogenization pressure of 200 bar and a homogenization time of 80 minutes, and then cool it to room temperature to obtain a nanoemulsion.

[0060] Example 4

[0061] This embodiment provides a nanoemulsion for preventing Alzheimer's disease, which includes, by weight, 10 parts of Acer truncatum seed oil, 5 parts of Xanthoceras sorbifolia oil, 0.04 parts of Tween 80, 0.45 parts of mono- and diglycerol fatty acid esters, 2.5 parts of phospholipids, 0.05 parts of sage extract, 0.10 parts of walnut peptide, 0.45 parts of fermented jujube seed powder, 2.5 parts of glycerol, 0.025 parts of vitamin E, and 78.885 parts of water.

[0062] Its preparation method is:

[0063] (1) Weighing ingredients: Weigh all materials according to the proportion and set aside; (2) Mix Acer truncatum seed oil, Xanthoceras sorbifolia oil, Tween 80, mono- and diglycerol fatty acid esters, phospholipids, glycerol and water, heat and stir at 65°C for 25 minutes to fully dissolve the ingredients, and obtain a crude emulsion A; (3) Cool the crude emulsion A to room temperature, add vitamin E and stir for 20 minutes to mix evenly to obtain a crude emulsion B; (4) Shear the crude emulsion B at room temperature at a speed of 7000 rpm for 15 minutes, add sage extract, walnut peptide and fermented jujube kernel powder during the shearing process, and add phosphate buffer to adjust the pH to 6.4 to obtain an emulsion C; (5) High-pressure homogenize the emulsion C at a homogenization pressure of 300 bar and a homogenization time of 40 minutes, and then cool it to room temperature to obtain a nanoemulsion.

[0064] Example 5

[0065] This embodiment provides a nanoemulsion for preventing Alzheimer's disease, which includes, by weight, 12 parts of Acer truncatum seed oil, 10 parts of Xanthoceras sorbifolia oil, 0.05 parts of Tween 80, 0.35 parts of mono- and diglycerol fatty acid esters, 3.0 parts of phospholipids, 0.06 parts of sage extract, 0.30 parts of walnut peptide, 0.50 parts of fermented jujube seed powder, 3 parts of glycerol, 0.03 parts of vitamin E, and 70.71 parts of water.

[0066] Its preparation method is:

[0067] (1) Weighing ingredients: Weigh all materials according to the proportion and set aside; (2) Mix Acer truncatum seed oil, Xanthoceras sorbifolia oil, Tween 80, mono- and diglycerol fatty acid esters, phospholipids, glycerol and water, heat and stir at 75°C for 15 minutes to fully dissolve the ingredients, and obtain a crude emulsion A; (3) Cool the crude emulsion A to room temperature, add vitamin E and stir for 15 minutes to mix evenly to obtain a crude emulsion B; (4) Shear the crude emulsion B at room temperature at a speed of 8000 rpm for 15 minutes, add sage extract, walnut peptide and fermented jujube kernel powder during the shearing process, and add phosphate buffer to adjust the pH to 6.4 to obtain an emulsion C; (5) High-pressure homogenize the emulsion C at a homogenization pressure of 350 bar and a homogenization time of 60 minutes, and then cool it to room temperature to obtain a nanoemulsion.

[0068] Comparative Example 1

[0069] This comparative example provides a nanoemulsion for preventing Alzheimer's disease, which differs from Example 1 only in that it does not include Xanthoceras sorbifolia oil, that is, it includes 17 parts by weight of Acer truncatum seed oil, 0.035 parts by weight of Tween 80, 0.25 parts by weight of mono- and diglycerol fatty acid esters, 2.0 parts by weight of phospholipids, 0.035 parts by weight of sage extract, 0.175 parts by weight of walnut peptide, 0.35 parts by weight of fermented jujube seed powder, 2 parts by weight of glycerol, 0.02 parts by weight of vitamin E, and 78.135 parts by weight of water.

[0070] The preparation method is the same as that of Example 1.

[0071] Comparative Example 2

[0072] This comparative example provides a nanoemulsion for preventing Alzheimer's disease, which differs from Example 1 only in that it does not include Acer truncatum seed oil, that is, it includes 17 parts by weight of Xanthoceras sorbifolia oil, 0.035 parts by weight of Tween 80, 0.25 parts by weight of mono- and diglycerol fatty acid esters, 2.0 parts by weight of phospholipids, 0.035 parts by weight of sage extract, 0.175 parts by weight of walnut peptide, 0.35 parts by weight of fermented jujube seed powder, 2 parts by weight of glycerol, 0.02 parts by weight of vitamin E, and 78.135 parts by weight of water.

[0073] The preparation method is the same as that of Example 1.

[0074] Comparative Example 3

[0075] This comparative example provides a nanoemulsion for preventing Alzheimer's disease, which differs from Example 1 only in that the functional ingredient does not include sage extract, that is, it includes 9 parts by weight of Acer truncatum seed oil, 8 parts by weight of Xanthoceras sorbifolia oil, 0.035 parts by weight of Tween 80, 0.25 parts by weight of mono- and diglycerol fatty acid esters, 2.0 parts by weight of phospholipids, 0.175 parts by weight of walnut peptide, 0.35 parts by weight of fermented jujube seed powder (wherein the total weight of walnut peptide and fermented jujube seed powder is the same as the total weight of sage extract, walnut peptide and fermented jujube seed powder in Example 1), 2 parts by weight of glycerin, 0.02 parts by weight of vitamin E, and 78.135 parts by weight of water.

[0076] The preparation method is the same as that of Example 1.

[0077] Comparative Example 4

[0078] This comparative example provides a nanoemulsion for preventing Alzheimer's disease, which differs from Example 1 only in that the functional ingredient does not include walnut peptide, that is, it includes 9 parts by weight of Acer truncatum seed oil, 8 parts by weight of Xanthoceras sorbifolia oil, 0.035 parts by weight of Tween 80, 0.25 parts by weight of mono- and diglycerol fatty acid esters, 2.0 parts by weight of phospholipids, 0.035 parts by weight of sage extract, 0.35 parts by weight of fermented jujube seed powder (wherein the total weight of sage extract and fermented jujube seed powder is the same as the total weight of sage extract, walnut peptide and fermented jujube seed powder in Example 1), 2 parts by weight of glycerin, 0.02 parts by weight of vitamin E, and 78.135 parts by weight of water.

[0079] The preparation method is the same as that of Example 1.

[0080] Comparative Example 5

[0081] This comparative example provides a nanoemulsion for preventing Alzheimer's disease, which differs from Example 1 only in that the functional ingredient does not include fermented jujube seed powder, that is, it includes 9 parts by weight of Acer truncatum seed oil, 8 parts by weight of Xanthoceras sorbifolia oil, 0.035 parts by weight of Tween 80, 0.25 parts by weight of mono- and diglycerol fatty acid esters, 2.0 parts by weight of phospholipids, 0.035 parts by weight of sage extract, 0.175 parts by weight of walnut peptide (wherein the total weight of sage extract and walnut peptide is the same as the total weight of sage extract, walnut peptide and fermented jujube seed powder in Example 1), 2 parts by weight of glycerin, 0.02 parts by weight of vitamin E, and 78.135 parts by weight of water.

[0082] The preparation method is the same as that of Example 1.

[0083] Comparative Example 6

[0084] This comparative example provides a nanoemulsion for preventing Alzheimer's disease, which differs from Example 1 only in that Xanthoceras sorbifolia oil is replaced by pine nut oil, that is, the nanoemulsion comprises 9 parts by weight of Acer truncatum seed oil, 8 parts by weight of pine nut oil, 0.035 parts by weight of Tween 80, 0.25 parts by weight of mono- and diglycerol fatty acid esters, 2.0 parts by weight of phospholipids, 0.035 parts by weight of sage extract, 0.175 parts by weight of walnut peptide, 0.35 parts by weight of fermented jujube seed powder, 2 parts by weight of glycerol, 0.02 parts by weight of vitamin E, and 78.135 parts by weight of water.

[0085] The preparation method is the same as that of Example 1.

[0086] Comparative Example 7

[0087] This comparative example provides a nanoemulsion for preventing Alzheimer's disease, which differs from Example 1 only in that mono- and diglycerol fatty acid esters are replaced by sucrose fatty acid esters (HLB value 9), namely, the nanoemulsion comprises 9 parts by weight of Acer truncatum seed oil, 8 parts by weight of Xanthoceras sorbifolia oil, 0.035 parts by weight of Tween 80, 0.25 parts by weight of sucrose fatty acid, 2.0 parts by weight of phospholipids, 0.035 parts by weight of sage extract, 0.175 parts by weight of walnut peptide, 0.35 parts by weight of fermented jujube seed powder, 2 parts by weight of glycerol, 0.02 parts by weight of vitamin E, and 78.135 parts by weight of water.

[0088] The preparation method is the same as that of Example 1.

[0089] Experimental Example 1 Observation of Nanoemulsion State

[0090] 30 mL of the nanoemulsion prepared in Examples 1-5 and Comparative Examples 1-7 was poured into a test cup and placed at room temperature for 30 days, and its state was observed. Figure 1 .

[0091] The nanoemulsions of Examples 1-5 were stable after 30 days without demulsification. This indicates that the oils, emulsifiers, and functional ingredients selected in the present invention work synergistically to stabilize the nanoemulsion system for preventing Alzheimer's disease. The nanoemulsions of Comparative Examples 1-6 were all stable after 30 days without demulsification. However, the nanoemulsion of Comparative Example 7 showed demulsification after 30 days, indicating that the combination of Tween 80, mono- and diglycerol fatty acid esters, and phospholipids as emulsifiers selected in the present invention with other components can better achieve the stability of the nanoemulsion system.

[0092] Experimental Example 2 Animal Experiment

[0093] D-galactose was purchased from Beijing Solaibao Technology Co., Ltd., product batch number: 0637; scopolamine hydrobromide injection was purchased from Suicheng Pharmaceutical Co., Ltd., product batch number: H41021048; malondialdehyde (MDA) assay kit (A003-1-1), superoxide dismutase (SOD) kit (A001-2-1), acetylcholinesterase (AcHE) assay kit (H529-1-1), acetylcholine transferase (ChAT) assay kit (A079-1-1) and total protein quantitative assay kit (BCA method, A045-4-1) were all purchased from Nanjing Jiancheng Bioengineering Institute.

[0094] To verify that the nanoemulsion of the present invention has the effect of preventing Alzheimer's disease, the following animal experiments are described:

[0095] Animals: C57BL / 6 mice, SPF grade, 5-month-old male, weighing (28±2) g.

[0096] Mice were fed with conventional feed and drinking water for adaptability feeding at room temperature for 1 week, and then randomly divided into 13 groups, 10 in each group, namely normal group, model group, embodiment 1-5 and comparative example 1-6. All animals were fed with common feed under uniform conditions and had free access to water. Except for the normal group injected with normal saline, the other groups were injected with D-galactose 140 mg / kg subcutaneously in the neck at the same time every day for 4 consecutive weeks, and scopolamine (2 mg / kg) was increased from the 3rd week for 2 consecutive weeks. A water maze experiment was performed in the 4th week of modeling, which took 1 week. After modeling was completed, administration was started, and the corresponding nanoemulsions of embodiment 1-5 and comparative example 1-6 were fed, and the model group and normal group were fed with equal amounts of normal saline, once a day, for 4 consecutive weeks.

[0097] The dosing regimen was to administer the drug by gavage once a day at a fixed time. The daily dose for humans was 100 mL, and the equivalent dose was calculated by body surface area ratio to be 13 mL / kg for mice. This dose was administered by gavage.

[0098] 2.1 Morris water maze test

[0099] Mice underwent a Morris water maze test once at week 4 of modeling and once after the end of drug administration. The Morris water maze primarily tests orientation and navigation and spatial exploration. The water maze pool is divided into four quadrants by equidistantly marked squares on the wall. A platform was placed in the center of one quadrant. The pool was filled with water maintained at approximately 23°C. White pure milk was evenly dispersed throughout the pool and stirred to minimize the influence of specific reference points. Reference points around the pool remained constant.

[0100] The navigation experiment is used to test the spatial learning and cognitive ability of mice after a period of training. Four times a day, the mice are placed in the water from the center of four equally divided quadrants with their heads facing the pool wall. The training records the time it takes for the mice to find the platform within 120 seconds (escape latency / duration). If the mouse does not find the platform within the specified time of 120 seconds, it is guided to the platform and stay for about 10 seconds to facilitate the mouse's memory. The escape latency at this time is recorded as 120 seconds. Each training session is separated by 40 seconds, and the experiment lasts for 6 days.

[0101] Based on the navigation experiment, the Space Exploration Laboratory removed the platform from the water maze on the seventh day and randomly placed mice in the water from the center of the quadrant. The number of times the mice crossed the original platform position within 120 seconds was recorded to assess their memory ability. The experimental results are shown in Table 1 below.

[0102] Table 1 Experimental results of the escape latency and number of times mice crossed the original platform after the model was established and after the drug administration

[0103]

[0104] Note: In the same column of the table, if there is one identical letter, the difference is not significant; if there are different letters, the difference is significant (P<0.05).

[0105] Result analysis:

[0106] The escape latency (duration) refers to the time required for a mouse to swim from the center of a specific quadrant into the water maze within 120 seconds to the water maze platform and stay on the platform for 10 seconds. It can test the learning and cognitive abilities of mice. The number of platform crossings refers to the number of times a mouse crosses the original platform position after being placed in the water maze from the midpoint of a specific quadrant within 120 seconds. It can test the memory ability of mice.

[0107] Compared with the normal group, the escape latency of mice in Examples 1-5, Comparative Examples 1-6 and the model group was significantly prolonged and the number of times they crossed the platform was significantly reduced after modeling, and the differences were statistically significant (P<0.05), indicating that the learning ability of mice in the model group decreased and the modeling was successful.

[0108] In Examples 1-5, after administration and modeling, the escape latency of the mice was shortened to varying degrees, and the number of times they crossed the platform increased to varying degrees, indicating that the nanoemulsion provided by the present invention can improve the learning ability of Alzheimer's disease mice.

[0109] After administration to the mice of Comparative Examples 1-6 and after modeling, the escape latency of the mice was shortened to varying degrees, and the number of times they crossed the platform increased to varying degrees, but the effect was not ideal. The difference between Comparative Examples 1-2, Comparative Example 6 and Example 1 is that the oils are different. As shown in Table 1, the escape latency of the mice after administration in Comparative Examples 1-2 and Comparative Example 6 was prolonged to varying degrees compared to Example 1, and the number of times they crossed the platform decreased to varying degrees, and there was a significant difference, indicating that the synergistic combination of Acer truncatum seed oil and Xanthoceras sorbifolia oil provided by the present invention can better improve the learning ability of mice, thereby improving the effect of Alzheimer's disease.

[0110] The difference between Comparative Examples 3-5 and Example 1 is that the composition of the functional ingredients is different. Compared with Example 1, the escape latency of the mice after administration of Comparative Examples 3-5 was significantly prolonged, and the number of times they crossed the platform was significantly reduced, and there was a significant difference, indicating that the functional ingredients can affect the learning ability of mice. Therefore, the functional ingredients sage extract, walnut peptide and fermented jujube seed powder provided by the present invention and the synergistic combination of oils can well improve the effect of Alzheimer's disease.

[0111] 2.2 Detection of antioxidant enzyme activity and cholinergic levels in brain tissue

[0112] After the water maze experiment, the mice were anesthetized and blood was collected by enucleation. This procedure adhered to animal welfare requirements. After blood collection, the blood tubes were placed in a centrifuge to prepare serum. The hippocampus tissue from the mouse brain was slowly rinsed in 5°C saline in an ice bath. The surface moisture was then blotted dry with filter paper. The hippocampus tissue was weighed and, at a weight-to-volume ratio of 1:9, 9 volumes of saline were added. Mechanical homogenization was performed in an ice bath at 2500 rpm for approximately 10 minutes. The supernatant was removed and used for later use. The liver tissue homogenate was extracted using the same method. SOD, MDA, AChE, and ChAT levels were determined according to the kit instructions. Protein content was determined using the BCA assay. Specific procedures were performed according to the instructions. The experimental results are shown in Tables 2-3 below.

[0113] Table 2 Effects of each group on brain tissue, liver tissue and serum SOD in Alzheimer's disease mice

[0114]

[0115] Note: In the same column of the table, if there is one identical letter, the difference is not significant; if there are different letters, the difference is significant (P<0.05).

[0116] Table 3 Effects of each group on brain tissue, liver tissue and serum MDA in Alzheimer's disease mice

[0117]

[0118] Note: In the same column of the table, if there is one identical letter, the difference is not significant; if there are different letters, the difference is significant (P<0.05).

[0119] Result analysis:

[0120] Alterations in lipid metabolism are believed to be highly correlated with the development of AD. Abnormal APP cleavage and Aβ formation, neuroinflammation, and abnormal oxidative stress in the nervous system caused by changes in lipid metabolism may be important aspects of the AD pathogenesis. Oxidative stress response is caused by an imbalance between the body's oxidative and antioxidant capacities, generating large amounts of peroxides and free radicals that exceed the body's tolerance, leading to immune cell infiltration and increased protease secretion, thus causing irreversible damage to cells. SOD is a key enzyme in the body that eliminates free radicals, and the level of SOD reveals, to a certain extent, the body's ability to scavenge oxygen free radicals. MDA is a metabolite in the body, and its level reveals, to a certain extent, the extent of damage to the body's cells caused by free radical destruction.

[0121] Compared with the model group, the SOD content in the brain tissue, liver tissue and serum of mice in Examples 1-5 was increased, and the MDA content was decreased, indicating that the nanoemulsion provided by the present invention can improve the effect of Alzheimer's disease from the perspective of neurooxidative stress.

[0122] Compared with Example 1, the SOD content in the brain tissue, liver tissue and serum of mice in Comparative Examples 1-2 and Comparative Example 6 was reduced, and the MDA content was significantly increased, indicating that the synergistic combination of oils provided by the present invention can better improve the body's oxidative stress abnormalities.

[0123] Compared with Example 1, the SOD content in the brain tissue, liver tissue and serum of mice in Comparative Examples 3-5 was reduced, and the MDA content was significantly increased, indicating that the functional ingredients and other ingredients have a certain synergistic effect, which can affect the effect of improving Alzheimer's disease. Therefore, the functional ingredients provided by the present invention are the best choice and can effectively improve Alzheimer's disease.

[0124] Table 4 Effects of each group on AChE and ChAT in brain tissue of Alzheimer's disease mice

[0125] Group AChE(U / mgprot) ChAT (U / g) Normal group 8.38±0.23h 141.29±4.33a Model Group 17.98±0.14a 71.22±3.13f Example 1 10.15±0.19g 115.49±3.93b Example 2 10.37±0.26g 113.34±3.25b Example 3 10.28±0.21g 112.63±3.41b Example 4 10.21±0.16g 114.91±3.51b Example 5 10.42±0.44g 111.95±1.46b Comparative Example 1 14.99±0.36c 90.51±3.53e Comparative Example 2 15.53±0.27b 88.92±3.40e Comparative Example 3 13.61±0.17e 99.79±2.68cd Comparative Example 4 13.45±0.11e 99.01±5.53cd Comparative Example 5 13.14±0.18f 100.69±3.65c Comparative Example 6 14.14±0.22d 95.83±3.74d

[0126] Note: In the same column of the table, if there is one identical letter, the difference is not significant; if there are different letters, the difference is significant (P<0.05).

[0127] Result analysis:

[0128] The cholinergic hypothesis is one of the key theories explaining the pathogenesis of neurological diseases, particularly Alzheimer's disease (AD). Its core concept is that decreased function of the central cholinergic system leads to cognitive impairment. Acetylcholine (ACh), secreted by cholinergic neurons in the cholinergic system, is a neurotransmitter directly linked to cognition and memory, and is directly involved in the storage and recall of information. ChAT, as the key enzyme for ACh synthesis and the enzyme responsible for its breakdown by AChE, regulates ACh metabolism and plays a crucial role in learning and memory. Acetylcholinesterase (AChE) is primarily found on the postsynaptic membrane of motor nerve terminals and at synapses in the brain, regulating neurotransmission. AChE breaks down acetylcholine throughout the body and is widely distributed in brain tissue, making it closely related to learning and memory. ChAT, a key enzyme for the synthesis of acetylcholine, is primarily found in cholinergic neurons. Its synthesis in the cell body is an important marker of cholinergic neuronal function.

[0129] Compared with the model group, the AChE content in the mouse brain tissue of Examples 1-5 was reduced, and the ChAT content was increased, indicating that the nanoemulsion prepared by the present invention can affect the AChE and ChAT contents in the mouse brain, thereby preventing Alzheimer's disease.

[0130] Compared with Example 1, the AChE content in the brain tissue of mice in Comparative Examples 1-6 increased, and the ChAT content decreased, and the decrease was significant, indicating that the oils and functional ingredients provided by the present invention can synergistically improve the AChE content and ChAT content in mice, thereby achieving the treatment of Alzheimer's disease.

[0131] In summary, the nanoemulsion provided by the present invention can prevent / alleviate Alzheimer's disease from two perspectives: lipid metabolism and cholinergic hypothesis.

[0132] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A nanoemulsion for preventing Alzheimer's disease, characterized in that: The invention comprises the following ingredients by weight: 11-22 parts of oils, 1.11-3.50 parts of emulsifiers, 1-3 parts of processing aids, 0.31-0.86 parts of functional ingredients, 0.01-0.03 parts of antioxidants and 70-90 parts of water; wherein the oils include Acer truncatum seed oil and Xanthoceras sorbifolia oil.

2. The nanoemulsion for preventing Alzheimer's disease according to claim 1, characterized in that The weight ratio of the Acer truncatum seed oil to the Xanthoceras sorbifolia oil is (6-12):(5-10).

3. The nanoemulsion for preventing Alzheimer's disease according to claim 1, characterized in that The functional ingredients include sage extract, walnut peptide and fermented jujube seed powder.

4. The nanoemulsion for preventing Alzheimer's disease according to claim 3, characterized in that The weight ratio of sage extract, walnut peptide and fermented jujube kernel powder in the functional ingredients is (0.01-0.06): (0.1-0.3): (0.2-0.5).

5. The nanoemulsion for preventing Alzheimer's disease according to claim 1, characterized in that The emulsifier comprises Tween 80, mono- and di-glycerol fatty acid esters and phospholipids; the weight ratio of Tween 80, mono- and di-glycerol fatty acid esters and phospholipids in the emulsifier is (0.01-0.05): (0.1-0.45): (1.0-3.0).

6. The nanoemulsion for preventing Alzheimer's disease according to claim 1, characterized in that The processing aid includes one or more of glycerol and ethanol; and the antioxidant is vitamin E.

7. The nanoemulsion for preventing Alzheimer's disease according to claim 1, characterized in that The invention comprises the following ingredients in parts by weight: 9 parts of Acer truncatum seed oil, 8 parts of Xanthoceras sorbifolia oil, 0.035 parts of Tween 80, 0.25 parts of mono- and diglycerol fatty acid esters, 2.0 parts of phospholipids, 0.035 parts of sage extract, 0.175 parts of walnut peptide, 0.35 parts of fermented jujube kernel powder, 2 parts of glycerol, 0.02 parts of vitamin E, and 78.135 parts of water.

8. A method for preparing the nanoemulsion for preventing Alzheimer's disease according to any one of claims 1 to 7, characterized in that: The steps include: (1) Mixing oil, emulsifier, processing aid and water, heating and stirring to fully dissolve the components, and obtaining a crude emulsion A; (2) Cooling the crude emulsion A to room temperature, adding an antioxidant and stirring to uniformly mix to obtain a crude emulsion B; (3) Shearing the crude emulsion B at room temperature, adding functional ingredients during the shearing process and adjusting the pH to 6-7, to obtain an emulsion C; (4) After high-pressure homogenization, cooling the emulsion C to room temperature to obtain a nanoemulsion.

9. The method for preparing the nanoemulsion for preventing Alzheimer's disease according to claim 8, characterized in that: The heating and stirring in step (1) is performed at a temperature of 60-75° C. for 15-30 minutes; The stirring time in step (2) is 10-20 min; The shearing time in step (3) is 15-25 min and the rotation speed is 6500-8000 rpm; In the step (4), the homogenization pressure is 200-400 bar, and the homogenization time is 20-80 min.

10. Use of the nanoemulsion according to any one of claims 1 to 7 and the nanoemulsion obtained by the preparation method according to any one of claims 8 to 9 in preparing a product for preventing Alzheimer's disease.

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