A nanostructured lipid carrier for Acer truncatum seed oil, its preparation method and application

Abstract

This invention discloses a nanostructured lipid carrier for Acer truncatum seed oil, its preparation method, and its applications. The invention involves dropwise adding an aqueous phase containing an emulsifier into an oil phase consisting of molten solid lipids and Acer truncatum seed oil. A primary emulsion is prepared using an ultrasonic cell disruptor, and the nanostructured lipid carrier is then prepared under homogenization pressure. The nanostructured lipid carrier significantly improves the stability, bioavailability, targeting ability, and long-lasting effect of Acer truncatum seed oil. The nanostructured lipid carrier prepared by this invention effectively enhances the antioxidant activity of Acer truncatum seed oil, strengthens its anti-aging effects, and the preparation process requires no high-temperature heating, exhibits stable performance, is simple, and has low production costs, making it suitable for large-scale industrial production applications.

Description

Technical Field

[0001] This invention belongs to the field of nanostructured lipid carrier technology, specifically relating to a nanostructured lipid carrier of Acer truncatum seed oil, its preparation method, and its application. Background Technology

[0002] *Acer truncatum* Bunge is a plant belonging to the genus *Acer* in the family Aceraceae. It is named for its winged fruits, which resemble the ancient Chinese ingot "yuanbao" (a type of gold ingot). Primarily found in the middle and lower reaches of the Yellow River and southern Northeast my country, it is not only endemic to my country but also an important resource plant with multiple uses, including edible, medicinal, ornamental, and industrial applications. *Acer truncatum* seed oil (ASO) is a functional oil extracted from the kernels of *Acer truncatum* seeds. It is a high-quality woody plant oil rich in unsaturated fatty acids, primarily oleic acid, linoleic acid, and a high content of nervonic acid. Nervonic acid can not only delay nerve cell aging and repair damage but also improve immunity, sleep quality, and lower blood lipids. *Acer truncatum* seed oil also contains abundant vitamin E and phytosterols; its vitamin E content is higher than that of common edible oils, exhibiting strong antioxidant properties that can scavenge free radicals and delay aging. In addition, studies have shown that Acer truncatum seed oil has anti-tumor, antibacterial, gut microbiota-regulating, immune-enhancing, cognitive-improving, inflammation-reducing, and myelin-regenerating effects in multiple sclerosis (MS) mice.

[0003] Nanostructured lipid carriers (NLCs) are a novel type of lipid carrier system. They utilize a mixture of lipids as a carrier, incorporating lipids that are liquid at room temperature into solid lipids, maintaining their solid state at room and body temperatures. These are nanoparticles with a particle size of less than 500 nm. NLCs exhibit excellent physical stability, providing stable storage conditions for the encapsulated active nutrients and reducing their oxidation. They can also mask unpleasant odors and tastes of the active nutrients, minimizing nutrient loss and improving user compliance. Furthermore, NLCs offer targeted and long-lasting effects. Due to their high bioavailability, high biocompatibility, and low toxicity, NLCs are widely used in the food and pharmaceutical industries.

[0004] Acer truncatum seed oil, a plant oil rich in natural active substances, is rich in various nutrients such as unsaturated fatty acids, linoleic acid, nervonic acid, and vitamin E. These components give Acer truncatum seed oil significant antioxidant properties, effectively scavenging free radicals in the body and delaying the aging process. However, Acer truncatum seed oil has low bioavailability in the intestines. Furthermore, its high content of unsaturated fatty acids makes it prone to oxidative deterioration under conditions of sufficient oxygen, light, and high temperature, resulting in unstable properties. This oxidative deterioration not only reduces its nutritional value but also produces substances harmful to the human body, leading to low safety. Therefore, improving the safety, stability, and bioavailability of Acer truncatum seed oil is an urgent problem to be solved.

[0005] Therefore, it is necessary to take certain measures to increase the stability of Acer truncatum seed oil in order to ensure product quality and safety, while also improving its bioavailability. Summary of the Invention

[0006] The primary objective of this invention is to provide a nanostructured lipid carrier for Acer truncatum seed oil to improve its safety, stability, and bioavailability.

[0007] Another objective of this invention is to provide a method for preparing a nanostructured lipid carrier of Acer truncatum seed oil that is simple to prepare and easy to implement.

[0008] Another objective of this invention is to provide an application of nanostructured lipids from Acer truncatum seed oil in Acer truncatum seed oil-related products.

[0009] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0010] A nanostructured lipid carrier for Acer truncatum seed oil, wherein the raw material formula of the nanostructured lipid carrier for Acer truncatum seed oil is: 3-5% Acer truncatum seed oil, 5-7% solid lipids, 2-4% emulsifier, and the balance being water.

[0011] Preferably, the raw material formula of the Acer truncatum seed oil nanostructure lipid carrier is: 5% Acer truncatum seed oil, 5% solid lipid, 4% emulsifier, and 86% water.

[0012] Preferably, in the raw material formulation of the Acer truncatum seed oil nanostructure lipid carrier, the solid lipid is selected from one or more combinations of glyceryl betaine (ATO888), glyceryl distearate (ATO5), cocoa butter, and palmitate.

[0013] More preferably, in the raw material formulation of the Acer truncatum seed oil nanostructure lipid carrier, the solid lipid is cocoa butter and palmitate.

[0014] Preferably, in the raw material formulation of the Acer truncatum seed oil nanostructured lipid carrier, the emulsifier is poloxamer 188.

[0015] The present invention discloses a method for preparing a nanostructured lipid carrier of Acer truncatum seed oil, comprising the following steps:

[0016] (1) Solid lipids and Acer truncatum seed oil are melted in a water bath and stirred evenly to obtain an oil phase solution;

[0017] (2) Add the emulsifier to water and melt it under water bath conditions, stirring until the emulsifier is dissolved to obtain an aqueous solution;

[0018] (3) Under magnetic stirring conditions, the aqueous phase solution in step (2) is added dropwise to the oil phase solution in step (1) to obtain a mixture;

[0019] (4) Place the mixture obtained in step (3) into an ultrasonic cell disruptor for ultrasonic treatment. Set the power of the ultrasonic cell disruptor to 200-600W and the ultrasonic treatment time to 4-12min to obtain colostrum.

[0020] (5) The colostrum obtained in step (4) is placed in a homogenizer for homogenization. The homogenization pressure is set to 250-1250 bar to obtain the Acer truncatum seed oil nanostructure lipid carrier.

[0021] Preferably, in step (4), the power of the ultrasonic cell disruptor is set to 300-500W and the ultrasonic time is 6-10min.

[0022] More preferably, in step (4), the power of the ultrasonic cell disruptor is set to 400W and the ultrasonic time is 8min.

[0023] Preferably, in step (5), the homogenization pressure is set to 1000 bar.

[0024] The nanostructured lipid carrier of Acer truncatum seed oil prepared in this invention is applied in anti-aging drugs.

[0025] The beneficial effects of this invention are as follows:

[0026] 1. This invention provides a nanostructured lipid carrier for Acer truncatum seed oil with a minimum particle size of 244.40 nm by employing ultrasonic cell disruption and high-pressure homogenization. This nanostructured lipid carrier for Acer truncatum seed oil has the advantages of good bioavailability, stability and targeting.

[0027] 2. Using *C. elegans* as a model, the results of *C. elegans* cell apoptosis staining and lifespan determination showed that the *Acer truncatum* seed oil nanostructured lipid carrier can reduce DNA damage in *C. elegans* and maintain cell viability and integrity to the maximum extent. Therefore, the *Acer truncatum* seed oil nanostructured lipid carrier has good stability and targeting properties, and can prolong the lifespan of *C. elegans*.

[0028] 3. The present invention, through the determination of antioxidant activity in Caenorhabditis elegans and the results of acute oxidative stress test, shows that the nanostructured lipid carrier of Acer truncatum seed oil can improve the activity of antioxidant enzymes in the body, enhance the antioxidant capacity of Caenorhabditis elegans, alleviate oxidative damage and aging of Caenorhabditis elegans, activate the potential of the antioxidant defense system in Caenorhabditis elegans, improve bioavailability in the body, and improve the stability of Acer truncatum seed oil.

[0029] 4. This invention demonstrates, through measurements of the motility and reproductive capacity of *C. elegans*, that the nanostructured lipid carrier of *Acer truncatum* seed oil can extend the lifespan of *C. elegans* without affecting its reproductive capacity, and significantly improve its motility. The nanostructured lipid carrier of *Acer truncatum* seed oil can maintain cell vitality, slow down the aging process, and prolong lifespan.

[0030] 5. This invention demonstrates through mortality tests on *C. elegans* that *Acer truncatum* seed oil and its nanostructured lipid carrier do not produce toxic effects on *C. elegans* at a concentration of 500 μg / mL, indicating that the nanostructured lipid carrier of *Acer truncatum* seed oil has good safety during use.

[0031] 6. The nanostructured lipid carrier for Acer truncatum seed oil provided by this invention has a simple process, low production cost, and can be applied on a large scale in industrial production. Attached Figure Description

[0032] Figure 1 The effect of ultrasonic time in a cell disruptor on the particle size of the nanostructured lipid carrier of Acer truncatum seed oil;

[0033] Figure 2 The effect of ultrasonic power of a cell disruptor on the particle size of nanostructured lipid carriers for Acer truncatum seed oil.

[0034] Figure 3 The effect of homogenization pressure on the particle size of nanostructured lipid carriers in Acer truncatum seed oil.

[0035] Figure 4 Transmission electron microscopy image of a nanostructured lipid carrier for Acer truncatum seed oil;

[0036] Figure 5 The effect of Acer truncatum seed oil nanostructured lipid carriers on the survival curve of Caenorhabditis elegans;

[0037] Figure 6 The effect of Acer truncatum seed oil nanostructured lipid carriers on oviposition of Caenorhabditis elegans;

[0038] Figure 7 The effect of Acer truncatum seed oil nanostructured lipid carriers on the motility of Caenorhabditis elegans;

[0039] Figure 8 The effect of Acer truncatum seed oil nanostructured lipid carriers on heat stress in Caenorhabditis elegans;

[0040] Figure 9 The effect of Acer truncatum seed oil nanostructured lipid carriers on acute oxidative stress in Caenorhabditis elegans;

[0041] Figure 10 The effect of Acer truncatum seed oil nanostructured lipid carrier on the antioxidant capacity of Caenorhabditis elegans in vivo;

[0042] Figure 11 The effect of Acer truncatum seed oil nanostructured lipid carrier on apoptosis in Caenorhabditis elegans cells. Detailed Implementation

[0043] The present invention will be further described below through specific embodiments, but these are not intended to limit the scope of protection of the present invention.

[0044] Example 1:

[0045] Formula: 5% Acer truncatum seed oil, 4% cocoa butter, 1% palmitate, 18.84% poloxamer, 86% water.

[0046] Prepare the above formula according to preparation methods 1-6.

[0047] Preparation method 1:

[0048] (1) Cocoa butter, palmitate palmitate and Acer truncatum seed oil are melted in a water bath and stirred evenly to obtain an oil phase solution;

[0049] (2) Add poloxamer 188 to water and melt it under water bath conditions, stirring until poloxamer 188 dissolves to obtain an aqueous solution;

[0050] (3) Under magnetic stirring conditions, the aqueous phase solution in step (2) is added dropwise to the oil phase solution in step (1) to obtain a mixture;

[0051] (4) Place the mixture obtained in step (3) into an ultrasonic cell disruptor for ultrasonic treatment. The power of the ultrasonic cell disruptor is set to 400W and the ultrasonic treatment time is 8min to obtain colostrum.

[0052] (5) The colostrum obtained in step (4) is placed in a homogenizer for homogenization. The homogenization pressure is set to 1000 bar to obtain the Acer truncatum seed oil nanostructure lipid carrier.

[0053] Preparation method 2:

[0054] (1) Cocoa butter, palmitate palmitate and Acer truncatum seed oil are melted in a water bath and stirred evenly to obtain an oil phase solution;

[0055] (2) Add poloxamer 188 to water and melt it under water bath conditions, stirring until poloxamer 188 dissolves to obtain an aqueous solution;

[0056] (3) Under magnetic stirring conditions, the aqueous phase solution in step (2) is added dropwise to the oil phase solution in step (1) to obtain a mixture;

[0057] (4) Place the mixture obtained in step (3) into an ultrasonic cell disruptor for ultrasonic treatment. The power of the ultrasonic cell disruptor is set to 200W and the ultrasonic treatment time is 4min to obtain colostrum.

[0058] (5) The colostrum obtained in step (4) is placed in a homogenizer for homogenization. The homogenization pressure is set to 250 bar to obtain the Acer truncatum seed oil nanostructure lipid carrier.

[0059] Preparation method 3:

[0060] (1) Cocoa butter, palmitate palmitate and Acer truncatum seed oil are melted in a water bath and stirred evenly to obtain an oil phase solution;

[0061] (2) Add poloxamer 188 to water and melt it under water bath conditions, stirring until poloxamer 188 dissolves to obtain an aqueous solution;

[0062] (3) Under magnetic stirring conditions, the aqueous phase solution in step (2) is added dropwise to the oil phase solution in step (1) to obtain a mixture;

[0063] (4) Place the mixture obtained in step (3) into an ultrasonic cell disruptor for ultrasonic treatment. The power of the ultrasonic cell disruptor is set to 300W and the ultrasonic treatment time is 6min to obtain colostrum.

[0064] (5) The colostrum obtained in step (4) is placed in a homogenizer for homogenization. The homogenization pressure is set to 500 bar to obtain the Acer truncatum seed oil nanostructure lipid carrier.

[0065] Preparation method 4:

[0066] (1) Cocoa butter, palmitate palmitate and Acer truncatum seed oil are melted in a water bath and stirred evenly to obtain an oil phase solution;

[0067] (2) Add poloxamer 188 to water and melt it under water bath conditions, stirring until poloxamer 188 dissolves to obtain an aqueous solution;

[0068] (3) Under magnetic stirring conditions, the aqueous phase solution in step (2) is added dropwise to the oil phase solution in step (1) to obtain a mixture;

[0069] (4) Place the mixture obtained in step (3) into an ultrasonic cell disruptor for ultrasonic treatment. The power of the ultrasonic cell disruptor is set to 400W and the ultrasonic treatment time is 8min to obtain colostrum.

[0070] (5) The colostrum obtained in step (4) is placed in a homogenizer for homogenization. The homogenization pressure is set to 750 bar to obtain the Acer truncatum seed oil nanostructure lipid carrier.

[0071] Preparation method 5:

[0072] (1) Cocoa butter, palmitate palmitate and Acer truncatum seed oil are melted in a water bath and stirred evenly to obtain an oil phase solution;

[0073] (2) Add poloxamer 188 to water and melt it under water bath conditions, stirring until poloxamer 188 dissolves to obtain an aqueous solution;

[0074] (3) Under magnetic stirring conditions, the aqueous phase solution in step (2) is added dropwise to the oil phase solution in step (1) to obtain a mixture;

[0075] (4) Place the mixture obtained in step (3) into an ultrasonic cell disruptor for ultrasonic treatment. Set the power of the ultrasonic cell disruptor to 500W and the ultrasonic treatment time to 10min to obtain colostrum.

[0076] (5) The colostrum obtained in step (4) is placed in a homogenizer for homogenization. The homogenization pressure is set to 1000 bar to obtain the Acer truncatum seed oil nanostructure lipid carrier.

[0077] Preparation method 6:

[0078] (1) Cocoa butter, palmitate palmitate and Acer truncatum seed oil are melted in a water bath and stirred evenly to obtain an oil phase solution;

[0079] (2) Add poloxamer 188 to water and melt it under water bath conditions, stirring until poloxamer 188 dissolves to obtain an aqueous solution;

[0080] (3) Under magnetic stirring conditions, the aqueous phase solution in step (2) is added dropwise to the oil phase solution in step (1) to obtain a mixture;

[0081] (4) Place the mixture obtained in step (3) into an ultrasonic cell disruptor for ultrasonic treatment. The power of the ultrasonic cell disruptor is set to 600W and the ultrasonic treatment time is 12min to obtain colostrum.

[0082] (5) The colostrum obtained in step (4) is placed in a homogenizer for homogenization. The homogenization pressure is set to 1250 bar to obtain the Acer truncatum seed oil nanostructure lipid carrier.

[0083] Example 2:

[0084] Formula: 1% Acer truncatum seed oil, 9% sorbic acid glycerides, 18.84% poloxamer, 86% water.

[0085] Example 3:

[0086] Formula: 2% Acer truncatum seed oil, 8% glyceryl sorbate, 18.84% poloxamer, 86% water.

[0087] Example 4:

[0088] Formula: 3% Acer truncatum seed oil, 7% sorbic acid glycerides, 18.84% poloxamer, 86% water.

[0089] Example 5:

[0090] Formula: 4% Acer truncatum seed oil, 6% sorbic acid glycerides, 18.84% poloxamer, 86% water.

[0091] Example 6:

[0092] Formula: 5% Acer truncatum seed oil, 5% sorbic acid glycerides, 18.84% poloxamer, 86% water.

[0093] Example 7:

[0094] Any of the formulations in Examples 2-6 were prepared according to the preparation methods 1-6 in Examples 1, except that the cocoa butter and palmitate in preparation methods 1-6 were replaced with glyceryl succinate in any of the formulations in Examples 2-6.

[0095] Example 8:

[0096] Formula: 1% Acer truncatum seed oil, 9% glyceryl distearate, 18.84% poloxamer, 86% water.

[0097] Example 9:

[0098] Formula: 2% Acer truncatum seed oil, 8% glyceryl distearate, 18.84% poloxamer, 86% water.

[0099] Example 10:

[0100] Formula: 3% Acer truncatum seed oil, 7% glyceryl distearate, 18.84% poloxamer, 86% water.

[0101] Example 11:

[0102] Formula: 4% Acer truncatum seed oil, 6% glyceryl distearate, 18.84% poloxamer, 86% water.

[0103] Example 12:

[0104] Formula: 5% Acer truncatum seed oil, 5% glyceryl distearate, 18.84% poloxamer, 86% water.

[0105] Example 13:

[0106] Any of the formulations in Examples 8-12 was prepared according to the preparation methods 1-6 in Examples 1, except that the cocoa butter and palmitate palmitate in preparation methods 1-6 were replaced with glyceryl distearate in any of the formulations in Examples 8-12.

[0107] Example 14:

[0108] Formula: 5% Acer truncatum seed oil, 5% cocoa butter, 18.84% poloxamer, 86% water.

[0109] Example 15:

[0110] Formula: 4% Acer truncatum seed oil, 6% cocoa butter, 18.84% poloxamer, 86% water.

[0111] Example 16:

[0112] Formula: 3% Acer truncatum seed oil, 7% cocoa butter, 18.84% poloxamer, 86% water.

[0113] Example 17:

[0114] Formula: 2% Acer truncatum seed oil, 8% cocoa butter, 18.84% poloxamer, 86% water.

[0115] Example 18:

[0116] Formula: 1% Acer truncatum seed oil, 9% cocoa butter, 18.84% poloxamer, 86% water.

[0117] Example 19:

[0118] Any of the formulations in Examples 14-19 was prepared according to the preparation methods 1-6 in Examples 1, except that the cocoa butter and palmitate palmitate in preparation methods 1-6 were replaced with cocoa butter in any of the formulations in Examples 14-19.

[0119] Example 20:

[0120] Formula: 5% Acer truncatum seed oil, 5% palmitate, 18.84% poloxamer, 86% water.

[0121] Example 21:

[0122] Formula: 4% Acer truncatum seed oil, 6% palmitate, 18.84% poloxamer, 86% water.

[0123] Example 22:

[0124] Formula: 3% Acer truncatum seed oil, 7% palmitate, 18.84% poloxamer, 86% water.

[0125] Example 23:

[0126] Formula: 2% Acer truncatum seed oil, 8% palmitate, 18.84% poloxamer, 86% water.

[0127] Example 24:

[0128] Formula: 1% Acer truncatum seed oil, 9% palmitate, 18.84% poloxamer, 86% water.

[0129] Example 25:

[0130] Any of the formulations in Examples 20-24 were prepared according to the preparation methods 1-6 in Examples 1, except that the cocoa butter and palmitate in preparation methods 1-6 were replaced with palmitate in any of the formulations in Examples 20-24.

[0131] Example 26:

[0132] Formula: 5% Acer truncatum seed oil, 1% cocoa butter, 4% palmitate, 18.84% poloxamer, 86% water.

[0133] Example 27:

[0134] Formula: 5% Acer truncatum seed oil, 2% cocoa butter, 3% palmitate, 18.84% poloxamer, 86% water.

[0135] Example 28:

[0136] Formula: 5% Acer truncatum seed oil, 3% cocoa butter, 2% palmitate, 18.84% poloxamer, 86% water.

[0137] Example 29:

[0138] Any of the formulations in Examples 26-28 were prepared according to the preparation methods 1-6 in Example 1.

[0139] Experimental example:

[0140] The inventors conducted extensive experimental research on the formula, process parameters, and efficacy of this invention, some of which are excerpted below:

[0141] 1. Solid lipid optimization experiment

[0142] Solid lipids possess excellent stability. Due to their high melting point and high crystallinity, they remain stable in a solid state at room temperature and are not prone to oxidation, hydrolysis, or other chemical reactions. Simultaneously, solid lipids exhibit good biocompatibility, lubricity, and spreadability, allowing for uniform distribution within products. Furthermore, solid lipids have an adjustable melting point range; by altering their chemical composition and crystal structure, the melting point can be precisely controlled, enabling them to adapt to the needs of different products. To improve the digestion and absorption of Acer truncatum seed oil nanostructured lipid carriers in the human body, the inventors of this invention conducted experimental screening of mixed lipids of four solid lipids—glyceryl betaine, glyceryl distearate, cocoa butter, and palmitate—with Acer truncatum seed oil. The aim is to prepare a Acer truncatum seed oil nanostructured lipid carrier with a melting point of 35–40℃. This temperature is close to the temperature of the human stomach, further facilitating its digestion and absorption in the human body. By observing the fusion of solid lipids with Acer truncatum seed oil, solid lipids that can be used to prepare Acer truncatum seed oil nanostructured lipid carriers with stable storage and precise melting point were selected.

[0143] Table 1 shows a solid lipid screening method applicable to nanostructured lipid carriers of Acer truncatum seed oil.

[0144]

[0145]

[0146] Experimental Results: As can be seen from Table 1, due to the addition of a solid lipid, it is impossible to obtain a solid lipid nanostructure lipid carrier for Acer truncatum seed oil with moderate hardness and a melting point temperature range of 35℃~40℃ that is conducive to human absorption. Moreover, the amount of Acer truncatum seed oil added is too low. Therefore, it is recommended to mix two solid lipids with low melting points (cocoa butter and palmitate) to increase the content of Acer truncatum seed oil for further investigation.

[0147] Table 2 shows two solid lipids suitable for screening nanostructured lipid carriers of Acer truncatum seed oil.

[0148] Case Solid lipid 1 / use amount g Solid lipids 2 / use amount g Liquid lipids / usage g room temperature 1 Cocoa butter --- 1g Palmitate palmitate---4g Acer truncatum seed oil --- 5g Softer 2 Cocoa butter --- 2g Palmitate palmitate---3g Acer truncatum seed oil --- 5g Softer 3 Cocoa butter --- 3g Palmitate palmitate---2g Acer truncatum seed oil --- 5g Softer 4 Cocoa butter --- 4g Palmitate palmitate --- 1g Acer truncatum seed oil --- 5g The temperature is suitable, and the firmness is just right.

[0149] Experimental results: As can be seen from Table 2, the composition formulation of Case 4 can be completely melted at 35℃~40℃. Therefore, cocoa butter and palmitate palmitate are preferred as solid lipids for the nanostructure lipid carrier of Acer truncatum seed oil, and subsequent experiments were conducted in this ratio.

[0150] 2. Experiment on optimizing the ratio of emulsifier to water

[0151] The nanostructured lipid carrier comprises a lipid phase and an aqueous phase. The lipid phase consists of solid and liquid lipids, while the aqueous phase consists of an emulsifier and water. The total mass of the lipid phase does not exceed 10% of the total mass of the nanostructured lipid carrier. Therefore, the aqueous phase comprises 90% of the total volume.

[0152] The commonly used emulsifier was chosen as poloxamer 188 because it has non-ionic properties, which can reduce interfacial tension and allow the oil and water phases to mix stably together to form an emulsion. It has excellent emulsifying properties, which enhance its stability.

[0153] Preparation method: Poloxamer 188 was added to water and stirred until dissolved. When the amount of poloxamer 188 added was 3%, a small number of oil droplets were still suspended on the surface of the nanostructured lipid carrier. When the amount added was 4%, no oil droplets were suspended on the surface of the nanostructured lipid carrier. Therefore, the preferred emulsifier ratio was 4% poloxamer 188 and water was 86%, and subsequent experiments were conducted using this ratio.

[0154] 3. Single-factor experiment on preparation process

[0155] Using liposome particle size and encapsulation efficiency as indicators, the effects of cell disruptor sonication time, sonication power, and homogenization pressure on the particle size and particle size distribution index (PDI) of Acer truncatum seed oil nanoliposome carriers were investigated through single-factor experiments.

[0156] Particle size: The particle size of the nanolipid carrier is determined using a particle size analyzer. The smaller the particle size of the nanolipid carrier, the better its absorption, distribution and metabolism in the human body.

[0157] Particle size distribution index (PDI): This coefficient is an indicator of whether the nanolipocarrier formulation is uniformly distributed. A PDI < 0.3 indicates that the nanolipocarrier formulation is uniformly distributed.

[0158] 3.1 Effect of cell disruptor sonication time on the particle size of Acer truncatum seed oil nanolipid carriers

[0159] Prescription: Take 5 parts of Acer truncatum seed oil, 4 parts of cocoa butter, 1 part of palmitate, 1884 parts of poloxamer, and 86 parts of water.

[0160] Preparation method: Cocoa butter, palmitate palmitate, and Acer truncatum seed oil were melted in a water bath and stirred until homogeneous to obtain an oil phase solution; Poloxamer 188 was added to water and melted in a water bath, stirred until Poloxamer 188 dissolved to obtain an aqueous phase solution; Under magnetic stirring, the aqueous phase solution was added dropwise to the oil phase solution to obtain a mixture; The resulting mixture was placed in an ultrasonic cell disruptor for ultrasonic treatment at a power of 300W for ultrasonic treatment times of 4 min, 6 min, 8 min, 10 min, and 12 min to obtain a colostrum; The obtained colostrum was homogenized in a homogenizer at a pressure of 750 bar to obtain the Acer truncatum seed oil nanostructured lipid carrier.

[0161] The particle size and particle size distribution index (PDI) of the nanostructured lipid carriers of Acer truncatum seed oil were determined. The experimental results are shown in […]. Figure 1 .

[0162] Experimental results: From Figure 1 It can be seen that the particle size change is relatively slow when the ultrasonic time is 4 min and 6 min, but decreases significantly at 8 min (P < 0.01). The particle size change is slow at 8 min, 10 min, and 12 min. Particle size distribution is described by the polydispersity index (PDI), which indicates the uniformity of nanoparticle distribution. The PDI fluctuates around 0.15 between 4 and 12 min. It is generally believed that smaller particles are more effective in achieving the desired target, with a PDI value of 0.1-0.25 indicating a narrow particle size distribution and 0.5 indicating a wide particle size distribution. Measurements of the prepared samples confirmed that the liposome distribution was narrow; therefore, a parameter indicating a smaller particle size should be selected, and an ultrasonic time of 8 min was chosen.

[0163] 3.2 Effect of Ultrasonic Power of Cell Disruptor on Particle Size of Acer truncatum Seed Oil Nanolipid Carrier

[0164] Prescription: Take 5 parts of Acer truncatum seed oil, 4 parts of cocoa butter, 1 part of palmitate, 1884 parts of poloxamer, and 86 parts of water.

[0165] Preparation method: Cocoa butter, palmitate palmitate, and Acer truncatum seed oil were melted in a water bath and stirred until homogeneous to obtain an oil phase solution; Poloxamer 188 was added to water and melted in a water bath, stirred until Poloxamer 188 dissolved to obtain an aqueous phase solution; Under magnetic stirring, the aqueous phase solution was added dropwise to the oil phase solution to obtain a mixture; The resulting mixture was placed in an ultrasonic cell disruptor for ultrasonic treatment with power settings of 200W, 300W, 400W, 500W, and 600W for 6 minutes to obtain a colostrum; The obtained colostrum was homogenized in a homogenizer at a pressure of 750 bar to obtain the Acer truncatum seed oil nanostructured lipid carrier.

[0166] The particle size and particle size distribution index (PDI) of the Acer truncatum seed oil nanolipid carrier were determined. The experimental results are shown in […]. Figure 2 .

[0167] Experimental results: From Figure 2 It can be seen that when the power increases from 200W to 400W, the particle size decreases significantly (P<0.05), while when the power increases to 400W, the particle size does not change significantly. Further increasing the power gradually increases the particle size (P<0.05). Therefore, 400W is selected as the optimal ultrasonic power parameter for preparation.

[0168] 3.3 Effect of Homogenization Pressure on the Particle Size of Acer truncatum Seed Oil Nanolipid Carriers

[0169] Prescription: Take 5 parts of Acer truncatum seed oil, 4 parts of cocoa butter, 1 part of palmitate, 1884 parts of poloxamer, and 86 parts of water.

[0170] Preparation method: Cocoa butter, palmitate palmitate, and Acer truncatum seed oil were melted in a water bath and stirred until homogeneous to obtain an oil phase solution; poloxamer 188 was added to water and melted in a water bath, stirred until poloxamer 188 dissolved to obtain an aqueous phase solution; the aqueous phase solution was added dropwise to the oil phase solution under magnetic stirring to obtain a mixture; the resulting mixture was placed in an ultrasonic cell disruptor for ultrasonic treatment at a power of 300W for 6 minutes to obtain a colostrum; the obtained colostrum was homogenized in a homogenizer at pressures of 250 bar, 500 bar, 750 bar, 1000 bar, and 1250 bar to obtain the Acer truncatum seed oil nanostructured lipid carrier.

[0171] The particle size and particle size distribution index (PDI) of the Acer truncatum seed oil nanolipid carrier were determined. The experimental results are shown below. Figure 3 .

[0172] Experimental results: From Figure 3 It can be seen that when the homogenization pressure increases from 250 bar to 1000 bar, the particle size gradually decreases (P < 0.01), while when the pressure increases to 1250 bar, the particle size increases significantly (P < 0.01). Therefore, when considering the particle size of liposomes, a homogenization pressure of 1000 bar is the best choice.

[0173] 3.4 Characterization of the properties of nanostructured lipid carriers in Acer truncatum seed oil

[0174] 3.4.1 Appearance and centrifugal stability

[0175] The Acer truncatum seed oil nanostructured lipid carrier prepared according to the formulation of Example 1 of this invention and preparation method 1 in Example 1 of this invention is a milky white liquid with a special aroma of cocoa butter and a uniform and delicate appearance. No layering or precipitation was observed when centrifuged at 5000 r / min for 5 min and at 10000 r / min for 3 min, indicating that it has good centrifugal stability.

[0176] 3.4.2 Microscopic morphological observation of the nanostructured lipid carrier of Acer truncatum seed oil

[0177] Preparation method: The Acer truncatum seed oil nanostructured lipid carrier prepared according to preparation method 1 in Example 1 of this invention was diluted 100 times with distilled water and then dropped onto a copper grid. After drying at room temperature, its morphology was observed and photographed using transmission electron microscopy (TEM). The results are shown below. Figure 4 .

[0178] from Figure 4 As can be seen from the transmission electron microscope, the microstructure of the Acer truncatum seed oil nanostructure lipid carrier is spherical and evenly distributed.

[0179] 3.4.3 Determination of particle size, polydispersity index, and zeta potential of the Acer truncatum seed oil nanostructured lipid carrier

[0180] The nanostructured lipid carrier of Acer truncatum seed oil prepared according to the formulation of Example 1 of this invention was diluted with 3 mL of purified water and measured using a Brookhaven Zeta potential and particle size analyzer.

[0181] Experimental results showed that the Acer truncatum seed oil nanostructured lipid carrier had a particle size of 244.40±4.46 nm and a PDI of 0.17±0.02, which met the expected requirements, i.e., a particle size of 100-500 nm. Oral administration of this material can improve drug stability. Furthermore, a particle size distribution with a PDI below 0.3 is beneficial for the long-term stability of the nanostructured lipid carrier, and the particle size distribution is uniform. The Zeta potential reflects the degree of repulsion between charged particles; the Zeta potential of ASO-NLC was -30.07±0.95 mV, which can provide sufficient repulsive force to maintain the stability of the nanostructured lipid carrier. These results indicate that the Acer truncatum seed oil nanostructured lipid carrier exhibits a normal, concentrated, and uniform particle size distribution, demonstrating its excellent stability and bioavailability.

[0182] 3.4.4 Determination of Encapsulation Efficiency

[0183] This invention uses a cell wall disruption-low speed centrifugation method to determine the encapsulation efficiency of the nanostructured lipid carrier of Acer truncatum seed oil.

[0184] Dissolve an appropriate amount of Acer truncatum seed oil in petroleum ether. Following the formulation of Example 1 of this invention, prepare the Acer truncatum seed oil nanostructured lipid carrier and blank lipid carrier using preparation method 1 in Example 1 of this invention. After demulsification with petroleum ether, dilute an appropriate amount of the supernatant and perform a full-wavelength UV scan (190-400 nm), selecting the UV measurement wavelength for Acer truncatum seed oil. Add 5 mL of Acer truncatum seed oil nanostructured lipid carrier to 20 mL of petroleum ether and sonicate for 8 min in an ultrasonic cell disruptor. Transfer the mixed solution to a centrifuge tube and sonicate for another 10 min to completely demulsify. Centrifuge at 4000 r / min for 5 min and collect the supernatant for later use. Perform the same procedure on another sample except for cell disruption. Simultaneously, treat the nanostructured lipid carrier without Acer truncatum seed oil in the same way to obtain two supernatants to eliminate matrix influence. The absorbance of the obtained supernatant was measured at a selected wavelength. A certain amount of Acer truncatum seed oil was weighed, dissolved, and diluted to volume to obtain a standard solution. 0, 2, 4, 6, 8, and 10 mL of this standard solution were then diluted to 10 mL to obtain a series of Acer truncatum seed oil standard solutions. The absorbance of each standard solution was measured at a selected wavelength. The calculation formula is shown in the formula below.

[0185]

[0186] Experimental results: Based on the ultraviolet scanning spectra of Acer truncatum seed oil and the matrix required for preparing the nanostructured lipid carrier at wavelengths of 190-400 nm, selective absorption was measured at a wavelength of 265 nm. According to the formula, the encapsulation rate of the Acer truncatum seed oil nanostructured lipid carrier prepared by this invention was calculated to be 96.95%. The results show that the carrier structure can greatly improve the absorption of Acer truncatum seed oil, thereby enhancing the anti-aging effect.

[0187] 4. Research on the application of Acer truncatum seed oil nanostructured lipid carriers in products for improving aging

[0188] This experiment uses *Caenorhabditis elegans* as a model to explore the anti-aging ability of nanostructured lipid carriers in Acer truncatum seed oil, providing a theoretical basis for the development of functionalized Acer truncatum seed oil products.

[0189] 4.1 Experimental Materials and Reagents

[0190] Wild-type C. elegans (the Bristol Strain N2), Fujian Shangyuan Biotechnology Co., Ltd.; Acer truncatum seed oil, Guizhou Xuande Biotechnology Co., Ltd.; Cocoa butter, Shanghai Yien Chemical Technology Co., Ltd.; Palmitate palmitate, Shanghai Yien Chemical Technology Co., Ltd.; Poloxamer 188, Shandong Yousuo Chemical Technology Co., Ltd.; NGM medium (20230910), Ruichu Biotechnology Jiangsu Co., Ltd.; Peptone (20230630), yeast extract (20230718), Beijing Aoboxing Biotechnology Co., Ltd.; Malondialdehyde (MDA) kit, glutathione peroxidase (GSH-Px) kit, catalase (CAT) kit, superoxide dismutase (SOD) kit, Nanjing Urban Construction Bioengineering Institute.

[0191] M9 buffer solution: 3g Na2HPO4, 1.5g KH2PO4, 2.5g NaCl, 0.125g MgSO4·7H2O, add water to 500mL, sterilize at 121℃ for 15min, prepare and use immediately.

[0192] Acetylcholine (ACh) kit, acetylcholinesterase (AChE) kit, superoxide dismutase (SOD) kit, and malondialdehyde (MDA) kit were all purchased from Nanjing Jiancheng Bioengineering Institute.

[0193] 4.2 Instruments and Equipment

[0194] SCIENTZ-ПD Ultrasonic Cell Disruptor, Ningbo Xinzhi Biotechnology Co., Ltd.; 3000puls High-Pressure Microfluidic Homogenizer, Gui'an New District Ruicheng Bioengineering Co., Ltd.; NanoBrook 90plus PALS Zeta Potential and Particle Size Analyzer, Brookhaven Instruments, Inc.; UV1800 UV-Vis Spectrophotometer, Shimadzu Corporation; MD50 Stereomicroscope, Guangzhou Mingmei Optoelectronic Technology Co., Ltd.; LDZX-50L Vertical High-Pressure Steam Sterilizer, Shanghai Shenan Medical Instrument Factory; SR205DU 0.00000g Balance, Mettler Toledo Group; NH-40BS Electric Thermostatic Incubator, Shanghai Lichen Bangxi Instrument Technology Co., Ltd.; ZWY-2101 Vertical Double-Layer Full-Temperature Shaker, Shanghai Zhicheng Analytical Instrument Manufacturing Co., Ltd.; Varioskan LUX Microplate Reader, Thermo Fisher Scientific (China) Co., Ltd.

[0195] 4.3 Experimental Methods

[0196] 4.3.1 Culture and passage of Caenorhabditis elegans

[0197] Preparation method: Caenorhabditis elegans in the oviposition period were selected and placed on Caenorhabditis elegans growth medium (NGM) coated with Escherichia coli OP50 and incubated at a constant temperature of 20℃ in a biochemical incubator for subsequent experiments.

[0198] 4.3.2 Synchronization of Caenorhabditis elegans

[0199] Preparation method: Synchronization culture of *C. elegans* was performed using sodium perchlorate bleaching. Oviposition-stage adults were washed into sterile EP tubes with M9 buffer solution. Lysis buffer was added to lyse the *C. elegans*, followed by shaking and centrifugation. The supernatant was removed, and the tubes were washed 2-3 times with M9 buffer solution. After centrifugation and discarding the supernatant, *C. elegans* from the bottom of the EP tube were dropped into the sterile NGM area. After approximately 48 hours, the eggs had largely developed into L4-stage adults, completing the synchronization process, which was then applied to subsequent experiments.

[0200] 4.3.3 Test on the lethality of *C. elegans* var. *truncatum* seed oil nanostructured lipid carrier.

[0201] Acer truncatum seed oil and Acer truncatum seed oil nanostructure lipid carrier were prepared into different concentrations (0, 0.1, 1, 10, 100, 200, 500 μg / mL), and cultured on NGM plates (without OP50) at 20℃ for 24 h to observe the lethality of Caenorhabditis elegans.

[0202] Experimental Results: After culturing *C. elegans* at 20℃ for 24 h on NGM plates (without OP50) with various concentrations of *Acer truncatum* seed oil and its nanostructured lipid carrier, no mortality was observed in any group. This indicates that *Acer truncatum* seed oil and its nanostructured lipid carrier do not produce toxic effects on *C. elegans* at a concentration of 500 μg / mL, demonstrating the good safety profile of the *Acer truncatum* seed oil nanostructured lipid carrier.

[0203] 4.3.4 Experimental Grouping

[0204] Preparation method: Caenorhabditis elegans was divided into a blank control group (NC), an ASO group (ASO-100) with an administration dose of 100 μg / mL, and high, medium, and low dose groups of ASO-NLC with administration doses of 50, 100, and 200 μg / mL (named ASO-NLC-50, ASO-NLC-100, and ASO-NLC-200, respectively), which were used in subsequent experiments.

[0205] 4.3.5 Experiment on the lifespan determination of *C. elegans* using *Acer truncatum* seed oil nanostructured lipid carrier

[0206] Preparation method: Sample groups were mixed with E. coli OP50 bacterial suspension and spread on NGM plates. 50 *C. elegans* worms were placed on each plate, with three replicates per group. The plates were incubated at 20°C. Synchronized L4-stage adults were picked and transferred to each plate, marked as day 0. For the first five days, *C. elegans* worms were transferred to new plates daily, and their survival was observed and recorded daily. After 5 days, *C. elegans* worms were transferred to new plates every other day, and the survival and death counts were recorded again, until all *C. elegans* worms in all groups were dead (death was defined as *C. elegans* showing no movement and no response after 10 seconds of platinum-gold stimulation).

[0207] Experimental results: Lifespan is the most important indicator for measuring aging in *C. elegans*. The effect of ASO-NLC on the lifespan of *C. elegans* is shown in Table 3. Figure 5 The results showed that ASO-NLC intervention significantly prolonged the lifespan of *C. elegans*. The low, medium, and high doses of ASO-NLC showed a dose-response relationship on the median and mean lifespan. The longest lifespan in all three ASO-NLC dose groups was more than 50.00% longer than that in the NC group, while the ASO group only saw a 14.29% increase. ASO-NLC was superior to the ASO group. The high-dose ASO-NLC group showed the most significant effect, with a 36.00% increase in mean lifespan compared to the control group.

[0208] Table 3. Effects of ASO-NLC on the lifespan of *C. elegans*

[0209] Group Median lifespan / d Average lifespan / d Maximum lifespan / d NC 10±0.6 7.5±1.1 14±1.1 ASO-100 11±0.7 9.6±0.9 <![CDATA[16±0.8 * ]]> ASO-NLC-50 10±0.4 <![CDATA[8.4±1.3 * ]]> <![CDATA[21±1.1 ** ]]> ASO-NLC-100 11±0.6 <![CDATA[9.7±1.1 ** ]]> <![CDATA[22±1.2 ** ]]> ASO-NLC-200 <![CDATA[13±0.5 * ]]> <![CDATA[10.2±1.2 ** ]]> <![CDATA[22±1.1 ** ]]>

[0210] 4.3.6 Determination of the reproductive capacity of *C. elegans* by *Acer truncatum* seed oil nanostructured lipid carrier

[0211] Preparation method: One *C. elegans* was selected from each group and cultured separately, with three replicates for each sample. Every 24 hours, the *C. elegans* was transferred to a new *C. elegans* culture medium until it lost its reproductive capacity. The transferred *C. elegans* were then incubated at 20°C for another 24 hours. The oviposition of the *C. elegans* was recorded daily, and the total number of ovipositions on all transferred *C. elegans* growth media was calculated to obtain the total number of ovipositions per *C. elegans*.

[0212] Experimental results: The oviposition rate of *C. elegans* is directly related to its reproductive capacity. Generally, reproductive capacity is negatively correlated with lifespan, but some reports indicate that *Dendrobium officinale* leaf polysaccharides and resveratrol can prolong the lifespan of *C. elegans* without affecting its reproductive capacity. Figure 6As shown, compared with the NC group, the oviposition rates of the ASO-100, ASO-NLC-50, and ASO-NLC-100 groups were slightly increased, but the differences were not statistically significant; while the ASO-NLC-200 group showed a 14% increase, which was statistically significant (P<0.05). This indicates that ASO-NLC can prolong the lifespan of Caenorhabditis elegans without affecting its reproductive capacity.

[0213] 4.3.7 Test on the effect of Acer truncatum seed oil nanostructured lipid carrier on the motility of Caenorhabditis elegans

[0214] Preparation method: Gently touch the body of *C. elegans* with a platinum wire needle. If the *C. elegans* exhibits continuous and coordinated sinusoidal movement within 30 seconds, it is recorded as rapid movement; otherwise, it is recorded as slow movement. Movement levels are divided into three levels: spontaneous and steady movement, labeled A; movement following stimulation in a non-sinusoidal trajectory, labeled B; and no forward movement, but a response to touch via head or tail swaying, labeled C.

[0215] Experimental results: Motor behavior is an indicator reflecting the basic function of the nervous system in *C. elegans*. The motor ability of *C. elegans* is directly related to its lifespan and can serve as an important indicator for evaluating the rate of aging in an organism. Figure 7 As shown, the motility of *C. elegans* gradually declines with aging. During the first 5 days of culture, ASO-NLC had no significant effect on the motility of *C. elegans* (P > 0.05). After 10 days of culture, compared with the NC group, all three dosage groups of ASO-100 and ASO-NLC improved the motility of *C. elegans* (P < 0.05 or P < 0.01). At 15 days of culture, the motility of *C. elegans* in the NC group was mainly at grades B and C, while the motility of *C. elegans* in the ASO and ASO-NLC intervention groups was significantly higher than that in the NC group, indicating that ASO-NLC can significantly improve the motility of *C. elegans* (P < 0.01), with the high-dose group showing the best effect.

[0216] 4.3.8 Heat stress test of *C. elegans* var. *truncatum* nanostructured lipid carrier using *Acer emblica* seed oil

[0217] Preparation method: After culturing synchronized Caenorhabditis elegans to the L4 stage, the samples were treated with different concentrations for 5 days in groups. Then, they were transferred to a 37℃ environment for 12 hours. The number of surviving and dying Caenorhabditis elegans per hour was calculated under an inverted fluorescence microscope, and survival curves were plotted until all Caenorhabditis elegans died.

[0218] Experimental Results: High temperatures can lead to metabolic disorders and enzyme inactivation within the body, resulting in the production of large amounts of reactive oxygen species (ROS) and causing oxidative stress. Oxidative stress is a state in which highly reactive molecules such as ROS accumulate excessively in the body, the degree of cellular oxidation exceeds the scavenging capacity of oxidants, and the oxidative and antioxidant systems become imbalanced, leading to cellular senescence and tissue damage. It is considered an important factor contributing to aging and disease. Therefore, this study investigated the resistance of *C. elegans* to acute oxidative damage through oxidative stress experiments. Heat stress experiment results. Figure 8 Compared with the control group, there were no significant differences among the groups in the early stage. After 4 hours, the survival rate of all ASO and ASO-NLC dosage groups was significantly prolonged (P<0.05), with the longest survival time reaching 10 hours, which was 1 hour longer than that of the NC group. This indicates that ASO-NLC can improve the heat stress resistance of Caenorhabditis elegans.

[0219] 4.3.9 Effects of Acer truncatum seed oil nanostructured lipid carrier on acute oxidative stress in Caenorhabditis elegans

[0220] Preparation method: Each group of *C. elegans* was transferred to new NGM medium. At this time, 10 μL of 30% hydrogen peroxide (v / v) was added to every 10 mL of NGM medium. The number of surviving *C. elegans* was recorded every 1 hour until all *C. elegans* died. Survival curves were plotted based on the survival and mortality times of *C. elegans*.

[0221] Experimental Results: Results of Acute Oxidative Stress Test Figure 9 The results showed no significant differences between the groups in the early stage compared with the NC group; however, after 3 hours, the survival rate of each ASO-NLC dose group was significantly prolonged (P<0.05), with the longest survival time being 9 hours, which was 3 hours longer than that of the NC group. This indicates that ASO-NLC can enhance the acute oxidative stress resistance of Caenorhabditis elegans.

[0222] 4.3.10 Assay for the antioxidant activity of Acer truncatum seed oil nanostructured lipid carrier against Caenorhabditis elegans in vivo

[0223] Preparation method: After the synchronized Caenorhabditis elegans reached the L4 stage, the Caenorhabditis elegans of each group were transferred to NGM medium. The Caenorhabditis elegans were washed into centrifuge tubes with M9 buffer, centrifuged and the supernatant was discarded. The Caenorhabditis elegans were resuspended in M9 buffer, frozen and ground, centrifuged, and the supernatant was collected. The enzyme activities were then detected by SOD, CAT, MDA and GSH-Px enzyme activity kits, respectively.

[0224] Experimental Results: The body's metabolic rate slows down with age. A large number of reactive oxygen species (ROS) trigger oxidative stress, damaging cellular macromolecules and accelerating the aging process, ultimately leading to a decline in health. CAT, SOD, and GSH-Px are the main antioxidant enzymes in *C. elegans*. Increasing the activity of these antioxidant enzymes can eliminate excess free radicals in *C. elegans*, thereby promoting oxidative balance. Figure 10 Compared with the control group, the low, medium, and high dose groups of ASO-NLC showed significantly increased GSH-Px activity (21.6%, 36.0%, and 48.8%, respectively); CAT activity (22.4%, 43.1%, and 58.6%, respectively); SOD activity (16.1%, 41.5%, and 75.9%, respectively); and decreased MDA activity (6.8%, 22.3%, and 34.9%, respectively). A dose-response relationship was observed among the three dose groups, and the antioxidant capacity of ASO-NLC was significantly superior to that of ASO. These results indicate that ASO-NLC can, to some extent, enhance the antioxidant capacity of *C. elegans*, alleviate oxidative damage and senescence in *C. elegans*, and has the potential to activate the antioxidant defense system in *C. elegans*.

[0225] 4.3.11 Assay for Apoptosis Staining of Caenorhabditis elegans Cells Using Acer truncatum Seed Oil Nanostructured Lipid Carrier

[0226] Preparation method: Synchronized *C. elegans* were collected and divided into groups. After culturing each group of *C. elegans* for 72 h, the *C. elegans* were collected and washed three times with M9 buffer. Apoptosis staining assay: *C. elegans* were transferred to 200 μL of acridine orange solution (25 μg / mL) and incubated in the dark at room temperature for 1 h. After washing off the stain with M9 buffer, *C. elegans* were transferred to blank NGM culture dishes, anesthetized with levamisole hydrochloride solution, placed on 3% agarose slides, and observed under a fluorescence microscope. The fluorescence intensity was photographed and calculated. The excitation and emission wavelengths were 485 nm and 530 nm, respectively.

[0227] Experimental Results: DNA damage is closely related to apoptosis. Acridine orange staining results can reflect the integration status of *C. elegans* DNA. This is because acridine orange can enter the cell membrane of apoptotic cells and bind to DNA. Cells appear green under fluorescent light. *C. elegans* worms in the NC group showed a bright green color, while *C. elegans* worms in all ASO and ASO-NLC dose groups showed a light green color overall, and the bright green area of ​​*C. elegans* worms was smaller than that in the NC group. Figure 11The relative fluorescence intensities of the ASO-NLC-50, ASO-NLC-100, and ASO-NLC-200 groups were significantly lower than those of the NC group, by 13.7%, 15.7%, and 17.0%, respectively (P < 0.01). The differences in acridine orange staining results indicate that ASO-NLC can alleviate DNA damage in *C. elegans*, thereby prolonging its lifespan.

[0228] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A nanostructured lipid carrier for Acer truncatum seed oil, characterized in that, The raw material formulation of the Acer truncatum seed oil nanostructure lipid carrier consists of the following components by weight percentage: 5% Acer truncatum seed oil, 5% solid lipids, 4% emulsifier, and 86% water; the solid lipids are cocoa butter and palmitate palmitate, with cocoa butter accounting for 4% by weight and palmitate palmitate accounting for 1% by weight; the emulsifier is poloxamer 188. The preparation method of the Acer truncatum seed oil nanostructured lipid carrier includes the following steps: (1) Solid lipids and Acer truncatum seed oil are melted in a water bath and stirred evenly to obtain an oil phase solution; (2) Add the emulsifier to water and melt it under water bath conditions, stirring until the emulsifier is dissolved to obtain an aqueous solution; (3) Under magnetic stirring conditions, the aqueous phase solution in step (2) is added dropwise to the oil phase solution in step (1) to obtain a mixture; (4) Place the mixture obtained in step (3) into an ultrasonic cell disruptor for ultrasonic treatment. Set the power of the ultrasonic cell disruptor to 200-600W and the ultrasonic treatment time to 4-12min to obtain colostrum. (5) The colostrum obtained in step (4) is placed in a homogenizer for homogenization. The homogenization pressure is set to 250-1250 bar to obtain the Acer truncatum seed oil nanostructure lipid carrier.

2. The nanostructured lipid carrier for Acer truncatum seed oil according to claim 1, characterized in that, In step (4) of the preparation method of the Acer truncatum seed oil nanostructure lipid carrier, the power of the ultrasonic cell disruptor is set to 300-500W and the ultrasonic time is 6-10min.

3. The nanostructured lipid carrier for Acer truncatum seed oil according to claim 2, characterized in that, The ultrasonic cell disruptor was set to 400W and the ultrasonic time was 8 minutes.

4. The nanostructured lipid carrier for Acer truncatum seed oil according to claim 1, characterized in that, In step (5) of the preparation method of the Acer truncatum seed oil nanostructure lipid carrier, the homogenization pressure is set to 1000 bar.

5. The application of the Acer truncatum seed oil nanostructured lipid carrier according to claim 1, characterized in that, The application of the Acer truncatum seed oil nanostructure lipid carrier in the preparation of drugs to improve aging.

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