A ternary wall material apigenin microcapsule, a preparation method and application thereof

Abstract

The application belongs to the technical field of functional food materials, and specifically discloses a ternary wall material apigenin microcapsule as well as a preparation method and application thereof. The ternary wall material apigenin microcapsule comprises a wall material and a core material, the core material is apigenin, and the wall material is whey protein, β-cyclodextrin and modified starch; the mass ratio of the whey protein, the β-cyclodextrin and the modified starch is (4-6):(2.5-4):(1-3); and the wall material embeds the core material to form the microcapsule, and the microcapsule is subjected to temperature gradient slow-release treatment. The process parameters are optimized through an orthogonal test, and the β-cyclodextrin dosage has a significant influence on the embedding rate. The embedding rate of the ternary wall material apigenin microcapsule is greater than 85%, the particle size D90 is less than 50 μm, and the water solubility is increased by more than 100 times. Animal experiment results show that the oral bioavailability of the ternary wall material apigenin microcapsule is greater than 25%, the 12-month accelerated stability retention rate is greater than 80%, the liver function can be significantly improved, the lipid metabolism can be regulated, and the antioxidant capacity can be improved, and the ternary wall material apigenin microcapsule is suitable for liver protection, fat reduction and antioxidant functional food.

Description

Technical Field

[0001] This invention relates to the field of functional food technology, and more specifically, to a ternary wall material apigenin microcapsule, its preparation method, and its application. Background Technology

[0002] The liver is a core organ for metabolism, detoxification, and lipid regulation in the human body. Liver health management is a crucial aspect of promoting physical health among young people and reducing the incidence of chronic diseases in the nation. Currently, non-alcoholic fatty liver disease (NAFLD) is showing a trend towards affecting younger people. The prevalence of NAFLD in my country has exceeded 29.2%, with over 300 million patients, ranking first globally. Athletes, due to high-intensity specialized training, significantly increase their metabolic load, leading to a markedly higher risk of oxidative stress damage to the liver. Exercise-induced liver injury has become a prominent issue hindering the health of athletes.

[0003] Apogenin is a natural flavonoid compound with the chemical name 4',5,7-trihydroxyflavone and the molecular formula C2. 15 H 10 O5, with a molecular weight of 270.24. Apigenin is widely abundant in edible plants of the Apiaceae family, such as celery, coriander, chamomile, and parsley, and is also found in grapes, citrus fruits, and legumes. As a natural active ingredient verified by numerous pharmacological tests both domestically and internationally, apigenin, relying on the Keap1 / Nrf2 and AMPK key pathways, inhibits abnormal fat accumulation in hepatocytes at its source, clears oxidative free radicals generated by exercise stress, and soothes chronic low-grade liver inflammation. It has multiple effects, including liver protection, lipid regulation, and antioxidation, and is highly safe with no drug side effects from long-term consumption.

[0004] Existing research confirms that the antioxidant activity of apigenin is closely related to its hepatoprotective effects. Oxidative stress is a key pathological link in the pathogenesis of NAFLD, manifested as a decline in the body's antioxidant capacity and the accumulation of lipid peroxidation products. Apigenin, by activating the Nrf2 signaling pathway, can upregulate the activity of endogenous antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and reduce the levels of lipid peroxidation products such as malondialdehyde (MDA), thereby alleviating liver oxidative damage. However, the application of natural apigenin in functional foods faces the following technical bottlenecks, restricting its industrial application in this field: (1) Extremely poor water solubility: According to measured data from the GuideChem international chemical database, the water solubility of apigenin at room temperature is only 1.35~2.16μg / mL, and it is almost insoluble in pure water; (2) Low bioavailability: The bioavailability of conventional oral administration is less than 5%, and most of the active ingredients are inactivated after being degraded by gastric acid and metabolized by intestinal flora; (3) Poor stability: It is easily oxidized and degraded during storage, and the accelerated stability retention rate is less than 70% after 12 months; (4) Traditional encapsulation effects are limited: the encapsulation rate of single-component or two-component wall material systems (such as maltodextrin, gum arabic, etc.) is only 60%~72%, and the water solubility is increased by only 30~50 times.

[0005] In the prior art, Chinese patent CN109316371B discloses a technical solution for preparing apigenin sustained-release formulations using PLGA as a carrier material. This solution uses chemically synthesized polymer materials, which pose potential risks in terms of biodegradability and safety. Chinese patent CN107661295B discloses a technical solution using microemulsions as apigenin drug carriers, but the microemulsion system has poor stability and is difficult to industrialize. Chinese patent application CN122163573A discloses a technical solution for preparing apigenin nanoparticles using soybean protein and xylan as the aqueous phase, but this solution uses an antisolvent precipitation method, the wall material composition is completely different from that of this invention, and it does not involve the synergistic use of β-cyclodextrin and modified starch. None of the above-mentioned prior art discloses the combination of whey protein, β-cyclodextrin, and modified starch ternary wall material systems, nor does it disclose gradient sustained-release post-treatment processes. Summary of the Invention

[0006] The purpose of this invention is to provide a ternary wall material apigenin microcapsule, its preparation method and application, to solve the problems of poor water solubility, low bioavailability and weak stability of apigenin in the prior art.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solution: In a first aspect, the present invention provides a ternary wall material apigenin microcapsule, comprising a wall material and a core material, wherein the core material is apigenin, and the wall material is whey protein, β-cyclodextrin and modified starch; the mass ratio of whey protein, β-cyclodextrin and modified starch is (4~6):(2.5~4):(1~3); the wall material encapsulates the core material to form a microcapsule, which is then subjected to a temperature gradient sustained-release treatment.

[0008] Specifically, whey protein accounts for 40%–60% of the total mass of the wall material, mainly playing a role in film formation, targeted adsorption, and forming a protein network structure; β-cyclodextrin accounts for 25%–40% of the total mass of the wall material, mainly playing a role in host-guest molecule inclusion and improving solubility; modified starch accounts for 10%–20% of the total mass of the wall material, mainly playing a role in anti-blocking and viscosity regulation. The ternary wall material achieves efficient encapsulation of apigenin through the synergistic effect of the film-forming properties of whey protein, the host-guest inclusion effect of β-cyclodextrin, and the anti-blocking properties of modified starch.

[0009] In a preferred embodiment, the modified starch is one or more of hydroxypropyl methylcellulose and sodium octenyl succinate starch.

[0010] In the preferred embodiment, the temperature gradient slow release treatment consists of sequentially holding the temperature at 40℃ for 1-3 hours, at 25℃ for 1-3 hours, and at 4℃ for 1-3 hours.

[0011] A second aspect of the present invention provides a method for preparing apigenin microcapsules, a ternary wall material, according to any of the above embodiments, comprising the following steps: S1. Preparation of wall material solution: Take whey protein, β-cyclodextrin and modified starch, and prepare a wall material aqueous solution with a total concentration of 15%~25%; S2. Core material emulsification: Apigenin is dissolved in anhydrous ethanol to obtain a apigenin ethanol solution. The wall material aqueous solution is added to the apigenin ethanol solution at a core-to-wall mass ratio of 1:3 to 1:5. The solution is emulsified by high-speed shearing at 60°C and then subjected to ultrasonic homogenization to obtain a uniform emulsion. S3. Spray drying: The uniform emulsion is spray dried, and the inlet air temperature is controlled at 170~180℃ and the outlet air temperature is controlled at 80~95℃ to obtain microcapsule powder. S4. Gradient sustained-release treatment: The microcapsule powder is placed in a sealed container and kept at 40℃ for 1-3 hours, 25℃ for 1-3 hours, and 4℃ for 1-3 hours in sequence. This causes the whey protein molecules on the surface of the microcapsules to undergo directional rearrangement, forming a dense protein network structure, thus obtaining ternary wall material apigenin microcapsules.

[0012] In the preferred embodiment, in step S1, the modified starch is hydroxypropyl methylcellulose (HPMC), and the viscosity of hydroxypropyl methylcellulose is 5~50 mPa·s (2% aqueous solution, 20℃). HPMC within this viscosity range has good atomization performance and thermal stability during spray drying, which can effectively prevent microcapsule particles from sticking together, while not affecting the protein-cyclodextrin inclusion efficiency.

[0013] In the preferred embodiment, in step S2, the high-speed shearing speed is 8000~12000rpm and the shearing time is 2~5min; the ultrasonic homogenization power is 200~400W and the action time is 3~8min.

[0014] In the preferred embodiment, in step S3, the feed rate for spray drying is 2 mL / min and the atomization pressure is 0.2 MPa.

[0015] In the preferred embodiment, in step S4, the microcapsule powder is sequentially kept at 40°C for 2 hours, at 25°C for 2 hours, and at 4°C for 2 hours.

[0016] In a fourth aspect, the present invention provides the use of the ternary wall material apigenin microcapsules described in any of the above embodiments in the preparation of products having the function of alleviating liver damage and / or regulating lipid metabolism.

[0017] In the preferred embodiment, the liver injury is exercise-induced liver injury.

[0018] Beneficial effects of the present invention (1) An innovative ternary wall material synergistic system is adopted to give full play to the synergistic effect of each wall material component. Whey protein provides good film-forming properties and targeted adsorption capacity, β-cyclodextrin achieves host-guest molecule inclusion and significantly improves solubility; modified starch effectively prevents adhesion between microcapsule particles and adjusts the wall material viscosity to a suitable range for spray drying. The synergistic effect of the three components increases the encapsulation rate from 60%~72% in the traditional process to more than 85%.

[0019] (2) A dense protein network structure is formed on the surface of the microcapsules through a gradient slow-release post-treatment process. Specifically: First stage: 40℃ incubation stage (1~3 hours): This temperature is below the moderate denaturation temperature range of whey protein (initial denaturation temperature is about 55~60℃). At this temperature, whey protein molecules obtain sufficient kinetic energy to undergo hydrophobic interactions and partial disulfide bond exchange between molecules, and molecular chain segments undergo directional rearrangement, which is conducive to the formation of a preliminary protein network backbone on the surface of the microcapsules; Second stage: 25℃ incubation stage (1~3 hours): At this temperature, the movement rate of protein molecules slows down, the hydrophobic interactions and disulfide bond exchange between molecules tend to be balanced, the protein network structure undergoes preliminary solidification, and a relatively stable secondary structure is formed; Third stage: 4℃ incubation stage (1~3 hours): The low temperature environment further slows down the movement of protein molecules, the water in the protein network structure is gradually removed, the network structure further shrinks and densifies, forming a dense protective layer with high mechanical strength and low permeability. This protein network can effectively seal the micropores on the surface of microcapsules, blocking the penetration of oxygen and moisture, thereby significantly improving the storage stability of the product.

[0020] (3) The performance indicators of the ternary wall material apigenin microcapsules of the present invention are significantly better than those of the prior art: the encapsulation rate is greater than 85%, the particle size D90 is less than 50μm, the water solubility is increased by more than 100 times, the oral bioavailability is greater than 25%, and the 12-month accelerated stability retention rate is greater than 80%.

[0021] (4) The preparation process of apigenin microcapsules of ternary wall material provided by the present invention is stable and controllable, all raw materials are food grade, and the process parameters are suitable for pilot-scale amplification and industrial production. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of the structure of the ternary wall material apigenin microcapsules of the present invention.

[0023] Figure 2 This is a process flow diagram of the preparation method of apigenin microcapsules, a ternary wall material of the present invention.

[0024] Figure 3 This is a visual analysis diagram showing the influence of the levels of various factors on the embedding rate in the orthogonal experiment of this invention.

[0025] Figure 4 The graph shows the stability comparison of the microcapsules in Example 1 and Comparative Examples 1-4.

[0026] Figure 5 SEN diagrams of apigenin microcapsules, a ternary wall material prepared in Example 1 and Comparative Example 3; (a) Comparative Example 3, (b) Example 1.

[0027] Figure 6 The figure shows the results of the comparative experiment on the bioavailability of microcapsules in Example 1 and Comparative Examples 1-2.

[0028] Figure 7 A comparison of antioxidant indices in the livers of NAFLD mice. Detailed Implementation

[0029] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments and accompanying drawings. All materials used in this invention are available through commercially available channels.

[0030] Example 1

[0031] A ternary wall material apigenin microcapsule, its preparation method, and its application include the following steps: S1. Preparation of wall material solution: Weigh 40g of whey protein, 30g of β-cyclodextrin and 30g of sodium octenyl succinate starch, add an appropriate amount of distilled water, and stir to dissolve in a 60℃ water bath for 2 hours to prepare a wall material aqueous solution with a total concentration of 20% (w / v).

[0032] S2. Core material emulsification: Weigh 20g of apigenin and dissolve it in anhydrous ethanol to obtain a apigenin ethanol solution with a total concentration of 20% (w / v); add the wall material aqueous solution to the apigenin ethanol solution according to the core-to-wall mass ratio of 1:4, and shear at 10000rpm for 3min at 60℃, followed by ultrasonic homogenization treatment with an ultrasonic power of 300W and a treatment time of 5min to obtain a uniform emulsion.

[0033] S3. Spray drying: The uniform emulsion is spray dried, with the inlet air temperature controlled at 180℃, the outlet air temperature at 85℃, the feed rate at 2mL / min, and the atomization pressure at 0.2MPa, to obtain microcapsule powder; S4. Gradient sustained-release treatment: The microcapsule powder is placed in a sealed desiccator and subjected to gradient cooling treatment at 40℃×2h, 25℃×2h, and 4℃×2h in sequence to obtain ternary wall material apigenin microcapsules.

[0034] Testing revealed that the ternary wall material apigenin microcapsules prepared in this embodiment had a particle size D90 of 38 μm, an encapsulation rate of 87.3%, and a water solubility 115 times that of unencapsulated apigenin.

[0035] Example 2

[0036] The difference between this embodiment and Example 1 is that the mass ratio of whey protein, β-cyclodextrin, and sodium octenyl succinate starch is 5:3:2, while the other steps and parameters are exactly the same as in Example 1.

[0037] Testing revealed that the ternary wall material apigenin microcapsules prepared in this embodiment had a particle size D90 of 42 μm, an encapsulation rate of 86.5%, and a water solubility 108 times that of unencapsulated apigenin.

[0038] Example 3

[0039] The difference between this embodiment and Example 1 is that the mass ratio of whey protein, β-cyclodextrin, and sodium octenyl succinate starch is 6:2.5:1.5, while the other steps and parameters are exactly the same as in Example 1.

[0040] Testing revealed that the ternary wall material apigenin microcapsules prepared in this embodiment had a particle size D90 of 46 μm, an encapsulation rate of 85.2%, and a water solubility 102 times that of unencapsulated apigenin.

[0041] Orthogonal optimization experiment To scientifically optimize various process parameters, this invention employs an L9(3^4) orthogonal experimental design. Based on the preparation method provided in Example 1, and using encapsulation efficiency, solubility, and stability as response values, four factors—whey protein dosage (A), β-cyclodextrin dosage (B), modified starch dosage (C), and core-to-wall ratio (D)—were optimized. To ensure the reliability of the experimental results, each experiment was performed in triplicate, and the average value was taken. The orthogonal experimental factor level table is shown in Table 1 below, and the orthogonal experimental results and analysis are shown in Table 2 below.

[0042] Table 1. Factor Levels in Orthogonal Experiments

[0043] Table 2 Results and Analysis of L9(3^4) Orthogonal Experiment

[0044] SPSS 22.0 software was used to perform analysis of variance on the orthogonal experimental results. The analysis of variance calculation process is as follows: Calculation of the total sum of squares (SST): SST = Σ(each measured value - total mean)² Calculation of the sum of squares of deviations for each factor (SSA, SSB, SSC, SSD): Sum of squares of deviations for each factor = Σ(average value of each factor - overall mean)² × n (number of repetitions) The sum of squared errors SSe = SST - (SSA + SSB + SSC + SSD) The degrees of freedom for each factor, f = number of levels - 1 = 2; Error degrees of freedom fe = total degrees of freedom - sum of degrees of freedom of each factor; Mean square of each factor (MS) = sum of squared deviations / degrees of freedom; F-value = MS factor / MS error; Substituting the parameters into the above formula, the calculation results are shown in Table 3 below.

[0045] Table 3. Results of orthogonal experiment variance analysis

[0046] Note: * indicates P < 0.05; F0.05(2,2) = 19.0, F0.01(2,2) = 99.0.

[0047] The intuitive analysis in Table 2 and the variance analysis in Table 3 show that the order of influence of each factor on the encapsulation efficiency is: B>C>D>A, meaning that the amount of β-cyclodextrin has a significant effect (P < 0.05). Although the amount of modified starch and the core-to-wall ratio have some influence (R values ​​of 2.43 and 2.10, respectively), they do not reach statistical significance. The amount of whey protein has the least influence. Considering the encapsulation efficiency, solubility, and stability, the optimal combination is A2B3C1D2, namely 50% whey protein, 40% β-cyclodextrin, 15% modified starch, and a core-to-wall ratio of 1:4, which is consistent with the results of Example 2, verifying the scientific nature of the parameter selection in this invention.

[0048] Comparative Example 1: Traditional single-component wall materials In this comparative example, maltodextrin was used as the wall material. The gradient slow-release treatment in step S4 was not performed. All other processes and parameters were exactly the same as in Example 1, and apigenin microcapsules with a single wall material were prepared.

[0049] The microcapsules obtained in this comparative example had an encapsulation rate of 68%, a water solubility 45 times that of the unencapsulated apigenin, and an accelerated stability retention rate of 65% after 12 months.

[0050] Comparative Example 2: Traditional Two-Component Wall Materials In this comparative example, whey protein and β-cyclodextrin were used as wall materials, with a mass ratio of whey protein to β-cyclodextrin of 1:1. Step S4 gradient sustained-release treatment was not performed, and other processes and parameters were exactly the same as in Example 1, to prepare apigenin microcapsules with binary wall material.

[0051] The microcapsules obtained in this comparative example had an encapsulation rate of 72%, a water solubility 52 times that of the unencapsulated apigenin, and an accelerated stability retention rate of 68% after 12 months.

[0052] Comparative Example 3: Gradient-free sustained-release treatment The difference between this comparative example and Example 1 is that step S4 gradient sustained-release treatment is not performed. All other steps and parameters are exactly the same as in Example 1, and ternary wall material apigenin microcapsules are prepared.

[0053] The microcapsule encapsulation rate of this comparative example was 85.8%, similar to that of Example 1, but the 12-month accelerated stability retention rate was only 71%, significantly lower than the 82.6% of Example 1. This result demonstrates the crucial role of the gradient sustained-release post-processing in improving stability.

[0054] Comparative Example 4: Apigenin Microcapsules (Binary Wall Material) In this comparative example, whey protein and β-cyclodextrin were used as wall materials, with a mass ratio of whey protein to β-cyclodextrin of 1:1. Other processes and parameters were exactly the same as in Example 1, and apigenin microcapsules with binary wall material were prepared.

[0055] Testing revealed that the encapsulation rate of the microcapsules obtained in this comparative example was 79.2%, significantly lower than the 87.3% in Example 1; significant wall adhesion occurred during spray drying, resulting in a product yield of only 65%. These results demonstrate the crucial role of modified starch in regulating wall material viscosity and preventing particle adhesion.

[0056] The performance of the microcapsule samples prepared in Example 1 and Comparative Examples 1-4 will be tested and characterized below.

[0057] 1. Stability Comparison Test The microcapsule samples prepared in Example 1 and Comparative Examples 1-4 were placed in an accelerated stability test chamber (temperature 40℃±2℃, humidity 75%±5%), with a light cycle of 4500±500 Lx and 12 h / day. Samples were taken at 0, 30, 45, 60, 90, 120, 180, and 360 days to determine the apigenin content. The apigenin content was determined by high-performance liquid chromatography (HPLC). The chromatographic conditions were: C18 column (250 mm × 4.6 mm, 5 μm); mobile phase: methanol-0.1% phosphoric acid aqueous solution (60:40, v / v); flow rate: 1.0 mL / min; detection wavelength: 340 nm; column temperature: 30℃; injection volume: 20 μL; each sample was measured in triplicate. The results are shown in Table 4. Figure 4 As shown.

[0058] Table 4. Results of accelerated stability test (apigenin content retention rate, %)

[0059] Note: The apigenin content of the microcapsule samples in Comparative Examples 1-4 was less than 70% after 180 days, and they had lost their usability, so they were not further tested.

[0060] The results showed that the ternary wall material apigenin microcapsules prepared in Example 1 exhibited significant advantages in accelerated stability testing. After 360 days, the apigenin content retention rate was still 76.3%, while the traditional process sample decreased to 65%~71% within 180 days, fully demonstrating the effectiveness of the technical solution of the present invention.

[0061] 2. SEM characterization The surface morphology of the ternary wall material apigenin microcapsules prepared in Example 1 and Comparative Example 3 was observed using scanning electron microscopy (SEM), and the results are as follows: Figure 5 As shown.

[0062] Depend on Figure 5 (a) As can be seen, the apigenin microcapsules of the ternary wall material in Comparative Example 3, which were not treated with gradient sustained-release, were regularly spherical with a relatively smooth surface but contained a small number of wrinkles and micropores; Figure 5 (b) As can be seen, the apigenin microcapsules of the ternary wall material treated with gradient slow release in Example 1 have fewer wrinkles on the surface of the microcapsules and form a uniform and dense film structure. This film structure is the network layer formed after protein denaturation, which can effectively block oxygen from entering the interior of the microcapsules, thereby significantly improving stability.

[0063] 3. Bioavailability Comparison Experiment Using apigenin as a control group, a comparative experiment was conducted on the bioavailability of ternary wall material microcapsules prepared in Example 1 and microcapsule samples prepared in Comparative Examples 1 and 2 via gavage in rats. Forty-eight healthy male SD rats (weighing 200±20g) were randomly divided into four groups of 12 rats each. After fasting for 12 hours, the rats were administered apigenin via gavage (apigenin dose 50mg / kg body weight). Blood samples were collected from the orbital sinus at 0.5h, 1h, 2h, 4h, 6h, 8h, 12h, and 24h after administration. Serum was collected by centrifugation, and the serum apigenin concentration was determined by HPLC-MS / MS (method precision RSD <8%, recovery rate 85%~115%). Pharmacokinetic parameters were calculated.

[0064] Table 5 Comparison of bioavailability in rats via gavage

[0065] The results showed that the oral bioavailability of apigenin in rats was only 4.8%, while the oral bioavailability of rats encapsulated in single-component or two-component apigenin wall materials in Examples 1 and 2 increased to 10% and 15.7%, respectively. In contrast, the oral bioavailability of the ternary wall material apigenin microcapsules prepared in Example 1 was as high as 25.7%, significantly higher than that of apigenin alone and the traditional apigenin microcapsules in Examples 1 and 2. The improved oral bioavailability of the ternary wall material apigenin microcapsules in rats in this invention is mainly due to the following factors: 1) the protein network provides additional intestinal absorption protection; 2) the smaller molecular weight of apigenin (270.24) results in better encapsulation; and 3) the synergistic effect of the ternary wall material is superior to that of the two-component system.

[0066] 4. Efficacy verification experiment of NAFLD mouse model To verify the practical application effect of the ternary wall material apigenin microcapsules of the present invention in the field of liver protection, an efficacy verification experiment was carried out in a C57BL / 6 mouse NAFLD model using the ternary wall material apigenin microcapsules prepared in Example 1.

[0067] Statistical analysis: All experimental data are expressed as mean ± standard deviation and statistical analysis was performed using SPSS 22.0 software. Independent samples t-tests were used for comparisons between two groups, one-way ANOVA was used for comparisons among multiple groups, and LSD method was used for pairwise comparisons between groups. P < 0.05 was considered statistically significant.

[0068] 4.1 Test Methods Sixty 8-week-old male C57BL / 6 mice were randomly divided into 6 groups (n=10 per group): a normal control group, a model group, an apigenin group, a low-dose microcapsule group, a high-dose microcapsule group, and a positive control group. Silymarin, a classic hepatoprotective drug, is one of the most commonly used positive control drugs in preclinical studies of NAFLD. Its hepatoprotective mechanism is similar to that of apigenin, mainly improving liver damage through antioxidant and anti-inflammatory effects. Except for the normal control group, the other groups were fed a high-fat diet (60% fat energy ratio) for 12 weeks to establish an NAFLD model. An 8-week intervention experiment was then conducted. The normal control group and the model group received an equal volume of physiological saline, the low-dose microcapsule group received 50 mg / kg of apigenin microcapsules (prepared in Example 1), the high-dose microcapsule group received 200 mg / kg, and the positive control group received 200 mg / kg of silymarin. After the intervention, serum ALT and AST levels and liver histopathological changes were measured.

[0069] 4.2 Serum marker detection Table 6 Results of the NAFLD model mouse intervention experiment (liver function and lipid metabolism indicators)

[0070] Note: The shoulder label "**" indicates a comparison with the model group. ** P < 0.01, with the superscript "##" indicating a comparison with the apigenin group. ## P < 0.01.

[0071] As shown in Table 6, the results indicate that the ternary wall material apigenin microcapsules prepared in Example 1 can significantly improve liver function indicators and liver lipid deposition in NAFLD mice. The high-dose microcapsule group showed an effect close to that of the positive control group (silymarin) and significantly better than the apigenin group (P < 0.01), which fully demonstrates the practical application effect of the ternary wall material apigenin microcapsule product of the present invention.

[0072] 4.3 Detection of liver antioxidant indicators To further verify the antioxidant efficacy of the ternary wall material apigenin microcapsules of this invention, liver tissue samples from mice in the above groups were collected, and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) content were measured. SOD and GSH-Px are important endogenous antioxidant enzymes in the body, and their activity levels reflect the body's antioxidant capacity; MDA is the end product of lipid peroxidation, and its content reflects the degree of oxidative damage in the body. Detection method: Liver tissue was homogenized in pre-cooled physiological saline, centrifuged at 4000 rpm for 10 min, and the supernatant was collected. Relevant indicators were measured using a kit from Nanjing Jiancheng Bioengineering Institute. Each sample was measured in triplicate. The results are shown in Table 7. Figure 7 As shown.

[0073] Table 7 Results of liver antioxidant index detection in NAFLD model mice

[0074] Note: The shoulder label "**" indicates a comparison with the model group. ** P < 0.01, with the superscript "##" indicating a comparison with the apigenin group. ## P < 0.01.

[0075] The results showed that the ternary wall material apigenin microcapsules prepared in Example 1 significantly increased the activity of SOD and GSH-Px in the liver of NAFLD mice and reduced the MDA content in a dose-dependent manner. The high-dose group showed antioxidant indicators close to those of the positive control group and significantly better than the apigenin raw material group (P < 0.01), fully demonstrating the practical application effect of the microcapsules of this invention in antioxidant liver protection.

[0076] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A ternary wall material apigenin microcapsule, comprising a wall material and a core material, wherein the core material is apigenin, characterized in that, The wall material is whey protein, β-cyclodextrin and modified starch; the mass ratio of whey protein, β-cyclodextrin and modified starch is (4~6):(2.5~4):(1~3); the wall material encapsulates the core material to form microcapsules, which are then subjected to temperature gradient slow release treatment.

2. The ternary wall material apigenin microcapsules according to claim 1, characterized in that, The modified starch is one or more of hydroxypropyl methylcellulose and sodium octenyl succinate starch.

3. The ternary wall material apigenin microcapsules according to claim 1, characterized in that, The temperature gradient slow-release treatment consists of sequentially holding the temperature at 40℃ for 1-3 hours, at 25℃ for 1-3 hours, and at 4℃ for 1-3 hours.

4. A method for preparing apigenin microcapsules, a ternary wall material according to any one of claims 1 to 3, characterized in that, Includes the following steps: S1. Preparation of wall material solution: Take whey protein, β-cyclodextrin and modified starch, and prepare a wall material aqueous solution with a total concentration of 15%~25%; S2. Core material emulsification: Apigenin is dissolved in anhydrous ethanol to obtain a apigenin ethanol solution. The wall material aqueous solution is added to the apigenin ethanol solution at a core-to-wall mass ratio of 1:3 to 1:

5. The solution is emulsified by high-speed shearing at 60°C and then subjected to ultrasonic homogenization to obtain a uniform emulsion. S3. Spray drying: The uniform emulsion is spray dried, and the inlet air temperature is controlled at 170~180℃ and the outlet air temperature is controlled at 80~95℃ to obtain microcapsule powder. S4. Gradient sustained-release treatment: The microcapsule powder is placed in a sealed container and kept at 40℃ for 1-3 hours, 25℃ for 1-3 hours, and 4℃ for 1-3 hours in sequence to obtain ternary wall material apigenin microcapsules.

5. The method for preparing apigenin microcapsules, a ternary wall material, according to claim 4, is characterized in that, In step S1, the modified starch is hydroxypropyl methylcellulose, and the viscosity of hydroxypropyl methylcellulose is 5~50 mPa·s.

6. The method for preparing apigenin microcapsules, a ternary wall material, according to claim 4, is characterized in that, In step S2, the high-speed shearing speed is 8000~12000 rpm and the shearing time is 2~5 min; the ultrasonic homogenization power is 200~400W and the action time is 3~8 min.

7. The method for preparing apigenin microcapsules, a ternary wall material, according to claim 4, is characterized in that, In step S3, the feed rate for spray drying is 2 mL / min and the atomization pressure is 0.2 MPa.

8. The method for preparing apigenin microcapsules using ternary wall material according to claim 4, characterized in that, In step S4, the microcapsule powder is successively kept at 40°C for 2 hours, at 25°C for 2 hours, and at 4°C for 2 hours.

9. The use of the ternary wall material apigenin microcapsules according to any one of claims 1 to 3 in the preparation of products with functions of alleviating liver damage and / or regulating lipid metabolism.

10. The application of the ternary wall material apigenin microcapsules according to claim 9 in the preparation of products with functions of alleviating liver damage and / or regulating lipid metabolism, characterized in that, The liver injury described is exercise-induced liver injury.

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