Modified Zein nano system for delivering seabuckthorn flavone as well as preparation method and application of modified Zein nano system

Abstract

According to the modified Zein nano system for delivering the hippophae rhamnoides flavone and the preparation method and application thereof, the hippophae rhamnoides flavone is loaded in modified zein and combined with anionic polysaccharide to form the TFH-D-Zein-AP nano system, the stability and solubility of the hippophae rhamnoides flavone in vivo can be improved, and therefore the bioavailability of the hippophae rhamnoides flavone is improved; the delivery efficiency of the seabuckthorn flavone is improved; meanwhile, the anti-inflammatory activity of the seabuckthorn flavone can be improved, so that the seabuckthorn flavone can play a role in vivo for a longer time, and a more effective and safer inflammation treatment means is provided.

Description

[Technical field] The invention belongs to the technical field of nano delivery systems, and in particular relates to a modified Zein nano system for delivering seabuckthorn flavonoids, and a preparation method and application thereof. [Background technology] Seabuckthorn flavonoids (TFH) are phytochemicals extracted from seabuckthorn berries, with the main components being quercetin, isorhamnetin, kaempferol and their glycoside derivatives. These components have significant antioxidant, anti-inflammatory, anti-cancer, antibacterial and antiviral biological activities, especially in relieving intestinal inflammation and repairing mucosal damage, and are beneficial to intestinal health.

[0003] However, the poor solubility and low bioavailability of seabuckthorn flavonoids limit their application in the field of functional foods and preventive medicine. Whether flavonoids can effectively function in the body depends on whether they can be fully absorbed and accurately delivered to the target tissue. Most flavonoid components in natural products are substrates of phase II metabolic enzymes or transporters, and are prone to phase II metabolic reactions, resulting in reduced bioavailability in the body. [Summary of the invention] The present invention aims to provide a modified Zein nanosystem for delivering seabuckthorn flavonoids, so as to solve the problems that seabuckthorn flavonoids have poor solubility and low bioavailability, and their application in the field of functional food and preventive medicine is limited. The present invention is achieved by the following technical solutions: A modified Zein nanosystem for delivering seabuckthorn flavonoids, wherein the modified Zein nanosystem for delivering seabuckthorn flavonoids is composed of modified zein (D-Zein), seabuckthorn flavonoids (TFH) and anionic polysaccharides (AP), and the weight ratio of the modified zein to seabuckthorn flavonoids is (13-16):1.

[0005] The present invention forms a TFH-D-Zein-AP nanosystem by loading seabuckthorn flavonoids into modified zein and combining it with anionic polysaccharides. First, the modified zein is used as a surfactant to improve the stability and solubility of seabuckthorn flavonoids in the body, thereby improving its bioavailability. The higher the ratio of D-Zein:TFH, the more conducive it is to the dissolution of TFH. However, when the ratio is too high, the stability of the nanosystem will deteriorate. The selection of a ratio range of (13-16):1 can ensure the dissolution effect of TFH and the stability of the system. Second, the use of anionic polysaccharides as stabilizers can improve the stability and dispersibility of nanoparticles, thereby improving the delivery efficiency of seabuckthorn flavonoids; at the same time, as a coating material, it can be physically combined with nanoparticles to form an interface film on the surface to promote the stability of the nanosystem. Third, the TFH-D-Zein-AP nanosystem formed by seabuckthorn flavonoids, modified zein and anionic polysaccharides provides a protective environment for seabuckthorn flavonoids, which can effectively protect seabuckthorn flavonoids from oxidation and degradation, which helps to maintain the anti-inflammatory activity of seabuckthorn flavonoids and enable them to play a role in the body for a longer time. In addition, this TFH-D-Zein-AP nanosystem can adjust the size and surface properties of nanoparticles by adjusting the condition parameters during the preparation process, making it possible to regulate the release rate of seabuckthorn flavonoids, thereby achieving a more precise therapeutic effect.

[0006] Specifically, by Figure 1 Fourier transform infrared spectroscopy showed that the formation of nanoparticles was mainly due to hydrogen bonding and electrostatic interactions between modified zein, TFH and pectin. Figure 2 As shown in the XRD spectrum, free TFH and the physical mixture with the same composition as the nanoparticles are in crystalline form, while the crystalline peak of TFH in the nanoparticles disappears, and after the nanoparticles are formed, they present an amorphous structure, indicating that TFH is well embedded in the nanodelivery system.

[0007] Preferably, the anionic polysaccharide is one of pectin, xanthan gum and chitosan.

[0008] Among them, both pectin and xanthan gum have excellent stability and dispersibility, good water solubility, and high biocompatibility and safety. In addition to basic properties, pectin also has a variety of biological activities such as immunomodulation and anti-oxidation. Xanthan gum has a unique helical copolymer structure, which enables it to form a stable network structure, support the morphology of solid particles, droplets and bubbles, and give it excellent emulsification stability and high suspension performance. It also has thixotropy and pseudoplasticity, which can improve the taste and rheology of the product.

[0009] Specifically, the preparation method of the pectin solution is: adding pectin powder into ultrapure water, heating in a water bath, stirring, rapidly cooling, stirring, and obtaining the pectin solution.

[0010] The preparation method of the xanthan gum comprises the following steps: adding xanthan gum powder into ultrapure water and stirring the water by magnetic force to obtain a xanthan gum solution.

[0011] The present invention uses anionic polysaccharides as stabilizers, which can improve the stability and dispersibility of nanoparticles, thereby improving the delivery efficiency of seabuckthorn flavonoids and ensuring that seabuckthorn flavonoids can be more evenly and effectively distributed in the body. At the same time, as a coating material, it can be physically combined with nanoparticles to form an interface film on the surface, thereby promoting the stability of the nanosystem.

[0012] Preferably, the method for preparing the modified zein comprises the following steps: Zein is dispersed in an alkaline aqueous solution and stirred at room temperature to obtain a 1% dispersion. The 1% dispersion is heated in a constant temperature water bath to obtain a modified solution. After the modified solution is cooled, it is dialyzed for 24 hours and freeze-dried to obtain the modified zein (D-Zein).

[0013] Among them, zein is the main storage protein in corn, accounting for about 50% of the total corn protein. It is a mixture of peptides with different molecular weights, solubility and charge. In its natural state, zein has limited solubility.

[0014] The present invention can transform the spatial configuration of Zein from a tightly folded state to a more open and flexible structure by deamidating Zein under alkaline heating conditions, thereby increasing its solubility and further improving its dispersibility in various solvents (such as alkaline aqueous solutions, ethanol solutions, etc.); it can enhance the interaction between D-Zein and seabuckthorn flavonoids, increase the loading amount and stability of seabuckthorn flavonoids; and it can also optimize the performance of the TFH-D-Zein-AP nanosystem, and improve the delivery efficiency and anti-inflammatory activity of seabuckthorn flavonoids.

[0015] In addition, the “1%” of the 1% dispersion means that the mass of zein accounts for 1% of the volume of the alkaline aqueous solution.

[0016] Preferably, the pH value of the alkaline aqueous solution is 12.

[0017] Among them, during the dispersion process of zein, it is necessary to ensure that the pH value of the dispersion is 12, thereby ensuring the smooth deamidation modification process of Zein, providing a good foundation for the subsequent nanosystem construction and seabuckthorn flavonoids delivery.

[0018] Preferably, the heating temperature is 70° C., and the heating time is 4 h, 6 h, 8 h, 10 h or 12 h.

[0019] The present invention also provides a method for preparing a modified Zein nanosystem for delivering seabuckthorn flavonoids.

[0020] The method for preparing the modified Zein nanosystem for delivering seabuckthorn flavonoids comprises the following steps: S1. Dissolving modified Zein: Add 70% ethanol aqueous solution into a reaction vessel, add modified Zein freeze-dried powder, stir, and adjust the pH value of the solution to neutral to obtain a modified Zein ethanol solution; S2. Loading seabuckthorn flavonoids: adding seabuckthorn flavonoids to the modified Zein ethanol solution in step S1, stirring to fully dissolve the seabuckthorn flavonoids, and obtaining a modified Zein ethanol solution loaded with seabuckthorn flavonoids; S3, anti-solvent precipitation: under continuous stirring, dispersing the modified Zein ethanol solution loaded with seabuckthorn flavonoids in step S2 into water, adjusting the pH value to obtain a dispersion, and continuing to stir the dispersion, evaporating the ethanol in the dispersion by a rotary evaporator, and then adding a corresponding volume of water to compensate for the evaporated ethanol volume; S4, stabilizing the system: injecting the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S3 into the anionic polysaccharide solution, stirring, evaporating the ethanol in the solution by a rotary evaporator, and then adding a corresponding volume of water to compensate for the evaporated ethanol volume to obtain a modified Zein nanosystem for delivering seabuckthorn flavonoids.

[0021] The present invention forms a TFH-D-Zein-AP nanosystem by loading seabuckthorn flavonoids in modified zein and combining it with anionic polysaccharides. First, 70% ethanol aqueous solution is used to completely dissolve D-Zein to obtain a uniform modified Zein ethanol solution, and the solution is adjusted to neutrality to ensure the stability of the protein and prevent it from deforming or aggregating, thereby laying a foundation for subsequent steps. Second, by loading seabuckthorn flavonoids in modified zein, the stability and solubility of seabuckthorn flavonoids in the body can be improved, thereby improving its bioavailability. Third, by utilizing the principle of anti-solvent precipitation, the modified zein loaded with seabuckthorn flavonoids is precipitated from the ethanol solution to form stable nanoparticles, thereby improving the bioavailability, delivery efficiency and anti-inflammatory activity of seabuckthorn flavonoids, and then ethanol is removed by evaporation, so that the nanoparticles are stably present in the aqueous phase, thereby improving the biocompatibility and safety of the nanosystem. Fourth, through the coating and stabilizing effects of the anionic polysaccharide, a TFH-D-Zein-AP nanosystem with good stability and dispersibility is formed.

[0022] Preferably, the stirring speed in step S1 is 500 rpm, and the stirring time is 1 hour, and the stirring speed in step S2 is 700 rpm, and the stirring time is 1 hour.

[0023] Preferably, the stirring speed in step S3 is 800 rpm, and the stirring time is 5 min; the stirring speed in step S4 is 800 rpm, and the stirring time is 1 h.

[0024] The present invention also provides an application of a modified Zein nanosystem for delivering seabuckthorn flavonoids.

[0025] The modified Zein nanosystem for delivering seabuckthorn flavonoids is used in the treatment of inflammation.

[0026] The beneficial effects of the present invention are as follows: The modified Zein nanosystem for delivering seabuckthorn flavonoids of the present invention is formed by loading seabuckthorn flavonoids into modified zein and combining it with anionic polysaccharides to form a TFH-D-Zein-AP nanosystem. First, the use of modified zein can improve the stability and solubility of seabuckthorn flavonoids in the body, thereby improving its bioavailability. Second, the use of anionic polysaccharides as stabilizers can improve the stability and dispersibility of nanoparticles, thereby improving the delivery efficiency of seabuckthorn flavonoids; at the same time, as a coating material, a protective layer is formed on the surface of the nanoparticles. Third, the TFH-D-Zein-AP nanosystem formed by seabuckthorn flavonoids, modified zein and anionic polysaccharides provides a protective environment for seabuckthorn flavonoids, which can effectively protect seabuckthorn flavonoids from oxidation and degradation, which helps to maintain the anti-inflammatory activity of seabuckthorn flavonoids and enable them to play a role in the body for a longer time.

[0027] The invention discloses a method for preparing a modified Zein nanosystem for delivering seabuckthorn flavonoids. First, a 70% ethanol aqueous solution is used to completely dissolve D-Zein to obtain a uniform modified Zein ethanol solution, and the solution is adjusted to neutrality to ensure the stability of the protein and prevent it from deforming or aggregating, thereby laying a foundation for subsequent steps. Second, by loading seabuckthorn flavonoids in modified zein, the stability and solubility of seabuckthorn flavonoids in the body can be improved, thereby improving its bioavailability. Third, by utilizing the principle of anti-solvent precipitation, the modified zein loaded with seabuckthorn flavonoids is precipitated from the ethanol solution to form stable nanoparticles, thereby improving the bioavailability, delivery efficiency and anti-inflammatory activity of seabuckthorn flavonoids, and then removing ethanol by evaporation to allow the nanoparticles to stably exist in the aqueous phase, thereby improving the biocompatibility and safety of the nanosystem. Fourth, by the coating and stabilizing effects of anionic polysaccharides, a TFH-D-Zein-AP nanosystem with good stability and dispersibility is formed.

[0028] The application of the modified Zein nanosystem for delivering seabuckthorn flavonoids of the present invention can promote the anti-inflammatory effect of seabuckthorn flavonoids by delivering seabuckthorn flavonoids through the nanosystem, and provide a more effective and safer means of treating inflammation.

Brief Description of the Drawings

[0031] The pectin solution is prepared by adding 0.1 g of pectin powder into 100 mL of ultrapure water, heating in a water bath at 70° C., stirring for 30 min, rapidly cooling, and stirring for 1 h to obtain the pectin solution.

[0032] The xanthan gum preparation method comprises the following steps: adding 0.1 g of xanthan gum powder into 100 mL of ultrapure water, and stirring the mixture with magnetic force for 1 hour to obtain a xanthan gum solution.

[0033] Example 1 The method for preparing the modified Zein nanosystem for delivering seabuckthorn flavonoids comprises the following steps: S1. Dissolving the modified Zein: Add 20 mL of 70% ethanol aqueous solution into a reaction container, add 0.4 g of the Z10 freeze-dried powder, stir at 500 rpm for 1 h, and adjust the pH value of the solution to neutral (pH=7) to obtain a modified Zein ethanol solution; S2. Loading seabuckthorn flavonoids: adding 0.031 g seabuckthorn flavonoids (Z10:TFH=13:1) to the modified Zein ethanol solution in step S1, stirring at 700 rpm for 1 hour to fully dissolve the seabuckthorn flavonoids, and obtaining a modified Zein ethanol solution loaded with seabuckthorn flavonoids; S3, anti-solvent precipitation: 4 mL of the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S2 was dispersed into 20 mL of water with continuous stirring at 800 rpm, and the pH value was adjusted to 5 to obtain a dispersion, and the dispersion was continued to be stirred at 800 rpm for 5 min, and the ethanol in the dispersion was evaporated by a rotary evaporator, and then a corresponding volume of water was added to compensate for the evaporated volume of ethanol; S4, stabilizing the system: inject 4 mL of the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S3 into 16 mL of pectin solution, stir at 800 rpm for 1 h, evaporate the ethanol in the solution by a rotary evaporator, and then add a corresponding volume of water to compensate for the evaporated ethanol volume to obtain the TFH-Z10-AP nanosystem.

[0034] Finally, the TFH-Z10-AP nanosystem was stored at 4° C. or freeze-dried for future use.

[0035] Example 2 The method for preparing the modified Zein nanosystem for delivering seabuckthorn flavonoids comprises the following steps: S1. Dissolving the modified Zein: Add 20 mL of 70% ethanol aqueous solution into a reaction container, add 0.4 g of the Z10 freeze-dried powder, stir at 500 rpm for 1 h, and adjust the pH value of the solution to neutral (pH=7) to obtain a modified Zein ethanol solution; S2. Loading seabuckthorn flavonoids: adding 0.027 g seabuckthorn flavonoids (Z10:TFH=15:1) to the modified Zein ethanol solution in step S1, stirring at 700 rpm for 1 hour to fully dissolve the seabuckthorn flavonoids, and obtaining a modified Zein ethanol solution loaded with seabuckthorn flavonoids; S3, anti-solvent precipitation: 4 mL of the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S2 was dispersed into 20 mL of water with continuous stirring at 800 rpm, and the pH value was adjusted to 5 to obtain a dispersion, and the dispersion was continued to be stirred at 800 rpm for 5 min, and the ethanol in the dispersion was evaporated by a rotary evaporator, and then a corresponding volume of water was added to compensate for the evaporated volume of ethanol; S4, stabilizing the system: inject 4 mL of the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S3 into 16 mL of pectin solution, stir at 800 rpm for 1 h, evaporate the ethanol in the solution by a rotary evaporator, and then add a corresponding volume of water to compensate for the evaporated ethanol volume to obtain the TFH-Z10-AP nanosystem.

[0036] Finally, the TFH-Z10-AP nanosystem was stored at 4° C. or freeze-dried for future use.

[0037] Example 3 The method for preparing the modified Zein nanosystem for delivering seabuckthorn flavonoids comprises the following steps: S1. Dissolving modified Zein: Add 20 mL of 70% ethanol aqueous solution into a reaction container, add 0.4 g of the Z8 freeze-dried powder, stir at 500 rpm for 1 h, and adjust the pH value of the solution to neutral (pH=7) to obtain a modified Zein ethanol solution; S2. Loading seabuckthorn flavonoids: adding 0.025 g seabuckthorn flavonoids (Z8:TFH=16:1) to the modified Zein ethanol solution in step S1, stirring at 700 rpm for 1 hour to fully dissolve the seabuckthorn flavonoids, and obtaining a modified Zein ethanol solution loaded with seabuckthorn flavonoids; S3, anti-solvent precipitation: 4 mL of the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S2 was dispersed into 20 mL of water with continuous stirring at 800 rpm, and the pH value was adjusted to 5 to obtain a dispersion, and the dispersion was continued to be stirred at 800 rpm for 5 min, and the ethanol in the dispersion was evaporated by a rotary evaporator, and then a corresponding volume of water was added to compensate for the evaporated volume of ethanol; S4, stabilizing the system: inject 4 mL of the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S3 into 16 mL of pectin solution, stir at 800 rpm for 1 h, evaporate the ethanol in the solution by a rotary evaporator, and then add a corresponding volume of water to compensate for the evaporated ethanol volume to obtain the TFH-Z8-AP nanosystem.

[0038] Finally, the TFH-Z8-AP nanosystem was stored at 4°C or freeze-dried for future use.

[0039] Example 4 The method for preparing the modified Zein nanosystem for delivering seabuckthorn flavonoids comprises the following steps: S1. Dissolving the modified Zein: Add 20 mL of 70% ethanol aqueous solution into a reaction container, add 0.4 g of the Z10 freeze-dried powder, stir at 500 rpm for 1 h, and adjust the pH value of the solution to neutral (pH=7) to obtain a modified Zein ethanol solution; S2. Loading seabuckthorn flavonoids: adding 0.031 g seabuckthorn flavonoids (Z10:TFH=13:1) to the modified Zein ethanol solution in step S1, stirring at 700 rpm for 1 hour to fully dissolve the seabuckthorn flavonoids, and obtaining a modified Zein ethanol solution loaded with seabuckthorn flavonoids; S3, anti-solvent precipitation: 4 mL of the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S2 was dispersed into 20 mL of water with continuous stirring at 800 rpm, and the pH value was adjusted to 5 to obtain a dispersion, and the dispersion was continued to be stirred at 800 rpm for 5 min, and the ethanol in the dispersion was evaporated by a rotary evaporator, and then a corresponding volume of water was added to compensate for the evaporated volume of ethanol; S4, stabilizing the system: inject 4 mL of the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S3 into 16 mL of xanthan gum solution, stir at 800 rpm for 1 h, evaporate the ethanol in the solution by a rotary evaporator, and then add a corresponding volume of water to compensate for the evaporated ethanol volume to obtain the TFH-Z10-AP nanosystem.

[0040] Finally, the TFH-Z10-AP nanosystem was stored at 4° C. or freeze-dried for future use.

[0041] The above-mentioned embodiments 1-4 were subjected to performance tests. The test conditions were as follows and the test data are shown in Table 1.

[0042] 1) Heat treatment NaCl was added to the modified Zein nanosystem for delivering seabuckthorn flavonoids, the final concentration of NaCl being 0-50 mM, and then a digital camera was used to take photos to record the overall appearance of the treated nanosystem, and to measure the average diameter, PDI and Zeta potential of the nanoparticles.

[0043] Among them, PDI is the polydispersity index, which is used to describe the uniformity of the size distribution of nanoparticles. When PDI is close to 0, it means that the size distribution of nanoparticles is very uniform, and when PDI is close to 1, it means that the size distribution of nanoparticles is very uneven.

[0044] Zeta potential is used to describe the surface charge density of nanoparticles and the interaction between nanoparticles and the surrounding environment. When the absolute value of Zeta potential is higher, it means that the repulsion between nanoparticles is stronger and the system is more stable. When the absolute value of Zeta potential is lower, it means that the repulsion between nanoparticles is weaker and the system is more unstable.

[0045] 2) Evaluation of the effect of promoting TFH absorption efficiency A: Simulated digestion Gastric phase: Free TFH powder, TFH-D-Zein-Ap and a physical mixture with the same composition as the nanosystem were dissolved in 20 mL of deionized water and magnetically stirred at 800 rpm for 1 h. 20 mL of simulated gastric fluid was added and stirring was continued. The pH value of the sample was then adjusted to 2.5, 0.064 g of pepsin was added, and the gastric digestion phase was started in an oscillating water bath at 37 °C. After 120 min, the pH was adjusted to 7.0 to terminate gastric digestion.

[0046] The preparation method of the simulated gastric fluid is as follows: 1 g NaCl and 3.5 mL hydrochloric acid are dissolved in 500 mL distilled water, and the pH is adjusted to 1.2.

[0047] Intestinal phase: After the gastric phase digestion is completed, add 4 mL of bile salt solution and 2.5 mL of pancreatic enzyme suspension, and then use a pH titrator to continue digestion at pH 7.0 for 4 hours at 50 rpm with magnetic stirring. After the small intestine digestion is completed, heat the sample in a constant temperature water bath at 90°C for 5 minutes to inactivate the pancreatic enzymes.

[0048] The bile salt solution and the pancreatic enzyme suspension were both prepared using 5 mM PBS.

[0049] B: Bioavailability determination The evaluation of the nano-delivery system in promoting the in vivo absorption of TFH was characterized by bioavailability. The above digestion solution was centrifuged and the supernatant was collected. After dilution with DMSO, the release amount of TFH was measured by a spectrophotometer. The calculation formula is as follows: Gastric phase: Bioavailability of TFH (%) = amount of TFH released after gastric digestion / amount of TFH added before digestion × 100%.

[0050] Intestinal phase: Bioavailability of TFH (%) = amount of TFH released after intestinal digestion / amount of TFH added before digestion × 100%.

[0051] 3) Evaluation of the promotion effect of TFH anti-inflammatory effect Blank control group: untreated mouse peritoneal primary macrophages.

[0052] Inflammation model group: Primary mouse peritoneal macrophages were stimulated with LPS.

[0053] Free TFH group: Seabuckthorn flavonoids were added 2 hours before LPS stimulation of mouse peritoneal primary macrophages.

[0054] TFH-Z10-AP group: The modified Zein nanosystem for delivering seabuckthorn flavonoids was added 2 hours before LPS stimulation of mouse peritoneal primary macrophages.

[0055] The above four groups of LPS-induced inflammatory cell models were treated at different times, and the relative expression levels of the circadian clock genes Bmal1 and Per2 were detected by RT-qPCR.

[0056] Table 1 Examples 1-4 Heat treatment data

[0057] Table 2 Bioavailability of TFH-Z10-AP nanosystem in Example 1

[0058] Table 3 Example 1 Regulation of TFH-Z10-AP nanosystem on inflammation-related circadian clock genes

[0059] Note: Different superscript letters in the expression level values ​​indicate significant differences between groups.

[0060] As can be seen from Table 1, the nanoparticles of the nanosystems of Examples 1-4 are all nearly circular in morphology, with particle sizes of 150.3, 169.2, 113.5 and 111.8 nm, respectively, PDI values ​​close to 0, Zeta potential absolute values ​​relatively high, and TFH loading rates above 83.61%.

[0061] This indicates that the nanosystem prepared by the anti-solvent precipitation method of the present invention has small nanoparticles, uniform distribution, good system stability, and high TFH loading rate.

[0062] Specifically, Figure 3 As shown in FIG. 1 , the particle size distribution of the nanoparticles of the TFH-Z10-AP nanosystem of Example 1 is a narrow single-peak distribution, with a particle size of 150.3 nm. Figure 4 As shown, the particle shape is nearly round and relatively uniform.

[0063] As shown in Table 2, in the gastric phase, the bioavailability of free TFH powder was only 1.28%, that of the physical mixture was 4.71%, and that of TFH released by the nanosystem was as high as 6.66%. In the intestinal phase, the bioavailability of TFH released by the nanosystem was 39.78%, which was still significantly higher than that of free TFH powder (20.34%) and physical mixture (23.73%).

[0064] This indicates that the nanosystem of the present invention has a significant promoting effect on the absorption efficiency of TFH in vivo.

[0065] As shown in Table 3, the expression level of Bmal1 in the inflammation model group (100+5.23 c ) was significantly lower than the blank control group (140+8.25 a ), indicating that the rhythmicity of the Bmal1 clock gene was disrupted in the inflammatory model. After adding free TFH to the inflammatory model, the expression level of Bmall increased to 112+4.33 b After adding TFH-Z10-AP, the expression level of Bmall increased significantly to 135+8.25 a The expression level of Bmall was close to that of the blank control group (140+8.25 a ).

[0066] The expression of Per2 gene in the inflammation model group, free TFH group and TFH-Z10-AP group showed a similar trend to that of Bmal1. After adding TFH-Z10-AP to the inflammation model, the expression level of Per2 was significantly increased to 140+3.21 a The Per2 expression level was also close to that of the blank control group (152+3.21 a ).

[0067] This indicates that TFH regulates the expression of circadian clock genes Bmall and Per2 in the process of exerting its anti-inflammatory effect, and the TFH-D-Zein-AP nanosystem can promote the anti-inflammatory effect of TFH.

[0068] The above examples are only used to further illustrate the technical content of the present invention, so that readers can understand it more easily, but they do not mean that the implementation of the present invention is limited to this. Any technical extension or re-creation made according to the present invention is protected by the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. A modified Zein nanosystem for delivering seabuckthorn flavonoids, characterized in that: The modified Zein nanosystem for delivering seabuckthorn flavonoids is composed of modified zein, seabuckthorn flavonoids and anionic polysaccharides, and the weight ratio of the modified zein to seabuckthorn flavonoids is (13-16):

1.

2. The modified Zein nanosystem for delivering seabuckthorn flavonoids according to claim 1, characterized in that: The anionic polysaccharide is one of pectin, xanthan gum and chitosan.

3. The modified Zein nanosystem for delivering seabuckthorn flavonoids according to claim 1, characterized in that: The preparation method of the modified zein comprises the following steps: The zein was dispersed in an alkaline aqueous solution and stirred at room temperature to obtain a 1% dispersion, the 1% dispersion was heated in a constant temperature water bath to obtain a modified solution, and after the modified solution was cooled, it was dialyzed for 24 hours and freeze-dried to obtain the modified zein.

4. The modified Zein nanosystem for delivering seabuckthorn flavonoids according to claim 3, characterized in that: The pH value of the alkaline aqueous solution is 12.

5. The modified Zein nanosystem for delivering seabuckthorn flavonoids according to claim 3, characterized in that: The heating temperature is 70° C., and the heating time is 4 h, 6 h, 8 h, 10 h or 12 h.

6. The method for preparing the modified Zein nanosystem for delivering seabuckthorn flavonoids according to claims 1-5, characterized in that: The following steps are involved: S1. Dissolving modified Zein: Add 70% ethanol aqueous solution into a reaction vessel, add modified Zein freeze-dried powder, stir, and adjust the pH value of the solution to neutral to obtain a modified Zein ethanol solution; S2. Loading seabuckthorn flavonoids: adding seabuckthorn flavonoids to the modified Zein ethanol solution in step S1, stirring to fully dissolve the seabuckthorn flavonoids, and obtaining a modified Zein ethanol solution loaded with seabuckthorn flavonoids; S3, anti-solvent precipitation: under continuous stirring, dispersing the modified Zein ethanol solution loaded with seabuckthorn flavonoids in step S2 into water, adjusting the pH value to obtain a dispersion, and continuing to stir the dispersion, evaporating the ethanol in the dispersion by a rotary evaporator, and then adding a corresponding volume of water to compensate for the evaporated ethanol volume; S4, stabilizing the system: injecting the modified Zein ethanol solution loaded with seabuckthorn flavonoids described in step S3 into the anionic polysaccharide solution, stirring, evaporating the ethanol in the solution by a rotary evaporator, and then adding a corresponding volume of water to compensate for the evaporated ethanol volume to obtain a modified Zein nanosystem for delivering seabuckthorn flavonoids.

7. The method for preparing the modified Zein nanosystem for delivering seabuckthorn flavonoids according to claim 6, characterized in that: The stirring speed in step S1 is 500 rpm and the stirring time is 1 hour. The stirring speed in step S2 is 700 rpm and the stirring time is 1 hour.

8. The method for preparing the modified Zein nanosystem for delivering seabuckthorn flavonoids according to claim 6, characterized in that: In step S3, the stirring speed is 800 rpm, and the stirring time is 5 min. In step S4, the stirring speed is 800 rpm, and the stirring time is 1 h.

9. Use of the modified Zein nanosystem for delivering seabuckthorn flavonoids according to claims 1-5 in the treatment of inflammation.

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