Preparation method and application of an exosome ferulic acid nanocomplex

Abstract

The present invention belongs to the field of biological carrier technology, and specifically relates to a method for preparing an exosome ferulic acid nanocomposite and its application. The present invention provides a method for preparing ferulic acid nanoparticles using exosomes, especially a preparation process for forming a nanocomposite of stress yeast exosomes and ferulic acid produced by stimulation. The method for preparing the exosome ferulic acid nanocomposite provided by the present invention has the advantages of high safety, low immune rejection, high cell fusion, high encapsulation rate and good stability. In addition, the stress yeast exosomes selected by the present invention can also effectively protect and enhance the anti-inflammatory and antioxidant effects of ferulic acid, so that the prepared exosome ferulic acid nanocomposite has strong anti-inflammatory, antioxidant and repair effects, and has good application prospects in the fields of cosmetics or medicine.

Description

Technical Field

[0001] The present invention belongs to the technical field of biological carriers, and particularly relates to a preparation method and application of an exosome ferulic acid nanocomplex. Background Art

[0002] Ferulic acid (FA) is a phenolic acid extracted from plant species such as Ferula, Angelica sinensis, Ligusticum chuanxiong, Cimicifuga foetida, etc., and is also one of the main active ingredients of these Chinese herbal medicines. Ferulic acid mainly exists in the cell walls of plants, mostly in the form of compounds. It is a cinnamic acid-type phenolic acid and one of the derivatives of cinnamic acid. Ferulic acid has a wide range of pharmacological effects and is clinically used to treat diseases such as hypertension, coronary heart disease, vascular embolism vasculitis, acute cerebral thrombosis, migraine, and atherosclerosis. With in-depth research in recent years, it has been found that ferulic acid also has many new pharmacological effects, such as inhibiting tyrosinase activity, absorbing ultraviolet rays, antioxidation, scavenging free radicals, and delaying skin aging. However, its low solubility in water, low bioavailability, and poor stability under physical and thermal stresses greatly limit the application of ferulic acid.

[0003] Exosomes refer to nanospheres with a bilayer membrane structure that are produced by cells for intercellular communication, with a diameter between 40 and 100 nm. They can carry nucleic acids, proteins, lipids, and enzymes. Exosomes from different sources have their specific surface molecules and play an important role in intercellular information transmission. Due to the small size of exosomes, they can reduce the clearance effect of the immune system and transport more drugs to the lesion site. Therefore, the potential function of exosomes as drug carriers can load compounds into the vesicle-like structure to achieve the functions of aqueous solution drug loading, increased transdermal and targeted delivery. At present, exosomes have been applied in the encapsulation of compounds, achieving good transdermal and delivery effects.

[0004] Patent document CN115105494A discloses an exosome-encapsulated glabridin with whitening efficacy, its preparation method and application. The exosome-encapsulated glabridin is prepared by mixing exosomes secreted by dermal fibroblasts with an alcohol solution dissolved with glabridin at a mass ratio of (1 - 5):(10 - 20). The exosome-encapsulated glabridin solves the problem that the effect of glabridin is limited in actual application. The exosome-encapsulated glabridin of dermal fibroblasts can assist glabridin to efficiently penetrate the melanocyte cell membrane, enhance the ability of glabridin to target melanocytes, play a role in inhibiting melanin synthesis, and thus effectively treat diseases such as abnormal pigmentation, and improve the efficiency and utilization rate of glabridin.

[0005] Patent document CN 116392601A discloses a preparation of resveratrol encapsulated in composite-modified exosomes and its preparation method. This preparation uses milk-derived exosomes to encapsulate resveratrol, and then cholesterol-RGD modification is carried out on the exosomes to obtain the preparation, making the preparation of resveratrol encapsulated in composite-modified exosomes have the advantages of strong availability, low cost, low toxicity, high yield, and suitability for large-scale production. By cholesterol-RGD modification of milk-derived exosomes, the bioavailability of encapsulated resveratrol is enhanced, and the efficacy of resveratrol can also be improved.

[0006] In view of the fact that current carrier technologies for ferulic acid, such as phospholipid complexes, nanoemulsions, cyclodextrin clathrates, hydrogels, molecular cages, etc., all have certain defects, including low encapsulation efficiency, low stability, decreased anti-inflammatory and antioxidant effects after encapsulation, immune rejection, etc. Therefore, it is an urgent problem to be solved at present to research and develop a ferulic acid encapsulation process with high biocompatibility, high encapsulation efficiency, and high antioxidant and anti-inflammatory effects. Summary of the Invention

[0007] To solve the defects of the prior art, the present invention provides a method for preparing ferulic acid nanoparticles using exosomes, especially a preparation method of forming a nano-complex by stress-induced yeast exosomes and ferulic acid. The purpose of the present invention is to provide a preparation method of exosome-ferulic acid nano-complex, especially using stress-induced yeast exosomes to obtain an exosome-ferulic acid nano-complex with high biocompatibility, high encapsulation efficiency, and high antioxidant and anti-inflammatory effects. Another purpose of the present invention is to provide the application of exosome-ferulic acid nano-complex in the preparation of cosmetics or drugs.

[0008] The present invention provides a preparation method of exosome-ferulic acid nano-complex, including the following steps:

[0009] Step S1. Add yeast-derived exosomes to phosphate buffered saline for resuspension, so that the mass percentage of exosomes in the suspension is 0.05 - 0.12%, to obtain a yeast-derived exosome solution; the concentration of the phosphate buffered saline is 0.01 mol / L;

[0010] Step S2. Add ferulic acid to an ethanol solution with a volume concentration of 75 - 95%, and stir evenly to obtain a ferulic acid solution;

[0011] Step S3. Mix the yeast-derived exosome solution prepared in Step S1 and the ferulic acid alcohol solution prepared in Step S2 evenly to obtain a mixture. Under the condition of ice bath, ultrasonically treat the mixture, then let it stand for 60 - 100 min at a temperature of 20 - 37°C, centrifuge, and take the supernatant to obtain the product.

[0012] Furthermore, the preparation method of the yeast-derived exosomes in Step S1 is:

[0013] Step A: Take the activated Pichia pastoris plate strain and inoculate it into a conical flask containing YEPD medium. Incubate it on a shaker at 28°C for 12 h to obtain the Pichia pastoris seed solution.

[0014] Step B: Inoculate the Pichia pastoris seed solution prepared in Step A into a conical flask containing fresh YEPD medium at an inoculation amount of 5%. Incubate it on a shaker at 28°C until the mid-logarithmic phase, then raise the temperature of the medium to 40°C and continue to incubate for 36 h until the stationary phase is reached to obtain the fermentation broth.

[0015] Step C: Centrifuge the fermentation broth prepared in Step B at 6000 - 7000 rpm for 8 - 10 min, collect the supernatant, then centrifuge the supernatant at 2 - 8°C and 5000 - 8000 g for 10 - 30 min, collect the supernatant, and filter the supernatant through a 0.2 μm sterile filter membrane to obtain the filtrate.

[0016] Step D: Centrifuge the filtrate prepared in Step C at 100000 - 120000 g for 60 - 80 min, discard the supernatant to obtain the product.

[0017] Furthermore, in Step S2, the concentration of the ferulic acid solution is 1 - 10 mg / mL.

[0018] Furthermore, in Step S3, the yeast-derived exosome solution and the ferulic acid solution are mixed evenly at a mass percentage of (80 - 95):(5 - 20).

[0019] Furthermore, the conditions for ultrasonic treatment in Step S3 are as follows: under the condition of ice bath, the ultrasonic power is 200 - 400 W, the working time is 2 - 10 s, the interval time is 2 - 34 s, and it is cycled 5 - 40 times. After the ice bath cools down for 3 - 10 min, ultrasonic treatment is carried out again.

[0020] Furthermore, the conditions for centrifugation in Step S3 are as follows: centrifuge at 2 - 4°C and 5000 - 10000 g for 10 - 15 min.

[0021] In addition, the present invention also claims the exosome-ferulic acid nanocomplex prepared by the method for preparing the exosome-ferulic acid nanocomplex.

[0022] In addition, the present invention also provides the application of the exosome-ferulic acid nanocomplex in the preparation of cosmetics or drugs.

[0023] Furthermore, the present invention also provides the application of the exosome-ferulic acid nanocomplex in the preparation of repair and antioxidant cosmetics.

[0024] At present, in order to improve the water solubility and stability of ferulic acid, the inventors consulted a large number of literature materials and for the first time tried to use exosomes to load ferulic acid to improve its performance. However, through experiments, it was found that the exosomes secreted by dermal fibroblasts and plant exosomes did not achieve the expected effect on the encapsulation rate of ferulic acid. Through continuous exploration and research, the inventors found that when Pichia pastoris encounters heat stress, the stress exosomes secreted by yeast cells due to stress responses have good anti-inflammatory and antioxidant effects. The inventors combined and encapsulated the Pichia pastoris stress exosomes with ferulic acid and unexpectedly found that the Pichia pastoris stress exosomes can not only effectively encapsulate ferulic acid, but also induce the expression of the protein content contained in the Pichia pastoris stress exosomes, and can further increase the encapsulation performance of ferulic acid. At the same time, the Pichia pastoris stress exosomes can also enhance the anti-inflammatory and antioxidant effects of ferulic acid, and can effectively solve the defects of low encapsulation rate, decreased stability and decreased anti-inflammatory and antioxidant functions existing in the current ferulic acid encapsulation technology.

[0025] In summary, compared with the prior art, the method for preparing the exosome-ferulic acid nanocomplex provided by the present invention has the advantages of high safety, low immune rejection, high cell fusion degree, high encapsulation rate and good stability. In addition, the stress yeast exosomes selected by the present invention can also effectively protect and enhance the anti-inflammatory and antioxidant effects of ferulic acid, so that the prepared exosome-ferulic acid nanocomplex has strong anti-inflammatory, antioxidant and repair effects, and the nanocomplex is easy to be industrially produced and applied, and has good application prospects in the fields of cosmetics or medicine. Description of the Drawings

[0026] Figure 1 Comparison diagram of the repair effects of ferulic acid and Example 2 on NIH-3T3 cells damaged by hydrogen peroxide. Detailed Embodiments

[0027] The present invention will be further described below through the description of specific embodiments, but this is not a limitation of the present invention. Those skilled in the art can make various modifications or improvements according to the basic idea of the present invention, but as long as they do not depart from the basic idea of the present invention, they are all within the scope of the present invention. The materials involved in the present invention are all food grades and can be obtained through commercial purchase or conventional technical means in the art. For example: the Pichia pastoris is purchased from Beijing Biovector Science Lab Co., Ltd., with the original number: KM-71, the brand: Biovector Science Lab, and the product number: bio-81782.

[0028] Example 1: Preparation method of exosomes derived from Pichia pastoris

[0029] Step A: Take the activated Pichia pastoris plate strain and inoculate it into a conical flask containing YEPD medium. Incubate it on a shaker at 28°C for 12 h to obtain the Pichia pastoris seed solution.

[0030] Step B: Inoculate the Pichia pastoris seed solution prepared in Step A into a conical flask containing fresh YEPD medium. The inoculation amount of the Pichia pastoris seed solution is 5%. Incubate it on a shaker at 28°C until the mid-log phase, then raise the temperature in the medium to 40°C and continue to incubate for 36 h to reach the stationary phase to obtain the fermentation broth.

[0031] Step C: Centrifuge the fermentation broth prepared in Step B at 6000 rpm for 8 min, collect the supernatant, then centrifuge the supernatant at 4°C and 6000 g for 20 min, collect the supernatant, and filter the supernatant through a 0.2-μm sterile filter membrane to obtain the filtrate.

[0032] Step D: Centrifuge the filtrate prepared in Step C at 120000 g for 80 min, discard the supernatant, and that's it.

[0033] Example 2: A method for preparing exosome ferulic acid nanocomposite

[0034] Step S1. Resuspend the Pichia pastoris-derived exosomes prepared in Example 1 in phosphate buffer solution with a concentration of 0.01 mol / L to make the mass percentage of exosomes in the suspension 0.1% to obtain the yeast-derived exosome solution.

[0035] Step S2. Add ferulic acid to ethanol solution with a volume concentration of 95% and stir evenly to make the ferulic acid concentration 1 mg / mL to obtain the ferulic acid solution.

[0036] Step S3. Take 91% of the yeast-derived exosome solution prepared in Step S1 and 9% of the ferulic acid alcohol solution prepared in Step S2 by mass percentage, mix them evenly to obtain a mixed solution. Under the condition of ice bath, ultrasonically treat the mixed solution. The conditions of the ultrasonic treatment are: ultrasonic power is 200 W, working time is 5 s, interval time is 10 s, cycle 8 times. After cooling in the ice bath for 5 min, perform ultrasonic treatment again, then let it stand at 37°C for 60 min, centrifuge at 4°C and 8000 g for 15 min, and take the supernatant, and that's it.

[0037] Example 3: A method for preparing exosome ferulic acid nanocomposite

[0038] Step S1. Resuspend the exosomes derived from Pichia pastoris prepared in Example 1 in a phosphate buffer solution with a concentration of 0.01 mol / L, such that the mass percentage of exosomes in the suspension is 0.08%, to obtain a yeast-derived exosome solution;

[0039] Step S2. Add ferulic acid to an ethanol solution with a volume concentration of 95%, stir evenly to make the concentration of ferulic acid 5 mg / mL, to obtain a ferulic acid solution;

[0040] Step S3. Take 95% of the yeast-derived exosome solution prepared in Step S1 and 5% of the ferulic acid alcohol solution prepared in Step S2 by mass percentage, mix them evenly to obtain a mixture. Under the condition of ice bath, ultrasonically treat the mixture. The conditions for the ultrasonic treatment are: ultrasonic power is 50 W, working time is 10 s, interval time is 2 s, cycle 40 times. After cooling in the ice bath for 10 min, perform ultrasonic treatment again. Then, let it stand at 25 °C for 80 min, centrifuge at 8 °C and 10,000 g for 10 min, and take the supernatant to obtain the product.

[0041] Example 4. A method for preparing an exosome-ferulic acid nanocomplex

[0042] Step S1. Resuspend the exosomes derived from Pichia pastoris prepared in Example 1 in a phosphate buffer solution with a concentration of 0.01 mol / L, such that the mass percentage of exosomes in the suspension is 0.12%, to obtain a yeast-derived exosome solution;

[0043] Step S2. Add ferulic acid to an ethanol solution with a volume concentration of 95%, stir evenly to make the concentration of ferulic acid 10 mg / mL, to obtain a ferulic acid solution;

[0044] Step S3. Take 84% of the yeast-derived exosome solution prepared in Step S1 and 16% of the ferulic acid alcohol solution prepared in Step S2 by mass percentage, mix them evenly to obtain a mixture. Under the condition of ice bath, ultrasonically treat the mixture. The conditions for the ultrasonic treatment are: ultrasonic power is 400 W, working time is 2 s, interval time is 34 s, cycle 5 times. After cooling in the ice bath for 3 min, perform ultrasonic treatment again. Then, let it stand at 20 °C for 100 min, centrifuge at 2 °C and 5,000 g for 10 min, and take the supernatant to obtain the product.

[0045] Comparative Example 1. Preparation of ferulic acid-β-cyclodextrin inclusion compound

[0046] Prepare the ferulic acid-β-cyclodextrin inclusion complex according to the optimal method in the reference (Wang Liping. Study on the preparation of ferulic acid-β-cyclodextrin inclusion complex and preservation of hairtail [D]. Zhejiang University, 2018.), that is, under the condition of a molar ratio of β-cyclodextrin to ferulic acid of 1:0.5, mix the ferulic acid aqueous solution and the β-cyclodextrin ethanol solution, stir for 24 h, keep it overnight in a refrigerator at 4 °C, and then dry it in an oven at 60 °C for 12 h to obtain it.

[0047] Comparative Example 2. Preparation of ferulic acid solid lipid nanoparticles

[0048] Prepare ferulic acid solid lipid nanoparticles according to the method in the reference (Gao Yimin, Zhang Zhen, Chen Peimin, et al. Comparison of the processes for preparing ferulic acid solid lipid nanoparticles by emulsion evaporation method and thin film-ultrasound method [J]. Food and Fermentation Industries, 2019).

[0049] Comparative Example 3. Preparation of exosome-ferulic acid nanocomposites

[0050] Prepare conventional yeast exosomes according to the method in the reference (Yu Qiang. Research on the extraction, characterization of tumor-related extracellular exosomes and their application methods based on aptamers [D]. Beijing: Beijing University of Chemical Technology, 2020.), and the remaining steps refer to Example 2 to obtain exosome-ferulic acid nanocomposites.

[0051] Experimental Example 1. Encapsulation efficiency test

[0052] 1. Experimental materials:

[0053] The exosome-ferulic acid nanocomposites prepared in Example 2, Example 3 and Example 4, the ferulic acid-β-cyclodextrin inclusion complex prepared in Comparative Example 1, the ferulic acid solid lipid nanoparticles prepared in Comparative Example 2, and the exosome-ferulic acid nanocomposites prepared in Comparative Example 3.

[0054] 2. Experimental method:

[0055] Take the exosome-ferulic acid nanocomposites prepared in Example 2, Example 3 and Example 4, the ferulic acid-β-cyclodextrin inclusion complex prepared in Comparative Example 1, the ferulic acid solid lipid nanoparticles prepared in Comparative Example 2, and the exosome-ferulic acid nanocomposites prepared in Comparative Example 3 for liquid chromatography analysis; take 10 mg of the sample to be tested, dissolve it in 1 mL of deionized water, ultrasonically dissolve it at room temperature for 10 min, and centrifuge it at 8000 r / min for 15 min. Take the supernatant, take 200 μL of the supernatant, dissolve it in 2 mL of deionized water, centrifuge it at 8000 g at 4 °C for 15 min, and then take the supernatant for liquid chromatography analysis to determine the content of the encapsulated ferulic acid. Calculate the encapsulation efficiency according to the following formula:

[0056] Encapsulation efficiency = Content of the encapsulated ferulic acid / Amount of ferulic acid input * 100%

[0057] HPLC analysis was performed using a Shimadzu high performance liquid chromatograph, and the detector was SPD-M20; chromatographic conditions: C18 column (4.6 mm * 150 mm; 5 μm), detection wavelength: 313 nm, column temperature: 35 °C, and the mobile phase was tested with methanol-0.1% formic acid water at 1 mL / min. The conditions were as follows: time program (gradient elution): 0 - 5 min, 10% methanol, 90% formic acid water; 20 min, 100% methanol, 0% formic acid water; 20 - 27 min, 100% methanol, 0% formic acid water.

[0058] Preparation of ferulic acid standard solution: Accurately weigh 0.0053 g of ferulic acid, dissolve it with methanol and make up the volume to 10 mL in a volumetric flask to form a stock solution (530 μg / mL). Respectively pipette 0.2, 0.4, 0.8, 1.6, and 3.2 mL, dilute with methanol and make up the volume to 10 mL in a volumetric flask to form a series of standard solutions with concentrations of 10.6, 21.2, 42.2, 84.8, and 169.6 μg / mL. Ferulic acid standard curve: y = 46048x + 51151, R 2 = 0.9993.

[0059] 3. Test results:

[0060] The test results are shown in Table 1.

[0061] Table 1 Test results of ferulic acid encapsulation rate

[0062] Group Wrapping rate (%) Example 2 89.33 Example 3 83.16 Example 4 80.09 Comparative Example 1 68.82 Comparative Example 2 52.76 Comparative Example 3 73.36

[0063] As can be seen from Table 1, the encapsulation rates of the exosome-ferulic acid nanocomposites prepared in Examples 2 - 4 of the present invention are all higher than those of Comparative Example 1, Comparative Example 2, and Comparative Example 3, indicating that the Pichia pastoris stress exosomes prepared in the present invention are more suitable for encapsulating ferulic acid than β-cyclodextrin, solid lipid nanoparticles, and conventional yeast exosomes, have a better encapsulation effect, and can effectively improve the stability of ferulic acid.

[0064] Test Example 2. DPPH free radical scavenging ability test

[0065] 1. Test materials:

[0066] Vitamin C, ferulic acid, exosome-ferulic acid nanocomposite prepared in Example 2, exosome-ferulic acid nanocomposite prepared in Comparative Example 3.

[0067] 2. Test method:

[0068] Dissolve DPPH in 95% ethanol to prepare a DPPH solution with a concentration of 0.04 mg / mL. Prepare solutions of vitamin C, ferulic acid, the exosome-ferulic acid nanocomplex prepared in Example 2, and the exosome-ferulic acid nanocomplex sample prepared in Comparative Example 3 at a concentration of 1 mg / mL, and dilute them into sample groups with concentrations of 5 μg / mL and 25 μg / mL (the complex concentration is calculated based on ferulic acid). Mix 100 μL of DPPH with 50 μL of each sample group, shake well, let stand at room temperature for 30 min, and then measure the absorbance at 519 nm, denoted as A1; the control group is to mix 100 μL of DPPH with 50 μL of 95% ethanol, measure the absorbance at 519 nm, denoted as A0; the blank group is to mix 100 μL of 95% ethanol with 50 μL of each sample group, measure the absorbance at 519 nm, denoted as A2.

[0069] The DPPH radical scavenging rate is calculated according to the following formula:

[0070]

[0071] 3. Test results:

[0072] The test results are shown in Table 2.

[0073] Table 2 Test results of DPPH radical scavenging ability

[0074]

[0075] As can be seen from Table 2, compared with the ferulic acid group and the Comparative Example 3 group, encapsulating ferulic acid with the Pichia pastoris stress exosomes prepared by the present invention can effectively improve the DPPH radical scavenging rate of the ferulic acid complex. When the ferulic acid concentration is 25 μg / mL, its DPPH radical scavenging rate is close to that of vitamin C at the same concentration, indicating that the exosome-ferulic acid nanocomplex prepared by the present invention has strong antioxidant effects.

[0076] Test Example Three: Cell repair test

[0077] 1. Test materials:

[0078] Ferulic acid (FA), the exosome-ferulic acid nanocomplex prepared in Example 2.

[0079] 2. Test method:

[0080] NIH-3T3 cells were cultured in complete DMEM medium (containing 10% fetal bovine serum and 1% penicillin-streptomycin). 100 μL of NIH-3T3 cell suspension at 5000 cells / well was seeded in each well of a 96-well plate. After culturing for 24 h, the medium was aspirated, and the cells were gently washed once with PBS solution. Then, 100 μL of 600 μmol / mL hydrogen peroxide solution was added to establish a model for 2 h. After that, the hydrogen peroxide solution was aspirated, and the cells were gently washed once with PBS solution. The drug treatment groups (ferulic acid group and Example 2 group) were added with 100 μL of each concentration of the prepared drug (the complex concentration was calculated based on ferulic acid), the model group and the blank control group (cells without model establishment) were added with complete medium, and a zero adjustment group containing only medium without cells was set. After incubating for 48 h, the solution was aspirated, and the cells were gently washed once with PBS. Then, 100 μL of MTT solution (0.5 mg / mL) was added. After 4 h, the MTT solution was aspirated, and 150 μL of DMSO solution was added. After 10 min, the OD value was measured at 570 nm using an enzyme-linked immunosorbent assay (ELISA) reader, and the relative cell survival rate was calculated.

[0081] The relative cell survival rate (%) = (OD 模型 / 给药 - OD 调零 ) / (OD 阴性对照 - OD 调零 ) × 100%;

[0082] 3. Test results:

[0083] The test results are as Figure 1 shown. It can be seen from Figure 1 the figure that for the cell survival rates of ferulic acid and Example 2, as the concentration of ferulic acid increases, the cell survival rate of Example 2 shows an obvious enhancing effect. When the concentration of ferulic acid is 100 μg / mL, the cell survival rate of ferulic acid is 69.06%, and the cell survival rate of Example 2 is 77.84%, with the cell survival rate increasing by 8.84%. When the concentration of ferulic acid is 200 μg / mL, the cell survival rate of ferulic acid is 78.45%, and the cell survival rate of Example 2 is 85.90%, with the cell survival rate increasing by 7.45%.

Claims

1. A method for preparing an exosome ferulic acid nanocomposite, characterized in that: The following steps are involved: Step S1. adding yeast-derived exosomes to a phosphate buffered saline solution and resuspending the exosomes so that the mass percentage of the exosomes in the suspension is 0.05-0.12%, thereby obtaining a yeast-derived exosome solution; Step S2. adding ferulic acid to an ethanol solution having a volume concentration of 75 to 95%, stirring uniformly to obtain a ferulic acid solution; Step S3. The yeast-derived exosome solution obtained in step S1 and the ferulic acid alcohol solution obtained in step S2 are uniformly mixed to obtain a mixed solution, and the mixed solution is subjected to ultrasonic treatment under ice bath conditions, and then allowed to stand at a temperature of 20 to 37° C. for 60 to 100 min, centrifuged, and the supernatant is obtained; The method for preparing yeast-derived exosomes in step S1 is: Step A, taking the activated Pichia pastoris plate bacteria, inoculating it into a conical flask filled with YEPD medium, and culturing it in a shaking incubator at a temperature of 28° C. for 12 h to obtain a Pichia pastoris seed solution; Step B, inoculating the Pichia pastoris seed liquid prepared in step A into a conical flask filled with fresh YEPD medium at an inoculation amount of 5%, culturing in a shaking incubator at a temperature of 28° C. to the mid-logarithmic phase, then raising the temperature in the medium to 40° C., and continuing to culture for 36 h to reach a stable phase, to obtain a fermentation broth; Step C, centrifuging the fermentation broth obtained in step B at a speed of 6000-7000 rpm for 8-10 min, collecting the supernatant, centrifuging the supernatant at a temperature of 2-8°C and 5000-8000 g for 10-30 min, collecting the supernatant, and filtering the supernatant through a 0.2 μm sterile filter membrane to obtain a filtrate; Step D: Centrifuge the filtrate obtained in step C at 100,000 to 120,000 g for 60 to 80 min, and discard the supernatant to obtain the product.

2. The method for preparing the exosome ferulic acid nanocomposite according to claim 1, characterized in that: The concentration of the ferulic acid solution in step S2 is 1-10 mg / mL.

3. The method for preparing the exosome ferulic acid nanocomposite according to claim 1, characterized in that: In step S3, the yeast-derived exosome solution and the ferulic acid solution are uniformly mixed in a mass percentage of (80-95):(5-20).

4. The method for preparing the exosome ferulic acid nanocomposite according to claim 1, characterized in that: The conditions for the ultrasonic treatment in step S3 are: under ice bath conditions, the ultrasonic power is 200-400 W, the working time is 2-10 s, the interval time is 2-34 s, the cycle is 5-40 times, and the ice bath is cooled for 3-10 min, and then the ultrasonic treatment is performed again.

5. The method for preparing the exosome ferulic acid nanocomposite according to claim 1, characterized in that: The centrifugal conditions in step S3 are: centrifugation at a temperature of 2-4°C and 5000-10000 g for 10-15 min.

6. The exosome ferulic acid nanocomplex prepared by the method for preparing the exosome ferulic acid nanocomplex according to any one of claims 1 to 5.

7. Use of the exosome ferulic acid nanocomplex as claimed in claim 6 in the preparation of cosmetics or medicines.

8. Use of the exosome ferulic acid nanocomplex as claimed in claim 6 in the preparation of repairing antioxidant cosmetics.

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