A method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis, and the extracted exosome-like nanovesicles and applications thereof

Abstract

The present invention relates to a method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis, and the extracted exosome-like nanovesicles and applications thereof. Specifically, the freeze-drying and enzymatic hydrolysis method for extracting exosome-like nanovesicles from Platycladus orientalis leaves provided by the present invention ensures a stable supply of raw materials, is easy to store for a long time, and significantly improves the extraction efficiency and economy, opening up a new path for the commercial production of exosome-like nanovesicles from plant sources. Through freeze drying and low-temperature enzymatic hydrolysis technology, the active ingredients in Platycladus orientalis leaves are retained to the maximum extent, and the fiber and polysaccharide colloid are effectively removed, thereby improving the purity and biological activity of the exosome-like nanovesicles. The method is simple, easy to operate, low in cost, and suitable for large-scale production. In addition, the present invention applies the extracted Platycladus orientalis leaves exosome-like nanovesicles to hair growth, especially for androgenic alopecia (AGA), providing a safe and efficient new treatment strategy, and is expected to lead a new trend in the preparation and application of plant exosome-like nanovesicles.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to a method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis, and the extracted exosome-like nanovesicles and applications thereof. Background Art

[0002] With the acceleration of social development, the pressure of work and study has increased significantly, and the problem of hair loss has become a widespread concern worldwide. According to the latest statistics, as of the end of 2021, the number of people with hair loss in my country has surged to 267 million. Among them, androgenetic alopecia (AGA), as the most common type of hair loss, not only has an incidence rate of 21% among men in my country, but also 6% among women. This trend is gradually spreading to young people, which has a serious impact on the quality of life and self-esteem of patients. Therefore, the clinical demand for effective treatment of AGA is becoming more and more urgent.

[0003] At present, drug therapy and hair transplant surgery are the two main means of treating AGA, but these methods have side effects to varying degrees, which limits their widespread application. In this context, traditional Chinese herbal medicine treatment strategies have gradually attracted the attention of researchers due to their advantages such as high compliance, less side effects, a wide range of effects and low prices. As a herbal medicine with a long history of hair growth, Platycladus orientalis leaves have a particularly significant application potential in the treatment of AGA. However, the current treatment of AGA with Platycladus orientalis leaves still faces challenges such as low solubility of active ingredients, limited efficacy due to insufficient bioavailability, and unclear specific mechanisms of action. It is urgent to explore a new Chinese herbal medicine improvement strategy with more precise effects and more in-depth research.

[0004] Exosomes, small vesicles secreted by cells, play an important role in intercellular communication and material transfer due to their rich bioactive molecules such as proteins, lipids and nucleic acids. Plant-derived exosome-like nanovesicles, as a cell-derived bilayer lipid vesicle, have similar structures and functions to mammalian exosomes and can be extracted from common fruits and vegetables. Studies have shown that plant exosome-like nanovesicles not only participate in the communication between plant cells, but also cross the mammalian barrier to achieve communication between plant and animal species. Compared with animal-derived exosome-like nanovesicles, plant exosome-like nanovesicles have multiple advantages such as wide sources, high extraction efficiency, excellent stability, low immunogenicity and economic feasibility. They have shown good therapeutic effects in the treatment of various diseases such as skin whitening, periodontitis, tumors and colitis.

[0005] However, most of the currently disclosed patented technical solutions for plant exosome-like nanovesicles rely on fresh plants as raw materials, and plant juice is obtained by squeezing, homogenization and other methods to extract exosome-like nanovesicles. The limitation of this method is that the supply of fresh plants is strictly restricted by season and region, and the quality of plants is closely related to the harvest season, resulting in significant differences in efficacy. In addition, fresh plants have a short shelf life and are prone to rot and deterioration. They need to be consumed within a short period of time or properly refrigerated and stored, and their active ingredients are easily destroyed during the storage process.

[0006] In view of the above challenges, the present invention aims to provide an improved method for extracting exosome-like nanovesicles from Platycladus orientalis leaves, which can effectively overcome the limitations of the prior art without sacrificing biological activity. By developing a mild treatment process suitable for drying Platycladus orientalis leaves, the integrity and activity of exosome-like nanovesicles can be preserved, thereby improving their potential for application in the treatment of diseases such as AGA. This method is not only expected to bring innovation to the treatment of hair loss, but also provides new ideas and references for the extraction of exosome-like nanovesicles from other plants. Summary of the invention

[0007] In view of this, the object of the present invention is to provide a method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis. The present invention also provides exosome-like nanovesicles extracted by this extraction method and applications thereof.

[0008] Specifically, the present invention provides a method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis, which is characterized by comprising the following steps:

[0009] Step 1: After fully enzymatically hydrolyzing the lyophilized powder of Platycladus orientalis leaves, the juice is collected to obtain a crude juice containing exosome-like nanovesicles of Platycladus orientalis leaves;

[0010] Step 2: Separate and purify the juice using one or a combination of differential centrifugation, fractionation purification based on particle size and molecular weight, and ultracentrifugation to obtain exosome-like nanovesicles derived from Platycladus orientalis leaves.

[0011] In some specific embodiments of the present invention, the specific operating steps of step 1 are: take the freeze-dried powder of Platycladus orientalis leaves, add a PBS solution containing pectinase and cellulase, and homogenize it into a paste-like semi-solid under low temperature; let the paste-like semi-solid stand at 2-5°C for 2-4 hours for enzymatic hydrolysis; soak the Platycladus orientalis leaf slurry after enzymatic hydrolysis in PBS buffer and mix it thoroughly, and filter out the crude juice containing Platycladus orientalis leaf exosome-like nanovesicles with a filter.

[0012] In some specific embodiments of the present invention, the total content of pectinase and cellulase in the PBS solution is 25% to 35%, wherein the mass ratio of the pectinase to the cellulase is 1:(0.5 to 2); the mass ratio of the dry powder to the PBS solution containing pectinase and cellulase is (300 to 200):1; the mass ratio of the Platycladus orientalis leaf slurry after enzymatic hydrolysis to the PBS buffer solution is 1 to (1:2), and the pore size of the filter is 120 mesh.

[0013] In some specific embodiments of the present invention, the specific preparation steps of the Platycladus orientalis leaf freeze-dried powder in step 1 are as follows: fresh Platycladus orientalis leaves are placed in liquid nitrogen for quick freezing or -80°C for 24 to 48 hours, and then freeze-dried at a temperature ≤45°C and a vacuum degree ≤10pa for 24 to 48 hours to obtain freeze-dried Platycladus orientalis leaves; and then ground into powder at 2 to 5°C to obtain Platycladus orientalis leaf freeze-dried powder.

[0014] 7. In some specific embodiments of the present invention, the specific operation steps of step 2 are: centrifuge the crude juice containing the exosome-like nanovesicles of Platycladus orientalis leaves at 1000×g for 1 minute, take the supernatant; centrifuge at 3000×g for 2 minutes, take the supernatant; then centrifuge at 10000×g for 3 minutes, take the supernatant; centrifuge the obtained supernatant at 150000×g for 4 minutes, then slowly aspirate the supernatant obtained by centrifugation, add PBS solution to resuspend the precipitate, and repeatedly blow and beat until the precipitate is completely dissolved. The obtained resuspension is centrifuged at 15000×g for 5 minutes, and the supernatant is filtered with a 0.2-0.5μm filter to prepare the exosome-like nanovesicles from Platycladus orientalis leaves.

[0015] In some specific embodiments of the present invention, t1=10; t2=30; t3=60; t4=60; t5=15.

[0016] The present invention also provides a Platycladus orientalis leaf exosome-like nanovesicle, wherein the Platycladus orientalis leaf exosome-like nanovesicle is extracted according to any of the methods for extracting Platycladus orientalis leaf exosome-like nanovesicle by freeze-drying and enzymatic hydrolysis.

[0017] In some specific embodiments of the present invention, the Platycladus orientalis leaf exosome-like nanovesicles are round or cup-shaped in appearance, and have a disc-shaped, concave structure, and the size of the nanovesicles is concentrated between 20 and 150 nm.

[0018] The present invention also provides the Platycladus orientalis leaf exosome-like nanovesicles extracted by the freeze-drying enzymatic hydrolysis method for extracting Platycladus orientalis leaf exosome-like nanovesicles or the use of the Platycladus orientalis leaf exosome-like nanovesicles in the preparation of medicines or daily chemical products that can be used to prevent or treat androgenic alopecia.

[0019] The present invention also provides a composition, medicine or daily chemical product that can be used to prevent or treat androgenic alopecia, which comprises the Platycladus orientalis leaf exosome-like nanovesicles extracted by the method of freeze-drying and enzymatic hydrolysis to extract Platycladus orientalis leaf exosome-like nanovesicles or the above-mentioned Platycladus orientalis leaf exosome-like nanovesicles.

[0020] In some specific embodiments of the present invention, the composition, medicine or daily chemical product for preventing or treating androgenic alopecia further comprises an auxiliary material or a pharmaceutically acceptable carrier.

[0021] Compared with the prior art, the present invention has the following significant advantages and effects:

[0022] (1) Highly efficient activity retention: By adopting freeze-drying and low-temperature enzymatic hydrolysis technology, the present invention can retain the natural active ingredients in Platycladus orientalis leaves to the maximum extent. Freeze-drying technology ensures that the biological activity of the active ingredients in the plant is not destroyed, while low-temperature enzymatic hydrolysis technology can efficiently remove impurities such as fiber and polysaccharide colloids in the plant, making the final product purer and rich in active ingredients.

[0023] (2) High solubility: The molecular weight of the freeze-dried Platycladus orientalis leaf material is small and highly water-soluble, making it easy to use directly or for subsequent processing. This high solubility makes the product more flexible and convenient during application.

[0024] (3) Long-term storage: Through freeze-drying technology, more than 95% to 96% of the water in fresh Platycladus orientalis leaves can be removed, so that the plant dry powder can be stored for a long time without deterioration. This greatly reduces the storage cost and difficulty of the product, while extending the service life of the product.

[0025] (4) Stable supply: Compared with fresh Platycladus orientalis leaves, the freeze-dried powder used in the present invention is not affected by the growth cycle and growth environment, and the supply is more stable. This enables the product to continuously and stably provide raw materials during the production process, ensuring the continuity and stability of the production line.

[0026] (5) Easy to store and transport: Since a large amount of water is removed from the freeze-dried powder, its volume and weight are greatly reduced, making it easier to store and transport. This not only reduces the cost of storage and transportation, but also improves the accessibility and market coverage of the product.

[0027] (6) High efficiency in keeping the active ingredients alive: Freeze-drying technology can ensure that the active ingredients in Platycladus orientalis leaves are not destroyed, while low-temperature enzymatic hydrolysis technology can effectively remove impurities such as fiber and polysaccharide colloids in the plant, which is conducive to the efficient extraction of purer plant exosome-like nanovesicles. This high efficiency in keeping the active ingredients alive and extracting technology has significantly improved the quality and purity of the final product.

[0028] (7) Efficient preparation: The method for extracting exosome-like nanovesicles provided by the present invention not only makes it easy to obtain raw materials, but also makes production more standardized. By optimizing the production process and technical parameters, an efficient and low-cost production method is achieved.

[0029] (8) Commercial feasibility: Due to the low production cost and stable and reliable product quality, the present invention lays a solid foundation for large-scale production and market promotion. This enables the product to be more widely used in the fields of medicine, health care products, cosmetics, etc., with broad market prospects and commercial value.

[0030] In summary, the patent of the present invention shows significant beneficial technical effects in maintaining plant active ingredients, improving solubility and long-term preservation, stable supply and easy storage, efficient preservation and extraction, efficient preparation and commercial feasibility. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0032] Figure 1 This is a transmission electron microscopy image characterizing the morphology and size of exosome-like nanovesicles derived from Platycladus orientalis leaves.

[0033] Figure 2 The size distribution of exosome-like nanovesicles derived from Platycladus orientalis leaves measured by dynamic light scattering.

[0034] Figure 3 It is the dosing scheme and grouping of experimental animals.

[0035] Figure 4 These are photos of hair regeneration in each group of mice.

[0036] Figure 5 It is a quantitative statistical chart of the number of days the skin of each group of mice turns darker.

[0037] Figure 6 It is a quantitative statistical graph of the weight changes of mice in each group.

[0038] Figure 7 are the trichoscopy images of mice in each group.

[0039] Figure 8 It is a quantitative statistical chart of hair coverage rate, an evaluation index of new hair quality in each group of mice.

[0040] Fig. 9It is a quantitative statistical chart of hair density, an evaluation index for the quality of new hair of each group of mice.

[0041] Fig.10 It is a quantitative statistical chart of hair diameter, an evaluation index for the quality of new hair of each group of mice. DETAILED DESCRIPTION

[0042] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention and should not be used to limit the protection scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work belong to the protection scope of the present invention.

[0043] Example 1: Preparation of Platycladus orientalis leaf freeze-dried powder

[0044] The preparation steps are as follows:

[0045] Material preparation: Collect fresh, uncontaminated Platycladus orientalis leaves.

[0046] Cleaning: Wash fresh arborvitae leaves thoroughly 7-8 times to remove surface dust and impurities.

[0047] Drain: After cleaning, drain most of the water for subsequent processing.

[0048] Freeze-drying: Freeze fresh Platycladus orientalis leaves at -80℃ for 12-48h, then freeze-dry them at a temperature ≤45℃ and a vacuum degree ≤10pa for 15-24h. After crushing, Platycladus orientalis leaf freeze-dried powder is obtained.

[0049] Example 2: Preparation of exosome-like nanovesicles (POELNVs) from Platycladus orientalis leaves

[0050] The preparation steps are as follows: take a certain amount of lyophilized powder of Platycladus orientalis leaves, add 30% PBS containing pectinase and cellulase (1:1) and homogenize it into a paste semi-solid at low temperature, and let the homogenate stand for 4 hours at low temperature for enzymatic hydrolysis, soak the Platycladus orientalis leaf slurry after enzymatic hydrolysis in 1200mL PBS buffer and mix it thoroughly, and filter out the crude juice containing Platycladus orientalis leaf exosome-like nanovesicles with a filter. The crude juice containing Platycladus orientalis leaf exosome-like nanovesicles is centrifuged at 1000×g for 10min, and the supernatant is taken; then centrifuged at 3000×g for 30min, and the supernatant is taken; then centrifuged at 10000×g for 60min, and the supernatant is taken; the supernatant obtained is centrifuged at 150000×g for 60min, and then the supernatant obtained by centrifugation is slowly aspirated, and when the supernatant is 0.5cm away from the bottom of the tube, 1mL PBS buffer is added to resuspend, and it is repeatedly blown until the precipitate is completely dissolved. The obtained resuspension was centrifuged at 15000×g for 15 min, and the supernatant was filtered with a 0.22 μm filter to obtain high-purity exosome-like nanovesicles from Platycladus orientalis leaves. The exosome-like nanovesicle solution was stored at -80°C for a long time or prepared into a freeze-dried powder for subsequent functional analysis and application.

[0051] Example 3: Characterization of exosome-like nanovesicles from Platycladus orientalis leaves

[0052] The prepared exosome-like nanovesicles derived from Platycladus orientalis leaves were fixed with 2% paraformaldehyde, negative contrast stained with 1.9% methylcellulose and 0.3% uranyl acetate, observed and micrographs were collected using a HITACHI HT-7700 electron microscope, and the size distribution of the exosome-like nanovesicles derived from Platycladus orientalis leaves was analyzed by dynamic light scattering using a Zetasizer Nano ZSE (Malvern Instrument, UK).

[0053] The results of ultrastructural and morphological analyses by transmission electron microscopy (respectively Figure 1 and Figure 2 The data shown in the figure show that the exosome-like nanovesicles isolated from Platycladus orientalis leaves have a typical round or cup-shaped appearance, and a disc-shaped, concave structure, similar to the exosome-like nanovesicles derived from mammalian cells; at the same time, it can be observed that the size of the nanovesicles is concentrated between 20-150nm, and the dynamic light scattering experiment also confirms the size distribution of the nanovesicles.

[0054] Example 4: Evaluation of the hair growth effect of Platycladus orientalis leaves-derived exosome-like nanovesicles on AGA mice

[0055] Establishment of the AGA mouse model and dosing regimen: Fifteen 6-week-old male C57BL / 6 mice were taken. After one week of adaptive feeding, they were anesthetized by intraperitoneal injection of tribromoethanol. Taking the dorsal line of the mouse's back as the midline, a 1cm×1cm area was shaved with a pet shaver, and then a depilatory cream was used for depilation for 3 minutes. Referring to existing literature, an AGA model induced by smearing testosterone was established, that is, from the first day after depilation (the day of depilation was positioned as day 0), 100 μL of 0.5% (w / v) testosterone solution was evenly applied to the depilated area on the back of the mouse every day for 28 consecutive days. The depilated mice were randomly divided into 3 groups, with 5 mice in each group. The grouping and treatment regimen were as follows. Figure 3 As shown: Model group: AGA modeling, no treatment; Platycladus orientalis leaf exosome-like nanovesicle injection group (POELNVs): at the same time as AGA modeling, 100 μL of Platycladus orientalis leaf exosome-like nanovesicle solution was intradermally injected on the 1st, 3rd, 5th, 7th, 9th, 11th, 13th, and 15th days after hair removal; Positive control minoxidil group (Minoxidil): at the same time as AGA modeling and starting from the first day after hair removal, 100 μL of commercially available minoxidil solution was evenly applied to the hair removal area on the back of mice every day for 13 consecutive days.

[0056] Hair regeneration investigation: Take photos to record the changes in the skin of the modeling area on the day of hair removal and on the 2nd, 8th, 10th, 12th, 14th, 16th, 18th, 20th, 22nd, 24th, 26th, and 28th days after hair removal. Figure 4 Count the days when the skin starts to darken, such as Figure 5 As shown, compared with the Moedel group, the number of days in which the skin in the hair removal area of ​​mice in the POELNVs group and the Minoxidil group turned darker was significantly reduced, while there was no significant difference between the POELNVs group and the Minoxidil group.

[0057] Evaluation of the quality of new hair: On the 28th day after hair removal, the ratio of the new hair coverage area to the total area of ​​the hair removal area was defined as the new hair coverage rate, and quantitative statistics were performed using Image J software. The hair density and hair diameter of the new hair were detected under a trichoscope. The trichoscope image is shown in Figure 2. Figure 7 As shown in Figure 2, the quantitative statistical results of new hair coverage, hair density and hair diameter are as follows: Figure 8-10 Compared with the Model group, the new hair coverage, new hair density and new hair diameter of the mice in the POELNVs group and the Minoxidil group increased significantly, and the new hair density of the mice in the POELNVs group was significantly higher than that in the Minoxidil group, but the new hair diameter of the mice in the POELNVs group was not as large as that in the Minoxidil group. According to existing literature, Minoxidil is currently the best drug for thickening hair.

[0058] Although the present invention is described using the above embodiments, it should be understood that the present invention may be further modified and varied without departing from the spirit of the present invention, and these modifications and variations are all within the scope of protection of the present invention.

Claims

1. A method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis, characterized in that: The steps include: Step 1: After fully enzymatically hydrolyzing the lyophilized powder of Platycladus orientalis leaves, the juice is collected to obtain a crude juice containing exosome-like nanovesicles of Platycladus orientalis leaves; Step 2: Separate and purify the juice using one or a combination of differential centrifugation, fractionation purification based on particle size and molecular weight, and ultracentrifugation to obtain exosome-like nanovesicles derived from Platycladus orientalis leaves.

2. The method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis as claimed in claim 1, characterized in that: The specific operation steps of step 1 are: taking lyophilized powder of Platycladus orientalis leaves, adding PBS solution containing pectinase and cellulase, and homogenizing into a paste-like semi-solid under low temperature; letting the paste-like semi-solid stand at 2-5°C for 2-4 hours for enzymatic hydrolysis; soaking the enzymatically hydrolyzed Platycladus orientalis leaf slurry in PBS buffer and mixing it thoroughly, and filtering out the crude juice containing Platycladus orientalis leaf exosome-like nanovesicles with a filter.

3. The method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis as claimed in claim 2, characterized in that: The total content of pectinase and cellulase in the PBS solution is 25% to 35%, wherein the mass ratio of the pectinase to the cellulase is 1:(0.5 to 2); the mass ratio of the dry powder to the PBS solution containing pectinase and cellulase is (300 to 200):1; the mass ratio of the Platycladus orientalis leaf slurry after enzymolysis to the PBS buffer solution is 1 to (1:2), and the pore size of the filter is 120 meshes.

4. The method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis as claimed in claim 1, characterized in that: The preparation steps of the Platycladus orientalis leaf freeze-dried powder in step 1 are as follows: fresh Platycladus orientalis leaves are placed in liquid nitrogen for quick freezing or -80°C for 24 to 48 hours, and then freeze-dried at a temperature ≤45°C and a vacuum degree ≤10pa for 24 to 48 hours to obtain freeze-dried Platycladus orientalis leaves; and then ground into powder at 2 to 5°C to obtain Platycladus orientalis leaf freeze-dried powder.

5. The method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis as claimed in claim 1, characterized in that: The specific operation steps of step 2 are as follows: the crude juice containing the exosome-like nanovesicles of Platycladus orientalis leaves is centrifuged at 1000×g for 1 minute, and the supernatant is taken; then the supernatant is centrifuged at 3000×g for 2 minutes, and the supernatant is taken; the supernatant is centrifuged at 10000×g for 3 minutes, and the supernatant is taken; the obtained supernatant is centrifuged at 150000×g for 4 minutes, and then the supernatant obtained by centrifugation is slowly aspirated, and the precipitate is resuspended by adding PBS solution, and the precipitate is repeatedly blown until the precipitate is completely dissolved. The obtained resuspension is centrifuged at 15000×g for 5 minutes, and the supernatant is filtered with a 0.2-0.5μm filter to prepare the exosome-like nanovesicles derived from Platycladus orientalis leaves.

6. The method for extracting exosome-like nanovesicles from Platycladus orientalis leaves by freeze-drying and enzymatic hydrolysis as claimed in claim 5, characterized in that: t1=10; t2=30; t3=60; t4=60; t5=15.

7. A Platycladus orientalis leaf exosome-like nanovesicle, characterized in that The Platycladus orientalis leaf exosome-like nanovesicles are extracted according to the method for extracting Platycladus orientalis leaf exosome-like nanovesicles by freeze-drying and enzymatic hydrolysis according to any one of claims 1 to 6.

8. The Platycladus orientalis leaf exosome-like nanovesicle according to claim 7, characterized in that The Platycladus orientalis leaf exosome-like nanovesicles have a round or cup-shaped appearance and a disc-shaped structure with concave edges. The size of the nanovesicles is concentrated between 20 and 150 nm.

9. Use of the Platycladus orientalis leaf exosome-like nanovesicles extracted by the method for extracting Platycladus orientalis leaf exosome-like nanovesicles by freeze-drying enzymatic hydrolysis as described in claims 1-6 or the Platycladus orientalis leaf exosome-like nanovesicles as described in any one of claims 6-7 in the preparation of drugs or daily chemical products that can be used to prevent or treat androgenic alopecia.

10. A composition, medicine or daily chemical product for preventing or treating androgenic alopecia, characterized in that: It comprises the Platycladus orientalis leaf exosome-like nanovesicles extracted by the method for extracting Platycladus orientalis leaf exosome-like nanovesicles by freeze-drying enzymatic hydrolysis as described in claims 1-6 or the Platycladus orientalis leaf exosome-like nanovesicles as described in any one of claims 7-8.

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