Polygonum multiflorum exosome-like vesicle and application thereof

Abstract

The invention relates to polygonum multiflorum exosome-like vesicles and application thereof, the polygonum multiflorum exosome-like vesicles are applied to preparation of antioxidant products, anti-inflammatory products and anti-ultraviolet products, the antioxidant products are products for removing one or more of DPPH free radicals, ABTS free radicals and hydroxyl free radicals, the anti-inflammatory products are products for inhibiting hyaluronidase, and the anti-ultraviolet products are products for removing one or more of DPPH free radicals, ABTS free radicals and hydroxyl free radicals. And the anti-ultraviolet product is a product capable of absorbing UVB. Compared with the prior art, the polygonum multiflorum exosome-like vesicles prepared by the method disclosed by the invention are low in cost and large in treatment capacity, and the obtained polygonum multiflorum exosome-like vesicles have excellent oxidation resistance, inflammation resistance and ultraviolet resistance.

Description

Technical Field

[0001] The present invention relates to the field of natural medicines, and in particular to the application of exosome-like vesicles of Polygonum multiflorum Background Art

[0002] Polygonum multiflorum is a traditional Chinese medicinal material rich in various bioactive components, and has been proven to have antioxidant, anti-inflammatory, anti-aging and other effects. Modern research shows that the main active components in Polygonum multiflorum can scavenge free radicals, inhibit inflammatory reactions, and reduce the damage of ultraviolet radiation to the skin. However, traditional extracts of Polygonum multiflorum may have problems such as poor stability and low permeability during application

[0003] Plant cells can release various vesicles with lipid bilayer membranes, and their diameter ranges from about 20 nm to 5 μm. These vesicles can be classified into exosomes, microvesicles, microparticles, etc. according to their functions, particle sizes and compositions. Due to their lipid bilayer structure, plant exosome-like vesicles can effectively encapsulate and deliver internal active components, improving their stability and bioavailability. At the same time, exosome-like vesicles derived from plants are safer than those derived from animals, with low toxicity and low immunogenicity. Recent research shows that plant exosome-like vesicles not only have signal transduction and regulation functions in organisms, but also show broad application potential in the fields of medicine and cosmetics

[0004] Although a large number of studies have been conducted on Polygonum multiflorum itself as a traditional Chinese medicinal material, the research on its exosome-like vesicles is relatively less, and it is still unknown whether it can replace the extract of Polygonum multiflorum Summary of the Invention

[0005] The purpose of the present invention is to provide an exosome-like vesicle of Polygonum multiflorum and its application to solve the defects existing in the application of the extract of Polygonum multiflorum, with the expectation of using the exosome-like vesicle of Polygonum multiflorum to replace the extract of Polygonum multiflorum with poor stability and low permeability in the application

[0006] The purpose of the present invention can be achieved by the following technical solutions

[0007] The technical solution of the present invention is to provide the application of exosome-like vesicles of Polygonum multiflorum in the preparation of any one or more of the following functional products a1) to a3):

[0008] a1) Antioxidant products

[0009] a2) Anti-inflammatory products

[0010] a3) Anti-ultraviolet products

[0011] In some specific embodiments, the antioxidant product is a product that scavenges any one or more of the following free radicals b1) to b3):

[0012] b1) Scavenging DPPH free radicals;

[0013] b2) Scavenging ABTS free radicals;

[0014] b3) Scavenging hydroxyl free radicals.

[0015] In some specific embodiments, the anti-inflammatory product is a product that inhibits hyaluronidase.

[0016] In some specific embodiments, the anti-UV product is a product that absorbs UVB.

[0017] In some specific embodiments, the particle size of the polygonum multiflorum exosome-like vesicles is (253 ± 15) nm, and the potential is (-15.85 ± 0.29) mV.

[0018] In some specific embodiments, the method for preparing the polygonum multiflorum exosome-like vesicles comprises the following steps:

[0019] S1. Add polygonum multiflorum tissue to PBS solution, break the cell wall and filter to obtain a filtrate;

[0020] S2. Centrifuge the filtrate obtained in step S1 at a low speed and collect the supernatant;

[0021] S3. Centrifuge the supernatant obtained in step S2 at a high speed and collect the supernatant;

[0022] S4. Filter the supernatant obtained in step S3 to obtain polygonum multiflorum exosome-like vesicles.

[0023] In some specific embodiments, in step S1, the mass ratio of the polygonum multiflorum tissue to PBS is 1:2.

[0024] In some specific embodiments, in step S1, the polygonum multiflorum tissue is polygonum multiflorum tuber.

[0025] In some specific embodiments, in step S2, the rotation speed of the low-speed centrifugation is 1000 - 6000g, and the centrifugation time is 40 - 55 min;

[0026] In step S3, the rotation speed of the high-speed centrifugation is 10000 - 14000g, and the centrifugation time is 50 - 60 min.

[0027] In some specific embodiments, in step S2, the low-speed centrifugation is divided into two stages. The rotation speed of the first stage of low-speed centrifugation is 1000 - 2000g, and the time is 10 - 15 min. After collecting the supernatant, the second stage of low-speed centrifugation is carried out;

[0028] In the second-stage low-speed centrifugation, the rotational speed is 4000 - 6000g and the time is 30 - 40 min, and the supernatant is collected.

[0029] In some specific embodiments, in steps S2 - S3, the temperature during centrifugation is 0 - 4°C.

[0030] In some specific embodiments, in step S4, the filtration is to filter the collected supernatant through a 0.22 - 0.25 μm filter membrane.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] (1) When preparing polygonum multiflorum exosome-like vesicles, the present invention reduces the centrifugation speed, reduces the risk of shear damage and vesicle rupture, makes the preparation cost of polygonum multiflorum exosome-like vesicles lower, can handle a larger volume, also improves the biological activity, and takes into account the ability to scavenge DPPH free radicals, ABTS free radicals, and hydroxyl free radicals in antioxidant activity.

[0033] (2) The polygonum multiflorum exosome-like vesicles prepared by the present invention retain the effects of polygonum multiflorum extract, have excellent antioxidant, anti-inflammatory, and anti-ultraviolet properties, and are expected to be prepared into antioxidant products, anti-inflammatory products, and anti-ultraviolet products and applied to fields such as cosmetics and medicine. Description of the Drawings

[0034] Figure 1 It is a test chart for anti-ultraviolet ability. Specific Embodiments

[0035] The present invention provides a method for preparing polygonum multiflorum exosome-like vesicles, including the following steps:

[0036] (1) According to the mass ratio of polygonum multiflorum tuber to PBS solution of 1:2, add polygonum multiflorum tuber to the PBS solution, perform cell wall breaking and filtration to obtain a filtrate;

[0037] (2) Centrifuge the filtrate obtained in step S1 at a rotational speed of 1000 - 2000g for 10 - 15 min, and collect the supernatant;

[0038] (3) Centrifuge the supernatant collected in step (2) at a rotational speed of 4000 - 6000g for 30 - 40 min, and collect the supernatant;

[0039] (4) Centrifuge the supernatant collected in step (3) at a rotational speed of 10000 - 14000g for 50 - 60 min, and collect the supernatant;

[0040] (5) Filter the supernatant collected in step (4) through a 0.22 - 0.25 μm filter membrane to obtain polygonum multiflorum exosome-like vesicles.

[0041] In the present invention, first, low-speed centrifugation is used to remove cells, cell debris, organelles, and large vesicles in the filtrate. The rotation speed of the low-speed centrifugation can be 1000 - 6000g, and it can also be carried out in stages, such as centrifuging successively at rotation speeds of 1000 - 2000g and 4000 - 6000g, so as to remove the aforementioned components of different sizes in sequence. Then, high-speed centrifugation is used to purify exosome-like vesicles. The rotation speed of the high-speed centrifugation is 10000 - 14000g. In different stages of the high-speed centrifugation, the same or increasing (preferably increasing) centrifugation rotation speeds can also be adopted. All centrifugation operations are carried out at 0 - 4°C, and the PBS solution can be pre-cooled at 4°C. The specific centrifugation rotation speeds, times, and temperatures in the above steps and stages can also be reasonably adjusted according to the volume and characteristics of different samples; however, those skilled in the art can undoubtedly understand that, in order to achieve the purpose of gradually removing debris of different sizes, the rotation speed of the subsequent centrifugation step should be equal to or higher than (preferably higher than) that of the previous centrifugation step.

[0042] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments. This embodiment is implemented on the premise of the technical solution of the present invention, and the detailed implementation manners and specific operation processes are given, but the protection scope of the present invention is not limited to the following embodiments.

[0043] In the following embodiments, unless otherwise specified for raw materials or treatment techniques, it indicates that they are all conventional commercially available raw material products or conventional treatment techniques in the art.

[0044] Example 1

[0045] This example provides a method for preparing polygonum multiflorum exosome-like vesicles, which includes the following steps:

[0046] (1) According to the mass ratio of polygonum multiflorum tuber to PBS solution of 1:2, add the washed polygonum multiflorum tuber to the PBS solution pre-cooled at 4°C in advance, carry out cell wall breaking and filtration to obtain a filtrate;

[0047] (2) Centrifuge the filtrate obtained in step S1 at a rotation speed of 1000g for 10 minutes, and collect the supernatant;

[0048] (3) Centrifuge the supernatant collected in step (2) at a rotation speed of 4000g for 30 minutes, and collect the supernatant;

[0049] (4) Centrifuge the supernatant collected in step (3) at a rotation speed of 10000g for 60 minutes, and collect the supernatant;

[0050] (5) Filter the supernatant collected in step (4) through a 0.22 - 0.25μm filter membrane to obtain polygonum multiflorum exosome-like vesicles.

[0051] The particle size of the polygonum multiflorum exosome-like vesicles collected is about 266.69 nm, and the potential is about -15.63 mV.

[0052] Example 2

[0053] This example provides a method for preparing polygonum multiflorum exosome-like vesicles, which includes the following steps:

[0054] (1) According to the mass ratio of polygonum multiflorum tuber to PBS solution of 1:2, add the cleaned polygonum multiflorum tuber to the PBS solution pre-cooled at 4 °C in advance, perform cell wall breaking and filtration to obtain a filtrate;

[0055] (2) Centrifuge the filtrate obtained in step S1 at a rotation speed of 1500 g for 10 min, and collect the supernatant;

[0056] (3) Centrifuge the supernatant collected in step (2) at a rotation speed of 4500 g for 30 min, and collect the supernatant;

[0057] (4) Centrifuge the supernatant collected in step (3) at a rotation speed of 12000 g for 60 min, and collect the supernatant;

[0058] (5) Filter the supernatant collected in step (4) through a 0.22 - 0.25 μm filter membrane to obtain polygonum multiflorum exosome-like vesicles.

[0059] The particle size of the polygonum multiflorum exosome-like vesicles collected is about 255.14 nm, and the potential is about -15.74 mV.

[0060] Example 3

[0061] This example provides a method for preparing polygonum multiflorum exosome-like vesicles, which includes the following steps:

[0062] (1) According to the mass ratio of polygonum multiflorum tuber to PBS solution of 1:2, add the cleaned polygonum multiflorum tuber to the PBS solution pre-cooled at 4 °C in advance, perform cell wall breaking and filtration to obtain a filtrate;

[0063] (2) Centrifuge the filtrate obtained in step S1 at a rotation speed of 2000 g for 10 min, and collect the supernatant;

[0064] (3) Centrifuge the supernatant collected in step (2) at a rotation speed of 5000 g for 30 min, and collect the supernatant;

[0065] (4) Centrifuge the supernatant collected in step (3) at a rotation speed of 14000 g for 60 min, and collect the supernatant;

[0066] (5) Filter the supernatant collected in step (4) through a 0.22 - 0.25 μm filter membrane to obtain polygonum multiflorum exosome-like vesicles.

[0067] The particle size of the polygonum multiflorum exosome-like vesicles collected was about 238.39 nm, and the potential was about -16.18 mV.

[0068] The polygonum multiflorum exosome-like vesicles prepared in Examples 1 to 3 were subjected to the following performance tests:

[0069] (1) Determination of antioxidant property:

[0070] (1-1) Determination of DPPH radical scavenging rate

[0071] Weigh 2 mg of DPPH powder and dissolve it in 40 mL of ethanol solution with a volume concentration of 95%. Ultrasonic for 15 min to make the DPPH solution evenly mixed. After ultrasonic treatment, take 2 mL of this solution and measure its absorbance at 517 nm. Adjust the absorbance to be between 1.2 and 1.3 to obtain the DPPH working solution.

[0072] Transfer 2 mL of the polygonum multiflorum exosome-like vesicle solution into a test tube, add 2 mL of the DPPH working solution, and label it as A1;

[0073] Take 2 mL of the polygonum multiflorum exosome-like vesicle solution and add 2 mL of PBS solution, and label it as A2;

[0074] Take 2 mL of PBS solution and add 2 mL of the DPPH working solution, and label it as A3;

[0075] Take 2 mL of PBS solution and label it as A0.

[0076] Mix each experimental group and react in the dark for 30 min. Measure the absorbance at a wavelength of 517 nm in parallel 3 times.

[0077] The scavenging ability of the polygonum multiflorum exosome-like vesicle solution to DPPH radicals, that is, the DPPH radical scavenging rate, is calculated by the following formula:

[0078]

[0079] In the formula: A1 is the absorbance of the DPPH working solution containing the polygonum multiflorum exosome-like vesicle solution; A2 is the absorbance of the PBS solution containing the polygonum multiflorum exosome-like vesicle solution; A3 is the absorbance of the PBS solution containing the DPPH working solution; A0 is the absorbance of the PBS solution.

[0080] The test data of the DPPH radical scavenging rate of the polygonum multiflorum exosome-like vesicles are shown in Table 1:

[0081] Table 1 DPPH radical scavenging rate

[0082] Item DPPH Free Radical Scavenging Rate / % Example 1 87.25±0.4 Example 2 87.55±0.3 Example 3 90.52±0.5

[0083] As can be seen from the data in Table 1, the exosome-like vesicles extracted from Polygonum multiflorum Thunb. of the present invention have excellent antioxidant effects, and among them, the exosome-like vesicles extracted in Example 3 have the strongest ability to scavenge DPPH free radicals. With the increase of the centrifugation speed, the particle size of the vesicles gradually decreases. The exosome-like vesicles of Polygonum multiflorum Thunb. prepared in Example 3 have the smallest particle size and the most stable potential. The small particle size means an increase in the relative surface area, increasing the contact opportunity between the antioxidant components carried by it and DPPH free radicals and improving the scavenging rate. A more negative potential indicates that the vesicles are more stable and are not easily aggregated or degraded during the experiment, which helps to maintain the activity of the antioxidant substances therein.

[0084] (1-2) Determination of ABTS free radical scavenging rate

[0085] Mix 5 mL of 7.4 mmol / L ABTS stock solution with 88 μL of 2.6 mmol / L K2S2O8 solution and let it stand for 12 - 16 hours. Take 0.4 mL of the above solution and measure the absorbance at 734 nm at room temperature, and dilute it with PBS solution to an absorbance of 0.7 ± 0.02 as the ABTS working solution.

[0086] Add 0.25 mL of the exosome-like vesicle solution of Polygonum multiflorum Thunb. to 1 mL of the ABTS working solution, denoted as A1;

[0087] Add 0.25 mL of the exosome-like vesicle solution of Polygonum multiflorum Thunb. to 1 mL of PBS solution, denoted as A2;

[0088] Add 0.25 mL of PBS solution to 1 mL of the ABTS working solution, denoted as A3;

[0089] Take 1.25 mL of PBS solution, denoted as A0;

[0090] Place each experimental group in the dark and let it stand at room temperature for 10 min. At room temperature, measure the absorbance at 734 nm wavelength in parallel for 3 times.

[0091] The scavenging ability of the exosome-like vesicle solution of Polygonum multiflorum Thunb. to ABTS free radicals, i.e., the ABTS free radical scavenging rate, is calculated by the following formula:

[0092]

[0093] In the formula: A1 is the absorbance of the ABTS working solution containing the exosome-like vesicle solution of Polygonum multiflorum Thunb.; A2 is the absorbance of the PBS solution containing the exosome-like vesicle solution of Polygonum multiflorum Thunb.; A3 is the absorbance of the PBS solution containing the ABTS working solution; A0 is the absorbance of the PBS solution.

[0094] The test data of the ABTS free radical scavenging rate of Polygonum multiflorum exosome-like vesicles are shown in Table 2:

[0095] Table 2 ABTS Free Radical Scavenging Rate

[0096] Item ABTS Free Radical Scavenging Rate / % Example 1 99.96±0.1 Example 2 99.95±0.1 Example 3 99.99±0.1

[0097] As can be seen from the data in Table 2, the Polygonum multiflorum exosome-like vesicles obtained by the extraction of the present invention have excellent antioxidant effects. Among them, the Polygonum multiflorum exosome-like vesicles obtained by the extraction of Example 3 have the strongest ability to scavenge ABTS free radicals. All three extraction methods can effectively scavenge ABTS free radicals, but the Polygonum multiflorum exosome-like vesicles prepared by Example 3 have better antioxidant performance. This is because the particle size of Example 3 is the smallest, the specific surface area of vesicles per unit mass is the largest, and the antioxidant active ingredients are more likely to contact ABTS+ free radicals, improving the scavenging rate. And its more negative potential indicates higher vesicle stability.

[0098] (1-3) Determination of Hydroxyl Radical Scavenging Rate

[0099] Absorb 1 mL of the Polygonum multiflorum exosome-like vesicle solution and add it successively to a test tube containing 2 mL of 2 mmol / L FeSO4 solution and 2 mL of 6 mmol / L salicylic acid solution. Shake well and add 3 mL of 1 mmol / L H2O2, denoted as A1;

[0100] Use 3 mL of deionized water instead of 3 mL of 1 mmol / L H2O2 as a blank control, denoted as A2;

[0101] Use 1 mL of deionized water instead of the Polygonum multiflorum exosome-like vesicle solution as a negative control, denoted as A0.

[0102] After heating at 37 °C for 30 min, adjust the wavelength of the spectrophotometer to 510 nm for detecting the absorbance value. The scavenging ability of the Polygonum multiflorum exosome-like vesicle solution for hydroxyl radicals, that is, the hydroxyl radical scavenging rate, is calculated by the following formula:

[0103]

[0104] In the formula: A0 is the absorbance value of the negative control group; A1 is the absorbance value of the sample group solution; A2 is the absorbance value of the blank control group.

[0105] The test data of the hydroxyl radical scavenging rate of Polygonum multiflorum exosome-like vesicles are shown in Table 3:

[0106] Table 3 Hydroxyl Radical Scavenging Rate

[0107]

[0108]

[0109] As can be seen from the data in Table 3, the exosome-like vesicles extracted from Polygonum multiflorum Thunb. of the present invention have excellent antioxidant effects, and among them, the exosome-like vesicles extracted in Example 2 have the strongest ability to scavenge hydroxyl radicals. The particle size of the exosome-like vesicles of Polygonum multiflorum Thunb. prepared in Example 2 is smaller than that in Example 1 but larger than that in Example 3, indicating that the antioxidant components (such as flavonoids, polyphenols, etc.) inside the exosome-like vesicles of Polygonum multiflorum Thunb. prepared in Example 2 are in a more favorable distribution state, carrying more water-soluble antioxidant components, and hydroxyl radicals have strong hydrophilicity, so the ability to scavenge hydroxyl radicals is improved.

[0110] (2) Determination of anti-inflammatory property (hyaluronidase inhibition rate):

[0111] Prepare 0.1M acetic acid buffer solution (prepared from 0.1M glacial acetic acid solution and 0.1M sodium acetate solution), 4000U / mL hyaluronidase solution, 12.5mM calcium chloride solution, sodium hyaluronate solution, 0.4M sodium hydroxide solution, acetylacetone solution (solvent is 0.5M sodium carbonate solution), and p-dimethylaminobenzaldehyde solution (solvent is 10M hydrochloric acid solution) for use.

[0112] Add each reagent according to Table 4, where the sample groups are the exosome-like vesicle solution of Polygonum multiflorum Thunb. and dipotassium glycyrrhizinate solution (DPG) respectively. The dipotassium glycyrrhizinate solution is used as a positive control for the inhibitory effect on hyaluronidase activity. After the reaction, measure the absorbance at 585nm with an enzyme-linked immunosorbent assay (ELISA) reader, and calculate the inhibitory ability of the exosome-like vesicle solution of Polygonum multiflorum Thunb. on hyaluronidase activity. Among them, three parallel experiments are set for each group.

[0113] Table 4 Hyaluronidase activity inhibition experiment

[0114]

[0115] The inhibition rate of the exosome-like vesicles of Polygonum multiflorum Thunb. on hyaluronidase can reflect the strength of its anti-inflammatory activity. The higher the inhibition rate, the stronger the anti-inflammatory activity. Calculate the inhibition rate of the exosome-like vesicle solution of Polygonum multiflorum Thunb. on hyaluronidase activity according to the following formula:

[0116]

[0117] In the formula: T is the absorbance value of the sample tube; T0 is the absorbance value of the sample control tube; C is the absorbance value of the negative control tube; C0 is the absorbance value of the blank control tube.

[0118] The test data of the inhibition rate of the exosome-like vesicles of Polygonum multiflorum Thunb. on hyaluronidase are shown in Table 5:

[0119] Table 5 Hyaluronidase inhibition rate

[0120] Item Hyaluronidase Inhibition Rate / % Example 1 70.1±0.5 Example 2 78.4±0.7 Example 3 91.4±0.4

[0121] As can be seen from the data in Table 5, the exosome-like vesicles of Polygonum multiflorum extracted by the present invention have excellent anti-inflammatory effects. Among them, the exosome-like vesicles of Polygonum multiflorum extracted in Example 3 have the strongest inhibitory ability against hyaluronidase. Since the exosome-like vesicles of Polygonum multiflorum prepared in Example 3 have smaller particle sizes and more negative potentials, they are vesicles with a larger specific surface area and greater stability, which provides more active sites to enhance the inhibitory effect on hyaluronidase.

[0122] (3) Determination of anti-ultraviolet ability

[0123] The ultraviolet spectrophotometry was used to measure the transmittance of the exosome-like vesicle solution of Polygonum multiflorum within the wavelength range of 200-500 nm, so as to evaluate the sunscreen performance of the exosome-like vesicles of Polygonum multiflorum.

[0124] The transmittances of the samples at 280, 290, 300, 310 and 320 nm in the UVB region and at 320, 330, 340, 350, 360, 370, 380, 390 and 400 nm in the UVA region were measured respectively.

[0125] The ultraviolet absorption rate was calculated by the following formula: ultraviolet absorption rate / % = (1 - transmittance) × 100%

[0126] Figure 1 is the absorption rate of the exosome-like vesicles of Polygonum multiflorum at the wavelengths in the UVB region and UVA region. As can be seen from Figure 1 It can be seen that the exosome-like vesicles of Polygonum multiflorum extracted by the present invention have a certain sunscreen effect in the UVB region, and the average absorption rate in the UVB region is 92%. Among them, the exosome-like vesicles of Polygonum multiflorum extracted in Example 3 have the highest absorption rate in the UVB region. This is because the exosome-like vesicles of Polygonum multiflorum prepared in Example 3 have smaller particle sizes, making their dispersibility better, increasing the absorption area of UVB. Moreover, the exosome-like vesicles of Polygonum multiflorum prepared in Example 3 may also be enriched with more components with UVB absorption ability such as flavonoids and polyphenols. In addition, the exosome-like vesicles of Polygonum multiflorum prepared in Example 3 also have a more stable surface potential, which may affect the optical properties of the vesicles and improve the absorption ability of ultraviolet light.

[0127] The above description of the embodiments is for the convenience of those of ordinary skill in the art to understand and use the invention. Those skilled in the art can obviously make various modifications to these embodiments easily and apply the general principles described herein to other embodiments without creative labor. Therefore, the present invention is not limited to the above embodiments, and the improvements and modifications made by those skilled in the art without departing from the scope of the present invention according to the disclosure of the present invention should be within the protection scope of the present invention.

Claims

1. Application of polygonum multiflorum exosome-like vesicles, characterized in that, The application of the polygonum multiflorum exosome-like vesicles in the preparation of any one or more of the following functional products a1) to a3): a1) Antioxidant products; a2) Anti-inflammatory products; a3) Anti-UV products.

2. The application according to claim 1, characterized in that, The antioxidant product is a product that scavenges any one or more of the following free radicals b1) to b3): b1) Scavenging DPPH free radicals; b2) Scavenging ABTS free radicals; b3) Scavenging hydroxyl free radicals.

3. The application according to claim 1, characterized in that, The anti-inflammatory product is a product that inhibits hyaluronidase.

4. The application according to claim 1, wherein The anti-UV product is a product that absorbs UVB.

5. The application according to claim 1, wherein The particle size of the polygonum multiflorum exosome-like vesicles is (253 ± 15) nm, and the potential is (-15.85 ± 0.29) mV.

6. The application according to claim 1, characterized in that, The preparation method of the polygonum multiflorum exosome-like vesicles includes the following steps: S1. Add polygonum multiflorum tissue to PBS solution, break the wall and filter to obtain a filtrate; S2. Centrifuge the filtrate obtained in step S1 at a low speed and collect the supernatant; S3. Centrifuge the supernatant obtained in step S2 at a high speed and collect the supernatant; S4. Filter the supernatant obtained in step S3 to obtain polygonum multiflorum exosome-like vesicles.

7. The application according to claim 6, characterized in that In step S1, the mass ratio of the polygonum multiflorum tissue to PBS is between 1:1 and 1:5, and the polygonum multiflorum tissue is polygonum multiflorum tuber.

8. The application according to claim 6, characterized in that, In step S2, the rotation speed of the low-speed centrifugation is 1000 - 6000g, and the centrifugation time is 40 - 55 min; In step S3, the rotation speed of the high-speed centrifugation is 10000 - 14000g, and the centrifugation time is 50 - 60 min; In steps S2 - S3, the temperature during centrifugation is 0 - 4°C.

9. The application according to claim 8, characterized in that, In step S2, the low-speed centrifugation is divided into two stages. The rotation speed of the first stage of low-speed centrifugation is 1000 - 2000g, and the time is 10 - 15 min. After collecting the supernatant, perform the second stage of low-speed centrifugation; The rotation speed of the second stage of low-speed centrifugation is 4000 - 6000g, and the time is 30 - 40 min, and collect the supernatant.

10. The application according to claim 6, wherein In step S4, the filtration is to filter the collected supernatant through a 0.22 - 0.25 μm filter membrane.

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