Plant nano-vesicle composition or freeze-dried powder rich in plant nano-vesicles and application of plant nano-vesicle composition or freeze-dried powder rich in plant nano-vesicles in preparation of cosmetics

Abstract

The invention relates to the technical field of biological medicines, and particularly discloses a plant nano-vesicle composition or freeze-dried powder rich in plant nano-vesicles and application of the plant nano-vesicle composition or the freeze-dried powder rich in the plant nano-vesicles in preparation of cosmetics. The plant nano-vesicle composition is prepared from peony nano-vesicles, rhizoma polygonati nano-vesicles and daisy flower nano-vesicles, wherein the weight ratio of the peony nano vesicles to the rhizoma polygonati nano vesicles to the daisy flower nano vesicles is (1-20): (1-10): (1-5). Researches show that the plant nano-vesicle composition disclosed by the invention has an effect of promoting proliferation of human keratinocytes; therefore, the plant nano-vesicle composition has important application value when being used as an active ingredient for preparing cosmetics or drugs with the effect of promoting skin barrier repair.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to a plant nanovesicle composition or a lyophilized powder rich in plant nanovesicles and application thereof in the preparation of cosmetics. BACKGROUND

[0002] With the rapid development of modern biotechnology and nanoscience, natural active ingredients from plants are increasingly widely used in the field of cosmetics. Traditional plant extracts, such as small molecule compounds such as polyphenols, flavonoids, and saponins, have certain antioxidant, anti-inflammatory, and whitening effects, but generally have poor stability, low transdermal absorption rate, and low bioavailability, which limits their in-depth application in high-end cosmetics. In recent years, researchers have found a class of natural vesicle structures with nanoscale in plant cells, which are called plant nanovesicles. This kind of vesicle is composed of a phospholipid bilayer and endogenously encapsulates various bioactive substances such as proteins, nucleic acids (miRNA, mRNA), lipids, and secondary metabolites, and is an important carrier for plant cell communication and material transfer.

[0003] Studies have shown that plant nanovesicles not only inherit the various biological activities of their parent plants, but also exhibit excellent biocompatibility, low immunogenicity, and good cell penetration ability due to their unique nanoscale structure and phospholipid bilayer membrane characteristics. These characteristics enable them to effectively cross the stratum corneum barrier and deliver the end-loaded active ingredients to the deep layers of the skin, thereby significantly improving the utilization efficiency of the active ingredients. In addition, plant nanovesicles themselves also have multiple physiological functions such as regulating skin cell metabolism, promoting collagen synthesis, and scavenging free radicals, showing great potential as a new type of active raw material for cosmetics.

[0004] Although plant nanovesicles have the above advantages, the efficacy of plant nanovesicles from a single source still needs to be further improved. Therefore, it is of great application value to provide a complex plant nanovesicle that can further improve efficacy. SUMMARY

[0005] In order to overcome at least one of the technical problems in the prior art, the present application provides a plant nanovesicle composition or a lyophilized powder rich in plant nanovesicles and application thereof in the preparation of cosmetics.

[0006] The technical solution of the present application is as follows:

[0007] The present application first provides a plant nanovesicle composition, characterized in that it comprises peony nanovesicles, polygonatum nanovesicles, and daisy flower nanovesicles.

[0008] Preferably, the weight ratio of peony nanovesicles, polygonatum nanovesicles, and daisy flower nanovesicles is 1-20:1-10:1-5.

[0009] Most preferably, the weight ratio of the peony nanovesicles, the rhizoma polygonati nanovesicles and the daisy flower nanovesicles is 10:7:3.

[0010] Preferably, the peony nanovesicles are prepared by the following method:

[0011] S11. The peony root bark is mixed with water, and then put into a cell disrupter to disrupt the cells, to obtain a disrupted slurry; the disrupted slurry is then pressed by a press to collect the pressed juice to obtain a peony root bark raw slurry;

[0012] S12. The peony root bark raw slurry is centrifuged to obtain supernatant and precipitate; the supernatant is concentrated and dried to obtain the peony nanovesicles.

[0013] Preferably, the weight ratio of the peony root bark to water in step S11 is 1:1-3.

[0014] Most preferably, the weight ratio of the peony root bark to water in step S11 is 1:2.

[0015] Preferably, the rotation speed of the cell disrupter in step S11 is 20000-40000 rpm; and the cell disruption time is 30-60 s.

[0016] Preferably, the centrifugation in step S12 is performed at a rotation speed of 5000-10000 rpm for 1-3 h.

[0017] Preferably, the preparation method of the peony nanovesicles further comprises an ultrafiltration step:

[0018] S13. The supernatant is first subjected to ultrafiltration using an ultrafiltration membrane with a diameter of 180-220 nm to obtain a first permeate and a first concentrated solution; the first permeate is then subjected to ultrafiltration using an ultrafiltration membrane with a diameter of 70-90 nm to obtain a second permeate and a second concentrated solution; and the second concentrated solution is concentrated and dried to obtain the peony nanovesicles.

[0019] Most preferably, the preparation method of the peony nanovesicles further comprises an ultrafiltration step:

[0020] S13. The supernatant is first subjected to ultrafiltration using an ultrafiltration membrane with a diameter of 200 nm to obtain a first permeate and a first concentrated solution; the first permeate is then subjected to ultrafiltration using an ultrafiltration membrane with a diameter of 80 nm to obtain a second permeate and a second concentrated solution; and the second concentrated solution is concentrated and dried to obtain the peony nanovesicles.

[0021] Preferably, the rhizoma polygonati nanovesicles are prepared by the following method:

[0022] S21. Take the rhizome of Polygonatum sibiricum and mix with water, then put into a cell wall breaking machine to break the cell wall, to obtain a broken cell wall slurry; then press the broken cell wall slurry through a press to collect the pressed juice to obtain a Polygonatum sibiricum raw slurry;

[0023] S22. Take the Polygonatum sibiricum raw slurry to centrifuge to obtain supernatant and precipitate; take the supernatant to concentrate and dry to obtain the Polygonatum sibiricum nanocapsule.

[0024] Preferably, the weight ratio of the rhizome of Polygonatum sibiricum to water in step S21 is 1:1-3.

[0025] Most preferably, the weight ratio of the rhizome of Polygonatum sibiricum to water in step S21 is 1:2.

[0026] Preferably, the rotating speed of the cell wall breaking machine in step S21 is 20000-40000 rpm; and the cell wall breaking time is 30-60 s.

[0027] Preferably, the centrifugation in step S22 is performed at a rotating speed of 5000-10000 rpm for 1-3 h.

[0028] Preferably, the preparation method of the Polygonatum sibiricum nanocapsule further comprises an ultrafiltration step:

[0029] S23. Take the supernatant to perform ultrafiltration through an ultrafiltration membrane with a diameter of 110-130 nm to obtain a first permeate and a first concentrated liquid; take the first permeate to perform ultrafiltration through an ultrafiltration membrane with a diameter of 40-60 nm to obtain a second permeate and a second concentrated liquid; take the second concentrated liquid to concentrate and dry to obtain the Polygonatum sibiricum nanocapsule.

[0030] Most preferably, the preparation method of the Polygonatum sibiricum nanocapsule further comprises an ultrafiltration step:

[0031] S23. Take the supernatant to perform ultrafiltration through an ultrafiltration membrane with a diameter of 120 nm to obtain a first permeate and a first concentrated liquid; take the first permeate to perform ultrafiltration through an ultrafiltration membrane with a diameter of 50 nm to obtain a second permeate and a second concentrated liquid; take the second concentrated liquid to concentrate and dry to obtain the Polygonatum sibiricum nanocapsule.

[0032] Preferably, the daisy flower nanocapsule is prepared by the following method:

[0033] S31. Take the daisy flower and mix with water, then put into a cell wall breaking machine to break the cell wall, to obtain a broken cell wall slurry; then press the broken cell wall slurry through a press to collect the pressed juice to obtain a daisy flower raw slurry;

[0034] S32. Take the daisy flower raw slurry to centrifuge to obtain supernatant and precipitate; take the supernatant to concentrate and dry to obtain the daisy flower nanocapsule.

[0035] Preferably, in step S31, the weight ratio of daisies to water is 1:1 to 3.

[0036] Most preferably, the weight ratio of daisies to water in step S31 is 1:2.

[0037] Preferably, in step S31, the speed of the blender is 20,000 to 40,000 rpm; the blending time is 30 to 60 seconds.

[0038] Preferably, the centrifugation mentioned in step S32 specifically refers to centrifugation at a speed of 5000 to 10000 rpm for 1 to 3 hours.

[0039] Preferably, the method for preparing daisy nanovesicles further includes an ultrafiltration step:

[0040] S33. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 40-60 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 10-30 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the daisy nanovesicles.

[0041] Most preferably, the method for preparing daisy nanovesicles further includes an ultrafiltration step:

[0042] S33. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 50 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 20 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the daisy nanovesicles.

[0043] The present invention also provides a freeze-dried powder rich in plant nanovesicles, which comprises the following components: peony nanovesicles, polygonatum nanovesicles, daisy nanovesicles, resveratrol, mannitol, and water.

[0044] Preferably, the freeze-dried powder rich in plant nanovesicles comprises the following components in parts by weight: 1-20 parts of peony nanovesicles; 1-10 parts of polygonatum nanovesicles; 1-5 parts of daisy nanovesicles; 0.5-5 parts of resveratrol; 2.5-10 parts of mannitol; and 50-100 parts of water.

[0045] Preferably, the resveratrol is supramolecular resveratrol.

[0046] Preferably, the mannitol is D-mannitol.

[0047] The present invention also provides a method for preparing a freeze-dried powder rich in plant nanovesicles, which includes the following steps: adding peony nanovesicles, polygonatum nanovesicles, daisy nanovesicles, resveratrol and mannitol to water, mixing evenly, and then freeze-drying to obtain the freeze-dried powder rich in plant nanovesicles.

[0048] The present invention also provides the application of the above-mentioned plant nanovesicle composition or freeze-dried powder rich in plant nanovesicles in the preparation of products that promote the proliferation of human keratinocytes.

[0049] The present invention also provides the application of the above-mentioned plant nanovesicle composition or freeze-dried powder rich in plant nanovesicles in the preparation of products that promote skin barrier repair.

[0050] Preferably, the product is a cosmetic or a pharmaceutical.

[0051] Beneficial effects: This invention provides a novel plant nanovesicle composition; studies have shown that the plant nanovesicle composition of this invention promotes the proliferation of human keratinocytes; therefore, using the plant nanovesicle composition of this invention as an active ingredient in the preparation of cosmetics or pharmaceuticals that promote skin barrier repair has significant application value.

[0052] Furthermore, the present invention also provides a freeze-dried powder rich in plant nanovesicles, providing more dosage form options for the application of plant nanovesicles. Detailed Implementation

[0053] The present invention will be further described in detail below with reference to specific embodiments, but the embodiments do not limit the present invention in any way.

[0054] Example 1: Preparation of plant nanovesicle composition

[0055] The plant nanovesicle composition is obtained by mixing peony nanovesicles, polygonatum nanovesicles and daisy nanovesicles in a weight ratio of 10:7:3.

[0056] The peony nanovesicles were prepared by the following method:

[0057] S11. Take peony root, add water and mix, then put it into a high-speed blender to break the cell wall and obtain a cell wall broken slurry; then press the cell wall broken slurry through a press and collect the juice to obtain peony root raw slurry; wherein, the weight ratio of peony root peel to water is 1:2; the speed of the high-speed blender is 30,000 rpm; the cell wall breaking time is 45 seconds.

[0058] S12. Centrifuge the original peony root bark pulp to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain peony nanovesicles; the centrifugation specifically refers to centrifugation at 8000 rpm for 2 hours.

[0059] The Polygonatum nanovesicles were prepared by the following method:

[0060] S21. Mix the rhizome of Polygonatum odoratum with water, then put it into a high-speed blender for blending to obtain a blended slurry; then press the blended slurry through a press to collect the juice to obtain the original pulp of Polygonatum odoratum rhizome; wherein, in step S21, the weight ratio of Polygonatum odoratum rhizome to water is 1:2; the speed of the high-speed blender is 30,000 rpm; and the blending time is 45 seconds.

[0061] S22. Centrifuge the original pulp of Polygonatum rhizome to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain Polygonatum nanovesicles; the centrifugation specifically refers to centrifugation at 8000 rpm for 2 hours.

[0062] The daisy-flower nanovesicles were prepared by the following method:

[0063] S31. Take daisies, add water and mix, then put them into a high-speed blender to break them down into a pulp; then press the pulp through a press and collect the juice to obtain daisy puree; wherein, the weight ratio of daisies to water is 1:2; the speed of the high-speed blender is 30,000 rpm; the breaking time is 45 seconds.

[0064] S32. Take the daisy pulp and centrifuge it to obtain the supernatant and precipitate; take the supernatant, concentrate and dry it to obtain daisy nanovesicles; the centrifugation specifically refers to centrifuging at a speed of 8000 rpm for 2 hours.

[0065] Example 2: Preparation of plant nanovesicle composition

[0066] The plant nanovesicle composition is obtained by mixing peony nanovesicles, polygonatum nanovesicles and daisy nanovesicles in a weight ratio of 10:7:3.

[0067] The peony nanovesicles were prepared by the following method:

[0068] S11. Take peony root, add water and mix, then put it into a high-speed blender to break the cell wall and obtain a cell wall broken slurry; then press the cell wall broken slurry through a press and collect the juice to obtain peony root raw slurry; wherein, the weight ratio of peony root peel to water is 1:2; the speed of the high-speed blender is 30,000 rpm; the cell wall breaking time is 45 seconds.

[0069] S12. Centrifuge the original peony root bark pulp to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain peony nanovesicles; the centrifugation specifically refers to centrifugation at a speed of 8000 rpm for 2 hours;

[0070] S13. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 200 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 80 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the peony nanovesicles.

[0071] The Polygonatum nanovesicles were prepared by the following method:

[0072] S21. Mix the rhizome of Polygonatum odoratum with water, then put it into a high-speed blender for blending to obtain a blended slurry; then press the blended slurry through a press to collect the juice to obtain the original pulp of Polygonatum odoratum rhizome; wherein, in step S21, the weight ratio of Polygonatum odoratum rhizome to water is 1:2; the speed of the high-speed blender is 30,000 rpm; and the blending time is 45 seconds.

[0073] S22. Centrifuge the raw pulp of Polygonatum rhizome to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain Polygonatum nanovesicles; the centrifugation specifically refers to centrifugation at 8000 rpm for 2 hours;

[0074] S23. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 120 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 50 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the Polygonatum nanovesicles.

[0075] The daisy-flower nanovesicles were prepared by the following method:

[0076] S31. Take daisies, add water and mix, then put them into a high-speed blender to break them down into a pulp; then press the pulp through a press and collect the juice to obtain daisy puree; wherein, the weight ratio of daisies to water is 1:2; the speed of the high-speed blender is 30,000 rpm; the breaking time is 45 seconds.

[0077] S32. Take the daisy flower pulp and centrifuge it to obtain the supernatant and precipitate; take the supernatant, concentrate and dry it to obtain daisy flower nanovesicles; the centrifugation specifically refers to centrifuging at a speed of 8000 rpm for 2 hours;

[0078] S33. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 50 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 20 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the daisy nanovesicles.

[0079] Comparative Example 1: Preparation of Plant Nanovesicle Composition

[0080] The plant nanovesicle composition is obtained by mixing peony nanovesicles, polygonatum nanovesicles and daisy nanovesicles in a weight ratio of 10:7:3.

[0081] The peony nanovesicles were prepared by the following method:

[0082] S11. Take peony root, add water and mix, then put it into a high-speed blender to break the cell wall and obtain a cell wall broken slurry; then press the cell wall broken slurry through a press and collect the juice to obtain peony root raw slurry; wherein, the weight ratio of peony root peel to water is 1:2; the speed of the high-speed blender is 30,000 rpm; the cell wall breaking time is 45 seconds.

[0083] S12. Centrifuge the original peony root bark pulp to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain peony nanovesicles; the centrifugation specifically refers to centrifugation at a speed of 8000 rpm for 2 hours;

[0084] S13. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 50 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 20 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the peony nanovesicles.

[0085] The Polygonatum nanovesicles were prepared by the following method:

[0086] S21. Mix the rhizome of Polygonatum odoratum with water, then put it into a high-speed blender for blending to obtain a blended slurry; then press the blended slurry through a press to collect the juice to obtain the original pulp of Polygonatum odoratum rhizome; wherein, in step S21, the weight ratio of Polygonatum odoratum rhizome to water is 1:2; the speed of the high-speed blender is 30,000 rpm; and the blending time is 45 seconds.

[0087] S22. Centrifuge the raw pulp of Polygonatum rhizome to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain Polygonatum nanovesicles; the centrifugation specifically refers to centrifugation at 8000 rpm for 2 hours;

[0088] S23. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 120 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 50 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the Polygonatum nanovesicles.

[0089] The daisy-flower nanovesicles were prepared by the following method:

[0090] S31. Take daisies, add water and mix, then put them into a high-speed blender to break them down into a pulp; then press the pulp through a press and collect the juice to obtain daisy puree; wherein, the weight ratio of daisies to water is 1:2; the speed of the high-speed blender is 30,000 rpm; the breaking time is 45 seconds.

[0091] S32. Take the daisy flower pulp and centrifuge it to obtain the supernatant and precipitate; take the supernatant, concentrate and dry it to obtain daisy flower nanovesicles; the centrifugation specifically refers to centrifuging at a speed of 8000 rpm for 2 hours;

[0092] S33. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 100 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 50 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the daisy nanovesicles.

[0093] Comparative Example 2: Preparation of Plant Nanovesicle Composition

[0094] The plant nanovesicle composition is obtained by mixing peony nanovesicles, polygonatum nanovesicles and daisy nanovesicles in a weight ratio of 10:7:3.

[0095] The peony nanovesicles were prepared by the following method:

[0096] S11. Take peony root, add water and mix, then put it into a high-speed blender to break the cell wall and obtain a cell wall broken slurry; then press the cell wall broken slurry through a press and collect the juice to obtain peony root raw slurry; wherein, the weight ratio of peony root peel to water is 1:2; the speed of the high-speed blender is 30,000 rpm; the cell wall breaking time is 45 seconds.

[0097] S12. Centrifuge the original peony root bark pulp to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain peony nanovesicles; the centrifugation specifically refers to centrifugation at a speed of 8000 rpm for 2 hours;

[0098] S13. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 100 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 50 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the peony nanovesicles.

[0099] The Polygonatum nanovesicles were prepared by the following method:

[0100] S21. Mix the rhizome of Polygonatum odoratum with water, then put it into a high-speed blender for blending to obtain a blended slurry; then press the blended slurry through a press to collect the juice to obtain the original pulp of Polygonatum odoratum rhizome; wherein, in step S21, the weight ratio of Polygonatum odoratum rhizome to water is 1:2; the speed of the high-speed blender is 30,000 rpm; and the blending time is 45 seconds.

[0101] S22. Centrifuge the raw pulp of Polygonatum rhizome to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain Polygonatum nanovesicles; the centrifugation specifically refers to centrifugation at 8000 rpm for 2 hours;

[0102] S23. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 50 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 20 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the Polygonatum nanovesicles.

[0103] The daisy-flower nanovesicles were prepared by the following method:

[0104] S31. Take daisies, add water and mix, then put them into a high-speed blender to break them down into a pulp; then press the pulp through a press and collect the juice to obtain daisy puree; wherein, the weight ratio of daisies to water is 1:2; the speed of the high-speed blender is 30,000 rpm; the breaking time is 45 seconds.

[0105] S32. Take the daisy flower pulp and centrifuge it to obtain the supernatant and precipitate; take the supernatant, concentrate and dry it to obtain daisy flower nanovesicles; the centrifugation specifically refers to centrifuging at a speed of 8000 rpm for 2 hours;

[0106] S33. Take the supernatant and perform ultrafiltration using an ultrafiltration membrane with a diameter of 100 nm to obtain a first permeate and a first concentrate; take the first permeate and perform ultrafiltration again using an ultrafiltration membrane with a diameter of 50 nm to obtain a second permeate and a second concentrate; take the second concentrate, concentrate and dry it to obtain the daisy nanovesicles.

[0107] Experimental example: HaCaT cell proliferation experiment

[0108] Human keratinocytes (HaCaT cells) were prepared into a cell suspension of 1×10⁵ cells / mL using complete culture medium. Then, 100 μL of the suspension was seeded into 96-well plates and cultured at 37°C and 5% CO₂ for 24 h. In the experimental group, 100 μg / mL of aqueous solution containing the test sample was added to each well for intervention. A blank group without the test sample solution was also included. After 48 h of intervention, the culture medium was discarded. 100 μL of CCK-8 reagent was added to each well and incubated at 37°C and 5% CO₂ for 1 h. The OD value at 450 nm was then measured using a microplate reader, and the cell viability of HaCaT cells was calculated. The results are shown in Table 1.

[0109] The samples to be tested are peony nanovesicles, polygonatum nanovesicles and daisy nanovesicles prepared by the method of Example 1, and plant nanovesicle compositions prepared by Examples 1-2 and Comparative Examples 1-2.

[0110] Table 1. Results of HaCaT cell survival rate assay

[0111]

[0112] As can be seen from the experimental results in Table 1, the survival rate of HaCaT cells in experimental groups 1-3 was higher than that in the blank group; this indicates that the peony nanovesicles, polygonatum nanovesicles and daisy nanovesicles prepared according to the method described in this invention have a certain effect on promoting the proliferation of HaCaT cells.

[0113] As can be seen from the experimental results in Table 1, the survival rate of HaCaT cells in experimental group 4 was significantly higher than that in experimental groups 1-3. This indicates that the plant nanovesicle composition obtained by combining peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles prepared according to the method of this invention has a significantly higher HaCaT cell proliferation-promoting effect than the individual peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles prepared according to the method of this invention. The combination of peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles prepared according to the method of this invention can produce a synergistic effect in promoting HaCaT cell proliferation.

[0114] As shown in Table 1, the HaCaT cell survival rate in experimental group 5 was significantly higher than that in experimental group 4. This indicates that the plant nanovesicle composition obtained by combining peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles after further ultrafiltration significantly enhanced the HaCaT cell proliferation promotion effect compared to the plant nanovesicle composition obtained by combining peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles without ultrafiltration.

[0115] As shown in Table 1, although the HaCaT cell survival rate in experimental groups 6 and 7 was improved compared to experimental group 4, the increase was not significant, and the increase was much smaller than that in experimental group 5. This indicates that ultrafiltration conditions are crucial in the preparation of peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles. Only by combining peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles obtained under the ultrafiltration conditions of this invention can the plant nanovesicle composition promoting HaCaT cell proliferation be further significantly enhanced. However, the plant nanovesicle composition obtained by combining peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles obtained under other ultrafiltration conditions does not achieve a further significant improvement in promoting HaCaT cell proliferation.

[0116] Example 3: Preparation of freeze-dried powder rich in plant nanovesicles

[0117] Based on 100 parts, the raw material composition by weight is as follows: 10 parts peony nanovesicles; 7 parts polygonatum nanovesicles; 3 parts daisy nanovesicles; 3 parts supramolecular resveratrol; 5 parts D-mannitol; the remainder is water.

[0118] Peony nanovesicles, Polygonatum nanovesicles, daisy nanovesicles, resveratrol, and mannitol are mixed evenly in water and then freeze-dried to obtain the freeze-dried powder rich in plant nanovesicles.

[0119] The freeze-drying parameters are as follows: Pre-freeze the product to -40±5℃ and hold for at least 180 minutes; First temperature increase: -15℃ for 60 minutes, hold at -15℃ for at least 180 minutes; Second temperature increase: -5℃ for 60 minutes, hold at -5℃ for at least 180 minutes; Third temperature increase: 5℃ for 60 minutes, hold at 5℃ for at least 180 minutes; Fourth temperature increase: 30℃ for 60 minutes, hold at 30℃ for at least 180 minutes. The vacuum level is maintained at 23±3 Pa.

Claims

1. A plant nanovesicle composition, characterized in that, It includes peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles.

2. The plant nanovesicle composition according to claim 1, characterized in that, The weight ratio of peony nanovesicles, polygonatum nanovesicles, and daisy nanovesicles is 1-20:1-10:1-5.

3. The plant nanovesicle composition according to claim 1, characterized in that, The peony nanovesicles were prepared by the following method: S11. Take peony root bark, add water and mix, then put it into a high-speed blender to break the cell wall and obtain a cell wall broken slurry; then press the cell wall broken slurry through a press and collect the juice to obtain peony root bark raw slurry. S12. Centrifuge the original pulp of peony root bark to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain peony nanovesicles.

4. The plant nanovesicle composition according to claim 1, characterized in that, The Polygonatum nanovesicles were prepared by the following method: S21. Take the rhizome of Polygonatum and mix it with water, then put it into a high-speed blender to break it down into a pulp; then press the pulp through a press and collect the juice to obtain the original pulp of Polygonatum rhizome. S22. Centrifuge the raw pulp of Polygonatum rhizome to obtain supernatant and precipitate; concentrate and dry the supernatant to obtain Polygonatum nanovesicles.

5. The plant nanovesicle composition according to claim 1, characterized in that, The daisy-flower nanovesicles were prepared by the following method: S31. Take daisy flowers, add water and mix, then put them into a high-speed blender to break them down into a pulp; then press the pulp through a press and collect the juice to obtain daisy flower puree. S32. Take the daisy flower pulp and centrifuge it to obtain the supernatant and precipitate; take the supernatant, concentrate and dry it to obtain daisy flower nanovesicles.

6. A freeze-dried powder rich in plant nanovesicles, characterized in that, It contains the following components: peony nanovesicles, polygonatum nanovesicles, daisy nanovesicles, resveratrol, mannitol, and water.

7. A freeze-dried powder rich in plant nanovesicles, characterized in that, It contains the following components in parts by weight: 1-20 parts of peony nanovesicles; 1-10 parts of polygonatum nanovesicles; 1-5 parts of daisy nanovesicles; 0.5-5 parts of resveratrol; 2.5-10 parts of mannitol; and 50-100 parts of water.

8. The method for preparing the freeze-dried powder rich in plant nanovesicles according to claim 6 or 7, characterized in that, Peony nanovesicles, Polygonatum nanovesicles, daisy nanovesicles, resveratrol, and mannitol are mixed evenly in water and then freeze-dried to obtain the freeze-dried powder rich in plant nanovesicles.

9. The use of the plant nanovesicle composition according to any one of claims 1 to 6 or the freeze-dried powder rich in plant nanovesicles in the preparation of products that promote the proliferation of human keratinocytes.

10. The use of the plant nanovesicle composition according to any one of claims 1 to 6 or the freeze-dried powder rich in plant nanovesicles in the preparation of products that promote skin barrier repair.

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