Application of ginseng exosomes in the preparation of nanomedicines for the treatment of diabetic ulcer vascular lesions

Abstract

The present invention discloses the application of ginseng exosomes in the preparation of nanomedicines for treating diabetic ulcer vascular lesions, and relates to the field of biomedicine. The present invention verifies through experiments that ginseng exosomes can reprogram the glycolysis process of vascular endothelial cells under high sugar conditions, and inhibit the expression of acetyl coenzyme A in cells, reduce the production of reactive oxygen species, and then promote the expression of nitric oxide synthase, thereby reducing or reversing cell apoptosis induced by high sugar. Ginseng exosomes are used as nanomedicines to prevent, treat or improve diabetic ulcer microangiopathy. The ginseng exosomes in the present invention have a simple preparation process, low cytotoxicity, high delivery efficiency, and good stability. They can be used as a new type of nanomedicine for the prevention, treatment or improvement of diseases related to diabetic vascular lesions.

Description

Technical Field

[0001] The present invention belongs to the field of biological medicines, and specifically relates to a new application of ginseng exosomes, and in particular to the application of ginseng exosomes in the preparation of nanomedicines for treating diabetic ulcer vascular lesions. Background Art

[0002] With the intensification of the aging process of the population and the changes in people's lifestyles, the occurrence and development of metabolic diseases such as diabetes, obesity and metabolic syndrome are increasing day by day, and have become a global public health issue. Among them, diabetes is a lifelong sugar metabolism disease caused by excessive blood sugar concentration, which causes abnormal tissue structure and organ function. According to the statistics of the International Diabetes Federation (IDF) in 2021, there are approximately 537 million adults (20-79 years old) suffering from diabetes worldwide. There are many complications of diabetes. Among them, long-term high blood sugar environment can easily cause vascular diseases such as retinopathy, arteriosclerosis, cardiovascular and small blood vessels, and most drug treatments have little effect after complications occur and are difficult to reverse. However, as of now, the blood sugar control rate of diabetes is only 49.4%, which makes it impossible for most drugs to play a therapeutic role from the fundamental problem of sugar metabolism regulation. Therefore, the development of new drugs is still an urgent problem and research hotspot in the field of anti-diabetes and its complications.

[0003] Among them, the chain reaction of endothelial cells caused by hyperglycemia metabolism disorder, which leads to insufficient angiogenesis, is the root cause of diabetic ulcer vascular lesions and angiogenesis disorder. Under normal circumstances, after glucose enters the cell, it undergoes glycolysis and mitochondrial metabolism to produce substances and energy supply to support various physiological functions of the cell; however, endothelial cells in a hyperglycemic environment will undergo a series of abnormal metabolic processes: 1. Excessive glucose metabolism intermediates, acetyl coenzyme A molecules, are converted into acylcarnitines, and the accumulation of acylcarnitines will cause cytotoxicity; 2. Excessive substrate load causes cell oxidative phosphorylation failure, ATP production loss and increased ROS levels, followed by mitochondrial failure; 3. Advanced glycation end products are formed, leading to changes or loss of protein function, promoting the release of ROS and triggering a series of changes in cell signal transduction; 4. Excess glucose is also metabolized through polyol and hexosamine pathways, which lead to increased ROS and inflammation levels, respectively. Therefore, how to effectively regulate glucose metabolism in endothelial cells in a hyperglycemic environment is a research hotspot in this field.

[0004] At present, in the treatment of vascular lesions of diabetic ulcers, especially the insufficient angiogenesis of diabetic ulcers, systemic treatment with oral hypoglycemic drugs is the most common, such as metformin, thiazolidinediones, and α-glucosidase inhibitors. However, long-term use can cause side effects such as weight gain, cardiovascular toxicity, liver toxicity, and gastrointestinal reactions, and the vascular treatment lacks specificity. In addition to systemic hypoglycemic treatment, oral or injectable antioxidant stress drugs (such as α-zinc sulfate), vasodilators (such as alprostadil), and local use of various growth factor preparations (such as VEGF) are the most common treatment methods. Although these drugs have certain effects and use values, they are generally difficult to solve the treatment problem from the perspective of the root cause of the disease, that is, endothelial cell glycolysis disorder, and have low bioavailability and single function, which limits their use.

[0005] Traditional Chinese medicine has thousands of years of clinical practice in the prevention and treatment of diabetes. After continuous development and innovation, it has gradually formed a systematic theoretical and academic system. A large number of preclinical studies and practices have shown that Chinese medicinal plants and their active ingredients have practical efficacy and great development prospects in regulating the treatment of glucose metabolism disorders. For example, extracts of Scrophulariaceae plants such as Rehmannia oligosaccharides, cinnamon polyphenols, leonurine, and the active ingredient of Coptis chinensis, berberine, have been shown to promote glucose metabolism by increasing the expression or activity of glucokinase (GK) mRNA; phoenix danshen polysaccharides, silkworm pupa oil, aloe extract, etc. can improve glucose metabolism disorders and insulin resistance; astragaloside IV and ginsenosides reduce glycogen degradation, reduce liver glucose production, and inhibit the over-activation of gluconeogenesis by inhibiting glycogen phosphorylase (GP) and acid enolpyruvate carboxykinase (PEPCK) in the glycolysis process, respectively, to play a hypoglycemic effect.

[0006] Exosomes are a type of spherical vesicles with a diameter of 30-150nm and a lipid bilayer membrane structure that are released by cells. They can be prepared on a large scale by density gradient centrifugation, tangential flow filtration, and other methods, and stored for a long time at low temperatures in the form of concentrated liquid or lyophilized powder. In addition to surface lipids, the proteins and nucleic acids (including mRNA, miRNA, lncRNA, and circle RNA) contained in exosomes are important functional substances, which play an important regulatory role in a variety of biological processes and disease treatments. Among them, exosomes extracted from traditional Chinese medicine plants not only have significant therapeutic activity, but also have low toxicity and side effects and low immunogenicity, and have become a new hot spot in the field of exosome research. Currently, grape-derived exosomes are undergoing clinical trials to treat oral mucositis caused by chemotherapy for head and neck cancer (NCT01668849); ginger and aloe-derived exosomes are undergoing clinical trials to treat polycystic ovary syndrome (NCT03493984); and the use of plant exosomes as nanocarriers to deliver curcumin to treat colon cancer has entered the clinical trial stage (NCT01294072) (https: / / beta.clinicaltrials.gov / ).

[0007] Although the effectiveness of exosome therapy and the clinical basis of Chinese medicinal plants in regulating glucose metabolism have a broad research basis, to date, there have been no reports on the use of plant exosomes to regulate the glucose metabolism process of endothelial cells in diabetes for the treatment of diabetic vascular disease, especially in the treatment of angiogenesis disorders in diabetic ulcers. Summary of the invention

[0008] In order to overcome the shortcomings of existing therapeutic drugs, the present invention provides a ginseng exosome nanomedicine that is efficient and safe for regulating the glucose metabolism process of vascular endothelial cells, and in particular provides the use of ginseng exosomes in the preparation of nanomedicines for treating diabetic vascular lesions. The present invention first discovered that ginseng exosomes have therapeutic ability for pathological vascular endothelial cells in a high-sugar environment, and can promote glucose metabolism by regulating their glycolysis process, while inhibiting the production of acetyl-CoA in cells, thereby downregulating ROS levels, and upregulating the expression of nitric oxide synthase, promoting the survival and proliferation of endothelial cells in high sugar, and restoring their normal physiological functions and activities. The present invention has important value and significance for the further development and application of various types of exosomes from traditional Chinese medicine plants, including ginseng exosomes.

[0009] To achieve the above object, the present invention adopts the following technical solution:

[0010] Application of ginseng exosomes in the preparation of nanomedicines for the treatment of diabetic vascular disease.

[0011] Furthermore, the ginseng exosomes are prepared and purified by ultracentrifugation, sucrose density gradient centrifugation, polymer precipitation or ultrafiltration combined with size exclusion chromatography.

[0012] Furthermore, the ginseng exosomes are prepared and purified by ultracentrifugation; when ultracentrifugation is used for extraction, the centrifugation sequence and conditions are: 2000-6000g for 20-50min, taking the supernatant; 7000-15000g for 1h, taking the supernatant; 100000-200000g for 1-3h, taking the precipitate as the extracted ginseng exosomes.

[0013] Furthermore, the active substances in the ginseng exosomes are derived from one or more of the proteins, lipids, mRNAs, and small RNAs contained therein; and the small RNAs are one or more of miRNAs, lncRNAs, and circle RNAs.

[0014] Furthermore, the diabetic vascular disease is microangiopathy and / or angiogenesis disorder in diabetic skin ulcers.

[0015] Furthermore, the nanomedicine is a preparation with ginseng exosomes as an active ingredient or a compound preparation with ginseng exosomes as a component.

[0016] Furthermore, the compound preparation is one or more of an ointment, a gel, and a patch.

[0017] The beneficial effects of the present invention include:

[0018] (1) The plant exosomes provided by the present invention can significantly reprogram the sugar metabolism process of vascular endothelial cells under high sugar or diabetes induction, upregulate the expression of genes PFKM, PGK1, and ENO1 to promote the glycolysis process, while reducing the production of acetyl-CoA, reducing the level of ROS in endothelial cells, and promoting the expression of nitric oxide synthase in endothelial cells.

[0019] (2) The plant exosomes provided by the present invention can significantly reduce vascular endothelial cell apoptosis, especially cell apoptosis induced by high sugar or in local skin ulcers of diabetic animals, reverse the damage of endothelial cells caused by high sugar treatment or diabetes, and promote cell proliferation.

[0020] (3) The plant exosomes provided by the present invention can help the local vascular endothelial cells of diabetic animal skin ulcers maintain a normal physiological state and promote angiogenesis and microvascular network formation in diabetic ulcers.

[0021] (4) The present invention is used to prepare nanomedicines for the prevention and treatment of diabetic vascular lesions, which can protect the health of diabetic patients and prolong their lifespan. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The particle size distribution diagram and potential value of ginseng exosomes obtained in one embodiment of the present invention;

[0023] Figure 2 This is a TEM image of ginseng exosomes obtained in one embodiment of the present invention;

[0024] Figure 3 This is a graph showing the effect of different concentrations of ginseng exosomes on the proliferation of vascular endothelial cells under normal sugar conditions in one embodiment of the present invention;

[0025] Figure 4 This is a graph showing the effect of 8ug / mL ginseng exosomes on the proliferation of vascular endothelial cells under high glucose conditions in one embodiment of the present invention;

[0026] Figure 5 This is a GSEA enrichment analysis diagram and differentially expressed genes of the glycolysis pathway in the genome of vascular endothelial cells before and after treatment with 8ug / mL ginseng exosomes in one embodiment of the present invention;

[0027] Figure 6 This is a Western blot analysis of the glycolytic genes PFKM, PGK1, and ENO1 in vascular endothelial cells before and after treatment with 8ug / mL ginseng exosomes in an embodiment of the present invention;

[0028] Figure 7 The glycolysis-related pathways and differentially expressed metabolites in the cell metabolomics before and after treatment with 8ug / mL ginseng exosomes in one embodiment of the present invention;

[0029] Figure 8 This is a graph showing the effect of 8ug / mL ginseng exosomes on cell expression of acetyl-CoA under high glucose conditions in one embodiment of the present invention;

[0030] Fig. 9 This is a graph showing the effect of 8ug / mL ginseng exosomes on cellular ROS levels under high sugar conditions in one embodiment of the present invention;

[0031] Fig.10 This is a graph showing the effect of 8ug / mL ginseng exosomes on the expression level of cellular nitric oxide synthase under high glucose conditions in one embodiment of the present invention;

[0032] Fig.11 This is a diagram showing the Western blot effect on the expression of glycolytic genes PFKM, PGK1, and ENO1 in the local tissue of skin ulcers after local application of ginseng exosomes to diabetic mice in one embodiment of the present invention;

[0033] Fig.12This is a graph showing changes in acetyl-CoA content in local tissues of skin ulcers after topical application of ginseng exosomes to diabetic mice in one embodiment of the present invention;

[0034] Fig.13 This is a graph showing changes in the content of nitric oxide synthase in the local tissue of skin ulcers after local application of ginseng exosomes to diabetic mice in one embodiment of the present invention;

[0035] Fig.14 This is a stereomicroscope picture of the formation of local microvascular network in skin ulcer after local application of ginseng exosomes to diabetic mice in one embodiment of the present invention. DETAILED DESCRIPTION

[0036] The present invention is further illustrated below by way of examples.

[0037] Example 1 Extraction of ginseng exosomes by ultracentrifugation

[0038] Ginseng was washed with deionized water, peeled, cut into pieces, and squeezed with a juicer. The obtained juice was centrifuged at 2000-6000g for 20-50min according to the characteristics of the medicinal parts of different Chinese medicinal plants, to remove the larger plant residues and take the supernatant; then centrifuged at 7000-15000g for 1h to remove the possible tiny plant fibers and debris, and take the supernatant; then centrifuged in a floor-standing ultracentrifuge (100000-200000g, 1-3h), the bottom sediment was taken, and resuspended with a small amount of phosphate buffer (PBS) to obtain ginseng exosome solution.

[0039] Example 2 Extraction of ginseng exosomes by sucrose density gradient centrifugation

[0040] Ginseng was washed with deionized water, peeled, cut into pieces, and juiced with a juicer. The resulting juice was frozen and centrifuged at 5000g-12000g for 1h, repeated twice, and the supernatant was taken, and then the supernatant was ultracentrifuged at 120000-180000g for 1-3h, the precipitate was taken, resuspended with PBS, and the resuspended solution was purified by sucrose density gradient centrifugation. Sucrose density gradient solutions with four different concentrations of 8%, 30%, 45% and 60% were prepared, and the resuspended solution was added to the top of the density gradient solution through a syringe, and then centrifuged in a floor-standing ultracentrifuge (100000-200000g, 1-3h). After centrifugation, the exosomes enriched between the 30 / 45% interface were recovered. The sucrose solution was then removed by a desktop ultracentrifuge (100,000-200,000 g, 1-3 h), and the bottom precipitate was resuspended with a small amount of PBS to obtain a purified ginseng exosome solution.

[0041] Example 3 Extraction of ginseng exosomes by polymer precipitation method

[0042] Ginseng was washed with deionized water, peeled, cut into pieces, squeezed with a juicer, and then purified by polymer precipitation. For polyethylene glycol (PEG)-based precipitation, a PEG-containing solution was mixed with plant juice at a volume ratio (v / v) of 1:5-1:1, and the mixture was incubated at 4°C overnight. The mixture was then centrifuged at 1000-3000g for 20-50 minutes at 4°C, and the supernatant was discarded and the precipitate containing exosome-like nanovesicles was resuspended. The exosome precipitate suspension was then centrifuged at 100000-1500000g for 1-4h using ultracentrifugation, the supernatant was removed, and the bottom precipitate was resuspended with a small amount of PBS to obtain a purified ginseng exosome solution.

[0043] Example 4 Extraction of ginseng exosomes by ultrafiltration combined with size exclusion chromatography

[0044] Ginseng was washed with deionized water, peeled, cut into pieces, squeezed with a juicer, and then purified by ultrafiltration combined with size exclusion chromatography. For size exclusion chromatography, a commercial column (Izon Science, Canterbury, New Zealand) was used for separation. First, the column was conditioned with 70 mL of PBS. Then 10 mL of transparent crude plant juice was loaded, and 10 mL of the eluate was collected as component 1. Then 20 mL of PBS was added to the column, and 20 mL of the eluate was collected as component 2. Finally, 50 mL of PBS was added to the column, and 20 mL of the eluate was collected as component 3. The abundance of plant exosomes in different components was analyzed, and the component with the highest abundance was selected and further concentrated using a 10,000 MWCO membrane to obtain a purified ginseng exosome solution.

[0045] Furthermore, in order to reduce the quality differences caused by the ginseng exosome extraction method, the ginseng exosomes used in the scanning electron microscopy characterization, quantitative and qualitative analysis of inclusions, cell experiments, and animal experiments in the following implementation cases 5-8 were all obtained by the ultracentrifugation method in Example 1.

[0046] Example 5 Characterization of ginseng exosomes and qualitative and quantitative analysis of their contents

[0047] Characterization of ginseng exosomes: The size and morphology of the ginseng exosomes prepared in Example 1 were observed using a transmission electron microscope. Figure 1 The particle size of the exosomes was measured using a Malvern particle size potential analyzer. Figure 2 .like Figure 1 As shown in Figure 2, the morphology observed under transmission electron microscopy is consistent with the characteristics of exosomes, showing regular spherical shapes, relatively uniform sizes, and a particle size of 117.7±6.3nm.

[0048] Separation of proteins contained in ginseng exosomes: The proteins contained in ginseng exosomes were separated and extracted with SDT (4% SDS, 100mMTris-HCl, pH7.6) buffer. 20μg of protein was taken from each sample, mixed with 5X loading buffer, boiled for 5min, separated with 4%-20% SDS-PAGE gel (constant pressure 180V, 45min), and stained with Coomassie blue R-250 to display protein bands.

[0049] Isolation and analysis of nucleic acids contained in ginseng exosomes: The total RNA of plant exosomes was isolated and purified using Trizol reagent (Invitrogen, Carlsbad, CA, USA), and the nucleic acids in plant exosomes, including mRNA and small RNA (miRNA, lncRNA, circle RNA), were isolated and analyzed through lysis, labeling, amplification, paired-end sequencing, and sequence mapping procedures.

[0050] Separation and analysis of lipids contained in ginseng exosomes: The lipids of G-Exos were extracted using the MTBE method. Briefly, the sample (100 μL) was spiked with internal lipid standards and then separated using reversed-phase liquid chromatography, and electrospray ionization (ESI) was used to detect positive and negative ions, and finally the lipid molecules contained in ginseng exosomes and internal standard lipid molecules were peak extracted and identified.

[0051] Example 6 Experiment on the effect of ginseng exosomes on cell proliferation in a high sugar environment

[0052] This example uses human umbilical vein endothelial cells (HUVECs) as a cell model to explore the therapeutic effects and mechanisms of ginseng exosomes on glucose metabolism disorders. HUVECs were derived from the Chinese Academy of Sciences (Shanghai) Cell Bank. HUVECs were cultured in RPMI 1640 medium containing 10% bovine serum (Gibco BRL), l-glutamine, penicillin (50U / mL) and streptomycin (50U / mL). Cells were stored at 37°C, 5% CO2.

[0053] In order to first explore the effect of different concentrations of ginseng exosomes on cell proliferation, HUVECs were inoculated in RPMI1640 medium and allowed to adhere overnight. The next day, the cells were cultured for 24 hours using RPMI 1640 alternative medium supplemented with different concentrations (0μg / mL, 0.25μg / mL, 0.5μg / mL, 1μg / mL, 2μg / mL, 4μg / mL, 8μg / mL, 10μg / mL, 12μg / mL) of plant exosomes. The cell proliferation rate was then determined using a cell counting kit (Beyotime). Briefly, 10μL CCK-8 (100μL culture medium) was added to each well. After incubation at 37°C for 2h, the absorbance at 450nm was measured using a BioTek Cytation3 celllimaging multimode reader. The results are shown in the following figure. Figure 3 As shown in the results, the cell proliferation rate increased with the increase of ginseng exosome administration concentration. When the concentration was greater than 8μg / mL, the proliferation rate did not change significantly. Therefore, the ginseng exosome concentration used in subsequent cell experiments was 8μg / mL.

[0054] In order to first explore the effect of ginseng exosomes on cell proliferation under high sugar conditions, the above experimental groups were replaced with normal group, high sugar group, high sugar + ginseng exosome group, and the other steps remained unchanged. The cell proliferation rate in different experimental groups was measured. In this example, the glucose content in the normal group was 16mmol / L, and the glucose content in the high sugar group and high sugar + ginseng exosome group was 45mmol / L. The results are shown in Figure 4 As shown, the results showed that the proliferation rate of the high glucose group was significantly lower than that of the normal group due to cell damage, while the proliferation rate of the high glucose + ginseng exosomes group was equivalent to that of the normal group, indicating that ginseng exosomes treatment effectively improved high glucose-induced cell apoptosis.

[0055] Example 7 Experiment on the effect of ginseng exosomes on intracellular genes PFKM, PGK1, ENO1, acetyl-CoA content, ROS expression, and nitric oxide synthase expression in a high-sugar environment

[0056] In order to first explore the effect of different concentrations of ginseng exosomes on cell proliferation, HUVECs were inoculated in RPMI1640 medium and allowed to adhere overnight. On the second day, RPMI1640 alternative medium with or without high sugar or ginseng exosomes was used for culture for 24 hours, that is, two experimental groups were set up: high sugar group and high sugar + ginseng exosome group, and the glucose content used was the same as in Example 6. Then, Western blot experiments, acetyl-CoA content determination kits, ROS fluorescent probe kits, and nitric oxide synthase expression determination kits were used to qualitatively and quantitatively analyze cells in different groups to determine the effects of ginseng exosomes on cellular ROS levels, gene and enzyme expressions in a high sugar environment. The results are shown in the figure. Figure 5,6,7,8,9,10. The results showed that the expression levels of PFKM, PGK1, and ENO1 genes in the ginseng exosome group were significantly higher than those in the control group ( Figure 5 ,6), the acetyl-CoA content was significantly lower than that of the control group ( Figure 7 ,8), and ROS expression was significantly lower than that in the control group ( Fig. 9 ), the expression of nitric oxide synthase was significantly higher than that of the control group ( Fig.10 ). This indicates that ginseng exosomes promoted the expression of cellular glycolysis-related genes, inhibited the expression of acetyl-CoA, reduced the level of cellular ROS, and increased the expression of nitric oxide synthase.

[0057] Example 8 Experiment on the effect of ginseng exosomes on the gene PFKM, PGK1, ENO1, acetyl-CoA content, and nitric oxide synthase expression in the ulcer skin tissue of diabetic animals

[0058] This example uses diabetic mice as an in vivo animal model, and creates a local full-skin tissue defect model to explore the mechanism of action of ginseng exosomes. The study used 8-12 week old male B6.BKS (D) -Lepr db / J (db / db) mice, the mice were generally anesthetized, and the wound was excised on the shaved back skin with a scalpel. The wound diameter was 12mm, and a 15mm diameter ring-mounted silicone splint was used to fix the wound to prevent wound contraction. The diabetic mice were randomly divided into 2 experimental groups. The wound was treated with 100μL PBS or ginseng exosome solution (100μg / mL-500μg / mL), respectively, as an animal control group and an animal dosing group, once every other day. During the wound healing period, the wound skin healing tissues of the two groups of mice were collected, and the tissues were ground or the skin sections were made. For tissue homogenate, Western blot experiment and acetyl-CoA content determination kit were used to determine the PFKM, PGK1, ENO1 gene expression and acetyl-CoA content in the homogenate; for skin sections, nitric oxide synthase staining kit was used to stain the skin sections to determine the nitric oxide synthase level in local tissues. The results are shown in Figure 2. Fig.11 ,12,13, the results showed that the expression levels of PFKM, PGK1, and ENO1 genes in the ulcer tissues of diabetic mice in the animal treatment group were significantly higher than those in the animal control group ( Fig.11 ), the acetyl-CoA content was significantly lower than that of the animal control group ( Fig.12 ), the expression of nitric oxide synthase was significantly higher than that of the control group ( Fig.13 ). This indicates that ginseng exosomes promoted the expression of glycolysis-related genes in local tissues of diabetic animal ulcers, inhibited the expression of acetyl-CoA, and increased the expression of nitric oxide synthase.

[0059] Example 9 Experiment on the Effect of Ginseng Exosomes on the Neovascularization and Formation of Skin Microvessels in Diabetic Animal Ulcers

[0060] The animal experiment scheme described in Example 8 was used to construct a diabetic mouse skin ulcer model, and the ginseng exosome administration scheme described in Example 8 was used for treatment. In the late stage of wound healing, the wound skin healing tissues of the two groups of mice were collected, and the microvessels in the skin tissue were imaged using a stereo microscope. The results are shown in Figure 2. Fig.14 As shown, the results showed that the density of microvascular network in the ulcer of diabetic mice in the animal administration group was significantly higher than that in the animal control group. This shows that local administration of ginseng exosomes significantly promoted angiogenesis and microvascular formation in the ulcer skin of diabetic animals.

Claims

1. Application of ginseng exosomes in the preparation of nanomedicines for treating diabetic vascular disease; the ginseng exosomes are obtained by ultracentrifugation, and the centrifugation sequence and conditions are as follows: centrifugation at 2000-6000g for 20-50min, taking the supernatant; centrifugation at 7000-15000g for 1h, taking the supernatant; centrifugation at 100000-200000g for 1-3h, taking the precipitate as the extracted ginseng exosomes; the diabetic vascular disease is microangiopathy and / or angiogenesis disorder in diabetic skin ulcers.

2. The use according to claim 1, characterized in that: The active substances in the ginseng exosomes are derived from one or more of the proteins, lipids, mRNAs, and small RNAs contained therein; the small RNAs are one or more of miRNAs, lncRNAs, and circleRNAs.

3. The use according to claim 1 or 2, characterized in that: The nano drug is a preparation with ginseng exosomes as an active ingredient or a compound preparation with ginseng exosomes as a component.

4. The use according to claim 3, characterized in that: The compound preparation is one or more of ointment, gel and patch.

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