CN116983333B

CN116983333B - Application of ginseng exosomes in the preparation of nanomedicines for the treatment of diabetic ulcer vascular lesions

Info

Publication number: CN116983333B
Application number: CN202311052083.4A
Authority: CN
Inventors: 彭丽华; 谈旻鸿
Original assignee: Zhejiang University ZJU
Current assignee: Zhejiang University ZJU
Priority date: 2023-08-21
Filing date: 2023-08-21
Publication date: 2025-01-28
Anticipated expiration: 2043-08-21
Also published as: CN116983333A

Abstract

The invention discloses application of ginseng exosomes in preparing nano-drugs for treating diabetic ulcer vascular lesions, and relates to the field of biological medicines. Experiments prove that the ginseng exosome can reprogram the glycolysis process of vascular endothelial cells under the condition of high sugar, and reduce the generation of active oxygen cluster substances by inhibiting the expression of acetyl CoA in the cells, thereby promoting the expression of nitric oxide synthase and reducing or reversing the apoptosis under the induction of high sugar. The ginseng exosome is used as nano medicine and has the functions of preventing, treating or improving diabetic ulcer microvasculopathy. The ginseng exosome has simple preparation process, low cytotoxicity, high delivery efficiency and good stability, and can be used as a novel nano medicine for preventing, treating or improving diabetes vascular lesion related diseases.

Description

Application of ginseng exosomes in preparing nano-medicament for treating diabetic ulcer vasculopathy

Technical Field

The invention belongs to the field of biological medicines, and particularly relates to a novel application of ginseng exosomes, in particular to an application of ginseng exosomes in preparing nano medicines for treating diabetic ulcer vascular lesions.

Background

With the progress of aging of population and the change of life style of people, the occurrence and development of metabolic diseases such as diabetes, obesity and metabolic syndrome are increasing, and become a global public health problem. Among them, diabetes is a life-long disease of sugar metabolism that causes abnormal tissue structure and organ functions due to excessive blood glucose concentration, and according to 2021 international diabetes union (International Diabetes Federation, IDF) statistics, it is shown that about 5.37 million adults (20-79 years) worldwide have diabetes. Diabetes has a great deal of complications, wherein long-term hyperglycemia is easy to cause retinopathy, arteriosclerosis, angiopathy such as cardiovascular diseases and small blood vessels, and most of medicaments have little treatment effect after the complications occur and are difficult to reverse. However, the blood sugar control rate of diabetes is only 49.4% so far, which makes a plurality of medicines unable to play a therapeutic role from the fundamental problem of sugar metabolism regulation, so the research and development of novel medicines are still the problems and research hot spots to be solved in the fields of anti-diabetes and its complications treatment.

Wherein, the vascular endothelial cell chain reaction caused by hyperglycemia metabolic disorder, thereby causing insufficient angiogenesis, is the root cause of diabetic ulcer vasculopathy and angiogenesis disorder. Under normal conditions, glucose enters the cell and is subjected to glycolysis and mitochondrial metabolism to produce substances and energy supply to support various physiological functions of the cell, while vascular endothelial cells in a hyperglycemic environment will undergo a series of abnormal metabolic processes 1. The excess sugar metabolic intermediate acetyl-CoA molecules are converted into acyl carnitine, accumulation of which can cause cytotoxicity 2. Excess substrate loading causes oxidative phosphorylation failure of the cell, loss of ATP production and increase of ROS levels, which subsequently lead to mitochondrial failure 3. Higher glycosylation end products are formed, resulting in altered or lost protein functions, promoting release of ROS and initiating a series of changes in cell signaling 4. Excess glucose is also metabolized via the polyol and hexosamine pathways, which respectively lead to increased ROS and inflammatory levels. It is therefore a focus of research in this area how to perform effective sugar metabolism modulation therapies on vascular endothelial cells in a hyperglycemic environment.

In the current treatment of diabetic ulcer vasculopathy, especially diabetic ulcer angiogenesis deficiency, oral hypoglycemic drugs are most commonly applied to systemic treatment, such as metformin, thiazolidinediones, alpha glycosidase inhibitors, but side effects such as weight gain, cardiovascular toxicity, hepatotoxicity, gastrointestinal reaction and the like can be caused by long-term use, and pertinence is lacking in vascular treatment, while the systemic hypoglycemic treatment is carried out, oral administration or injection of antioxidant stress drugs (such as alpha-zinc sulfate), vasodilating drugs (such as alprostadil) and local application of various growth factor preparations (such as VEGF) are the most common treatment means at present, and although the drugs have certain effects and use values, the drugs are difficult to solve the treatment problem from the aspect of disease occurrence root, namely endothelial cell glycolysis disorder, and have low bioavailability and single function, so that the application is limited.

The traditional Chinese medicine for preventing and treating diabetes mellitus has clinical practice for thousands of years, and a theoretical academic system of the system is gradually formed through continuous development and innovation. A great number of preclinical researches and practices show that the traditional Chinese medicine plants and the active ingredients thereof have practical curative effects and great development prospects in regulating the treatment of the diseases of the glycometabolism disorder, for example, the active ingredients of radix rehmanniae oligosaccharides, cinnamon polyphenol, leonurine, coptis chinensis and the like extracted from the figwort family plants are proved to be capable of promoting the glucose metabolism by increasing the expression or activity of Glucose Kinase (GK) mRNA, the polysaccharide of the cortex moutan, silkworm chrysalis oil, aloe extract and the like can be used for improving the glycometabolism disorder and insulin resistance, and astragaloside and ginsenoside respectively reduce the degradation of glycogen, reduce the generation of liver glucose and inhibit the overactivation of gluconeogenesis by inhibiting Glycogen Phosphorylase (GP) and acid enol type pyruvate carboxykinase (PEPCK) in the glycolysis process.

Exosomes are spherical vesicles released by cells and have a diameter of 30-150nm and a lipid bilayer membrane structure, can be prepared on a large scale by means of density gradient centrifugation, tangential flow filtration and the like, and are stored for a long time at low temperature in the form of concentrated solution or freeze-dried powder. Besides surface lipid, proteins and nucleic acids (including mRNA, miRNA, lncRNA and CIRCLE RNA) contained in exosomes are important functional substances, and have important regulation and control effects on various biological processes and disease treatment. The exosomes extracted from the traditional Chinese medicine plants have remarkable therapeutic activity, small toxic and side effects and low immunogenicity, and become a new hot spot for research in the field of exosomes. Currently, grape-derived exosomes are being developed for clinical trials to treat oral mucositis (NCT 01668849) caused by chemotherapy of head and neck cancer, ginger and aloe-derived exosomes are being clinically tested to treat polycystic ovary syndrome (NCT 03493984), and the use of plant exosomes as nanocarriers to deliver curcumin to treat colon cancer has entered the clinical trial stage (NCT 01294072) (https:// beta.

Although the effectiveness of exosome treatment and the clinical basis of regulating glycometabolism by Chinese medicinal plants have wide research foundation, the application of the plant exosome to the treatment of diabetes vascular lesions by regulating endothelial cell glycometabolism in diabetes is not reported so far, and particularly the application of the plant exosome to the treatment of angiogenesis disorder at diabetes ulcers is not yet involved.

Disclosure of Invention

In order to overcome the defects of the existing therapeutic drugs, the invention provides a ginseng exosome nano-drug for efficiently and safely regulating the glucose metabolic process of vascular endothelial cells, and particularly provides application of the ginseng exosome in preparing the nano-drug for treating diabetic vascular lesions. The invention discovers for the first time that the ginseng exosome has therapeutic capability on the pathologic vascular endothelial cells in the high-sugar environment, can promote glucose metabolism by regulating the glycolysis process, inhibit the production of acetyl-CoA in cells, further down regulate the ROS level, up regulate the expression of nitric oxide synthase, promote the survival and proliferation of the endothelial cells in the high-sugar environment, and restore the normal physiological functions and activities of the endothelial cells. The invention has important value and significance for further development and application of various traditional Chinese medicine plant exosomes including ginseng exosomes.

In order to achieve the above purpose, the invention adopts the following technical scheme:

Application of Ginseng radix exosome in preparing nanometer medicine for treating diabetic vasculopathy is provided.

Further, the ginseng exosomes are purified by adopting an ultracentrifugation method, a sucrose density gradient centrifugation method, a polymer precipitation method or an ultrafiltration combined size exclusion chromatography method.

Further, the ginseng exosomes are purified by adopting an ultracentrifugation method, and when the ginseng exosomes are extracted by adopting the ultracentrifugation method, the centrifugation sequence and conditions are that the supernatant is taken at 2000-6000g for 20-50min, the supernatant is taken at 7000-15000g for 1h, the supernatant is taken at 100000-200000g for 1-3h, and the sediment is taken to obtain the extracted ginseng exosomes.

Further, the active substances in the ginseng exosomes are derived from one or more of protein, lipid, mRNA and small RNA contained in the ginseng exosomes, and the small RNA is one or more of miRNA, lncRNA and CIRCLE RNA.

Further, the diabetic vasculopathy is a microangiopathy and/or an angiogenic disorder in a diabetic skin ulcer.

Furthermore, the nano-drug is a preparation taking the ginseng exosome as an active ingredient or a compound preparation taking the ginseng exosome as a component.

Further, the compound preparation is one or more of ointment, gel and patch.

The beneficial effects of the invention include:

(1) The plant exosome provided by the invention can obviously reprogram the glucose metabolism process of vascular endothelial cells under the induction of high glucose or diabetes, up-regulate the expression of genes PFKM, PGK1 and ENO1 to promote the glycolysis process, reduce the production of acetyl coenzyme A, reduce the ROS level in the endothelial cells and promote the expression of nitric oxide synthase by the endothelial cells.

(2) The plant exosome provided by the invention can obviously reduce vascular endothelial cell apoptosis, especially apoptosis under the induction of high sugar or on partial skin ulcer of diabetic animals, reverse damage of high sugar treatment or diabetes to endothelial cells and promote cell proliferation.

(3) The plant exosome provided by the invention can help the local vascular endothelial cells of the skin ulcers of the diabetic animals to maintain normal physiological states, and promote angiogenesis and microvascular network formation at the diabetic ulcers.

(4) The invention is used for preparing the nano-medicament for preventing and treating diabetic vascular lesions, and has the effects of guaranteeing the health of diabetics and prolonging the service life of diabetics.

Drawings

FIG. 1 shows a particle size distribution diagram and potential values of ginseng exosomes obtained according to an embodiment of the present invention;

FIG. 2 is a TEM image of an exosome of the ginseng according to one embodiment of the present invention;

FIG. 3 is a graph showing the effect of different concentrations of ginseng exosomes on vascular endothelial cell proliferation under normal sugar conditions according to one embodiment of the present invention;

FIG. 4 is a graph showing the effect of 8ug/mL ginseng exosomes on vascular endothelial cell proliferation under high sugar conditions in one embodiment of the present invention;

FIG. 5 is a graph of GSEA enrichment analysis of glycolytic pathways in vascular endothelial cell genomes before and after treatment of 8ug/mL of ginseng exosomes according to an embodiment of the present invention and differentially expressed genes;

FIG. 6 is a diagram showing Western blot analysis of glycolytic genes PFKM, PGK1, ENO1 in vascular endothelial cells before and after 8ug/mL of exosome treatment in accordance with an embodiment of the present invention;

FIG. 7 shows glycolytic related pathways and differentially expressed metabolites in cell metabolomics before and after treatment with 8ug/mL of ginseng exosomes according to an embodiment of the present invention;

FIG. 8 is a graph showing the effect of 8ug/mL ginseng exosomes on the expression of acetyl-CoA in cells under high sugar conditions in an embodiment of the present invention;

FIG. 9 is a graph showing the effect of 8ug/mL ginseng exosomes on cellular ROS levels under high sugar conditions in an embodiment of the present invention;

FIG. 10 is a graph showing the effect of 8ug/mL ginseng exosomes on cellular nitric oxide synthase expression levels under high sugar conditions in an embodiment of the present invention;

FIG. 11 is a graph showing the effect of Western blot on the expression of glycolytic genes PFKM, PGK1 and ENO1 in local tissues of skin ulcers after topical application of ginseng exosomes in diabetic mice according to an embodiment of the present invention;

FIG. 12 is a graph showing the variation of acetyl-CoA content in skin ulcer local tissue after topical application of ginseng exosomes to diabetic mice in accordance with an embodiment of the present invention;

FIG. 13 is a graph showing the variation of nitric oxide synthase content in skin ulcer local tissue after topical application of ginseng exosomes to diabetic mice in accordance with an embodiment of the present invention;

fig. 14 is a photograph of a microscope of a diabetic mouse with a local micro-vascular network formed by skin ulcers after the application of the ginseng exosomes in a local manner according to an embodiment of the present invention.

Detailed Description

The invention is further illustrated by means of examples.

EXAMPLE 1 extraction of Ginseng exosomes by ultracentrifugation

Washing Ginseng radix with deionized water, peeling, cutting, and squeezing with juicer. Centrifuging the obtained juice at different Chinese medicinal plant medicinal parts at different speed for 20-50min with table-type refrigerated centrifuge to remove larger plant residues, collecting supernatant, centrifuging at 7000-15000g for 1 hr to remove possible tiny plant fiber and debris, collecting supernatant, centrifuging at floor-type ultracentrifuge (100000-200000 g,1-3 hr), collecting bottom precipitate, and re-suspending with small amount of Phosphate Buffer (PBS) to obtain Ginseng radix exosome solution.

EXAMPLE 2 sucrose Density gradient centrifugation method for extracting Ginseng radix exosomes

Washing Ginseng radix with deionized water, peeling, cutting, and squeezing with juicer. The obtained juice is frozen and centrifuged for 1 hour under the condition of 5000g-12000g, the supernatant is taken after repeating twice, then the supernatant is ultracentrifuged for 1-3 hours under 120000-180000g, the precipitate is taken, PBS is used for resuspension, and the heavy suspension is purified by a sucrose density gradient centrifugation method. Sucrose density gradient solutions were prepared with four different concentrations of 8%,30%,45% and 60%, and the heavy suspension was added by syringe over the density gradient solution and then centrifuged in a floor ultracentrifuge (100000-200000 g,1-3 h). After centrifugation, the exosomes enriched between the 30/45% interfaces were recovered. Then, the sucrose solution was removed by a table ultracentrifuge (100000-200000 g,1-3 h), and the bottom pellet was resuspended with a small amount of PBS to obtain a purified ginseng exosome solution.

EXAMPLE 3 extraction of Ginseng exosomes by Polymer precipitation

Washing Ginseng radix with deionized water, peeling, cutting, squeezing juice with juicer, and purifying by polymer precipitation. For polyethylene glycol (PEG) -based precipitation, PEG-containing solutions were mixed with plant juice at a volume ratio (v/v) of 1:5-1:1, and the mixture was incubated overnight at 4 ℃. The mixture was then centrifuged at 1000-3000g for 20-50 min at 4 ℃ and the supernatant was discarded before resuspension of the pellet containing exosome-like nanovesicles. And centrifuging the exosome precipitate suspension with ultracentrifugation at 100000-1500000g for 1-4 hr, removing supernatant, and resuspending the bottom precipitate with small amount of PBS to obtain purified Ginseng radix exosome solution.

EXAMPLE 4 extraction of Ginseng exosomes by Ultrafiltration in combination with size exclusion chromatography

Washing Ginseng radix with deionized water, peeling, cutting, squeezing juice with juicer, and purifying by ultrafiltration combined with size exclusion chromatography. For size exclusion chromatography, separation was performed using a commercial chromatographic column (Izon Science, canterburg, new Zealand). The column was first conditioned with 70mL PBS. Then 10mL of clear crude extract plant juice was loaded and 10mL of eluent was collected as fraction 1. Then 20mLPBS was added to the column and 20mL of eluent was collected as fraction 2. Finally, 50mL of PBS was added to the column, and 20mL of the eluate was collected as fraction 3. Analyzing the abundance of plant exosomes in different components, selecting the component with the highest abundance, and further concentrating by using 10000MWCO film to obtain purified ginseng exosome solution.

Further, in order to reduce the quality difference caused by the extraction method of ginseng exosomes, the following examples 5 to 8 were obtained by the ultracentrifugation method in example 1, which are used for scanning electron microscope characterization, quantitative qualitative analysis of contents, cell experiments, and animal experiments.

Example 5 characterization of Ginseng exosomes and qualitative and quantitative analysis of their inclusion

Characterization of ginseng exosomes the size and morphology of the ginseng exosomes prepared in example 1 were observed using transmission electron microscopy, as shown in fig. 1. The particle size of the exosomes was measured using a malvern particle size potentiometer, as shown in fig. 2. As shown in figures 1 and 2, the observed forms under the transmission electron microscope all accord with the characteristics of exosomes, are in regular spherical shapes, have uniform sizes and have the particle size of 117.7+/-6.3 nm.

Isolation of proteins contained in Ginseng radix exosomes the proteins contained in Ginseng radix exosomes were isolated and extracted with SDT (4% SDS,100mM Tris-HCl, pH 7.6) buffer. 20 μg of protein was taken from each sample, mixed with 5 Xloading buffer, boiled for 5min, separated on 4% -20% SDS-PAGE gel (constant pressure 180V,45 min) and stained with Coomassie blue R-250 to reveal protein bands.

Isolation analysis of nucleic acids contained in Ginseng exosomes Total RNA from plant exosomes was isolated and purified using Trizol reagent (Invitrogen, carlsbad, calif., USA) and nucleic acids including mRNA and small RNA (miRNA, lncRNA, CIRCLE RNA) from plant exosomes were analyzed by cleavage, labeling, amplification, sequencing at paired ends, sequence mapping procedures.

Separation and analysis of lipid in ginseng exosome, extracting G-Exos lipid by MTBE method. Briefly, samples (100 μl) were labeled with a lactone standard, then separated by reverse phase liquid chromatography, positive and negative ions were detected by electrospray ionization (ESI), and finally peak extraction and identification of lipid-containing molecules and internal standard lipid molecules in ginseng exosomes were performed.

Example 6 experiments on the Effect of Ginseng exosomes on cell proliferation in high sugar environments

In this example, the therapeutic effect and mechanism of ginseng exosomes on glucose metabolism disorders were investigated using Human Umbilical Vein Endothelial Cells (HUVECs) as a cell model. HUVECs are derived from the cell bank of the national academy of sciences (Shanghai). 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 ℃,5% co ₂.

To first explore the effect of varying concentrations of ginseng exosomes on cell proliferation, HUVECs were inoculated in RPMI 1640 medium and allowed to adhere overnight. The following day, cells were cultured using RPMI 1640 supplemented with different concentrations of plant exosomes (0 μg/mL,0.25 μg/mL,0.5 μg/mL,1 μg/mL,2 μg/mL,4 μg/mL,8 μg/mL,10 μg/mL,12 μg/mL) instead of medium for 24h. Cell proliferation rate was then determined using a cell counting kit (Beyotime). Briefly, 10. Mu.L of CCK-8 (100. Mu.L of medium) was added to each well. After incubation at 37 ℃ for 2h, absorbance at 450nm was measured with BioTek Cytation3, 3 CELLLIMAGING multimode reader. As a result, as shown in FIG. 3, the cell proliferation rate increased with the increase of the administration concentration of ginseng exosomes, and when the concentration was more than 8 μg/mL, the proliferation rate did not significantly change, so that the concentrations of ginseng exosomes used in the subsequent cell experiments were all 8 μg/mL.

In order to first explore the influence of ginseng exosomes on cell proliferation under high sugar conditions, the experimental groups are replaced by normal groups, high sugar groups and high sugar+ginseng exosomes groups for experiments, the rest steps are unchanged, and the cell proliferation rates in different experimental groups are measured. In this example, the glucose content in the normal group was 16mmol/L, and the glucose content in the high sugar group and the high sugar+ginseng exosome group was 45mmol/L. The results are shown in fig. 4, and the results show that the proliferation rate of the high sugar group is obviously lower than that of the normal group due to the damage of cells, and the proliferation rate of the high sugar and ginseng exosome group is equivalent to that of the normal group, which indicates that the ginseng exosome treatment effectively improves the apoptosis induced by the high sugar.

Example 7 experiments of the influence of Ginseng exosomes on intracellular genes PFKM, PGK1, ENO1, acetyl-CoA content, ROS expression, nitric oxide synthase expression in high sugar environments

To first explore the effect of varying concentrations of ginseng exosomes on cell proliferation, HUVECs were inoculated in RPMI 1640 medium and allowed to adhere overnight. The following day, two experimental groups, a high sugar group, a high sugar + ginseng exosome group, were set up for 24 hours using RPMI 1640 replacement medium with or without the addition of high sugar or ginseng exosome, using the same glucose content as in example 6. And then carrying out qualitative and quantitative analysis on different groups of cells by using a Western blot experiment, an acetyl coenzyme A content measurement kit, an ROS fluorescent probe kit and a nitric oxide synthase expression measurement kit respectively so as to determine the influence of ginseng exosomes on the ROS level, genes and enzyme expression of the cells in a high-sugar environment. The results are shown in FIGS. 5,6,7,8,9, and 10. The results showed that the levels of PFKM, PGK1, ENO1 gene expression in cells of the ginseng exosome group were significantly higher than those of the control group (fig. 5, 6), the acetyl-coa content was significantly lower than that of the control group (fig. 7, 8), ROS expression was significantly lower than that of the control group (fig. 9), and nitric oxide synthase expression was significantly higher than that of the control group (fig. 10). The ginseng exosomes are shown to promote the expression of genes related to cell glycolysis, inhibit the expression of acetyl coenzyme A, reduce the level of cellular ROS and improve the expression of nitric oxide synthase.

Example 8 experiments of the influence of the exosomes of Ginseng on the expression of the genes PFKM, PGK1, ENO1, acetyl-CoA content, and nitric oxide synthase in ulcerated skin tissue of diabetic animals

In this example, a diabetic mouse was used as an in vivo animal model, and a partial whole skin tissue defect model was produced, and the mechanism of action of ginseng exosomes was investigated. Study Using 8-12 week old male B6.BKS (D) -Lepr db/J (db/db) mice, the mice were general anesthetized, the wound was excised with a scalpel on the back skin where the hair was shaved, the wound diameter was 12mm, and wound contraction was prevented by securing the wound with a 15mm diameter ring-packed silica gel splint. Diabetic mice were randomly divided into 2 experimental groups. The wound surface was treated with 100. Mu.L of PBS or ginseng exosome solution (100. Mu.g/mL-500. Mu.g/mL), respectively, as animal control group and animal administration group, once every other day. During wound healing, two groups of mouse wound skin healing tissues were collected, tissue ground or skin cut. For tissue homogenates, PFKM, PGK1, ENO1 gene expression and acetyl CoA content in the homogenates were determined using Western blot experiments, acetyl CoA content assay kit, and for skin sections, the cut skin was stained using nitric oxide synthase staining kit to determine nitric oxide synthase levels in local tissues. The results are shown in FIGS. 11,12 and 13, and the results show that the expression level of PFKM, PGK1 and ENO1 genes in the ulcerous tissues of the diabetic mice in the animal administration group is obviously higher than that in the animal control group (FIG. 11), the content of acetyl-CoA is obviously lower than that in the animal control group (FIG. 12), and the expression of nitric oxide synthase is obviously higher than that in the animal control group (FIG. 13). The ginseng exosome promotes the expression of genes related to glycolysis of local tissues of ulcers of diabetic animals, inhibits the expression of acetyl coenzyme A and improves the expression of nitric oxide synthase.

Example 9 experiments of the Effect of ginseng exosomes on the development of ulcerated skin micro-blood vessels in diabetic animals

A diabetic mouse skin ulcer model was constructed using the animal protocol described in example 8 and treated using the ginseng exosome dosing regimen described in example 8. In the later stage of wound healing, two groups of mouse wound skin healing tissues are respectively collected, and a body type microscope is used for imaging micro blood vessels in the skin tissues. The results are shown in fig. 14, and the results show that the micro-vascular network density at the ulcer of the diabetic mice in the animal administration group is significantly higher than that in the animal control group. It is shown that topical administration of ginseng exosomes significantly promotes angiogenesis and microvascular formation at the ulcerated skin of diabetic animals.

Claims

1. The application of ginseng exosomes in preparing nano-medicine for treating diabetic vasculopathy is that the ginseng exosomes are obtained by adopting an ultracentrifugation method, the centrifugation sequence and conditions are that centrifugation is carried out for 20-50min under 2000-6000g, supernatant is taken, centrifugation is carried out for 1h under 7000-15000g, supernatant is taken, centrifugation is carried out for 1-3 h under 100000-200000 g g, sediment is taken, and the extracted ginseng exosomes are obtained, and the diabetic vasculopathy is microvasculopathy and/or angiogenesis disorder in diabetic skin ulcer.

2. The use according to claim 1, wherein the active substance in the ginseng exosomes is derived from one or more of proteins, lipids, mRNA, small RNAs contained therein, the small RNAs being one or more of miRNA, lncRNA and CIRCLE RNA.

3. The use according to claim 1 or 2, wherein the nano-drug is a preparation with ginseng exosomes as active ingredients or a compound preparation with ginseng exosomes as constituent ingredients.

4. The use according to claim 3, wherein the compound preparation is one or more of an ointment, a gel, a patch.