Application of human umbilical cord-derived mesenchymal stem cell apoptotic vesicles in preparation of drugs for treating uveitis

Abstract

The application discloses application of human umbilical cord-derived mesenchymal stem cell apoptosis vesicles in preparation of drugs for treating uveitis, and relates to the technical field of biological medicine.The application first applies human umbilical cord-derived mesenchymal stem cell apoptosis vesicles to preparation of drugs for treating uveitis, and the human umbilical cord-derived mesenchymal stem cell apoptosis vesicles can significantly improve eye inflammation and retinal tissue damage of a mouse with uveitis, significantly reduce the proportion of Th1 and Th17 proinflammatory cells in a spleen, and increase the proportion of Treg anti-inflammatory cells.Compared with stem cell exosomes, the human umbilical cord-derived mesenchymal stem cell apoptosis vesicles can more effectively reduce inflammatory cell infiltration in a vitreous cavity and a retinal layer, relieve retinal folds and detachment, reduce damage to photoreceptor cells, reduce fundus clinical scores and retinal pathological scores, and the immunoregulation effect is significantly better than that of stem cell exosomes, and the human umbilical cord-derived mesenchymal stem cell apoptosis vesicles overcome technical bottlenecks of storage, transportation and batch consistency of living cell preparations, and provide a safer, more stable and more efficient new strategy for uveitis treatment.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the application of apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells in the preparation of drugs for treating uveitis. Background Technology

[0002] Uveitis is an inflammation of the uvea (including the iris, ciliary body, and choroid) of the eye and is one of the leading causes of blindness. Uveitis can be caused by a variety of factors, including infection, immune-mediated diseases, trauma, metabolic disorders, and other systemic diseases (such as rheumatoid arthritis and ankylosing spondylitis). Clinically, treatment for uveitis usually relies on medication, primarily corticosteroids and immunosuppressants. However, long-term use of immunosuppressants and corticosteroids can lead to a range of side effects, such as immunosuppression, osteoporosis, and diabetes. Furthermore, some patients do not respond well to traditional treatments, making the disease difficult to control and potentially leading to complications such as glaucoma, cataracts, and retinal damage. [3] With a deeper understanding of the pathogenesis of uveitis, more and more research is focusing on immunomodulatory treatment strategies. The core pathogenesis of uveitis is an excessive inflammatory response triggered by abnormal activation of the immune system. [4] Immunomodulation has become a core research direction in the treatment of this disease. Mesenchymal stem cells (MSCs), due to their immunomodulatory properties, have provided a new approach for cell therapy of uveitis, and their derived extracellular vesicle preparations have become a research hotspot in recent years.

[0003] Mesenchymal stem cells (MSCs) are pluripotent adult stem cells with multipotent differentiation potential and self-replicating biological characteristics. They were first isolated from bone marrow by Friedenstein et al. in 1976 and later formally named by Caplan. While existing technologies have documented the role of human umbilical cord-derived MSCs in the treatment of uveitis, live cell therapy with MSCs carries risks of tumorigenesis and embolism, high immunogenicity, and difficulties in preservation, transportation, and batch-to-batch consistency, limiting its application. Furthermore, although there is exploration of stem cell exosomes for inflammatory diseases, their efficacy in treating uveitis is limited, failing to effectively alleviate severe ocular inflammation and retinal tissue damage.

[0004] Therefore, existing technologies still need improvement and development. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide the application of human umbilical cord-derived mesenchymal stem cell apoptotic vesicles in the preparation of drugs for treating uveitis, aiming to provide a new approach to the treatment of uveitis.

[0006] The technical solution of the present invention is as follows: Firstly, it provides the application of apoptotic vesicles from human umbilical cord-derived mesenchymal stem cells in the preparation of drugs for treating uveitis.

[0007] A preferred technical solution, the method for preparing the apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells, includes: Stimulation with astrocytocin induced apoptosis in human umbilical cord mesenchymal stem cells. After washing away the apoptotic human umbilical cord mesenchymal stem cells, centrifuge at 600-1000g for 5-15 minutes, then centrifuge at 1800-2200g for 5-15 minutes, and collect the supernatant; Centrifuge the supernatant at 15000~20000g for 20~40min and collect the precipitate; The precipitate was resuspended in phosphate buffer, centrifuged at 20,000-22,000g for 30-40 minutes, and the supernatant was discarded to obtain the apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells.

[0008] In a preferred embodiment, the drug is a drug for treating uveitis by inhibiting the differentiation of Th1 / Th17 pro-inflammatory cells and / or promoting the differentiation of Treg anti-inflammatory cells.

[0009] In a second aspect, a drug for treating uveitis is provided, the drug comprising: apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells.

[0010] In a preferred embodiment, the drug further includes pharmaceutically acceptable excipients.

[0011] In a further preferred embodiment, the pharmaceutically acceptable excipients include one or more of the following: pharmaceutically acceptable excipients, pharmaceutically acceptable additives, and pharmaceutically acceptable adjuvants.

[0012] In a preferred embodiment, when the drug is applied, the therapeutic dose of apoptotic vesicles from human umbilical cord-derived mesenchymal stem cells is (1-8) × 10⁻⁶. 9 One vesicle per time.

[0013] In a preferred embodiment, the drug is administered every 6-8 days.

[0014] Beneficial effects: This invention provides the application of human umbilical cord-derived mesenchymal stem cell apoptotic vesicles in the preparation of drugs for treating uveitis. Its advantages are: (1) Pharmacological activity: Animal experiments have confirmed that hUCMSCs-apoVs can significantly improve ocular inflammation and retinal tissue damage in EAU mice. Compared with stem cell exosomes, it can more effectively reduce the infiltration of inflammatory cells in the vitreous cavity and retinal layer, alleviate retinal folds and detachment, reduce photoreceptor cell damage, and reduce fundus clinical scores and retinal pathological scores. (2) Immunomodulation: hUCMSCs-apoVs have a strong immunosuppressive function. By secreting cytokines and immunomodulatory substances, it can significantly reduce the proportion of Th1 and Th17 pro-inflammatory cells in the spleen and increase the proportion of Treg anti-inflammatory cells. The immunomodulatory effect is significantly better than that of stem cell exosomes. (3) Pharmaceutical characteristics: It has no cell structure, thus avoiding the tumorigenicity and embolism risks of live cell therapy; it can easily penetrate the intraocular tissue barrier at the nanoscale and has good targeted distribution; it has strong stability and long shelf life, making it easy to standardize production and quality control, thus solving the technical bottlenecks of preservation, transportation and batch consistency of MSCs live cell preparations. Attached Figure Description

[0015] Figure 1 This is a diagram showing the results of fundus clinical manifestations in different treatment groups in a mouse model of uveitis.

[0016] Figure 2 This is a diagram showing the retinal pathological manifestations of different treatment groups in a mouse model of uveitis.

[0017] Figure 3 This is a graph showing the changes in the proportion of immune cells in different treatment groups during a mouse model of uveitis. Detailed Implementation

[0018] This invention provides the application of apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells in the preparation of drugs for treating uveitis. To make the objectives, technical solutions, and effects of this invention clearer and more explicit, the invention is further described in detail below.

[0019] Apoptotic vesicles (apoVs) are bilayer lipid vesicles produced during the apoptosis of MSCs. They inherit the molecular characteristics and biological functions of parent cells and play an important role in immune regulation and tissue regeneration. Stem cell exosomes are another type of extracellular vesicle secreted by MSCs and are also being explored for application in the treatment of inflammatory diseases.

[0020] Compared to live MSCs and stem cell exosomes, human umbilical cord mesenchymal stem cell apoptotic vesicles (hUCMSCs-apoVs) have superior clinical application potential: their cellular structure avoids the tumorigenic and embolic risks associated with live cell therapy, significantly improving safety; their nanoscale structure allows for easy penetration of the intraocular tissue barrier, resulting in better targeting; they exhibit extremely low immunogenicity, allowing for repeated administration without immune rejection; and they demonstrate strong stability, long shelf life, and ease of standardized production and quality control. In the treatment of uveitis, experimental results from this invention show that its anti-inflammatory and tissue-protective effects are significantly superior to stem cell exosomes, and its preparation method is simpler, lower in cost, and easier to scale up. Therefore, hUCMSCs-apoVs provide a safer, more stable, and more efficient novel strategy for the treatment of uveitis.

[0021] Based on this, embodiments of the present invention provide the application of apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells in the preparation of drugs for treating uveitis.

[0022] Specifically, addressing the technical problems of significant side effects and poor response in some patients with existing uveitis treatments, as well as the limited therapeutic effects of extracellular vesicle preparations such as stem cell exosomes, this invention provides the application of human umbilical cord-derived mesenchymal stem cell apoptotic vesicles (hUCMSCs-apoVs) in the treatment of uveitis. These vesicles exert immunomodulatory effects by inhibiting the differentiation of Th1 and Th17 pro-inflammatory cells and promoting the differentiation of Treg anti-inflammatory cells. In an experimental autoimmune uveitis mouse model (EAU), they exhibit significantly better anti-inflammatory and retinal protective effects than stem cell exosomes, with fewer side effects and higher efficacy, overcoming the limitations of traditional clinical treatments and showing excellent clinical application prospects.

[0023] In some embodiments, the method for preparing the apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells includes: (1) Stimulate the human umbilical cord mesenchymal stem cells with astronein to induce apoptosis of the human umbilical cord mesenchymal stem cells; (2) After washing the apoptotic human umbilical cord mesenchymal stem cells, centrifuge at 600-1000g for 5-15 min, then centrifuge at 1800-2200g for 5-15 min and collect the supernatant; (3) Centrifuge the supernatant at 15000~20000g for 20~40min and collect the precipitate; (4) After resuspending the precipitate in phosphate buffer, centrifuge at 20,000 to 22,000 g for 30 to 40 min, discard the supernatant, and obtain the apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells.

[0024] In a more specific embodiment, in step (1), the concentration of staurosporine (STS) is 400-600 nM, and the stimulation time is 5-7 hours. Preferably, the concentration of staurosporine is 500 nM, and the stimulation time is 6 hours. Specifically, for example, 4 mL of STS can be added to a 10 cm petri dish, and 10 mL of STS can be added to a 15 cm petri dish.

[0025] In a more specific embodiment, step (2) is preferably: after washing the apoptotic human umbilical cord mesenchymal stem cells, centrifuge at 700-900g for 8-12 min, then centrifuge at 1900-2100g for 8-12 min, and collect the supernatant; for example, the first centrifugation speed can be 700g, 750g, 800g, 850g or 900g, and the centrifugation time can be 8 min, 9 min, 10 min, 11 min or 12 min; the second centrifugation speed can be 1900g, 1950g, 2000g, 2050g or 2100g, and the centrifugation time can be 8 min, 9 min, 10 min, 11 min or 12 min; but not limited to this.

[0026] In a more specific embodiment, step (3) is preferably: centrifuging the supernatant at 16000~18000g for 25~35min and collecting the precipitate; for example, the centrifugation speed can be 16000g, 16500g, 17000g, 17500g or 18000g, and the centrifugation time can be 25min, 28min, 30min, 32min or 35min, but is not limited thereto.

[0027] In a more specific embodiment, step (4) is preferably: after resuspending the precipitate in phosphate buffer, centrifuge at 20500~21500g for 35~45min, discard the supernatant, and obtain the apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells; for example, the centrifugation speed can be 20500g, 20700g, 21000g, 21200g or 21500g, and the centrifugation time can be 35min, 38min, 40min, 42min or 45min, but is not limited thereto.

[0028] In a more specific embodiment, the method for preparing the apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells includes: (1) Add 400-600 nM astrococcus to human umbilical cord mesenchymal stem cells for stimulation for 5-7 hours to induce apoptosis of the human umbilical cord mesenchymal stem cells; (2) After washing the apoptotic human umbilical cord mesenchymal stem cells, centrifuge at 700-900g for 8-12 min, then centrifuge at 1900-2100g for 8-12 min and collect the supernatant; (3) Centrifuge the supernatant at 16000~18000g for 25~35min and collect the precipitate; (4) After resuspending the precipitate in phosphate buffer (PBS), centrifuge at 20500~21500g for 35~45min, discard the supernatant, and obtain the apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells.

[0029] In some embodiments, the drug is a drug that treats uveitis by inhibiting the differentiation of Th1 / Th17 pro-inflammatory cells and / or promoting the differentiation of Treg anti-inflammatory cells, but is not limited thereto. Specifically, Th1 refers to helper T cell type 1, Th17 refers to helper T cell type 17, and Treg refers to regulatory T cells.

[0030] This invention also provides a drug for treating uveitis, the drug comprising: apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells.

[0031] In some embodiments, the drug further includes pharmaceutically acceptable excipients.

[0032] The term "pharmaceutical acceptable" refers to a substance that does not alter the main pharmacological effects or physicochemical properties of the active component in a drug, possesses relative safety, can be applied to the body without producing adverse biological reactions, and does not interact adversely with other components in the drug.

[0033] In some more specific embodiments, the pharmaceutically acceptable excipient includes one or more of pharmaceutically acceptable excipients, pharmaceutically acceptable additives, and pharmaceutically acceptable adjuvants.

[0034] In some embodiments, when the drug is applied, the therapeutic dose of apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells is (1-8) × 10⁻⁶. 9 One vesicle / time, preferably (3-7) × 10 9 One vesicle / time, for example, 3 × 10 9 1 vesicle / time, 4×10 9 1 vesicle / time, 5×10 9 1 vesicle / time, 6×10 9 One vesicle / time or 7×10 9 One vesicle per time, but not limited to this.

[0035] In some embodiments, the drug is administered at a frequency of (6-8) days per administration, for example, every 6 days, every 7 days or every 8 days, but is not limited thereto.

[0036] In some embodiments, the route of administration of the drug includes one or more combinations of intravenous injection, intravenous infusion, intraperitoneal injection, intramuscular injection, subcutaneous injection, and perfusion, but is not limited thereto.

[0037] The present invention will be further described below through specific embodiments.

[0038] Example 1 I. Preparation process of apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells (hUCMSCs-apoVs): 1. Staurosporine (STS) stimulates cell shrinkage: Add 4 mL of STS to each 10 cm cell culture dish of human umbilical cord mesenchymal stem cells (hUCMSCs) for about 6 hours (10 mL of STS to each 15 cm cell culture dish) to induce apoptosis. 2. Centrifugation after rinsing: Rinse apoptotic cells and supernatant into centrifuge tubes, centrifuge at 4°C, first at 800g for 10 min, then at 2000g for 10 min; 3. Collect the supernatant: After centrifugation, collect the supernatant and aliquot it into 1.5mL EP tubes (840μL×2 per EP tube). 4. Centrifugation: Centrifuge at 4℃ and 17500g for 30 minutes; 5. Resuspension: Discard the supernatant and resuspend the precipitate in PBS filtered through a 0.22 μm filter membrane; 6. Centrifugation: Centrifuge at 4℃ and 21000g for 30 min, then resuspend the precipitate in PBS filtered through a 0.22μm filter membrane to obtain hUCMSCs-apoVs suspension.

[0039] II. Preparation process of human umbilical cord-derived mesenchymal stem cell exosomes (hUCMSCs-EVs): 1. Human umbilical cord mesenchymal stem cells were routinely cultured until the cell confluence reached 95%, then the culture medium was replaced with serum-free medium and cultured for another 48 hours. The cell supernatant was then collected.

[0040] At 2.4℃, differential ultracentrifugation was used to remove impurities in stages: centrifugation at 800g for 10 min, centrifugation at 2000g for 10 min, and centrifugation at 20000g for 30 min, retaining only the supernatant each time; 3. The obtained clear supernatant was centrifuged at 120,000g for 120 min. The precipitate at the bottom of the tube was the exosomes derived from umbilical cord mesenchymal stem cells. After resuspending in sterile PBS, hUCMSCs-EVs suspension was obtained and stored at low temperature.

[0041] III. Experiment of the mouse model of uveitis (EAU) This experiment included an apoptotic vesicle treatment group, a control group, and a stem cell exosome treatment group, with 5 mice in each group. The therapeutic effects of hUCMSCs-apoVs and their superiority over exosomes were compared and verified.

[0042] 1. Establishment of the EAU model: IRBP1-20 peptides (Interphotoreceptor retinoid-binding protein, IRBP, the specific sequence of peptides 1-20 is GPTHLFQPSLVLDMAKVLLD (SEQ ID NO.1)) were mixed with complete Freund's adjuvant (CFA) using an ultrasonic homogenizer to prepare a homogeneous emulsion. The resulting mixture was stored on ice and immediately used for animal immunization. Each mouse received a subcutaneous injection of 200 μL of the emulsion (containing 150 μg IRBP1-20) at the base of the tail and on the lateral thighs. To enhance the antigen-specific immune response, each mouse was intraperitoneally injected with 0.5 μg of pertussis toxin (PTX) on days 0 and 1 post-immunization.

[0043] 2. Dosage regimen: Starting on day 7 of modeling, the apoptotic vesicle treatment group received a tail vein injection of 200 μL hUCMSCs-apoVs suspension (containing 5 × 10⁻⁶ mcg / mL hUCMSCs-apoVs suspension). 9 The treatment group received an equal volume and quantity of hUCMSCs-EVs suspension via tail vein injection once a week; the control group received 200 μL of PBS solution via tail vein injection once a week.

[0044] 3. Detection indicators and methods: (1) Clinical scoring of fundus: On day 21 after modeling and drug administration, mice were anesthetized by intraperitoneal injection of 10 mg / mL sodium pentobarbital solution (150 μL / mouse). The fundus was observed and photographed under a small animal retinal imaging instrument. The EAU mouse fundus clinical scoring criteria in Table 1 were used to score the degree of ocular inflammation in the three groups of mice.

[0045] Table 1. Clinical scoring criteria for fundus examination in EAU mice

[0046] (2) Hematoxylin-eosin (HE) sections and pathological scores of eyeballs: On day 21 after modeling, mice were anesthetized and euthanized by cervical dislocation. Eyeballs were placed in fixative and fixed at room temperature for more than 24 hours. After dehydration and paraffin embedding, 3 μM thick tissue sections were prepared along the corneal-optic nerve axis and stained with hematoxylin-eosin (HE). The sections were photographed by an inverted microscope and pathological scores were performed according to the EAU mouse retinal histological scoring criteria in Table 2. The retinal tissue damage and inflammatory infiltration of the three groups of mice were compared.

[0047] Table 2. Histological scores of EAU mice retina

[0048] (3) Flow cytometry detection of the proportion of immune cells in the spleen: 21 days after administration, splenic lymphocytes of three groups of mice were isolated, 1 μL of phorbol ester (PMA) and ionomycin were added and mixed and cultured for 1 h, and then 1 μL of Brayfieldsin A (BFA) was added and mixed and cultured for another 4 h. The cells were collected into flow cytometry tubes, incubated with mouse anti-CD4 flow cytometry antibody, and after membrane rupture and fixation, anti-IL-17, IFN-γ, and FOXP3 flow cytometry antibodies were added and incubated. The proportions of Th1, Th17, and Treg cells in the three groups of mice were detected and compared.

[0049] (III) Experimental Results 1. Fundus clinical manifestations: Mice in the control group and stem cell exosome treatment group showed severe retinitis and retinal vasculitis in the fundus, characterized by extensive inflammatory cell infiltration, widespread retinal fold detachment, and photoreceptor cell damage. Mice in the apoptotic vesicle treatment group showed only mild retinitis and vasculitis in the fundus, with significantly reduced inflammation. Their clinical scores were significantly lower than those in the control group and exosome treatment group (n=5). See Figure 1 .Depend on Figure 1 It can be seen that the ocular anti-inflammatory effect of apoptotic vesicles (hUCMSCs-apoVs) is significantly better than that of exosomes.

[0050] 2. Retinal pathological manifestations: Apoptotic vesicle treatment significantly reduced vitreous cavity inflammatory cell infiltration in EAU mice, alleviated retinal folding and detachment, and reduced photoreceptor cell damage; compared with the stem cell and exosome treatment group, the apoptotic vesicle treatment group showed less retinal tissue damage, lower pathological scores, and significantly better tissue protection than exosomes (n=5), see [link to relevant documentation]. Figure 2 .Depend on Figure 2 It can be seen that apoptotic vesicles (hUCMSCs-apoVs) have a significantly better protective effect on retinal tissue than exosomes.

[0051] 3. Changes in the proportion of immune cells: Apoptotic vesicle therapy significantly reduced the proportion of Th1 and Th7 pro-inflammatory cells and increased the proportion of Treg anti-inflammatory cells in the spleen of EAU mice. Its corrective effect on immune imbalance was significantly superior to that of stem cell exosomes (n=5), confirming its stronger immunomodulatory activity. (See...) Figure 3 .

[0052] In summary, this invention provides the application of apoptotic vesicles from human umbilical cord-derived mesenchymal stem cells in the preparation of drugs for treating uveitis, and its key innovation lies in: (1) First discovery and verification: Apoptotic vesicles derived from human umbilical cord mesenchymal stem cells can be used as a novel active ingredient for the treatment of uveitis, and its therapeutic effect is significantly better than that of stem cell exosomes currently being studied. This fills the gap in the application of apoptotic vesicles in the treatment of uveitis and solves the technical problem of the limited efficacy of stem cell exosomes.

[0053] (2) Unique multi-target immunomodulatory mechanism: The apoptotic vesicle effectively corrects the immune imbalance of uveitis by synergistically inhibiting the differentiation of pro-inflammatory Th1 / Th17 cells and promoting the differentiation of anti-inflammatory Treg cells. Its immunomodulatory activity is significantly stronger than that of stem cell exosomes, which explains its excellent therapeutic effect from a mechanistic perspective.

[0054] (3) It has multiple superior characteristics: Compared with MSCs live cells, the apoptotic vesicles have no cellular structure, avoiding tumorigenicity and embolism risks, and are safer; the nanoscale can easily penetrate the intraocular tissue barrier, and the targeting is better; the immunogenicity is extremely low, and it can be repeatedly administered without immune rejection; it has strong stability and long shelf life, and is easy to standardize production and quality control, which solves the clinical translation bottleneck of MSCs live cell preparations; compared with stem cell exosomes, its comprehensive efficacy in anti-inflammatory, tissue protection and immune regulation is better, and the preparation method is simpler, the cost is lower and it is easy to scale up production, so it has higher clinical application value.

[0055] It should be understood that the application of the present invention is not limited to the examples above. Those skilled in the art can make improvements or modifications based on the above description, and all such improvements and modifications should fall within the protection scope of the appended claims.

Claims

1. Application of apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells in the preparation of drugs for treating uveitis.

2. The application according to claim 1, characterized in that, The method for preparing apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells includes: Stimulation with astrocytocin induced apoptosis in human umbilical cord mesenchymal stem cells. After washing away the apoptotic human umbilical cord mesenchymal stem cells, centrifuge at 600-1000g for 5-15 minutes, then centrifuge at 1800-2200g for 5-15 minutes, and collect the supernatant; Centrifuge the supernatant at 15000~20000g for 20~40min and collect the precipitate; The precipitate was resuspended in phosphate buffer, centrifuged at 20,000-22,000g for 30-40 minutes, and the supernatant was discarded to obtain the apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells.

3. The application according to claim 1, characterized in that, The drug is a drug that treats uveitis by inhibiting the differentiation of Th1 / Th17 pro-inflammatory cells and / or promoting the differentiation of Treg anti-inflammatory cells.

4. A drug for treating uveitis, characterized in that, The drug includes: apoptotic vesicles of human umbilical cord-derived mesenchymal stem cells.

5. The drug according to claim 4, characterized in that, The drug also includes pharmaceutically acceptable excipients.

6. The drug according to claim 5, characterized in that, The pharmaceutically acceptable excipients include one or more of the following: pharmaceutically acceptable excipients, pharmaceutically acceptable additives, and pharmaceutically acceptable adjuvants.

7. The drug according to claim 4, characterized in that, When the drug is applied, the therapeutic dose of apoptotic vesicles from human umbilical cord-derived mesenchymal stem cells is (1-8) × 10⁻⁶. 9 One vesicle per time.

8. The drug according to claim 4, characterized in that, When the drug is used, the frequency of administration is (6-8) days / time.

Images