Mev and use thereof in preparation of drug for preventing and / or treating gastric ulcer

Abstract

Provided are a milk extracellular vesicle (mEV) and a use thereof in preparation of a drug for preventing and / or treating gastric ulcer. The mEV comprises a protein component beneficial to gastric mucosa reconstruction and immunoregulation and an miRNA closely related to immunoregulation and anti-infection functions.

Description

mEV and its application in preparing medicine for preventing and / or treating gastric ulcer Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to milk extracellular vesicles (milkEV, mEV) and their use in preparing drugs for preventing and / or treating gastric ulcers. Background Art

[0002] Clinical treatments for gastric ulcers primarily fall into three categories: suppressing gastric acid, suppressing Helicobacter pylori, and protecting the gastric mucosa. Medications that suppress gastric acid include 1) antacids (aluminum hydroxide, hydrotalcite, calcium carbonate, and magnesium oxide, among others); 2) H2 receptor antagonists (famotidine, roxatidine, lafutidine, and nizatidine, among others); and 3) proton pump inhibitors (lansoprazole, rabeprazole, omeprazole, pantoprazole, and esomeprazole). Medications that suppress Helicobacter pylori include clarithromycin, metronidazole, and amoxicillin. Gastric mucosal protectants include potassium citrate, sucralfate, colloidal bismuth tartrate capsules, and teprenone.

[0003] Due to the frequent occurrence of gastric ulcers caused by modern high pressure, fast pace of life and unhealthy eating habits, the cumulative side effects and safety risks brought about by long-term use of small molecule chemical drugs cannot be ignored.

[0004] Researchers are beginning to explore biotechnology approaches, such as cell therapy, to prevent or treat gastric ulcers. Cells and cell derivatives offer safety and significant efficacy. For example, culturing endothelial progenitor cells and extracting extracellular vesicles from the culture medium can be used to treat bleeding disorders, including gastric ulcers, by injection.

[0005] Currently, problems with existing cell therapy technologies include:

[0006] ① The source of endothelial progenitor cells is limited by ethics and cost, making it difficult to scale up and culture;

[0007] ② Due to the limitation of the culture scale of EPCs, the production scale of EPC exosomes is also limited and the cost is high;

[0008] ③ The existing technology requires intravenous administration, which has poor clinical compliance for general gastric ulcer patients;

[0009] ④ Progenitor cells are pluripotent stem cells with a certain long-term proliferation capacity. Their exosomes may carry bioactive components that promote cell differentiation and proliferation, which may pose a carcinogenic risk after injection into the body;

[0010] Therefore, it is of great practical significance to develop a safe, effective and clinically convenient biotechnology product for the prevention and treatment of gastric ulcers.

[0011] Summary of the Invention

[0012] In view of this, the present invention uses skimmed fresh milk and cheese whey as raw materials to extract milk extracellular vesicles (milk EVs, mEVs) for the prevention and treatment of gastric ulcers.

[0013] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0014] In a first aspect, the present invention provides the use of mEVs in the preparation of a medicament for preventing and / or treating gastric ulcers.

[0015] In some specific embodiments of the present invention, the method for preparing mEV comprises the following steps:

[0016] Step 1: Take the raw material after pretreatment, filter it through a 400-mesh mesh, and then filter it through a 0.8 / 0.45 μm capsule filter to collect the filtrate;

[0017] Step 2: The filtrate was concentrated by 300 kD tangential flow membrane, and the filtrate was washed and the solution was changed to 1× PBS;

[0018] Step 3: The concentrated sample was purified using Capto Core 700 (Cytiva) to remove free impurities;

[0019] Step 4: Concentrate to the target volume or concentration using a 750 kD tangential flow hollow fiber column.

[0020] In some specific embodiments of the present invention, the raw material includes skim fresh milk or cheese whey;

[0021] When the raw material is skim milk, the pretreatment includes preparing a 3M ammonium sulfate solution, slowly adding an equal volume of milk to the ammonium sulfate solution while stirring, to a final concentration of 1.5M ammonium sulfate; and leaving it at room temperature for 3 hours to allow precipitation to form.

[0022] When the raw material is cheese whey, the pretreatment includes centrifuging the cheese whey at 5500 rpm for 10 minutes and removing the floating fat layer.

[0023] In some specific embodiments of the present invention, the mEV comprises one or more of glycoproteins, EV marker proteins, apolipoproteins, immune-related proteins or cytokines.

[0024] In some embodiments of the invention, the mEV comprises:

[0025] One or a combination of two or more of GLYCAM1, BTN1A1, LGB, MFGE8, CD36, XDH, ABCG2, FABP3, SLC34A2, MUC15, FASN, IDH1, CD9, ENPP3, MUC1, APOA1, ANXA5, RAB18, BPTI, PIGR, PPIA, PLIN2, GDI2, NT5E, ALB, EZR, HSPA8, RAC1, CYB5R3, RAB7A, RAB1B, TLR2, CDC42, CD63, CD46, ARHGDIA, EEF1A1, APOE, RHOA, GPD1, EEF2, or PTPA.

[0026] In some embodiments of the invention, the mEV comprises:

[0027] The proteins contained in the mEVs and their abundance ratios are shown in FIG5 .

[0028] In some embodiments of the invention, the mEV comprises:

[0029] The abundance ratios of proteins contained in the mEVs are shown in FIG6 .

[0030] In some embodiments of the present invention, the mEV further comprises:

[0031] (I), APOA4, CSN3, LALBA, PEBP1, SOD1, LDHB, ANXA2, or MYO1C, or a combination of two or more thereof; and / or

[0032] (II), TF, LTF, FOLR1, CD14, CIB1, VPS25 or GNAS, or a combination of two or more thereof.

[0033] In some embodiments of the present invention, the mEV further comprises:

[0034] The abundance ratios of proteins contained in the mEVs are shown in Figure 7 ;

[0035] or

[0036] The abundance ratios of proteins contained in the mEVs are shown in FIG8 .

[0037] In some embodiments of the present invention, the mEV further comprises:

[0038] The abundance ratios of proteins contained in the mEVs are shown in FIG9 .

[0039] or

[0040] The abundance ratios of proteins contained in the mEVs are shown in FIG10 .

[0041] In a second aspect, the present invention further provides mEV, wherein the preparation method of mEV comprises the following steps:

[0042] Step 1: Take the raw material after pretreatment, filter it through a 400-mesh mesh, and then filter it through a 0.8 / 0.45 μm capsule filter to collect the filtrate;

[0043] Step 2: The filtrate was concentrated by 300 kD tangential flow membrane, and the filtrate was washed and the solution was changed to 1× PBS;

[0044] Step 3: The concentrated sample was purified using Capto Core 700 (Cytiva) to remove free impurities;

[0045] Step 4: Concentrate to the target volume or concentration using a 750 kD tangential flow hollow fiber column.

[0046] In some specific embodiments of the present invention, the raw material includes skim fresh milk or cheese whey;

[0047] When the raw material is skim milk, the pretreatment includes preparing a 3M ammonium sulfate solution, slowly adding an equal volume of milk to the ammonium sulfate solution while stirring, to a final concentration of 1.5M ammonium sulfate; and leaving it at room temperature for 3 hours to allow precipitation to form.

[0048] When the raw material is cheese whey, the pretreatment includes centrifuging the cheese whey at 5500 rpm for 10 minutes and removing the floating fat layer.

[0049] In some embodiments of the present invention, the mEV comprises one or more of glycoproteins, EV marker proteins, apolipoproteins, immune-related proteins or cytokines.

[0050] In some embodiments of the invention, the mEV comprises:

[0051] One or a combination of two or more of GLYCAM1, BTN1A1, LGB, MFGE8, CD36, XDH, ABCG2, FABP3, SLC34A2, MUC15, FASN, IDH1, CD9, ENPP3, MUC1, APOA1, ANXA5, RAB18, BPTI, PIGR, PPIA, PLIN2, GDI2, NT5E, ALB, EZR, HSPA8, RAC1, CYB5R3, RAB7A, RAB1B, TLR2, CDC42, CD63, CD46, ARHGDIA, EEF1A1, APOE, RHOA, GPD1, EEF2, or PTPA.

[0052] In some embodiments of the invention, the mEV comprises:

[0053] The abundance ratios of proteins contained in the mEVs are shown in FIG5 .

[0054] In some embodiments of the invention, the mEV comprises:

[0055] The abundance ratios of proteins contained in the mEVs are shown in FIG6 .

[0056] In some embodiments of the present invention, the mEV further comprises:

[0057] (I), APOA4, CSN3, LALBA, PEBP1, SOD1, LDHB, ANXA2, or MYO1C, or a combination of two or more thereof; and / or

[0058] (II), TF, LTF, FOLR1, CD14, CIB1, VPS25 or GNAS, or a combination of two or more thereof.

[0059] In some embodiments of the present invention, the mEV further comprises:

[0060] The abundance ratios of proteins contained in the mEVs are shown in FIG7 .

[0061] or

[0062] The abundance ratios of proteins contained in the mEVs are shown in FIG8 .

[0063] In some embodiments of the present invention, the mEV further comprises:

[0064] The abundance ratios of proteins contained in the mEVs are shown in FIG9 .

[0065] or

[0066] The abundance ratios of proteins contained in the mEVs are shown in FIG10 .

[0067] In a third aspect, the present invention also provides a drug comprising the mEV and pharmaceutically acceptable excipients.

[0068] In a fourth aspect, the present invention also provides a drug combination comprising the mEV and any other active ingredients.

[0069] In a fifth aspect, the present invention also provides a method for using mEV, which comprises administering the mEV to a patient.

[0070] In a sixth aspect, the present invention also provides a method for using the drug, which involves administering the drug to a patient.

[0071] In a seventh aspect, the present invention also provides a method for using the drug combination, wherein the drug combination is administered to a patient.

[0072] In an eighth aspect, the present invention further provides a method for treating gastric ulcer, comprising administering to the patient any one or more of the following:

[0073] (I), the mEV;

[0074] (II), the drug; or

[0075] (III), the drug combination.

[0076] In their research, the applicant discovered that milk extracellular vesicles (mEVs) are rich in mucins and other protein components that are beneficial for gastric mucosal reconstruction and immune regulation. They also contain small nucleic acids (miRNAs) that are closely related to immune regulation and anti-infection functions. Therefore, the applicant has developed a new technology using purified mEVs for the prevention and treatment of gastric ulcers.

[0077] The beneficial effects of the present invention include but are not limited to:

[0078] ① mEV can be extracted from a variety of dairy products such as fresh milk, raw cow's milk, cheese whey, etc., with abundant content and unlimited preparation scale;

[0079] ② The mEV production process does not involve in vitro cell culture, and the production cost is low;

[0080] ③mEV can be taken orally;

[0081] ④ mEV has been verified by the long-term consumption of milk by the general public and poses no safety risks.

[0082] ⑤ BRIEF DESCRIPTION OF THE DRAWINGS

[0083] Figure 1 shows the mEV particle concentration and particle size (NanoFCM) in Example 1;

[0084] Figure 2 shows the morphology of mEVs in Example 1 (transmission electron microscopy);

[0085] Figure 3 shows the mEV particle concentration and particle size (NanoFCM) in Example 2;

[0086] Figure 4 shows the morphology of mEVs in Example 2 (transmission electron microscopy);

[0087] Figures 5 to 11 show the abundance ratios of proteins contained in mEVs. DETAILED DESCRIPTION

[0088] The present invention discloses mEV and its use in the preparation of a drug for preventing and / or treating gastric ulcers. Those skilled in the art can refer to the contents of this article and appropriately improve the process parameters. It should be noted in particular that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0089] The raw materials and reagents used in the mEV provided by the present invention and its application in the preparation of a medicament for preventing and / or treating gastric ulcers are all commercially available. The present invention is further described below with reference to the following examples:

[0090] Explanation of terms:

[0091] Gastric ulcers: Gastric ulcers are chronic sores that develop between the pyloric and cardia sections of the stomach. Peptic ulcers are a common and frequently occurring disease worldwide. An estimated 10% of the global population will develop this disease at some point in their lifetime. Gastric ulcers can occur at any age, but are more common in middle-aged and elderly people, and are more common in men. The pathogenesis of gastric ulcers is an imbalance between the erosive effects of gastric acid and pepsin and their defenses. Gastric acid digests the gastric mucosa, and combined with HP (Helicobacter pylori) infection, the gastric mucosa's protective barrier is weakened, leading to damage and erosion, resulting in ulcers.

[0092] Extracellular vesicles / nanovesicles: Extracellular vesicles are natural nanoscale vesicles (30-150nm) produced and released by cells. They contain genetic information, such as nucleic acids and proteins, from the parent cell. After being bound or internalized by recipient cells, they release active molecules to activate downstream reactions, thus playing a role in intercellular communication within the body. As a natural intercellular communication tool, extracellular vesicles can be engineered to load active molecules within the extracellular vesicles and then be delivered to the recipient cells to exert their activity. In particular, they can be used to directly deliver transcription factor proteins to stem cells during induced differentiation, avoiding the traditional method of overexpressing transcription factors through viral infection and other genetic modification, which can cause changes in the cell genome and the subsequent risk of tumorigenesis.

[0093] Milk extracellular vesicles (mEVs): Milk extracellular vesicles (mEVs) are biological nanoparticles composed of a phospholipid bilayer membrane that can be extracted in large quantities from milk. Their natural function involves delivering bioactive molecules (proteins, small nucleic acids, etc.) from the mother (cow) to the offspring (calf) through the gastrointestinal tract. Studies have demonstrated that mEVs can withstand damage from digestive enzymes, low pH, and other environmental factors within the digestive tract. Due to the widespread consumption of milk, the long-term exposure of most people to mEVs (through milk consumption) ensures that oral administration of mEVs in humans is safe and tolerable.

[0094] The present invention will be further described below in conjunction with the embodiments:

[0095] Example 1 Preparation of mEVs from skim milk by ammonium sulfate precipitation and validation in a rat gastric ulcer model

[0096] mEV therapeutic effects

[0097] Objective: To prepare mEVs from skim milk using ammonium sulfate precipitation and to verify the therapeutic effect of mEVs in a rat gastric ulcer model.

[0098] method:

[0099] mEV preparation:

[0100] A 3 M ammonium sulfate solution was prepared by slowly adding an equal volume of milk to the ammonium sulfate solution with stirring, to a final ammonium sulfate concentration of 1.5 M. The solution was allowed to stand at room temperature for 3 h. After precipitation, the solution was filtered through a 400-mesh cloth and then through a 0.8 / 0.45 μm capsule filter. The supernatant was concentrated using a 300 kD tangential flow membrane filter and washed with 1× PBS. The concentrated sample was purified using Capto Core 700 (Cytiva) to remove free proteins. Finally, the sample was concentrated to the target volume or concentration using a 750 kD tangential flow hollow fiber column. After preparation, the mEV particle concentration and size were determined using NanoFCM (Xiamen Fuliu). The morphology of A1 mEVs was determined using transmission electron microscopy. Protein distribution and quantification in mEVs were determined using proteomics analysis (Shanghai Huaying Biotechnology).

[0101] Animal Models:

[0102] SD rats were randomly divided into 6 groups according to their body weight and given drugs by gavage. The blank control group (group A, healthy rats), model control group (group B, gastric ulcer modeling, gavage with normal saline), positive drug control group (group C, gastric ulcer modeling, gavage with positive control drug rabeprazole sodium 20 mg / kg), low-dose milk exosome group (group D, gavage with 10 11 p / kg mEV), medium-dose milk exosome group (E group, gavage administered 10 12 p / kg mEV), high-dose milk exosome group (F group, gavage administered 10 13 p / kg mEV).

[0103] Dosage frequency: qd×5 (administered continuously for 5 days after model establishment).

[0104] Modeling: Before modeling, rats were fasted for 16 hours but not water. They were fixed on a wire mesh board in supine position (except for the blank control group) and immersed in a constant temperature box (20±1℃). The water level was maintained at the xiphoid process of the sternum.

[0105] Evaluation: After administration, animals were euthanized by CO2, and laparotomy was performed. The pylorus and cardia were ligated, and 10 mL of 2% formalin was injected into the stomach for fixation for 1 hour. The stomach wall was incised along the greater curvature and flattened. The number of ulcers was measured under a dissecting microscope, and the ulcer index was calculated. This was converted to the ulcer inhibition rate (%). Ulcer inhibition rate (%) = (average UI of the model group - average UI of the drug group) / average UI of the model group × 100%.

[0106] result:

[0107] 1. Characterization

[0108] mEV particle concentration and particle size (NanoFCM): as shown in Figure 1.

[0109] mEV morphology (transmission electron microscopy): as shown in Figure 2.

[0110] 2. Proteomic analysis

[0111] It contains a large amount of glycoproteins, with the highest content of Glycosylation-dependent cell adhesion molecule 1, which is involved in the attachment and signal transduction between tissue cells. The content of EV marker protein CD9 is also relatively rich. In addition, it is rich in apolipoproteins, immune-related proteins, etc.

[0112] Table 1 Accession numbers, gene names, and protein function descriptions of the top 50 most abundant proteins in A1 mEVs

[0113] The abundance ratios of proteins contained in the mEVs are shown in FIG6 .

[0114] 3. Gastric ulcer inhibition effect

[0115] Compared with the model group, different doses of mEV had a certain inhibitory effect on gastric ulcer in a dose-dependent manner.

[0116] Table 2 Gastric mucosal injury index in different drug groups

[0117] *: There is a significant difference compared with the model control group (P<0.05)

[0118] Table 3 Ulcer inhibition rate (%) of different drug groups calculated according to Table 2

[0119] (UI of drug-treated group - UI of model control group) / UI of model control group

[0120] Example 2 Preparation of mEVs from cheese whey and verification of the effect of mEVs in preventing gastric ulcers in a rat gastric ulcer model

[0121] Objective: To prepare mEVs from cheese whey and verify the preventive effect of mEVs on gastric ulcer in a rat gastric ulcer model.

[0122] method:

[0123] mEV preparation

[0124] Cheese whey was centrifuged at 5500 rpm for 10 minutes, the floating fat layer was removed, and the sample was filtered through a 400-mesh mesh and then filtered through a 0.8 / 0.45 μm capsule filter. The supernatant was concentrated using a 300 kD tangential flow membrane and washed with 1× PBS. The concentrated sample was purified using Capto Core 700 (Cytiva) to remove free proteins. Finally, the sample was concentrated to the target volume or concentration using a 750 kD tangential flow hollow fiber column. After preparation, the mEV particle concentration and size were determined using NanoFCM (Xiamen Fuliu); the mEV morphology was determined using transmission electron microscopy; and the protein distribution and quantification in the mEVs were determined using proteomics analysis (Shanghai Huaying Biotechnology).

[0125] Animal models

[0126] Preventive administration: administration before and during modeling. Other procedures are the same as in Example 1.

[0127] result:

[0128] 1. Characterization:

[0129] mEV particle concentration and particle size (NanoFCM): as shown in Figure 3.

[0130] mEV morphology (transmission electron microscopy): as shown in Figure 4.

[0131] 2. Proteomic analysis

[0132] It contains a large amount of glycoproteins, the highest content of which is lactadhesive (a glycoprotein with anti-diarrhea effect). The EV marker proteins CD9, CD81, and CD63 are also relatively rich, as well as some immune-related proteins, apolipoproteins and cytokines.

[0133] Table 4 Accession numbers, gene names, and protein function descriptions of the top 50 most abundant proteins in mEVs

[0134] The abundance ratios of proteins contained in the mEVs are shown in FIG11 .

[0135] 3. Gastric ulcer inhibition effect

[0136] Different doses of mEV have different degrees of inhibitory effect on gastric ulcer. 13 The 20mg / kg particles / kg administration group and the positive drug rabeprazole sodium 20mg / kg both produced significant ulcer inhibition effects.

[0137] Table 5 Gastric mucosal injury index in different drug groups

[0138] *: There is a significant difference compared with the model control group (P<0.05)

[0139] Table 6 Ulcer inhibition rate (%) of different drug administration groups calculated according to Table 5

[0140] (UI of drug-treated group - UI of model control group) / UI of model control group

[0141] The above is a detailed introduction to the mEV provided by the present invention and its use in the preparation of a drug for preventing and / or treating gastric ulcers. This article uses specific examples to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. It should be pointed out that for those skilled in the art, without departing from the principles of the present invention, the present invention can also be improved and modified, and these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. Use of mEV in the preparation of drugs for preventing and / or treating gastric ulcers.

2. The use according to claim 1, characterized in that The mEV preparation method comprises the following steps: Step 1: Take the raw material after pretreatment, filter it through a 400-mesh cloth, and then filter it through a 0.8 / 0.45 μm capsule filter to collect the filtrate; Step 2: The filtrate was concentrated by 300 kD tangential flow membrane bag, and the filtrate was washed and the solution was changed to 1× PBS; Step 3: The concentrated sample was purified using Capto Core700 (Cytiva) to remove free impurities; Step 4: Concentrate to the target volume or concentration using a 750 kD tangential flow hollow fiber column.

3. The use according to claim 2, characterized in that The raw materials include skimmed fresh milk or cheese whey; When the raw material is skim fresh milk, the pretreatment includes preparing a 3M ammonium sulfate solution, slowly adding an equal volume of milk to the ammonium sulfate solution while stirring, and the final concentration of ammonium sulfate is 1.5M; leaving it at room temperature for 3 hours to allow precipitation to form; When the raw material is cheese whey, the pretreatment includes centrifuging the cheese whey at 5500 rpm for 10 minutes and removing the floating fat layer.

4. The use according to any one of claims 1 to 3, characterized in that The mEV contains one or more of glycoproteins, EV marker proteins, apolipoproteins, immune-related proteins or cytokines.

5. The use according to claim 4, characterized in that The mEV comprises: One or a combination of two or more of GLYCAM1, BTN1A1, LGB, MFGE8, CD36, XDH, ABCG2, FABP3, SLC34A2, MUC15, FASN, IDH1, CD9, ENPP3, MUC1, APOA1, ANXA5, RAB18, BPTI, PIGR, PPIA, PLIN2, GDI2, NT5E, ALB, EZR, HSPA8, RAC1, CYB5R3, RAB7A, RAB1B, TLR2, CDC42, CD63, CD46, ARHGDIA, EEF1A1, APOE, RHOA, GPD1, EEF2 or PTPA.

6. The use according to claim 5, characterized in that The mEV comprises:

7. The use according to claim 5, characterized in that The mEV comprises:

8. The use according to any one of claims 5 to 7, characterized in that The mEV further comprises: (I), APOA4, CSN3, LALBA, PEBP1, SOD1, LDHB, ANXA2 or MYO1C, or a combination of two or more thereof; and / or (II), TF, LTF, FOLR1, CD14, CIB1, VPS25 or GNAS, or a combination of two or more thereof.

9. The use according to claim 8, characterized in that The mEV further comprises: or 10. The use according to claim 8, characterized in that The mEV further comprises: or 11. mEV, characterized in that The mEV preparation method comprises the following steps: Step 1: Take the raw material after pretreatment, filter it through a 400-mesh cloth, and then filter it through a 0.8 / 0.45 μm capsule filter to collect the filtrate; Step 2: The filtrate was concentrated by 300 kD tangential flow membrane bag, and the filtrate was washed and the solution was changed to 1× PBS; Step 3: The concentrated sample was purified using Capto Core700 (Cytiva) to remove free impurities; Step 4: Concentrate to the target volume or concentration using a 750 kD tangential flow hollow fiber column.

12. The mEV of claim 11, wherein The raw materials include skimmed fresh milk or cheese whey; When the raw material is skim fresh milk, the pretreatment includes preparing a 3M ammonium sulfate solution, slowly adding an equal volume of milk to the ammonium sulfate solution while stirring, and the final concentration of ammonium sulfate is 1.5M; leaving it at room temperature for 3 hours to allow precipitation to form; When the raw material is cheese whey, the pretreatment includes centrifuging the cheese whey at 5500 rpm for 10 minutes and removing the floating fat layer.

13. The mEV according to claim 11 or 12, characterized in that The mEV contains one or more of glycoproteins, EV marker proteins, apolipoproteins, immune-related proteins or cytokines.

14. The mEV of claim 13, wherein The mEV comprises: One or a combination of two or more of GLYCAM1, BTN1A1, LGB, MFGE8, CD36, XDH, ABCG2, FABP3, SLC34A2, MUC15, FASN, IDH1, CD9, ENPP3, MUC1, APOA1, ANXA5, RAB18, BPTI, PIGR, PPIA, PLIN2, GDI2, NT5E, ALB, EZR, HSPA8, RAC1, CYB5R3, RAB7A, RAB1B, TLR2, CDC42, CD63, CD46, ARHGDIA, EEF1A1, APOE, RHOA, GPD1, EEF2 or PTPA.

15. The mEV according to claim 13 or 14, characterized in that The mEV comprises:

16. The mEV according to claim 13 or 14, characterized in that The mEV comprises:

17. The mEV according to any one of claims 13 to 16, characterized in that The mEV further comprises: (I), APOA4, CSN3, LALBA, PEBP1, SOD1, LDHB, ANXA2 or MYO1C, or a combination of two or more thereof; and / or (II), TF, LTF, FOLR1, CD14, CIB1, VPS25 or GNAS, or a combination of two or more thereof.

18. The mEV of claim 17, wherein The mEV further comprises: or 19. The mEV of claim 17, wherein The mEV further comprises: or 20. A drug, characterized in that Comprising the mEV as described in any one of claims 11 to 19 and a pharmaceutically acceptable excipient.

21. A pharmaceutical combination, characterized in that Comprising the mEV according to any one of claims 11 to 19 and any other effective ingredients.

22. A method for using mEV, characterized in that: The mEV according to any one of claims 11 to 19 is administered to a patient.

23. A method of using a drug, characterized in that: Administering the medicament of claim 20 to a patient.

24. A method of using a drug combination, characterized in that: The pharmaceutical combination of claim 21 is administered to a patient.

25. A method for treating gastric ulcer, characterized in that: The patient is administered any one or more of the following: (I) The mEV according to any one of claims 11 to 19; (II), the medicament according to claim 20; or (III) The pharmaceutical combination according to claim 21.

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