Application of capillary artemisia-derived extracellular vesicle-like nanoparticles in preparation of medicines for preventing or treating liver diseases
By extracting and studying extracellular vesicle-like nanoparticles derived from Yinchen, the problem of lack of special drugs for alcoholic liver disease in the prior art was solved, and the application of the nanoparticles in the treatment of liver disease was realized, showing its in-depth analysis of the nanoparticles in the treatment of liver disease. Significant effects in improving liver damage and reducing inflammation.
Patent Information
- Application Number
- CN202411714518.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-27
- Publication Date
- 2025-02-18
AI Technical Summary
The prior art lacks specific drugs for alcoholic liver disease, and the feasibility and biological activity of extracellular vesicle-like nanoparticles extracted from Yinchen have not been fully studied.
By creatively extracting and purifying extracellular vesicle-like nanoparticles derived from Yinchen and studying their physiological effects, it was found that they can be internalized and absorbed by normal human liver cells, have good liver protection effects, can improve liver damage and reduce inflammation Factor expression level and protects mitochondria in liver tissue.
Extracellular vesicle-like nanoparticles derived from Yinchen have shown good anti-alcohol liver protection effects in animal experiments, which can effectively improve liver damage, reduce inflammation, and protect liver tissue. It is suitable for the preparation of anti-alcohol liver protection drugs.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of biomedicine and relates to the application of extracellular vesicle-like nanoparticles derived from Artemisia capillaris in the preparation of medicines for preventing or treating liver diseases. Background Art
[0002] Alcoholic liver disease is a common chronic disease worldwide. It is a liver damage disease caused by long-term excessive drinking, including alcoholic fatty liver, alcoholic hepatitis, alcoholic cirrhosis and liver cancer. Excessive drinking leads to alcoholic fatty liver. If not treated in time, liver cells will undergo necrosis and subsequently develop into alcoholic hepatitis. Alcoholic hepatitis is a necrotizing inflammatory process. Fibrosis occurs on the basis of liver cell necrosis, forming abnormal liver cell nodules surrounded by fibers, which develop into alcoholic cirrhosis. Among patients with alcoholic hepatitis, 40% develop cirrhosis and eventually even liver cancer.
[0003] As the daily alcohol intake increases, the proportion of deaths caused by alcoholic liver disease also increases. Swedish researchers compared the mortality rate of biopsy-confirmed alcoholic liver disease groups across the country and proved that the risk of death in patients with biopsy-confirmed alcoholic liver disease increased nearly five times compared to the general population. The risk of death in patients with alcoholic liver disease without cirrhosis is also increased, which once again proves the need to increase vigilance in the management of these individuals. In this situation, the study of targeted treatment options is imminent. At present, the main clinical treatment measures for alcoholic liver disease are alcohol withdrawal, nutritional support, and the administration of anti-inflammatory and liver protection drugs. There is a lack of specific drugs for alcoholic liver disease.
[0004] Plant-derived vesicle-like nanoparticles (PDVLNs) are plant components with a phospholipid bilayer structure, containing a variety of functional molecules such as RNA, DNA, lipids, and proteins. Chinese herbal medicine-derived extracellular vesicle-like particles (CHM-EVLP) have a lipid bilayer structure. After being secreted, they can recognize and bind to the surface of receptor cells, and interact with target cells to trigger functional responses of target cells. In addition, CHM-EVLP contains a variety of small molecule compounds, nucleic acids, and protein components, which give it pharmacological activities similar to those of the source plants. Therefore, CHM-EVLP can be used as a new medium for studying the pharmacological effects of traditional Chinese medicine, and explain the biological functions of traditional Chinese medicine from the perspective of molecular biology.
[0005] As a traditional Chinese medicine, Artemisiae Scopariae has low cost and good safety. It has a long history of application in sobering up and protecting the liver. At present, no team has studied Artemisiae Scopariae Herba-derived EV-like particles (AH-EVLP) and successfully extracted Artemisiae Scopariae-derived vesicle-like nanoparticles and studied their efficacy. Whether it is feasible to extract extracellular vesicles from Artemisiae Scopariae and whether the extracted Artemisiae Scopariae-derived extracellular vesicle-like nanoparticles have biological activity is still unknown. Therefore, it is meaningful to provide a method for extracting high-purity and high-quality extracellular vesicles from the traditional Chinese medicine Artemisiae Scopariae, and to deeply study the functions of the extracted extracellular vesicle-like nanoparticles to explore its application potential in the treatment of diseases. Summary of the invention
[0006] In view of the deficiencies of the prior art, the object of the present invention is to provide an application of extracellular vesicle-like nanoparticles derived from Artemisia capillaris in the preparation of a drug for preventing or treating liver diseases.
[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:
[0008] In a first aspect, the present invention provides the use of extracellular vesicle-like nanoparticles derived from Artemisia capillaris in the preparation of a drug for preventing or treating liver diseases.
[0009] The present invention creatively extracts and purifies extracellular vesicles derived from the traditional Chinese medicine Artemisia capillaris, and studies its physiological effects. It is found that the extracellular vesicles can be fully internalized and absorbed by normal human liver cells. Animal experiments have proved that the extracellular vesicles have good liver protection effects, can effectively improve liver damage, reduce the expression level of inflammatory factors, and have a protective effect on mitochondria in liver tissue. The extracellular vesicles can be used to prepare alcohol-detoxifying and liver-protecting drugs.
[0010] Preferably, the liver disease comprises alcohol-induced liver disease.
[0011] Preferably, the alcohol-induced liver disease includes alcoholic hepatitis, alcoholic cirrhosis or alcoholic fatty liver.
[0012] In a second aspect, the present invention provides the use of extracellular vesicle-like nanoparticles derived from Artemisia capillaris in the preparation of a drug having the effects of sobering up and protecting the liver.
[0013] Preferably, the Artemisia capillaris-derived extracellular vesicle-like nanoparticles are prepared by a preparation method comprising the following steps:
[0014] (1) filtering the juice of the Artemisia capillaris raw material, and concentrating the filtrate to obtain a concentrated solution;
[0015] (2) centrifuging the concentrate and collecting the supernatant;
[0016] (3) Centrifuge the supernatant, collect the precipitate, and resuspend the precipitate to obtain the product.
[0017] Preferably, the centrifugation temperature in step (2) is 0-4°C, the rotation speed is 9000-11000g, and the time is 25-35min.
[0018] The temperature can be selected from 0℃, 0.5℃, 1℃, 1.5℃, 2℃, 2.5℃, 3℃, 3.5℃, 4℃, etc., the rotation speed can be selected from 9000g, 9200g, 9500g, 9800g, 10000g, 10200g, 10500g, 10800g, 11000g, etc., and the time can be selected from 25min, 26min, 27min, 28min, 29min, 30min, 31min, 32min, 33min, 34min, 35min, etc. Other specific point values within the above numerical range can be selected, so they will not be elaborated here.
[0019] Preferably, the centrifugation temperature in step (3) is 0-4°C, the rotation speed is 120000-150000g, and the time is 60-80min.
[0020] The temperature can be selected from 0°C, 0.5°C, 1°C, 1.5°C, 2°C, 2.5°C, 3°C, 3.5°C, 4°C, etc., the rotation speed can be selected from 120000g, 122000g, 125000g, 128000g, 130000g, 132000g, 135000g, 138000g, 140000g, 142000g, 145000g, 148000g, 150000g, etc., and the time can be selected from 60min, 62min, 64min, 66min, 68min, 70min, 72min, 74min, 76min, 78min, 80min, etc. Other specific point values within the above numerical range can be selected, so they will not be elaborated here.
[0021] Preferably, the volume of the concentrate is 5-10% of the volume of the filtrate, for example 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, etc. Other specific points within the above numerical range can be selected and will not be described here one by one.
[0022] Preferably, the resuspension is followed by filtering using a 0.22 μm filter.
[0023] Preferably, the dosage form of the drug includes any one of tablets, capsules, pills, powders, solutions, aerosols or sprays.
[0024] Preferably, the drug further comprises a pharmaceutically acceptable excipient.
[0025] Preferably, the pharmaceutically acceptable excipients include any one of a diluent, a dispersant, a binder, a filler, a thickener, a lubricant, a pH adjuster, a taste masking agent, a colorant, an antioxidant or an antibacterial agent, or a combination of at least two thereof.
[0026] Compared with the prior art, the present invention has the following beneficial effects:
[0027] The present invention relates to the application of extracellular vesicle-like nanoparticles derived from Artemisia capillaris in the preparation of drugs for protecting the liver. The present invention creatively extracts and purifies extracellular vesicles derived from the traditional Chinese medicine Artemisia capillaris, studies its physiological effects, and finds that the extracellular vesicles can be fully internalized and absorbed by normal human liver cells. Animal experiments have proved that the extracellular vesicles have good effects of sobering up and protecting the liver, effectively improve liver damage, reduce the expression level of inflammatory factors, and have a protective effect on mitochondria in liver tissue. The extracellular vesicles can be used to prepare drugs for sobering up and protecting the liver. BRIEF DESCRIPTION OF THE DRAWINGS
[0028] Figure 1 This is the extraction flow chart of AH-EVLP.
[0029] Figure 2A This is the observation result of AH-EVLPS under transmission electron microscope.
[0030] Figure 2B is the particle size distribution result of AH-EVLPS.
[0031] Figure 3A is the particle size distribution of AH-EVLP in PBS.
[0032] Figure 3B is the particle size distribution of AH-EVLP in alcohol.
[0033] Figure 3C is the particle size distribution of AH-EVLP in artificial gastric fluid.
[0034] Figure 3D is the particle size distribution of AH-EVLP in artificial small intestinal fluid.
[0035] Figure 3E It is the particle size distribution of AH-EVLP in the mixture of artificial gastric juice and small intestinal juice.
[0036] Figure 3F This is the particle size result diagram of AH-EVLP in different environments.
[0037] Figure 4 This is a diagram of AH-EVLP being absorbed by normal human liver cells.
[0038] Figure 5A This is a graph showing the effect of alcohol concentration on cell activity.
[0039] Figure 5B This is a graph showing the effect of AH-EVLP concentration on cell activity.
[0040] Figure 5C This is the result of AH-EVLP reversing the decrease in cell activity caused by alcohol.
[0041] Fig. 6A This is the drunkenness score result chart.
[0042] Figure 6B This is the result chart of the soberness score.
[0043] Figure 7 This is a graph showing the results of ALT levels in serum.
[0044] Figure 8 This is a graph showing the results of ALP content in serum.
[0045] Fig. 9 This is the result graph of AST content in serum.
[0046] Fig.10 This is the GO enrichment factor map in the liver of each group of mice.
[0047] Fig.11 This is the KEGG enrichment factor map in the liver of each group of mice.
[0048] Fig.12 is a heat map of differentially expressed genes in the livers of each group of mice.
[0049] Fig.13 It is the volcano plot of differentially expressed genes in each group of mice.
[0050] Fig.14A This is the result diagram of TNF signaling pathway enrichment analysis.
[0051] Fig. 14B This is the result diagram of NF-κB signaling pathway enrichment analysis.
[0052] Fig. 14C This is the result diagram of MAPK signaling pathway enrichment analysis.
[0053] Fig.14D This is the result diagram of IL-17 signaling pathway enrichment analysis.
[0054] Fig.14E This is the result of enrichment analysis of Toll-like receptor signaling pathway.
[0055] Fig.14F This is the result diagram of PI3K-AKT signaling pathway enrichment analysis.
[0056] Fig.15It is the relative expression result of inflammation-related genes in liver tissue of mice with acute alcoholic hepatitis model.
[0057] Fig.16 These are electron micrographs of mitochondria in liver tissues of different groups of acute alcoholic hepatitis model mice.
[0058] Fig.17 These are HE-stained sections of liver tissues of different groups of acute alcoholic hepatitis model mice. DETAILED DESCRIPTION
[0059] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.
[0060] Example 1
[0061] Extraction and identification of Artemisia capillaris-derived vesicle-like nanoparticles (AH-EVLPS):
[0062] 1. The extraction process of AH-EVLPS is as follows Figure 1 The specific extraction method is as follows:
[0063] (1) Take 150 g of Chinese medicinal herb Artemisia capillaris, wash it with sterile water, squeeze out the juice, collect the supernatant in a clean 5L beaker after filtering, and filter the supernatant with a 0.22 μm filter; collect the filtered liquid in a clean beaker, and filter and concentrate it 20 times with a tangential flow filtration system;
[0064] (2) The concentrated liquid was collected in a clean 50 mL centrifuge tube and centrifuged at 10,000 g for 30 min at 4 °C to remove dead cells, detached vesicles and apoptotic bodies; the supernatant was collected in a 50 mL centrifuge tube and filtered with a 0.22 μm diaphragm vacuum pump to sterilize the supernatant; the filtered liquid was collected in a centrifuge tube dedicated to an ultra-high speed centrifuge and centrifuged at 135,000 g for 70 min at 4 °C, the supernatant was discarded, the precipitate was resuspended in pre-cooled 1× PBS buffer, and filtered using a 0.22 μm disposable syringe filter. Finally, the AH-EVLPS was collected in a sterilized EP tube and stored at -80 °C.
[0065] 2. Characterization of AH-EVLPS
[0066] (1) The morphology of AH-EVLPS was observed under a transmission electron microscope. Figure 2A As shown: the surface is concave, showing a typical cup-shaped and disc-shaped vesicle structure, the cell membrane structure is clearly visible, the membrane boundary is clear and sharp, the staining background is clean, and the contrast is obvious.
[0067] (2) The particle size distribution of AH-EVLPS was analyzed using a nanoparticle tracer analyzer. Figure 2B As shown: the particle size is 67.75±3.50nm, between 0-300nm, which is consistent with the extracellular vesicle particle size range reported in the literature, and the concentration is 4.8×10 11 Pieces / mL.
[0068] Example 2
[0069] Stability test of Artemisia capillaris-derived vesicle-like nanoparticles (AH-EVLPS)
[0070] Test method: The AH-EVLP prepared in Example 1 was mixed with PBS solution, 1M alcohol solution, artificial gastric juice (SGF, pH 1.2) and artificial small intestinal fluid (SIF, pH 6.8) at a ratio of 1:100, and then allowed to stand for 30 minutes. The particle size distribution, average particle size and particle number of AH-EVLP in the several mixed solutions were detected by NanoFCM nanoflow cytometer.
[0071] The results are shown in Figure 3. After AH-EVLP was treated with PBS solution, 1M alcohol solution, artificial gastric juice, artificial small intestinal juice, and a mixture of artificial gastric juice and small intestinal juice (the volume ratio of artificial gastric juice and artificial small intestinal juice was 1:1), the particle size distribution did not change significantly, indicating that AH-EVLP can exist stably in alcohol, artificial gastric juice, and artificial small intestinal juice and can be administered orally.
[0072] Example 3
[0073] Internalization and absorption test of Artemisia capillaris-derived vesicle-like nanoparticles (AH-EVLPS):
[0074] The full internalization and absorption of AH-EVLPS by normal human liver cells is the prerequisite for the subsequent effect of AH-EVLPS on normal human liver cells. Therefore, before conducting subsequent experiments, we must first verify whether AH-EVLPS can be internalized and absorbed by normal human liver cells.
[0075] 1. Fluorescence microscopy to detect the internalization and absorption of AH-EVLPS by normal human liver cells
[0076] 1.1 Preparation of Dil-AH-EVLPS solution
[0077] The number of particles is 1×10 10AH-EVLPS was added to 1 mL with 1× PBS buffer, mixed well, and 10 μL DiI dye was added. Labeling was carried out at 37°C in the dark for 30 minutes. Centrifuge at 4°C, 100,000×g for 70 minutes, discard the supernatant, resuspend the precipitate with 1× PBS buffer, and centrifuge 3 times under the same centrifugation conditions to wash the unlabeled AH-EVLPS and excess DiI dye. Finally, resuspend the precipitate with 100 μL PBS buffer and collect it in an EP tube, recorded as DiI+AH-EVLPS, and use it immediately or store it in a -80°C refrigerator.
[0078] 1.2 Fluorescence microscopy to detect the internalization and absorption of AH-EVLPS by normal human liver cells
[0079] The labeled AH-EVLPS were co-cultured with normal human hepatocytes for 2h, 4h and 8h. Then, the internalization and absorption of AH-EVLPS by normal human hepatocytes were detected using a fluorescence microscope. Figure 4 As shown, the results showed that Dil-AH-EVLPS could be internalized and absorbed by normal human hepatocytes, and the degree of internalization increased with the prolongation of the co-incubation time of Dil-AH-EVLPS. Most of the internalized and absorbed Dil-AH-EVLPS was located in the cytoplasm of normal human hepatocytes.
[0080] The above results indicate that AH-EVLPS can be fully absorbed by normal human liver cells.
[0081] Example 4
[0082] In vitro experimental effects of Artemisia capillaris-derived vesicle-like nanoparticles (AH-EVLPS)
[0083] Preparation of Artemisia Capillaris water extract: mix Artemisia Capillaris with PBS, squeeze the juice, and filter to obtain the extract.
[0084] Test method:
[0085] The CCK8 method was used to investigate the effects of alcohol and AH-EVLP on the activity of HepG2 cells: HePG2 cells were seeded in 96-well plates at a concentration of 10,000 cells / well. Different concentrations of AH-EVLPS and alcohol were co-cultured with normal human liver cells for 24 hours, and 110 μL of CCK8 solution was added to each well under light-proof conditions, and the wells were incubated in an incubator for 2 hours. Finally, the absorbance of each well was detected by an ELISA reader.
[0086] The results are shown in Figure 5. When the alcohol concentration reached 4%, the cell activity decreased significantly (p < 0.01). 11Therefore, after the cells were incubated with 4% alcohol for 24 hours, different concentrations of AH-EVLP were added and cultured for another 24 hours, during which alcohol was added every 12 hours, and then the cell activity was detected using the CCK8 kit. The results showed that AH-EVLP had a significant reversal effect on the decrease in cell activity caused by alcohol (p<0.05), and this effect was dose-dependent and superior to the water extract group (AE). The mass of Artemisia capillaris used in the water extract group was the same as that of 1×10 9 The quality of Artemisia capillaris raw materials used in the two groups was the same.
[0087] Example 5
[0088] In vivo experimental effects of Artemisia capillaris-derived vesicle-like nanoparticles (AH-EVLPS)
[0089] 1. Test of the effect of AH-EVLPS on alcohol detoxification and liver protection
[0090] Eight-week-old male C57BL / 6J mice were divided into blank group (NC), model group (MOD), AH-EVLPS high-dose group (AH-EVLPH), AH-EVLPS low-dose group (AH-EVLPL), and Metadoxine group, with 6 mice in each group. Except for the blank group, the other groups were intragastrically administered with 6 g / kg of alcohol to establish the mouse model of acute alcohol poisoning. 30 minutes after the completion of the acute alcohol poisoning modeling, each mouse in the model group was intragastrically administered with 100 μL PBS, and each mouse in the AH-EVLPS high-dose group was intragastrically administered with 100 μL of PBS with a particle count of 2×10 7 AH-EVLPS, the number of particles in each mouse in the low-dose group of AH-EVLPS was 1×10 7 AH-EVLPS; each mouse in the metadoxine group was intragastrically administered with 100 μL of metadoxine capsule solution with a concentration of 0.0065 g / mL.
[0091] Test method:
[0092] By observing the righting reflex reaction of each group of mice, the drunkenness of each group of mice was obtained according to the scoring criteria in Table 1. The results are shown in Figure 6. As can be seen from Figure 6, the AH-EVLPS obtained in the present application can significantly improve the drunkenness and soberness scores of mice.
[0093] Table 1
[0094] Fraction Intoxication time scoring criteria Sobering time scoring criteria 0 0 min < Drunkenness time ≤ 5 min die 1 5 min < Drunkenness time ≤ 10 min 8h≤sobering time 2 10 min < Drunkenness time ≤ 30 min 3h≤Decline time<8h 3 30 min < Drunkenness time ≤ 60 min 1h≤Decline time<3h 4 60min < Drunkenness time 0h≤Decline time<1h 5 Not drunk Not drunk
[0095] A fully automatic biochemical analyzer was used to detect the levels of ALT, AST and ALP in mouse serum.
[0096] The results are as follows Figure 7-9As shown, serum biochemistry results showed that AH-EVLP (high dose) could significantly reduce the levels of ALT, AST and ALP in serum compared with the model group at 1h, 4h, 8h and 24h after modeling, indicating that AH-EVLP can improve alcohol-induced liver injury in mice.
[0097] 2. AH-EVLPS in vivo anti-inflammatory level test
[0098] Transcriptome sequencing analysis was performed on mice 24 hours after acute alcohol gavage. The results of differential gene statistics and cluster analysis showed that there were certain differences in gene expression between the model group and the blank group, as well as between the AH-EVLP high-dose group and the model group, and the cluster analysis showed significant differences in genes. Figure 10-13 As shown in the figure, compared with the model group, the number of differentially expressed genes in the AH-EVLP group was 2999, of which 2120 were upregulated and 879 were downregulated. It is worth noting that compared with the model group, genes such as Bcl3, Nfkb2, Nfkbia, and Il1b were significantly downregulated in the AH-EVLP group, indicating that AH-EVLP can regulate inflammation. GO functional enrichment and KEGG pathway enrichment were used to analyze the differentially expressed genes. GO enrichment results showed that the differentially expressed genes were mainly enriched in biological functions such as transmembrane transport, biological processes, inflammatory response, peroxisomes, metabolic processes, fatty acid synthesis and metabolism, mitochondria, mitochondrial depolarization, and mitochondrial DNA replication. KEGG pathways were mainly enriched in signaling pathways such as MAPK, TNF, PI3K-AKT, IL-17, NF-B, AMPK, and Toll-like receptors.
[0099] 3. RT-qPCR method to verify the expression of genes related to inflammation, apoptosis, lipid metabolism and immune response
[0100] 0.1 g of the mouse liver was taken and ground at 70 Hz for 50 s. The liver tissue RNA was extracted using an RNA extraction kit, and cDNA was obtained using a reverse transcription kit. The expression of genes related to inflammation, apoptosis, lipid metabolism and immune response was detected by RT-qPCR. It was found that AH-EVLPS can significantly inhibit the expression levels of IL-1b, Cd14, Ripk4, IL-1r1, IL-17ra, Tnfaip3, Tnfrsf1b, Nfkb2 and Nfkbia genes, and significantly upregulated the expression levels of Acp2 and Mkks genes. The results are shown in Figures 14-15.
[0101] 4. AH-EVLPS has a protective effect on mitochondria in liver tissue in vivo
[0102] We used transmission electron microscopy to observe the ultrastructural changes of mitochondria in the liver of mice 24 hours after intragastric administration of alcohol. Fig.16As shown in the figure, the results showed that the mitochondrial cristae in the model group were disordered, the inner mitochondrial membrane was dissolved, the mitochondria were swollen and vacuolated; the mitochondrial structure in the AH-EVLP group was relatively complete, with no obvious swelling and vacuolation. This shows that AH-EVLP can maintain the integrity of mitochondrial structure in acute alcoholic liver injury.
[0103] 5. AH-EVLP has the effect of reducing liver inflammatory infiltration in vivo
[0104] The livers of mice in the blank group, model group and AH-EVLPS group (high dose) were stained with HE. The staining results are shown in Fig.17 As shown in the results, 24 hours after intragastric administration of alcohol, obvious inflammatory infiltration was observed in the liver of mice in the model group, while no obvious pathological changes were observed in the liver of mice in the AH-EVLP group. This result further indicates that AH-EVLP can protect the liver from alcohol damage.
[0105] The applicant declares that the present invention uses the above-mentioned embodiments to illustrate the application of the extracellular vesicle-like nanoparticles derived from Artemisia capillaris in the preparation of drugs for preventing or treating liver diseases, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.
[0106] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.
[0107] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.
Claims
1. Application of extracellular vesicle-like nanoparticles derived from Artemisia capillaris in the preparation of drugs for preventing or treating liver diseases.
2. Application of extracellular vesicle-like nanoparticles derived from Artemisia capillaris in the preparation of drugs with alcohol-relieving and liver-protecting effects.
3. The use according to claim 1 or 2, characterized in that: The extracellular vesicle-like nanoparticles derived from Artemisia capillaris are prepared by a preparation method comprising the following steps: (1) filtering the juice of the Artemisia capillaris raw material, and concentrating the filtrate to obtain a concentrated solution; (2) centrifuging the concentrate and collecting the supernatant; (3) Centrifuge the supernatant, collect the precipitate, and resuspend the precipitate to obtain the product.
4. The use according to claim 3, characterized in that: The centrifugation temperature in step (2) is 0-4°C, the rotation speed is 9000-11000g, and the time is 25-35min.
5. The use according to claim 3 or 4, characterized in that: The centrifugation temperature in step (3) is 0-4°C, the rotation speed is 120000-150000g, and the time is 60-80min.
6. The use according to any one of claims 3 to 5, characterized in that: The volume of the concentrate is 5-10% of the volume of the filtrate.
7. The use according to any one of claims 3 to 6, characterized in that: The resuspension further includes filtering using a 0.22 μm filter.
8. The use according to any one of claims 1 to 7, characterized in that: The dosage form of the drug includes any one of tablets, capsules, pills, powders, solutions, aerosols or sprays.
9. The use according to any one of claims 1 to 8, characterized in that: The drug also includes pharmaceutically acceptable excipients.
10. The use according to claim 9, characterized in that: The pharmaceutically acceptable excipients include any one of diluents, dispersants, adhesives, fillers, thickeners, lubricants, pH regulators, taste masking agents, colorants, antioxidants or antibacterial agents, or a combination of at least two thereof.