Method for reducing or inhibiting inflammatory expression, microvesicle composition, method for preparing same, and use thereof

Abstract

Provided is a microvesicle composition for reducing or inhibiting inflammatory expression, a method for preparing same, and use thereof. The composition comprises a mesenchymal stem cell microvesicle and a plant cell microvesicle. The plant is selected from any one of lonicera, schisandra, zingiber, astragalus, ophiopogon, panax, and momordica, or a combination thereof.

Description

A method for reducing or inhibiting inflammatory expression, microvesicle composition, preparation method and application thereof Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to a method for reducing or inhibiting inflammatory expression, a microvesicle composition capable of reducing or inhibiting inflammatory expression, a preparation method of the microvesicle composition, and applications thereof. Background Art

[0002] Currently, drugs such as anti-allergic drugs or hormone drugs are commonly used clinically to treat respiratory inflammation. However, these drugs may cause secondary damage to the lungs and may also cause comprehensive suppression of the human immune system. Stem cells and stem cell microvesicles, because they carry some immune factors and can inhibit pro-inflammatory factors such as TNF-α, are widely used as a biotechnology. However, stem cells cannot be atomized, and ethical issues regarding stem cells always exist, which limits their application. Stem cell microvesicles can be atomized and have some of the effects of stem cells. Therefore, some medical institutions now use stem cell microvesicles to treat, intervene early, or prevent some respiratory diseases, or reduce infection efficiency.

[0003] However, there are the following problems with stem cell microvesicle atomization: 1) Umbilical cord stem cells are mainly used on the market. Due to the instability of the umbilical cord, the miRNA that inhibits inflammation in the stem cell microvesicles cannot be stably expressed, so the inhibitory effect of different batches of microvesicles on inflammation varies greatly; 2) Even umbilical cord stem cell microvesicles from the same batch have many main ingredients, such as anti-apoptotic factors, chemokines, vascular growth factors, etc., and their functions are complex, and the inflammatory effect of inhibiting the TNF-α pathway is effective. Summary of the Invention

[0004] In view of the above-mentioned defects of the prior art, the present invention provides a method for reducing or inhibiting inflammatory expression, wherein plant cell microvesicles are added to the system containing mesenchymal stem cell microvesicles. The plant cell microvesicles have an enhancing effect on the effect of mesenchymal stem cell microvesicles in reducing inflammatory expression; to this end, the present invention also provides a microvesicle composition with the ability to reduce or inhibit inflammatory expression, wherein the microvesicle composition is a mixture of mesenchymal stem cell microvesicles and plant cell microvesicles. The mesenchymal stem cell microvesicles and the plant cell microvesicles work synergistically with each other to achieve an excellent effect of inhibiting inflammation; to this end, the present invention also provides a preparation method and application of the microvesicle composition with the ability to reduce or inhibit inflammatory expression.

[0005] In a first aspect, the present invention provides a method for reducing or inhibiting inflammatory expression, comprising adding plant cell microvesicles to a mesenchymal stem cell microvesicle system, wherein the ratio of the mesenchymal stem cell microvesicles to the plant cell microvesicles is 10:1-25:1.

[0006] As a preferred embodiment, the mesenchymal stem cell microvesicles are human-derived umbilical cord mesenchymal stem cell microvesicles.

[0007] As a preferred embodiment, the plant is a plant with medicinal and edible properties.

[0008] Plants with both medicinal and edible properties are edible and have medicinal properties. For example, cloves, a common spice, also have medicinal properties; sword beans, edible as a vegetable, also have medicinal properties.

[0009] As a more preferred embodiment, the plant is selected from any one or more combinations of honeysuckle, schisandra chinensis, ginger, astragalus, ophiopogon japonicus, and ginseng.

[0010] As a more preferred embodiment, the plant is honeysuckle or ginseng.

[0011] In a second aspect, the present invention provides the use of plant cell microvesicles in enhancing the ability of mesenchymal stem cell microvesicles to inhibit inflammatory expression, wherein the ratio of the mesenchymal stem cell microvesicles to the plant cell microvesicles is 10:1-25:1.

[0012] In a third aspect, the present invention provides a microvesicle composition capable of reducing or inhibiting inflammatory expression, comprising mesenchymal stem cell microvesicles and plant cell microvesicles, wherein the ratio of the mesenchymal stem cell microvesicles to the plant cell microvesicles is 10:1-25:1.

[0013] As a preferred embodiment, the concentration of the mesenchymal stem cell microvesicles is 2×10 9 -5×10 9 pieces / mL.

[0014] As a preferred embodiment, the concentration of the plant cell microvesicles is 1×10 8 -2×10 8 pieces / mL.

[0015] As a preferred embodiment, the average size of the mesenchymal stem cell microvesicles is 132.9 nm.

[0016] As a preferred embodiment, the average size of the plant cell microvesicles is 154.7 nm.

[0017] In a fourth aspect, the present invention provides a method for preparing a microvesicle composition capable of reducing or inhibiting inflammatory expression. The microvesicle composition is a mixture of mesenchymal stem cell microvesicles and plant cell microvesicles.

[0018] The fifth aspect of the present invention provides the use of a microvesicle composition having the ability to reduce or inhibit inflammatory expression in the preparation of a drug for treating respiratory diseases, brain diseases, intestinal diseases, and autoimmune diseases.

[0019] As a preferred embodiment, the microvesicle composition is used to reduce or inhibit the inflammatory expression of the TNF-α pathway.

[0020] TNF-α is a pro-inflammatory cytokine involved in normal inflammatory and immune responses, synergistically regulating the production of other cytokines, cell survival, and cell death to coordinate tissue homeostasis. TNF promotes inflammatory responses, which not only contribute to respiratory illnesses but also to numerous clinical issues associated with autoimmune diseases. Furthermore, dysregulated TNF production is associated with a variety of human diseases, including brain and intestinal disorders. Reducing or inhibiting inflammatory expression of the TNF-α pathway could be used to treat these respiratory, brain, intestinal, and autoimmune diseases.

[0021] As a preferred embodiment, the respiratory diseases include pneumonia, asthma, allergic rhinitis, and chronic pharyngitis.

[0022] As a preferred embodiment, the brain diseases include Alzheimer's disease, cerebral infarction, and major depression.

[0023] As a preferred embodiment, the intestinal diseases include inflammatory bowel disease, and the autoimmune diseases include rheumatoid arthritis and ankylosing spondylitis.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) The method of reducing or inhibiting inflammatory expression of the present invention is to add plant cell microvesicles to the system of mesenchymal stem cell microvesicles. The plant cell microvesicles have the effect of enhancing the inhibitory effect of mesenchymal stem cell microvesicles on inflammatory expression. Under the premise of no significant increase in cost, the inflammatory factors of the TNF-α pathway are significantly inhibited.

[0026] (2) The present invention has a microvesicle composition that reduces or inhibits inflammatory expression. The composition is formed by mixing mesenchymal stem cell microvesicles with plant cell microvesicles. The mesenchymal stem cell microvesicles and the plant cell microvesicles work together to reduce inflammatory factors by 75%. Compared with the use of mesenchymal stem cell microvesicles or plant cell microvesicles alone, it has a significant inflammatory effect of inhibiting the TNF-α pathway and can be used to treat respiratory diseases, brain diseases, intestinal diseases, and autoimmune diseases.

[0027] (3) The present invention has a microvesicle composition that reduces or inhibits inflammatory expression and has little effect on macrophage viability. The addition of the microvesicle composition does not affect cell viability while inhibiting inflammatory factors, and the cell state remains good.

[0028] The concept, specific structure and technical effects of the present invention will be further described below in conjunction with the accompanying drawings to fully understand the purpose, characteristics and effects of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] FIG1 is a particle size distribution diagram of mesenchymal stem cell microvesicles;

[0030] FIG2 is a diagram showing the particle size distribution of plant cell microvesicles;

[0031] FIG3 is a diagram showing the effect of mesenchymal stem cells and / or plant cell microvesicles on the activity of macrophages;

[0032] Figure 4 shows the absorption of mesenchymal stem cell / plant cell microvesicles by macrophages (72 hours); Figures 4-1, 4-2, and 4-3 show the absorption of macrophages in groups A2, B2, and C2, respectively;

[0033] FIG5 shows the results of mesenchymal stem cell and / or plant cell microvesicles inhibiting TNF-α secretion. DETAILED DESCRIPTION

[0034] In order to make the technical means, creative features, objectives and effects of the invention easier to understand, the invention is further described below with reference to specific diagrams. However, the invention is not limited to the following implementation cases.

[0035] It should be noted that the structures, proportions, sizes, etc. illustrated in the drawings in this specification are only used to match the contents disclosed in the specification so that people familiar with this technology can understand and read them. They are not used to limit the conditions under which the present invention can be implemented. Therefore, they have no substantive technical significance. Any modification of the structure, change in the proportion relationship or adjustment of the size should still fall within the scope of the technical content disclosed in the present invention without affecting the efficacy and purpose that can be achieved by the present invention.

[0036] Respiratory diseases are the most common illnesses in daily life. Their clinical manifestations include cough, accompanied by headache, chills, fever and general discomfort. Currently, they are mainly treated with anti-allergic drugs or hormone drugs, which may cause secondary damage to the lungs or overall suppression of human immunity.

[0037] To address this issue, some researchers are considering using stem cells or their exosomes. Stem cells carry some immune factors and have a certain inhibitory effect on pro-inflammatory factors such as TNF-α. However, ethical issues have plagued stem cell research, and stem cells cannot be aerosolized, making it difficult to rapidly suppress respiratory inflammation. However, stem cell exosomes can be absorbed by the body through aerosolization. Therefore, some medical institutions are using aerosolized stem cell exosomes for the treatment, early intervention, or prevention of certain respiratory diseases.

[0038] Exosomes are small membrane vesicles containing complex RNA and proteins, ranging in diameter from 30 to 100 nm. They were first discovered in sheep reticulocytes in 1983 and named "exosomes" by Johnstone in 1987. Exosomes are secreted by a variety of cells under both normal and pathological conditions. Exosomes primarily originate from multivesicular bodies formed by the invagination of intracellular lysosomal microsomes. These vesicles are released into the extracellular matrix after the outer membrane of the multivesicular body fuses with the cell membrane. Exosomes contain a variety of biomolecules, such as mRNA, miRNA, proteins, and lipids, which can be delivered to recipient cells, thereby altering their physiological or pathological functions. However, exosomes are monolayered and easily degraded, which to some extent limits their usefulness.

[0039] Microvesicles are vesicles formed by direct shedding of cells after budding and fusion with the cell membrane. Microvesicles and exosomes contain similar substances, but differ in that exosomes have a single-layer membrane structure, are 30-100 nm in size, and are easily degraded, whereas microvesicles have a double-layer membrane structure, are 100-300 nm in size, are highly protective, and are less susceptible to degradation. Therefore, microvesicles have been widely studied in the biotechnology field as an alternative to exosomes.

[0040] There are still the following problems with stem cell exosomes or microvesicles in the treatment of respiratory inflammation: 1) The stem cells on the market are mainly umbilical cord stem cells. Due to the instability of the umbilical cord, the miRNA that inhibits inflammation in the stem cell exosomes or microvesicles cannot be stably expressed, and the anti-inflammatory effects of different batches vary significantly; 2) The inflammatory effect of umbilical cord stem cells from the same batch on the TNF-α pathway is also very limited.

[0041] Traditional Chinese medicine plants, especially those with medicinal and edible properties, are ineffective against many viruses and bacteria harmful to humans due to their evolutionary divergence from the human body. Some plant-derived miRNA fragments are beneficial to human lung cells and macrophages. This application aims to investigate whether stem cell microvesicles can be combined with plant cell microvesicles to achieve a stable anti-inflammatory effect.

[0042] When mixing stem cell microvesicles with plant cell microvesicles, compatibility between the two needs to be considered. Not all plant cell microvesicles enhance the inhibitory effect of stem cell microvesicles on inflammatory expression when mixed with stem cell microvesicles. Plant cell microvesicles carry their own unique bioactive substances. Lipids, proteins, miRNAs, and secondary metabolites are important components of microvesicles. The composition of microvesicles is unique to each plant. For example, grapefruit microvesicles contain curcumin and enzymolysates; citrus lemon microvesicles contain phenolic acids, coumarins, carboxylic acids, amino acids, and microorganisms; and carrot microvesicles contain phenolic compounds, carotenoids, polyacetylenes, and ascorbic acid.

[0043] From the mixed screening experiment of medicinal and edible plants and stem cell microvesicles, it can be seen that when the protein content of plant cell microvesicles is too low, there are problems with compatibility with stem cell microvesicles, or they cannot enhance the effect, or even weaken the effect of inhibiting inflammatory expression. In addition, the content and type of plant-derived miRNA also have a significant impact on the inhibition of inflammatory expression. The more human miRNA a plant expresses, the more obvious the effect of inhibiting inflammatory expression. Combined with experimental data, when choosing honeysuckle, schisandra chinensis, ginger, astragalus, ophiopogon japonicus or ginseng, they can all achieve good results.

[0044] TNF, as a cytokine, is used by the immune system for cell signaling. If macrophages (certain white blood cells) detect an infection, they release TNF to alert other immune system cells as part of the inflammatory response. This application evaluates the effectiveness of plant cell microvesicles and / or stem cell microvesicles in inhibiting inflammatory expression by characterizing the expression of TNF-α.

[0045] In addition, since the expression of TNF-α factor is associated with a variety of diseases, evaluating the expression of TNF-α factor can also indirectly reflect the therapeutic effect of plant cell microvesicles and / or stem cell microvesicles in related diseases.

[0046] In the following examples, GRNA is ginseng microvesicles; MSC-MV is human umbilical cord mesenchymal stem cell microvesicles.

[0047] Example 1 Preparation of microvesicle composition

[0048] 1. Preparation of Mesenchymal Stem Cell Microvesicles

[0049] Cell source: human umbilical cord mesenchymal stem cells.

[0050] Seed cell source: ATCC seed bank.

[0051] Culture medium for stem cell culture: Beijing Youkang, product number NC0103+NC0103.S.

[0052] Digestive enzyme: Beijing Youkang, product number NC1004.1.

[0053] Stem cell culture flasks: NUNC brand T175 flask, item number 159910, T75 culture flask, item number 156499.

[0054] Other pipettes, centrifuge tubes, etc. are all from the NUNC brand under Thermo Fisher Scientific.

[0055] The preparation of stem cell microvesicles is carried out in the following steps:

[0056] S1, the purchased primary cells (P1) 1.0×10 6 / ml was washed once with stem cell culture medium by centrifugation at a centrifugal force of 500g for 5 minutes. The washed cells were resuspended in 12ml stem cell culture medium in a stem cell culture bottle and cultured in an incubator at 37°C and 5% carbon dioxide concentration;

[0057] S2. When the cells grow to about 85% confluency in step S1, they are passaged into three stem cell culture flasks at a ratio of 1:6. 25 ml of culture medium is added to each flask and cultured in an incubator at 37°C and 5% carbon dioxide concentration.

[0058] S3, the same subculture ratio was continued to P4, a total of 108 bottles were seeded, and finally about 2500 ml of P4 supernatant was harvested;

[0059] S4, take 270 ml of the harvested P4 supernatant, centrifuge at 12000 g, remove the precipitate after centrifugation, filter the supernatant through a 0.22 μm needle filter, and collect about 250 ml of the filtrate;

[0060] S5. Ultracentrifuge or electrostatic separation is performed on the 250 ml stem cell supernatant after centrifugation to extract and purify human umbilical cord mesenchymal stem cell microvesicles. The purified stem cell microvesicles are measured for particle size by NTA, labeled, and stored in an ultra-low temperature freezer until ready for use.

[0061] The particle size diagram of the obtained mesenchymal stem cell microvesicles is shown in Figure 1. As can be seen from Figure 1, the average diameter of the microvesicles is 132.9 nm, which exceeds 100 nm. Due to their large size, they contain relatively rich miRNAs and have a double-layer membrane structure, which is not easily degraded.

[0062] 2. Preparation of plant cell microvesicles

[0063] The preparation method of plant cell microvesicles comprises the following steps:

[0064] S1. Clean the plant samples by washing them with tap water to remove any surface sediment, then washing them twice with purified water and once with ultrapure water; cut the samples into slices of approximately 1.5 mm uniform thickness;

[0065] S2. Clean the juicer. Soak the stainless steel juicer in 0.1 mol / L sodium hydroxide solution for 30 minutes in advance to achieve the effect of preliminary removal of endotoxins; then rinse it with ultrapure water three times. Take the last cleaning solution and test it with pH test paper. If it is neutral, it can be used.

[0066] S3. Weigh 40 g of the sliced ​​sample, add 250 mL of 0.9% medical-grade saline or PBS solution, and soak in a juicer for 30 min. Then, start the juice extraction and filter through disposable sterile gauze to collect approximately 240 mL of the filtrate.

[0067] S4. The filtrate collected in S3 was divided into 50 mL centrifuge tubes and centrifuged. First, centrifuge at 2000 g for 20 minutes to remove the bottom sediment. The upper liquid was collected and centrifuged a second time at 12000 g for 30 minutes. Repeat the centrifugation twice (12000 g for 30 minutes) according to the sedimentation at the bottom of the liquid until there is no obvious sediment at the bottom. Then collect 220 mL of supernatant.

[0068] S5. Take the supernatant after centrifugation and sterilize it by filtering it with a 0.22 μm syringe filter to collect about 200 ml of the filtrate;

[0069] S6. Take 40 mL of the filtrate after filtration, and freeze the remaining sample in an ultra-low temperature freezer for later use; dilute the 40 mL sample with 0.9% medical saline or PBS to 160 mL, and extract the plant cell microvesicles by density gradient centrifugation or electrostatic separation to obtain 2 mL of plant cell microvesicle suspension;

[0070] S7. The collected microvesicles were counted using Zetaview's NTA, and then placed in an ultra-low temperature freezer for later use.

[0071] The particle size distribution of the obtained plant cell microvesicles is shown in FIG2 . As can be seen from FIG2 , the average diameter of the microvesicles is 154.7 nm, which is greater than 100 nm. The microvesicles have a double-layer membrane structure and are not easily degraded.

[0072] 3. Preparation of microvesicle composition

[0073] 2×10 9 -5×10 9 / mL mesenchymal stem cell microvesicles and 1×10 8 -2×10 8 / mL plant cell microvesicles are mixed to obtain a microvesicle composition.

[0074] Example 2 Effects of Mesenchymal Stem Cells and / or Plant Cell Microvesicles on Macrophage Viability

[0075] To compare the effects of mesenchymal stem cell microvesicles, plant cell microvesicles, or the combination of mesenchymal stem cell microvesicles and plant cell microvesicles on macrophage viability, the following groups were used:

[0076] Table 1

[0077] MSC-MV--10 9 The particle number refers to the concentration of MSC-MVs at 1×10 9 cells / mL; MSC-MV--10 10 The particle number refers to the concentration of MSC-MVs at 1×10 10 gRNA-10 8 The particle number refers to the concentration of gRNA at 1×10 8 cells / mL; MSC-MV-10 9 +gRNA-10 8 The system contains 1×10 9 MSC-MVs and 1×10 8 gRNA per mL.

[0078] The specific steps are as follows:

[0079] 1) Obtain macrophages from healthy human volunteers and resuscitate them;

[0080] 2) After digesting, pipetting and resuspending the cells, 1×10 5 Cells / mL were seeded in 96-well plates, 100 μL of cell suspension was added to each well, 3 parallel wells were set up for each group, and the plates were cultured in a 37°C, 5% CO2 incubator until the cells attached to the wall;

[0081] 3) Add microvesicles according to different groups, set corresponding blank zero wells (i.e., wells with only the corresponding culture medium but no cells), and incubate for 0 h, 48 h, 72 h, and 120 h;

[0082] 4) Add 10 μL of CCK-8 solution to each well and incubate in the incubator for 1 hour;

[0083] 5) Use a microplate reader to measure the absorbance at 450 nm, with cells in group A1 as the control group and the blank well as the blank, and calculate the cell survival rate of the drug according to the formula;

[0084] Cell survival rate % = (experimental group-blank) / (control group-blank)×100%.

[0085] After macrophages were treated with microvesicles in each group, the changes in cell viability are shown in Figure 3. As shown in Figure 3, mesenchymal stem cell microvesicles, plant cell microvesicles, or a combination of mesenchymal stem cell microvesicles and plant cell microvesicles had little effect on macrophage viability, and the cells in each group were in good condition.

[0086] Example 3 Absorption of mesenchymal stem cell / plant cell microvesicles by macrophages

[0087] In order to compare the uptake of mesenchymal stem cell microvesicles or plant cell microvesicles by macrophages, the following groups were divided:

[0088] Table 2

[0089] The concentration of GRNA in group B2 was 1×10 10 The concentration of MSC-MVs in group C2 was 1×10 9 pieces / mL.

[0090] 1) Obtain macrophages from healthy human volunteers, resuscitate them, and condition them to serve as recipient cells.

[0091] 2) Groups A2, B2, and C2 were mixed with PKH26, respectively, and incubated in the dark for 10 minutes to remove excess PKH26;

[0092] 3) Add the labeled microvesicles to the recipient cells and incubate at 37°C in the dark for 4 hours;

[0093] 4) Aspirate and discard the cell supernatant and wash 2-3 times with PBS;

[0094] 5) Add 4% paraformaldehyde and fix at room temperature for 30 minutes;

[0095] 6) Aspirate the paraformaldehyde fixative and wash 2-3 times with PBS;

[0096] 7) Draw circles with a histochemical pen and wash with PBS.

[0097] 8) Dilute red-light phalloidin-TRITC (or green-light phalloidin-488) at a dilution of 1:300 in 1% BSA to prepare a working solution. Incubate at 37°C for 1–1.5 hours or at 4°C overnight in the dark.

[0098] 9) Wash with PBS three times, 5 minutes each time, add DAPI to stain the nucleus, incubate at room temperature in the dark for 20-30 minutes, and then wash with PBS.

[0099] 10) Seal the slides with an anti-fluorescence quenching mounting medium, observe and photograph under a microscope, and observe the uptake of the stem cell / plant cell microvesicles by macrophages.

[0100] The absorption of microvesicles by macrophages in each group is shown in Figure 4 . As can be seen from Figure 4 , the cells are in good condition, and at 72 hours, the absorption of plant cell and mesenchymal stem cell microvesicles by macrophages in groups B2 and C2 can be seen. Therefore, mesenchymal stem cells and / or plant cell microvesicles can be used to evaluate the inhibition of TNF-α factor secretion.

[0101] Example 4 Detection of TNF-α secretion

[0102] In order to compare the inhibitory effects of mesenchymal stem cells and / or plant cell microvesicles on TNF-α secretion, the following groups were divided:

[0103] Table 3

[0104] The operation is as follows:

[0105] 1) Prepare macrophages and plate them one day in advance, and culture them for 24 hours;

[0106] 2) Add 100 ng / mL LPS and incubate at 37°C for 24 h;

[0107] 3) Add corresponding microvesicles according to different groups and incubate in a 37°C constant temperature incubator for 3 days;

[0108] 4) Centrifuge at 1000 g for 20 min, carefully collect the cell supernatant, and measure immediately to avoid freezing and thawing;

[0109] 5) Use TNF-α Elisa kit to detect the secretion of TNF-α factor.

[0110] The results of TNF-α factor secretion detection in each group are shown in Figure 5. It can be seen from Figure 5 that the TNF-α concentration in group B3 increased significantly compared with group A3, indicating that LPS effectively stimulated the secretion of TNF-α factor; after adding GRNA, MSC-MV and GRNA+MSC-MV to the LPS+macrophage system, the secretion of TNF-α showed a downward trend, and the E3 group (added with GRNA+MSC-MV) decreased most significantly, decreasing by about 75% compared with group B3. When GRNA (group C3) or MSC-MV (group D3) was added alone, the TNF-α concentration decreased by at most 40%, indicating that the combination of GRNA and MSC-MV has a significant inflammatory effect of inhibiting the TNF-α pathway.

[0111] The preferred embodiments of the present invention have been described in detail above. It should be understood that numerous modifications and variations based on the concepts of the present invention are possible without inventive effort by those skilled in the art. Therefore, any technical solution that can be derived by one skilled in the art through logical analysis, reasoning, or limited experimentation based on the concepts of the present invention and the prior art should be within the scope of protection defined by the claims.

Claims

1. A method for reducing or inhibiting inflammatory expression, characterized in that: Plant cell microvesicles are added to the mesenchymal stem cell microvesicle system, with the dosage ratio of the mesenchymal stem cell microvesicles to the plant cell microvesicles being 10:1-25:

1.

2. The method for reducing or inhibiting inflammatory expression according to claim 1, wherein: The plants are selected from any one or more combinations of honeysuckle, schisandra chinensis, ginger, astragalus, ophiopogon japonicus, ginseng, and bitter melon.

3. Application of plant cell microvesicles in enhancing the inhibitory effect of mesenchymal stem cell microvesicles on inflammatory expression, characterized in that: The dosage ratio of the mesenchymal stem cell microvesicles to the plant cell microvesicles is 10:1-25:

1.

4. A microvesicle composition capable of reducing or inhibiting inflammatory expression, characterized in that: The invention comprises mesenchymal stem cell microvesicles and plant cell microvesicles, wherein the dosage ratio of the mesenchymal stem cell microvesicles to the plant cell microvesicles is 10:1-25:

1.

5. The microvesicle composition having the ability to reduce or inhibit inflammatory expression according to claim 4, wherein: The concentration of the mesenchymal stem cell microvesicles is 2×10 9 -5×10 9 pieces / mL.

6. The microvesicle composition having the ability to reduce or inhibit inflammatory expression according to claim 4, wherein: The concentration of the plant cell microvesicles is 1×10 8 -2×10 8 pieces / mL.

7. The microvesicle composition for reducing or inhibiting inflammatory expression according to claim 4, wherein: The average size of the mesenchymal stem cell microvesicles is 132.9 nm.

8. The method for preparing the microvesicle composition having the function of reducing or inhibiting inflammatory expression according to any one of claims 4 to 7, characterized in that: The mesenchymal stem cell microvesicles and the plant cell microvesicles are mixed.

9. Use of the microvesicle composition having the ability to reduce or inhibit inflammatory expression according to any one of claims 4 to 7 in the preparation of a drug for treating respiratory diseases, brain diseases, intestinal diseases, and autoimmune diseases.

10. The use according to claim 9, characterized in that The microvesicle composition is used to reduce or inhibit the inflammatory expression of the TNF-α pathway.

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