CN112675203A - Application of cell-derived exosome in preparation of biological preparation for treating asthma and/or pulmonary fibrosis - Google Patents
Application of cell-derived exosome in preparation of biological preparation for treating asthma and/or pulmonary fibrosis Download PDFInfo
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Abstract
The invention discloses application of exosome derived from cells in preparation of a biological preparation for treating asthma and/or pulmonary fibrosis. The biological preparation can be prepared into aerosol, spray, oral preparation or injection, and the exosome can also be used for drug loading, surface modification or excipient. In vitro cell experiments are carried out, and the in vitro cell experiments show that the umbilical cord mesenchymal stem cell exosome extracted and purified by the method can repair damaged lung epithelial cells, can inhibit the lung epithelial cells from secreting inflammatory factors, and can inhibit the proliferation of lung fibroblasts, so that the exosome has the potential of treating asthma and/or pulmonary fibrosis, and can be applied to preparation of biological preparations for treating asthma and/or pulmonary fibrosis.
Description
Technical Field
The invention relates to the field of biological medicines, in particular to application of exosomes derived from cells in preparation of biological preparations for treating asthma and/or pulmonary fibrosis.
Background
The lung area of human is 35-100 square meters, which is responsible for inhaling oxygen and discharging carbon dioxide. During normal breathing, the lung surfaces will be exposed to microorganisms, air pollution, occupational chemicals and dust. All of these constitute risk factors for acute and chronic lung disease. A recent report by the world health organization has demonstrated that hundreds of millions of people in all ages worldwide suffer from chronic respiratory disease and rank pulmonary disease as the third leading cause of death worldwide, second only to cardiovascular disease and cancer.
Asthma (Aasthma) is a heterogeneous disease characterized by chronic airway inflammation and airway hyperresponsiveness. The main features include chronic inflammation of the airways, the high reactivity of the airways to various stimuli, variable reversible airflow limitation, and a series of changes in airway structure, i.e., airway remodeling, with the course of the disease. The clinical manifestations are recurrent wheezing, shortness of breath, chest distress or cough, which usually attacks or aggravates at night and in the morning, and most patients can relieve themselves or relieve after treatment. The typical symptoms of asthma are paroxysmal expiratory dyspnea with whitish sound, which can be accompanied by shortness of breath, chest distress or cough, the symptoms can be attacked within minutes, last for hours to days, and can be relieved or relieved automatically after being treated by an antiasthmatic medicament, and the attack or aggravation at night and early morning is an important clinical characteristic of asthma.
Pulmonary Fibrosis (RF) is a chronic, progressive and destructive interstitial lung disease, belonging to the category of fibroproliferative diseases. The main pathological features are the proliferation of fibroblasts, the aggregation of a large amount of extracellular matrixes, and the final-stage change of a large group of lung diseases characterized by inflammatory injury and tissue structure destruction, namely, structural abnormality caused by abnormal repair after normal alveolar tissues are damaged. When pulmonary fibrosis progresses, dyspnea also occurs at rest, and progressive dyspnea may occur in patients with severe pulmonary fibrosis. Other symptoms include dry cough and hypodynamia. Although dyspnea exists in the early stage of onset, the X-ray chest radiograph is probably basically normal, and diffuse reticular or nodular shadows appear in the middle and middle lung fields, so that pleural effusion, thickening or calcification are occasionally seen. Severe consequences of lung tissue fibrosis lead to structural changes in normal lung tissue and loss of function. A large amount of fibrous tissue without gas exchange function replaces alveoli, resulting in the inability of oxygen to enter the blood. The patient is not smooth in breathing, is lack of oxygen, is acidosis, loses labor force, lives by a breathing machine, and finally fails and dies.
Currently, there are very limited biological agents that can be used to treat asthma and/or pulmonary fibrosis. Clinically, glucocorticoids, immunosuppressants, cytotoxic biologics and anticoagulants are commonly used for treatment. Although these biological agents can slow down the decline of the lung function, they cannot reverse the progress of the disease, and the drugs or biological agents have many side effects on the human body, such as irritation to the gastrointestinal tract, nausea, dyspepsia and other symptoms, skin diseases such as skin allergy and damage to the liver function. Lung transplantation is a relatively effective means for treating pulmonary fibrosis, but the source of healthy lung donors is limited, so that the wide application of the lung donors is restricted, and immune rejection is overcome after the lung transplantation, wherein the survival period is less than 5 years.
At present, a large number of preclinical experiments show that the Mesenchymal Stem Cells (MSCs) can effectively improve pulmonary fibrosis diseases, and have various pharmacological effects including anti-inflammatory, immunoregulatory, regenerative, angiogenesis promoting, anti-fibrosis and other properties, so that the stem cells provide a novel pulmonary fibrosis treatment method. It is currently widely believed that the efficacy of stem cells has mainly three mechanisms of action, (1) homing ability, i.e., the selective migration of cells to damaged tissues by chemoattraction mediated by chemokines, cell surface receptors, integrins, or selectins; (2) differentiating into new different types of cells to replace the damaged cells; (3) substances capable of modulating cellular responses are secreted by paracrine action. However, many studies now show that the number of stem cells injected into the body is very low for effective transplantation and differentiation, which indicates that stem cells most likely act through a paracrine mechanism, so more and more researchers are focusing on the secretory component of stem cells, such as soluble factors like cytokines, chemokines and growth factors, and cellular microvesicles and exosomes, which are the focus of researchers in recent years.
The exosome is a small vesicle with a phospholipid bilayer structure secreted by living cells, has the diameter of 30-150nm and the density of 1.13-1.19g/mL, and can be present in various body fluids such as serum, plasma, saliva, urine, ascites, spinal fluid, milk and the like. Exosomes contain a variety of biomolecules, such as mRNA, miRNA, proteins, lipids, etc., that can be delivered to recipient cells, thereby altering the physiological or pathological function of the recipient cells. In recent years, exosomes have attracted considerable attention as an intercellular information transfer tool and a biomarker for various diseases, and have potential for application and development in the fields of biomedicine and disease diagnosis.
Disclosure of Invention
Therefore, the invention provides an application of the exosome derived from the cell in preparing a biological preparation for treating asthma and/or pulmonary fibrosis.
In order to achieve the above purpose, the invention provides the following technical scheme:
according to an aspect of the present invention, there is provided a use of a cell-derived exosome in the preparation of a biological agent for treating asthma and/or pulmonary fibrosis.
Further, the biological preparation can be prepared into aerosol, spray, oral preparation or injection.
Further, the exosomes can be used for drug loading, surface modification or excipient.
In another aspect of the present invention, there is provided a cell-derived exosome for use in preparing a biological agent for treating asthma and/or pulmonary fibrosis, the exosome being extracted by a method comprising ultracentrifugation, density gradient centrifugation, ultrafiltration, polymer precipitation, tangential flow filtration, immunocapture, affinity chromatography, microfluidics or size exclusion.
Further, the exosome has the effect of promoting lung epithelial cell repair.
Furthermore, the exosome can inhibit inflammatory factors produced by lung epithelial cells induced by LPS.
Further, the exosome can inhibit the proliferation of human embryonic lung fibroblasts by regulating the expression of an apoptotic protein.
Further, the cell sources are plants, microorganisms, animals and humans, including body fluids, tissue fluids, somatic cells, stem cells and cells induced to differentiate by the stem cells.
Furthermore, the exosome has the potential of treating asthma and/or pulmonary fibrosis, and can be applied to preparation of biological preparations for treating asthma and/or pulmonary fibrosis.
The invention has the following advantages:
the exosome can be used for treating pulmonary fibrosis diseases in various modes such as aerosol, spray, oral preparations, injection preparations and the like, and can also be prepared into a biological preparation by methods such as drug loading, surface modification or excipient and the like, so that the toxic and side effects of the biological preparation can be greatly reduced in the process of treating asthma and/or pulmonary fibrosis, and the problems of irritation of the traditional biological preparation to gastrointestinal tract, skin allergy, damage to liver function and the like are avoided.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
The structures, ratios, sizes, and the like shown in the present specification are only used for matching with the contents disclosed in the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions that the present invention can be implemented, so that the present invention has no technical significance, and any structural modifications, changes in the ratio relationship, or adjustments of the sizes, without affecting the effects and the achievable by the present invention, should still fall within the range that the technical contents disclosed in the present invention can cover.
FIG. 1 is a block diagram of the present invention FIG. 1 is a diagram of a dual tangential flow filtration system provided by the present invention, wherein 1 is a first storage tank, 2 is a first peristaltic pump, 3 is a first filtration device, 4 is a second storage tank, 5 is a second filtration device, and 6 is a second peristaltic pump;
FIG. 2 is a microscope observation image of umbilical cord mesenchymal stem cells provided by the present invention;
FIG. 3 is a diagram of the flow detection result of the umbilical cord mesenchymal stem cell specific protein CD34 provided by the invention;
FIG. 4 is a diagram of the flow detection result of the umbilical cord mesenchymal stem cell specific protein CD45 provided by the invention;
FIG. 5 is a diagram of the flow detection result of the umbilical cord mesenchymal stem cell specific protein CD73 provided by the invention;
FIG. 6 is a diagram of the flow detection result of the umbilical cord mesenchymal stem cell specific protein CD90 provided by the invention;
FIG. 7 is a diagram of the flow detection result of the umbilical cord mesenchymal stem cell specific protein CD105 provided by the invention;
FIG. 8 is an electron micrograph of double TFF purified exosomes provided by the present invention;
FIG. 9 is a graph of the nanoparticle size of double TFF purified exosomes provided by the present invention;
FIG. 10 is a result chart of the Western Blot provided by the invention for identifying CD9 protein of exosomes after double TFF purification;
FIG. 11 is a result chart of the Western Blot provided by the invention for identifying CD63 protein of exosomes after double TFF purification;
FIG. 12 is a graph showing the results of Western Blot for identifying ALIX protein of exosomes purified by double TFF provided by the present invention;
FIG. 13 is a graph showing the effect of different exosome concentrations on lung epithelial cell proliferative capacity provided by the present invention;
FIG. 14 is a graph showing the results of the content of IL-6, an inflammatory factor produced by lung epithelial cells induced by LPS;
FIG. 15 is a graph showing the results of the content of IL-8, an inflammatory factor produced by lung epithelial cells induced by LPS;
FIG. 16 is a graph showing the results of the content of inflammatory factor TNF- α induced by LPS in lung epithelial cells according to the present invention;
FIG. 17 is a graph showing the results of the effect of different exosome concentrations provided by the present invention on the proliferative capacity of human embryonic lung fibroblasts;
FIG. 18 is a result chart of apoptotic proteins Bcl-2 and Bax in human embryonic lung fibroblast strains detected by WB method, wherein A is a PBS buffer solution comparison chart, B is an exosome result chart extracted by adding dual TFF method, and C is an exosome result chart extracted by adding ultracentrifugation method.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
EXAMPLE 1 preparation and culture of Stem cells
1. Extraction and preparation of umbilical cord mesenchymal stem cells
Collecting umbilical cord of newborn from operating room, washing umbilical cord with PBS buffer solution, removing blood vessel with scissors and forceps, stripping Fahrenheit gelatin tissue, and sufficiently cutting tissue to 1mm3Tissue mass of size. And (3) uniformly spreading the tissue blocks in a culture dish, and placing the tissue blocks in an incubator for culture for 1h under the culture conditions: 5% CO2The temperature was 37 ℃. When the tissue block adheres to the wall firmly, adding a proper amount of stem cell culture medium, and placing the mixture in an incubator for culture under the culture conditions: 5% CO2The temperature was 37 ℃. After the cell adherence condition is good, slightly absorbing the culture medium, slightly rinsing the culture medium by using the stem cell culture medium once, adding 10mL of fresh stem cell culture medium, and placing the culture medium in an incubator for culture under the culture conditions: 5% CO2The temperature was 37 ℃. Digesting with digestive juice when the culture dish is full of 70-85% adherent cells, adding stop solution to stop digestion when the cells begin to fall off and float, slightly blowing to form single cell suspension by using a pipette, centrifuging at 1000rpm for 10min, removing supernatant, and finally re-suspending with 10mL of fresh stem cell culture medium to obtain the primary umbilical cord mesenchymal stem cells (P)0) Fig. 2 shows the microscope observation picture of the primary umbilical cord mesenchymal stem cells.
2. Identification of umbilical cord mesenchymal stem cells
Obtaining primary stem cell P0Then, sterile PBS buffer was added to adjust the cell concentration to 1X 106and/mL, taking five 200 mu L cell suspensions, respectively adding 5 mu L of fluorescently-labeled monoclonal antibody CD34, 5 mu L of fluorescently-labeled monoclonal antibody CD45, 5 mu L of fluorescently-labeled monoclonal antibody CD73, 5 mu L of fluorescently-labeled monoclonal antibody CD90 and 5 mu L of fluorescently-labeled monoclonal antibody CD105, mixing uniformly, incubating at room temperature in the dark for 20min, and detecting in a flow cytometer, wherein the flow detection result graphs of umbilical cord mesenchymal stem cell specific proteins CD34, CD45, CD73, CD90 and CD105 are respectively shown in figures 3, 4, 5, 6 and 7.
3. Culture of umbilical cord mesenchymal stem cells
Placing the umbilical cord mesenchymal stem cells into a stem cell culture medium for culture, wherein the culture conditions are as follows: containing 5% CO2And when the temperature is 37 ℃ and the adherent cell is fully paved by 90 percent, centrifuging and collecting the cell culture supernatant.
4. Pretreatment of umbilical cord mesenchymal stem cell culture supernatant
Adding the cell culture supernatant into a centrifuge tube, centrifuging for 5-50min at 12000g of 5000-.
Example 2 extraction, purification and characterization of exosomes
1. Double tangential flow filtration method for extracting exosome
(1) Adding sterile water into the first storage tank 1 and the second storage tank 4, respectively opening the peristaltic pumps (the first peristaltic pump 2 and the second peristaltic pump 6) and flushing the system;
(2) adding 4L of pretreated sample solution into a first storage tank 1, opening a first peristaltic pump 2 to circulate the system, concentrating and filtering the pretreated sample solution by a first filtering device 3 provided with a tangential flow filter membrane F-1, and enabling the solution containing exosomes to enter a second storage tank 4; when the liquid volume in the first storage tank 1 is the minimum operation volume, 2L of sterile PBS buffer solution is added to continue concentration and filtration to the minimum operation volume;
(3) when the liquid volume in the second storage tank 4 exceeds 400mL, the second peristaltic pump 6 is started to circulate the system, and the solution containing the exosome passes through a second filtering device 5 provided with a tangential flow filter membrane F-2 to be concentrated and filtered, so that small molecular substances are removed;
(4) and when the liquid volume in the second storage tank 4 is the minimum operation volume, adding 2000mL of PBS buffer solution into the second storage tank 4, continuing to concentrate and filter until the solution in the second storage tank 4 is 200mL, and collecting the solution to obtain the exosome extracted and purified by the dual tangential flow filtration system.
2. Ultracentrifugation method for extracting exosome
And adding the pretreated sample into an ultracentrifuge tube, centrifuging for 2h at 100000g at 4 ℃, and carrying out heavy suspension by using 1mL of eluent to obtain the ultracentrifuge separation and purification exosome.
3. Identification of exosomes
The exosomes after collection were passed through an electron microscope as shown in fig. 8; a nanoparticle size tracer analyzer, as shown in fig. 9; the specific protein CD9, as shown in figure 10; the specific protein CD63 is shown in figure 11; the specific protein ALIX, shown in FIG. 12, identifies exosomes.
Example 3 cell function experiment
1. Pulmonary epithelial cell repair
Preparing human normal lung epithelial cells (BEAS-2B) into cell suspension at 5 × 103The density of each well is inoculated on a 96-well plate, the plate is put into an incubator to be cultured for 24 hours, the culture solution is replaced by stem cell culture medium containing 250 mu mol/L hydrogen peroxide to culture cells for 5 hours, after the culture is finished, the cell culture medium is replaced, and 10 mu L of umbilical cord mesenchymal stem cell exosomes (1 multiplied by 10) with different concentrations are added into the culture medium6-32×106particles/mL), with 10. mu.L of PBS buffer as a negative control, incubation was continued for 24h, adding 20. mu.L of MTT solution (5mg/mL) per well. Incubation was continued for 4h, the culture was terminated and the culture supernatant from the wells was carefully aspirated. Add 150. mu.L DMSO per well and shake for 10 min. OD values were measured at 450nm with a microplate reader, 6 replicates per group, and the average was taken.
Experimental results FIG. 13 shows that compared with the control group, the exosomes extracted and purified by the double TFF method and the ultracentrifugation method can remarkably promote the proliferation of the normal lung epithelial cells (BEAS-2B) of the human, the activity is better along with the increase of the exosome concentration, and the optimal concentration of the exosomes is 8 × 106particles/mL, from the figureIt was also observed that exosomes extracted by the double TFF method had a better proliferation effect on human normal lung epithelial cells (BEAS-2B) than the ultracentrifugation method.
2. Inhibition of inflammatory factors of lung epithelial cells
Preparing human normal lung epithelial cells (BEAS-2B) into cell suspension at 5 × 103The density of each well is inoculated on a 96-well plate, the plate is put into an incubator to be cultured for 24 hours, and 10 mu L of umbilical cord mesenchymal stem cell exosome (1 multiplied by 10) with different concentrations is added at the same time when the culture solution is replaced by cell culture medium containing 10ug/mL LPS6-32×106particles/mL) was cultured for 12 hours with 10. mu.L of PBS as a negative control, and after completion of the culture, the culture supernatant was collected and the expression levels of IL-6, IL-8 and TNF-. alpha.in the culture were measured by ELISA kit.
As shown in FIGS. 14, 15 and 16, the umbilical cord mesenchymal stem cell exosomes can inhibit inflammatory factors IL-6, IL-8 and TNF-alpha secreted by lung epithelial cells (BEAS-2B) induced by LPS, and the activity of inhibiting the inflammatory factors is better as the exosome concentration is increased, and the optimal concentration of exosomes is 8 × 106particles/mL, and the exosomes extracted by the double TFF method are slightly superior to those extracted by the ultracentrifugation method. The experiment can prove that the umbilical cord mesenchymal stem cell exosome has the capacity of down-regulating the lung epithelial cell to secrete inflammatory factors, and can be prepared into a biological preparation for relieving asthma.
3. Human embryonic lung fibroblast inhibition
Taking human embryonic lung fibroblast (MRC-5), digesting and resuspending to prepare the product with the density of 5 multiplied by 103Inoculating the strain in a 96-well plate, culturing in an incubator for 24h, changing the cell culture medium, and adding 10 μ L of umbilical cord mesenchymal stem cell exosomes (1 × 10) with different concentrations into the culture medium6-32×106particles/mL), 10. mu.L of PBS buffer was used as a negative control, and after further incubation for 24h, 20. mu.L of MTT solution (5mg/mL) was added to each well. Incubation was continued for 4h, the culture was terminated and the culture supernatant from the wells was carefully aspirated. Add 150. mu.L DMSO per well and shake for 10 min. Detecting OD value at 450nm with enzyme labeling instrument, repeating each group for 6 times, averaging, and calculating proliferation inhibition rate. Proliferation inhibition (%) [ 1- (OD biologics-OD blank)/(O)D control-OD blank group)]×100％。
Experimental result figure 17 shows that umbilical cord mesenchymal stem cell exosome can obviously inhibit proliferation of human embryonic lung fibroblast (MRC-5) and has obvious dose-dependent effect, wherein the optimal exosome concentration is 8.0 multiplied by 106particles/mL. The purified exosome extracted by the dual TFF method and the ultracentrifugation method has the effect of inhibiting the proliferation of MRC-5 cells, and the effect of inhibiting the proliferation of the MRC-5 cells by the exosome extracted and purified by the dual TFF method is better than that of the ultracentrifugation method, so that the strong centrifugal force of the ultracentrifugation method possibly destroys part of the membrane structure of the exosome, and finally influences the biological function of the exosome, and the mode of extracting the exosome by the dual TFF method is milder, so that the biological activity of the exosome is better ensured.
4. Expression of apoptosis proteins Bcl-2 and Bax in human embryonic lung fibroblasts (MRC-5)
The proteins Bcl-2 and Bax are the most important cytokines for regulating and controlling MRC-5 cell apoptosis, Bcl-2 can inhibit MRC-5 cell apoptosis, and Bax promotes MRC-5 cell apoptosis.
Taking human embryonic lung fibroblast (MRC-5), digesting and resuspending to prepare the product with the density of 5 multiplied by 103Inoculating the strain in a 96-well plate, culturing in an incubator for 24h, changing the cell culture medium, and adding 10 μ L of umbilical cord mesenchymal stem cell exosomes (1 × 10) with different concentrations into the culture medium6-32×106particles/mL), using 10 mu L of PBS buffer solution as a negative control, continuously culturing for 24h, collecting cells, adding lysis solution for lysis, adding a proper amount of loading buffer into each collected protein sample, and heating in a boiling water bath to fully denature the protein. Performing SDS-PAGE electrophoresis on a denatured protein sample, transferring a membrane, sealing by 5% of sealing solution, incubating overnight at 4 ℃ by using primary antibodies (Bcl-2 and Bax) prepared in a proper proportion, incubating for 1h at room temperature by using prepared enzyme-labeled secondary antibodies in a proper proportion, placing the protein membrane in ECL luminescent liquid for reaction for 2min, and placing the protein membrane in a chemiluminescence imaging system for chromogenic imaging.
The experimental result is shown in figure 18, and the umbilical cord mesenchymal stem cell exosome has a more obvious influence on the expression of apoptosis proteins Bcl-2 and Bax in human embryonic lung fibroblasts (MRC-5). The exosomes extracted by the two methods can be found to be capable of remarkably reducing Bcl-2 protein expression and up-regulating Bax protein expression, and the figure shows that the exosomes extracted by the double TFF method have slightly stronger influence on the Bcl-2 and Bax protein expression level than the ultracentrifugation method, and the conclusion is consistent with the experimental result of the biological activity of the exosomes.
The exosome can be used for treating pulmonary fibrosis diseases in various modes such as aerosol, spray, oral preparations, injection preparations and the like, and can also be prepared into a biological preparation in modes such as drug loading, surface modification or excipient and the like, so that the toxic and side effects of the biological preparation can be greatly reduced in the process of treating asthma and/or pulmonary fibrosis, and the problems of irritation of the traditional biological preparation to gastrointestinal tract, skin allergy, damage to liver function and the like are avoided.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (9)
1. Application of cell-derived exosome in preparation of biological preparation for treating asthma and/or pulmonary fibrosis is provided.
2. Use of the stem cell-derived exosome according to claim 1 in the preparation of a biological formulation for treating asthma and/or pulmonary fibrosis, wherein the dosage form of the biological formulation comprises an aerosol, a spray, an oral formulation or an injectable formulation.
3. Use of stem cell-derived exosomes according to claim 1 in the preparation of a biological preparation for the treatment of asthma and/or pulmonary fibrosis, wherein the exosomes are capable of being used for drug loading, surface modification or excipient.
4. A cell-derived exosome for use in the preparation of a biological preparation for the treatment of asthma and/or pulmonary fibrosis, characterised in that the exosome is extracted by a method comprising ultracentrifugation, density gradient centrifugation, ultrafiltration, polymer precipitation, tangential flow filtration, immunocapture, affinity chromatography, microfluidics or size exclusion.
5. The cell-derived exosome for use in the preparation of a biological agent for the treatment of asthma and/or pulmonary fibrosis according to claim 4, characterised in that it has an effect of promoting lung epithelial cell repair.
6. The cell-derived exosome for use in preparing a biological agent for treating asthma and/or pulmonary fibrosis according to claim 4, wherein said exosome is capable of inhibiting inflammatory factors produced by lung epithelial cells induced by LPS.
7. The cell-derived exosome for use in preparing a biological agent for treating asthma and/or pulmonary fibrosis according to claim 4, wherein the exosome is capable of inhibiting proliferation of human embryonic lung fibroblasts by modulating expression of apoptotic proteins.
8. The exosome for preparing a cell-derived exosome for treating asthma and/or pulmonary fibrosis biological agent according to claim 4, wherein the cell-derived exosome is derived from plants, microorganisms, animals and humans, including body fluids, tissue fluids, somatic cells, stem cells and cells induced to differentiate by stem cells.
9. The cell-derived exosome for use in the preparation of a biological agent for treating asthma and/or pulmonary fibrosis according to claim 4, wherein the exosome has potential for treating asthma and/or pulmonary fibrosis and can be used in the preparation of a biological agent for treating asthma and/or pulmonary fibrosis.
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