KR20200115422A - Pulmonary fibrosis mouse model without GOLGA2 gene - Google Patents

Abstract

The present invention relates to a lung fibrosis mouse model in which a GOLGA2 gene is knocked out, a composition for manufacturing a lung fibrosis mouse model, a method for manufacturing a lung fibrosis mouse model, and a method for inhibiting lung fibrosis or screening a therapeutic agent. When the lung fibrosis mouse model in which a GOLGA2 gene is knocked out according to the present invention is used, a lung fibrosis model which did not exist can be obtained, and a model in which fibrosis is induced in an entire area of a lung due to genetic mutation without a serious lesion of lung tissue can be used. The model can be used in various ways as a model for study of association among abnormal morphology of a Golgi body, autophagocytosis, and liver fibrosis. The model can be used as an animal model for screening the therapeutic agent of genetic-based lung fibrosis rather than chemical-based lung fibrosis. The composition for manufacturing a lung fibrosis mouse model comprises embryonic stem cells in which exons 1-26 of the GOLGA2 gene are deleted.

Description

GOLGA2 gene knockout lung fibrosis mouse model {Pulmonary fibrosis mouse model without GOLGA2 gene}

The present invention relates to a lung fibrosis mouse model in which the GOLGA2 gene is knocked out, a composition for preparing the lung fibrosis mouse model, a method for preparing a lung fibrosis mouse model, and a screening method for suppressing lung fibrosis or a therapeutic agent.

Autophagy or self-eating is a process in which proteins, lipids and organelles in cells that are not needed or damaged are removed by lysosomes. It is playing an important role as a recycling system that supplies energy to maintain homeostasis and establishes the foundation for new organisations. In relation to this autophagy, the Golgi apparatus is known to provide a double sequestering membrane (phagophore) that is generated at the initial stage of assembly of autophagosomes.

On the other hand, pulmonary fibrosis is a disease of unknown cause characterized by chronically progressive fibrosis of the lung interstitial. The disease is mainly confined to the lungs and exhibits histologically characteristic normal interstitial pneumonia (UIP) findings. Although the prevalence of this disease varies from report to report, it is known to be 2-29 out of 100,000 population, and it is designated as a rare and intractable disease in Korea. The clinical course of pulmonary fibrosis is diverse, and it is a fatal disease with an average survival period of less than 2-3 years after diagnosis due to respiratory failure, which generally progresses to respiratory failure due to a slow progression of lung function. For this reason, many efforts have been made to identify the exact cause and etiology of pulmonary fibrosis and to develop a therapeutic agent according to it, but there is no clear treatment yet.

There is no existing model that caused lung fibrosis, and there are models for other tissues, such as liver fibrosis, but this is a chemical-based fibrosis model, which has a problem in that it causes serious lesions in the tissue area that the chemical touches. In addition, the relationship between the Golgi apparatus, autophagy and pulmonary fibrosis, and which genes are involved in these phenomena, has not been identified, and specific studies are required.

Therefore, the present inventors completed the present invention by confirming that the GOLGA2 gene knockout mouse present in the cis-Golgi apparatus, which is the formation surface of the Golgi apparatus, causes the collapse of the Golgi apparatus, induces autophagy, and causes lung fibrosis. Was done.

Accordingly, an object of the present invention is to provide a GOLGA2 gene knocked out, lung fibrotic mouse model, a composition for preparing the lung fibrotic mouse model, a method for preparing the lung fibrotic mouse model, and a method for screening lung fibrosis inhibition or a therapeutic agent.

In order to achieve the above object, the present invention provides a lung fibrosis mouse model in which the GOLGA2 gene is knocked out.

In addition, the present invention provides a composition for preparing a lung fibrosis mouse model, comprising embryonic stem cells in which GOLGA2 gene exons 1 to 26 are deleted.

In addition, the present invention provides a method for producing a lung fibrosis mouse model comprising; knocking out the GOLGA2 gene.

In addition, the present invention comprises the steps of (S1) treating a candidate material in the lung fibrotic mouse model; And (S2) determining a substance to inhibit or treat pulmonary fibrosis in a lung fibrosis mouse model treated with the candidate substance; comprising, a method for screening a pulmonary fibrosis inhibitory or therapeutic agent.

Using the GOLGA2 gene knockout lung fibrosis mouse model according to the present invention, a lung fibrosis model that did not exist can be obtained, and a model in which fibrosis is induced in the entire region of the lung due to genetic mutation without serious lesion of the lung tissue It has the advantage of being able to use it. In addition, the model can be used in various ways as a model for the study of the association between abnormal morphology of the Golgi apparatus, autophagy, and pulmonary fibrosis. Can be used as an animal model.

1 is a diagram showing the location of the GOLGA2 gene and a schematic diagram of the embryonic stem cells used to make a GOLGA2 knockout mouse.
2 is a diagram showing the results of confirming that the GOLGA2 protein is removed from Golgi by GOLGA2 knockout in a hepatocyte cell line.
3 is a diagram showing the results confirming that the increased autophagy action in the GOLGA2 knockout hepatocyte line decomposes fat droplets formed by free fatty acid (FFA).
Figure 4 is GOLGA2 A diagram showing the results confirming that the expression of the autophagy marker proteins SQSTM1, LC3I and LC3ll is increased in the knockout liver cell line.
5 is a diagram showing the results of genotyping of normal, heterozygous and GOLGA2 knockout mice.
Figure 6 is GOLGA2 A diagram showing the results of selecting GOLGA2 knockout mice through the analysis of the expression of GOLGA2 mRNA in the tissues of the knockout mice.
7 is a normal, heterozygous and GOLGA2 It is a diagram showing a picture comparing the size of a knockout mouse.
8 is a normal, heterozygous and GOLGA2 It is a diagram showing the size comparison graph of the knockout mouse.
9 is a normal, heterozygous and GOLGA2 It is a diagram showing a weight comparison graph of a knockout mouse.
Figure 10 is male GOLGA2 It is a diagram showing the results of confirming the decrease in organ size of the knockout mouse.
11 is GOLGA2 It is a diagram showing the results of confirming that LC3B, an autophagy marker protein, is highly activated in specific cells of the lung through immunofluorescence staining analysis of knockout mouse lung tissue.
12 is a diagram showing the result of activation of the autophagy marker protein LC3B in type 2 alveolar cells of GOLGA2 knockout mice.
13 is GOLGA2 A diagram showing the result of confirming that the number of discharged lamellar bodies increases through electron microscopic image analysis of knockout mouse lungs.
14 is a diagram showing the results of confirming that there is a lack of surfactant that maintains the alveoli in the lungs of GOLGA2 knockout mice through lipid scanning analysis of the lungs using MALDI-TOF.
15 is a diagram showing the results of confirming that a fibrotic lesion was found in the entire lung through histological analysis of the lungs of GOLGA2 knockout mice.
16 is a diagram showing the mechanism of manufacturing a lung tissue fibrosis mouse model by GOLGA2 gene knockout.

The present invention provides a lung fibrosis mouse model in which the GOLGA2 gene is knocked out.

The GOLGA2 gene of the present invention is a gene of NCBI Gene ID 99412 and is a gene belonging to chromosome 2 and exists in various species. Specifically, the GOLGA2 gene may be a polynucleotide consisting of the nucleotide sequence of NCBI Reference Sequence NM_133852. In addition, the GOLGA2 gene is a gene present in the cis-Golgi body, which is toward the formation surface of the Golgi body.

The mouse model may be a GOLGA2 protein knocked out in the Golgi apparatus. This causes the Golgi apparatus to collapse and induce autophagy. In addition, the mouse model may have an increased autophagy action in the lung. The mouse model may have increased expression of LC3B, an autophagy marker protein, in the lung.

In addition, the lung may be type 2 alveolar cells, and type 2 alveolar cells are cells that secrete lamellar bodies. In the GOLGA2 gene knockout mouse model of the present invention, the lamellar body of the type 2 peptic cells is discharged, and the lamellar body contains a surfactant as a main component, and the surfactant serves to maintain the alveoli.

In addition, in the mouse model, dipalmitoylphosphatidylcholine (DPPC) may be reduced in the lung, preferably dipalmitoyllecithin may be reduced. The DPPC is a surfactant, which is a main component of the lamellar body, and when DPPC decreases due to the discharge of the lamellar body of the type 2 fecal cell, a problem of irreversibly clogging the alveoli occurs, which may be the cause of lung fibrosis.

The mouse model may be made of embryonic stem cells in which exons 1 to 26 of the GOLGA2 gene are deleted, but is not limited thereto. In addition, the embryonic stem cells may be injected into the blastocyst of a mouse.

The GOLGA2 gene knockout mouse model of the present invention is a lung fibrosis model that did not exist in the past, and through the mouse model of the present invention, GOLGA2 protein is knocked out from the Golgi apparatus by GOLGA2 knockout, and autophagy is increased in the lung, and lung fibrosis It can be confirmed that is triggered. In addition, when the GOLGA2 gene knockout lung fibrosis mouse model according to the present invention is used, there is an advantage in that a model in which fibrosis is induced in the entire region of the lung due to genetic mutation can be used without serious lesion of the lung tissue. In addition, the model can be used in various ways as a model for the study of the association between abnormal morphology of the Golgi apparatus, autophagy, and pulmonary fibrosis. Can be used as an animal model.

In addition, the present invention provides a composition for preparing a lung fibrosis mouse model, comprising embryonic stem cells in which GOLGA2 gene exons 1 to 26 are deleted. The composition may preferably be a medium composition, and the "culture media" refers to a medium capable of supporting cell growth and survival in vitro, and is suitable for culturing cells. Includes all of the media. The medium may contain essential and non-essential amino acids, vitamins, energy sources, lipids, and trace elements necessary for minimal proliferation and/or survival of cells.

In addition, the present invention provides a method for producing a lung fibrosis mouse model comprising; knocking out the GOLGA2 gene. The knockout may be performed by a conventional method known in the art, and preferably may be performed using a GOLGA2 CRISPR/ Cas9 system, but is not limited thereto. When the lung fibrosis mouse model is prepared through the step of knocking out the GOLGA2 gene, a lung fibrosis model in which fibrosis is induced in the entire lung area due to genetic mutation can be obtained without serious lesion of the lung tissue. In addition, it is possible to obtain a genetically based lung fibrosis model rather than a chemical based lung fibrosis.

In addition, the present invention comprises the steps of (S1) treating a candidate material in the lung fibrotic mouse model; And (S2) determining a substance to inhibit or treat pulmonary fibrosis in a lung fibrosis mouse model treated with the candidate substance; comprising, a method for screening a pulmonary fibrosis inhibitory or therapeutic agent.

The step (S1) is a step of treating a candidate material in a lung fibrosis mouse model knocked out of the GOLGA2 gene of the present invention. The "candidate" is a substance expected to treat, prevent or improve lung fibrosis, and is a chemical substance, oligonucleotide, peptide, gene, protein, food, formulation, compound, composition, medicine, nutritional supplement, health care product Or it includes without limitation substances such as beverages. In addition, the "treatment" may be a method of administering a candidate substance to the lung fibrotic mouse model of the present invention, but is not limited thereto. As the "administration method", an appropriate method may be selected from conventional administration methods such as oral administration, intravenous injection, subcutaneous administration, intraperitoneal administration, etc., and the dosage of the candidate substance is the administration method, the weight and condition of the experimental animal, or prior It can be appropriately selected according to the experimental results, but is not limited thereto.

The step (S2) is a step of selecting a substance that has been confirmed to improve lung fibrosis among the candidate substances treated in step (S1) to determine as a substance to inhibit or treat lung fibrosis. The "selection" is a result of comparing the result of measuring the degree of lung fibrosis of the mouse model of lung fibrosis of the present invention before treatment of the candidate substance and the measurement result after treatment of the candidate substance. This can be achieved by selecting a candidate material that has reduced.

By using the method of screening for pulmonary fibrosis suppression or treatment using the lung fibrosis mouse model knocked out of the GOLGA2 gene of the present invention, it is possible to screen the treatment using a model in which fibrosis is induced in the entire region of the liver due to genetic mutation. There is an advantage that there is. In addition, it is possible to screen for therapeutic agents for genetic-based lung fibrosis rather than chemical-based lung fibrosis.

Hereinafter, the present invention will be described in more detail based on examples. The examples are only for describing the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the invention is not limited by these examples according to the gist of the present invention.

Statistics of each experiment in the examples were used as ± S.D, and were recorded as p <0.05 (* or #), p <0.01 (** or ##).

Experimental example 1. Liver cell line culture and GOLGA2 Knockout Cell line production

The Chang cell line, which is a normal human hepatocyte line, was cultured in a 37°C, 5% CO ₂ incubator with Dulbecco's modified Eagle medium/high-glucose medium (WELGENE Inc.), fetal bovine serum 10%, antibiotics 10%, and Heps 20% added.

In addition, the GOLGA2 knocked out Chang cell line was constructed with the GOLGA2 CRISPR/ Cas9 system.

Experimental example 2. GOLGA2 gene Knockout Mouse production

GOLGA2 1-26 exon-deleted embryonic stem cells were purchased from the Wellcome Trust Sanger Institute and injected into the blastocyst of C57B/L6J-type mice to obtain heterozygous offspring.Through genotyping analysis of offspring through inbreeding of the heterozygous mice. GOLGA2 knockout mice were selected.

PCR primers used for the genotyping analysis are as follows. GOLGA2 KO-F (5'-CTTCGTATAATGTATGCTATACGAAGTTATGCTAGC-3') and Reg- GOLGA2 -R(5'-ACTGAGCCGGTGAAAACTTAGAAGC-3') were used to check GOLGA2 (-/-) knockout, and Mus GOLGA2 Exon 27 wt-F (5'-AAGACTGGCGGCCAAAGCC-3') and Reg- GOLGA2 -R were used to identify GOLGA2 (+/+).

Figure 1 shows the location of the GOLGA2 gene and a schematic diagram of the embryonic stem cells used to make the GOLGA2 knockout mouse.As shown in Figure 1, the GOLGA2 gene belongs to the chromosome 2 and GOLGA2 exon 1-26 of the used embryonic stem cells It was confirmed that the bun was deleted.

Experimental example 3. Immunity Fluorescence method ( Immunofluorescence ) analysis

The Chang cell line was cultured on an 8-well chamber slide for culture, and then fixed with 3% paraformaldehyde at 4° C. for 10 minutes. Then, 0.25% Triton® X-100 was treated for 5 minutes, followed by blocking treatment with 3% bovine serum albumin for 30 minutes, and the primary antibody was treated at 4°C for one day, and then the fluorescently labeled secondary antibody was treated. I did. After that, the liver tissue was sectioned with 4 μm of paraffin block and transferred to the slide. After removing the paraffin with xylene, hydration, boiled and cooled with retrival buffer for 10 minutes to save antigen-antibody reaction. Analyzed by.

Experimental example 4. Mass Spectrometry

Lung mass analysis of the GOLGA2 gene knockout mice prepared in Experimental Example 2 was performed using MALDI-TOF.

Experimental example 5. Immunoblot (Western Blot) analysis

For immunoblot analysis, the first cultured Chang cell line was applied ³ ×10 ^{5 3} wells to a ⁶ well plate, and the media was removed after 24 hours of culture. After washing with 1X PBS, 20 mM Tris-HCl (pH 7.5), 250 mM NaCl, 3 mM EDTA, 3 mM EGTA, 0.5% NP40, 10 mM beta-glycerophosphate, 50 mM fluorine Cells were lysed with a lysis buffer made of sodium fluoride, 0.2 mM sodium vanadate, 10 mM PNPP, 5 mM PMSF, and 10 μg/ml leupeptin. After measuring the protein concentration of the dissolved protein by Bradford protein assay (Bio-Rad, Hercules, CA), 40ug was quantified, mixed with Laemmli sample buffer according to the concentration, boiled, loaded on SDS-PAGE, and transferred. blocking) and TBS-T washes, and then the primary antibody was added and reacted overnight. Thereafter, after washing with TBS-T the next day, the rabbit secondary antibody was incubated for 1 hour, washed with TBS-T 3 times for 20 minutes, and developed into a film. Mouse lung tissue was frozen at ultra-low temperature, crushed, and then obtained and analyzed in the same manner as above.

Experimental example 6. Histology and immunohistochemistry ( Immunohistochemistry ) Characteristic analysis

The lung tissue of the GOLGA2 gene knockout mouse was fixed in 10% formalin and then paraffin-embedded, and the tissue block was sectioned with 4 μm and transferred to a silane-coated slide. Tissue slides from which paraffin was removed for histological analysis were stained with hematoxylin & eosin (H&E) or with Sirius Red reagent to observe collagen deposition. In addition, for immunohistochemistry analysis, paraffin-removed tissue sections were boiled in retrival buffer, treated with 5% hydrogen peroxide for 15 minutes to remove endogenous peroxidase, and washed with TBS buffer containing tween 20. And blocked for 1 hour. Subsequently, after treatment with the first and second antibodies, they were washed again, colored with a DAB substrate solution, and quickly washed in sterilized water. For counter staining, it was stained with hematoxylin. After that, it was observed under a tissue microscope through alcoholation and mounting.

Experimental example 7. Transmission electron microscope TEM ) analysis

For TEM analysis, lung tissue was fixed overnight at 4°C with 2.5% glutaraldehyde buffer containing 1% osmium tetroxide, followed by alcohol treatment, and propylene oxide epon resin was applied to the tissue. After infiltrating and hardening overnight at 70° C., ultra-thin sliced at 40 nm, placed on a copper grid, and observed using JEM1010 TEM (JEOL).

Example 1. In liver cell lines GOLGA2 Knockout Autophagy Confirm that it causes action

Experimental Example In the GOLGA2 between knockout cell line prepared in 1 to determine the effect of the knockout on GOLGA2 was subjected to immunofluorescence (Immunofluorescence) analysis were also shown in 2 the results for it.

As shown in FIG. 2, when the GOLGA2 gene was knocked out in the liver cell line, it was confirmed that the GOLGA2 protein was removed from the Golgi, thereby confirming the GOLGA2 protein knockout in the Golgi apparatus in the GOLGA2 knockout cell line. Blue represents the nucleus.

In addition, in order to confirm that autophagy is induced in the GOLGA2 knockout liver cell line, the normal hepatocyte cell line (Chang) and the GOLGA2 knockout liver cell line were treated with 250uM free fatty acid (FFA) for 24 hours and then triglycerides ( neutral lipid) staining was used to confirm the fat droplets, and the results are shown in FIG. 3. In FIG. 3, blue represents a nucleus.

As shown in FIG. 3, it was confirmed that fat droplets formed by FFA were degraded due to an increase in autophagy in the GOLGA2 knockout liver cell line. Therefore, it was confirmed that the size of fat droplets decreased due to an increase in autophagy in the GOLGA2 knockout hepatocyte line.

In addition, in order to confirm the expression of autophagy marker proteins SQSTM1, LC3I and LC3ll in the GOLGA2 knockout liver cell line, an immunoblot analysis was performed and the results are shown in FIG. 4.

As shown in FIG. 4, it was confirmed that SQSTM1, LC3I and LC3II, known as autophagy marker proteins, were increased in the GOLGA2 knockout liver cell line.

Therefore, it was confirmed through the above results that when the GOLGA2 gene was knocked out, the GOLGA2 protein was knocked out in the Golgi apparatus and autophagy increased.

Example 2. GOLGA2 gene Knockout Mouse characterization

In order to examine the characteristics of the GOLGA2 gene knockout mouse produced in Experimental Example 2, the following analysis was performed.

* Example 2-1. Genotyping analysis

A polymerase chain reaction (PCR) was performed to select the GOLGA2 gene knockout mice prepared in Experimental Example 2, and the genotypic characteristics of normal, heterozygous and GOLGA2 knockout mice were analyzed through PCR. Indicated.

As shown in FIG. 5, as a result of confirming GOLGA2 knockout mice through PCR, mice in which only the GOLGA2 KO band was observed were selected as GOLGA2 knockout mice.

Example 2-2. mRNA Expression analysis

In order to confirm that all of the GOLGA2 genes were knocked out in the tissues of the selected GOLGA2 knockout mice, mRNA expression was analyzed and the results are shown in FIG. 6.

As shown in Fig. 6, it was confirmed that GOLGA2 was completely knocked out in all organs.

Example 2-3. Mouse size comparison analysis

Pictures and graphs are shown in Figs. 7 and 8, respectively, by comparing the sizes of normal, heterozygous and GOLGA2 knockout mice.

As shown in FIG. 7, it was confirmed that the size of the GOLGA2 knockout mouse was smaller than that of the normal mouse, especially in males.

In addition, as shown in the graph of FIG. 8, it was confirmed that only GOLGA2 knockout mice significantly decreased in size compared to normal regardless of male and female (n=3).

Example 2-4. Mouse weight comparison analysis

The weights of normal, heterozygous and GOLGA2 knockout mice were measured, and a comparative graph is shown in FIG. 9.

As shown in FIG. 9, it was confirmed that the females did not change the weight of each group, and the weights of the heterozygous and GOLGA2 knockout mice in males significantly decreased compared to normal mice (n=3).

Example 2-5. Knockout Organ size analysis in mice

The organ sizes of the GOLGA2 knockout mice are compared and shown in FIG. 10.

As shown in FIG. 10, it was confirmed that the organ size of the male GOLGA2 knockout mouse was reduced compared to that of the male normal mouse.

Example 3. GOLGA2 Knockout In the lungs of a mouse GOLGA2 Knockout Autophagy Confirmation of causing action

Immunofluorescence staining analysis was performed to confirm whether the GOLGA2 knockout induces autophagy in the actual lung through the GOLGA2 gene knockout mouse prepared in Experimental Example 2, and the results are shown in FIG. 11.

As shown in FIG. 11, it was confirmed that the autophagy marker protein LC3B staining in the lung tissue of GOLGA2 knockout mice was highly activated in specific cells of the lung. In addition, the number was found to be less than 10% of the total lung, and it was confirmed that these cells were highly likely to be alveolar macrophages or type 2 alveolar cells.

In addition, immunofluorescence staining analysis was performed to confirm the activation of autophagy in type 2 alveolar cells of GOLGA2 knockout mice, and the results are shown in FIG. 12.

As shown in Fig. 12, proSP-C is a marker protein of type 2 alveolar cells, and LC3B, an autophagy marker protein, was confirmed to be activated in type 2 alveolar cells of knockout mice.

In addition, the results of electron microscopy image analysis of the lungs of GOLGA2 knockout mice are shown in FIG. 13.

As shown in FIG. 13, it was confirmed that type 2 alveolar cells secrete lamellar vesicles, but in type 2 alveolar cells of GOLGA2 knockout mice, lamellar vesicles were discharged and the number of discharged lamellar vesicles increased. This lamellar body is mainly composed of DPPC (dipalmitoylphosphatidylcholine), which is the main component of alveolar surfactant, and it was confirmed that the possibility of abnormality in the surfactant system was increased through this.

In addition, the results of lipid scanning analysis were shown in FIG. 14 by performing mass analysis of mouse lungs using MALDI-TOF.

As shown in FIG. 14, it was confirmed that the lipid of PC(32:0) + Na was significantly reduced in knockout mice in the lung imaging analysis using a mass spectrometer. This lipid is DPPC, which is the main component of surfactant, and it was confirmed that there is a lack of surfactant that maintains alveoli in the lungs of GOLGA2 knockout mice.

Therefore, it was confirmed through the above results that GOLGA2 knockout activates autophagy in the lungs of GOLGA2 knockout mice and lacks a surfactant that maintains alveoli in type 2 alveolar cells.

Example 4. GOLGA2 Knockout Through the mouse In pulmonary fibrosis GOLGA2 Knockout Check the impact

Histological analysis was performed to confirm how the GOLGA2 knockout affects lung fibrosis in the actual lung through the GOLGA2 gene knockout mouse prepared in Experimental Example 2, and the results are shown in FIG. 15.

As shown in FIG. 15, it was confirmed that a number of macrophages (Macrophage, black arrowhead in FIG. 15) were deposited in the alveoli of GOLGA2 knockout mice during H&E staining, and that a fibrotic lesion was found in Sirius red staining (FIG. 15) Red arrow).

Therefore, through the above results, it was confirmed that GOLGA2 knockout activates autophagy and induces lung fibrosis in the lungs of GOLGA2 knockout mice.

Example 5. GOLGA2 gene Knocked out Lung tissue Establish a fibrotic mouse model

Through the above examples, it was confirmed that the deletion of the GOLGA2 gene, a Golgi body protein, induces autophagy, and in particular, decomposes intracellular lipids by autophagy in type 2 pulmonary cells containing many lipids. In addition, it was confirmed that the secretion of surfactant was inhibited in type 2 alveolar cells, causing fibrosis of lung tissue. Therefore, through this, finally established a lung tissue fibrosis mouse model knocked out the GOLGA2 gene, which is shown in FIG. 16.

Claims (6)

A composition for preparing a lung fibrosis mouse model comprising embryonic stem cells in which GOLGA2 gene exons 1 to 26 are deleted.
The method of claim 1,
The composition for preparing the mouse model,
A composition for producing a lung fibrotic mouse model, characterized in that knocking out the GOLGA2 protein in the Golgi apparatus of the mouse model.
The method of claim 1,
The composition for preparing the mouse model,
A composition for preparing a lung fibrotic mouse model, characterized in that to enhance the autophagy action in the lung of the mouse model.
The method of claim 3,
The lungs,
A composition for producing a lung fibrotic mouse model, characterized in that the alveolar cells.
The method of claim 1,
The composition for preparing the mouse model,
A composition for preparing a lung fibrotic mouse model, characterized in that to enhance the expression of the LC3B protein in the lung of the mouse model.
The method of claim 1,
The composition for preparing the mouse model,
A composition for preparing a lung fibrosis mouse model, characterized in that reducing DPPC (dipalmitoylphosphatidylcholine) in the lung of a mouse model.

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