Construction method and application of coal silicosis mouse model

Abstract

The invention belongs to the technical field of medical animal model construction, and discloses a coal silicosis mouse model construction method and application. According to the method, a coal-silicon mixed dust suspension containing 15%, 25% and 50% of free silicon dioxide is prepared, and the concentration is 50 mg / ml; the method comprises the following steps: hanging an anesthetized healthy adult (6-8 weeks) C57BL / 6N male mouse on a fixing device, instilling 50 microliters of coal and silicon mixed dust suspension once per week by adopting an oropharynx inhalation method for 4 weeks continuously, and establishing a coal silicosis model; applying equal-volume normal saline to the control group; respectively observing for 28 days, 56 days and 112 days, taking mouse lung tissues for pathological analysis and fibrosis index detection, and evaluating the model preparation effect. The method is simple to operate and low in cost, and the established coal silica dust mouse model is stable, efficient, repeatable and noninvasive.

Description

Technical Field

[0001] The present invention belongs to the technical field of constructing medical animal models, and particularly relates to a method for constructing a coal workers' pneumoconiosis mouse model and its application. Background Art

[0002] Coal workers' pneumoconiosis (CWP) is a disease mainly characterized by pulmonary fibrosis caused by long-term inhalation of respirable coal mine dust, which seriously endangers the health of coal miners. Anthracosilicosis is the most common type of CWP (accounting for more than 80% of CWP), with a rapid progression and severe harm. Its pathological changes combine the characteristics of silicosis and coal workers' pneumoconiosis. However, due to the unclear pathogenesis of anthracosilicosis, there is still a lack of effective treatment methods in clinical practice. Most treatments are mainly symptomatic support treatments, which cannot fundamentally prevent the occurrence and development of anthracosilicosis. Therefore, establishing an anthracosilicosis model has important theoretical significance and practical value for in-depth study of the pathogenesis and intervention strategies of anthracosilicosis.

[0003] Due to the differences in the distribution of coal-bearing strata, lithological combinations during coal-forming periods, and coal quality characteristics, the content of free silica (SiO2) in coal dust varies greatly in different regions. Due to the differences in the content of free SiO2 in coal dust, the incidence of coal workers' pneumoconiosis in each coal mine is also different. In the prior art, some used coal dust from a certain group's coal mining face (containing 5.3% free SiO2) to prepare coal-silicone dust with 25% high free SiO2 content as the medium dose, 15% as the low dose, and 50% as the high dose for pre-experiment on C57BL / 6N mice. The mice were exposed to dust once a week for 4 weeks. As a result, varying degrees of pulmonary fibrosis changes occurred in each dose group at 28 days, and the fibrosis changes were obvious at 56 days. Based on this, the low-dose coal-silicone dust containing 15% free SiO2 was selected as the experimental dust.

[0004] The pathological features of anthracosilicosis are typical coal-silicosis nodules, with the center composed of collagen fibers, and a large number of dust cells and coal dust deposits around. In the early stage, varying degrees of interstitial fibrosis appear in the alveolar septum and around the bronchi, and perifocal emphysema can be seen around. In the late stage, multiple nodules fuse to form massive fibrosis (PMF). Due to the complex etiology and slow progression of anthracosilicosis, there is a lack of early screening methods and it is even more impossible to observe its evolution process. Therefore, establishing a stable and easy-to-implement anthracosilicosis animal model helps to explore the pathogenesis of anthracosilicosis and provides technical support for disease prevention, treatment, and new drug research and development. Summary of the Invention

[0005] To overcome the problems existing in the related technologies, the disclosed embodiments of the present invention provide a method for constructing a coal-silicosis mouse model and its application. The purpose of the present invention is to establish a coal-silicosis mouse model by inducing different proportions of silica in coal dust by oral and pharyngeal autonomous inhalation in the non-tracheal exposure method, so as to provide technical support for further analyzing the pathogenesis of coal worker's pneumoconiosis and screening therapeutic drugs.

[0006] The technical solution is as follows: A method for constructing a coal-silicosis mouse model, the method comprising:

[0007] S1, fully grinding coal dust and silica dust, and preparing a coal-silica mixed dust suspension with different free silica contents;

[0008] S2, hanging anesthetized healthy adult male mice on a fixing device, and using the method of oral and pharyngeal inhalation to drip a quantitative coal-silica mixed dust suspension. The control group is given an equal volume of normal saline to establish a coal-silicosis mouse model;

[0009] S3, taking mouse lung tissue for pathological analysis and detection of fibrosis indexes to evaluate the modeling effect of the coal-silicosis mouse model.

[0010] In step S1, the coal-silica mixed dust suspension is a mixed dust suspension containing 15% silica, with a concentration of 50 mg / mL.

[0011] In step S1, the coal-silica mixed dust suspension is a mixed dust suspension containing 25% silica, with a concentration of 50 mg / mL.

[0012] In step S1, the coal-silica mixed dust suspension is a mixed dust suspension containing 50% silica, with a concentration of 50 mg / mL.

[0013] In step S2, 50 μL of the mixed dust suspension is respectively dripped by the method of oral and pharyngeal inhalation once a week for four consecutive weeks.

[0014] In step S3, on the 28th day after the first modeling, mouse lung tissue is taken for pathological analysis and detection of lung fibrosis indexes.

[0015] In step S3, on the 56th day after the first modeling, mouse lung tissue is taken for pathological analysis and detection of lung fibrosis indexes.

[0016] In step S3, on the 112th day after the first modeling, mouse lung tissue is taken for pathological analysis and detection of lung fibrosis indexes.

[0017] Another object of the present invention is to provide a coal-silicosis mouse model for verifying the pathological characteristic information of coal worker's pneumoconiosis. This application uses the coal-silicosis mouse model constructed by the above-mentioned method for constructing a coal-silicosis mouse model.

[0018] Another object of the present invention is to provide a coal-silicosis mouse model for screening therapeutic drugs for coal workers' pneumoconiosis. This application uses the coal-silicosis mouse model constructed by the method for constructing a coal-silicosis mouse model described above.

[0019] Another object of the present invention is to provide a coal-silicosis mouse model for detecting pulmonary fibrosis indexes in mouse lung tissue. This application uses the coal-silicosis mouse model constructed by the method for constructing a coal-silicosis mouse model described above.

[0020] Combining all the above technical solutions, the beneficial effects of the present invention are as follows:

[0021] First, the present invention prepares a coal-silicosis mixed dust suspension containing 15%, 25%, and 50% free silica, with a concentration of 50 mg / ml; anesthetizes healthy adult (6-8 weeks old) C57BL / 6N male mice and hangs them on a fixing device, and instills 50 μl of the coal-silicosis mixed dust suspension by the method of oropharyngeal inhalation, once a week for 4 consecutive weeks to establish a coal-silicosis model; the control group is given an equal volume of normal saline; observe for 28 days, 56 days, and 112 days respectively, take the mouse lung tissue for pathological analysis and detection of fibrosis indexes, and evaluate the effect of model preparation. This method is simple to operate, low in cost, the established coal-silicosis mouse model is stable and efficient, can be repeated multiple times, and is non-invasive. The present invention provides a good experimental animal model for further studying the pathogenesis of coal workers' pneumoconiosis and drug screening.

[0022] Second, after the technical solution of the present invention is transformed, it can provide a stable, efficient, and non-invasive research model for the pathogenesis of coal workers' pneumoconiosis, drug screening, etc., and the social benefits brought are immeasurable; the coal-silicosis model can be sold as a commodity, which can bring rich economic benefits and has potential commercial value. Previous studies on coal workers' pneumoconiosis have mainly considered the role of coal dust, so it is difficult to observe the pathological manifestations of coal-silicosis. Since more than 80% of coal workers' pneumoconiosis in clinical practice is coal-silicosis, the present invention has successfully constructed a coal-silicosis model using coal-silicosis mixed dust, overcoming the technical bias in the establishment of previous coal workers' pneumoconiosis models, and opening up a new technical method for the prevention and treatment of coal workers' pneumoconiosis. Description of the Drawings

[0023] The drawings here are incorporated into the specification and form a part of this specification, showing embodiments consistent with the present disclosure, and are used together with the specification to explain the principles of the present disclosure;

[0024] Figure 1 is a flow chart of the method for constructing a coal-silicosis provided by an embodiment of the present invention;

[0025] Figure 2 is a coal-silicosis model constructed with coal-silicosis dust with different free SiO2 contents (HE staining, 200×);

[0026] Figure 3Coal-silicosis models constructed for coal dust containing different free SiO2 contents (Masson staining, 200×);

[0027] Figure 4 Pathological scores of lung tissues of coal-silicosis models constructed for coal dust containing different free SiO2 contents;

[0028] Figure 5 Collagen volume (%) of lung tissues of coal-silicosis models constructed for coal dust containing different free SiO2 contents;

[0029] Figure 6 Expressions of type I collagen (COL1) and α-smooth muscle actin (α-SMA) in lung tissues of coal-silicosis models constructed for coal dust containing different free SiO2 contents (laser confocal microscope). Specific embodiments

[0030] To make the above objects, features and advantages of the present invention more obvious and understandable, the specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Many specific details are set forth in the following description in order to fully understand the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without departing from the connotation of the present invention. Therefore, the present invention is not limited by the specific embodiments disclosed below.

[0031] Example 1. The present invention uses the improved oropharyngeal inhalation method of the research group to instill a coal-silica mixed dust suspension to establish a coal-silicosis mouse model, providing an economical, stable, reliable and non-invasive method for constructing a coal-silicosis mouse model for the mechanism exploration of coal worker's pneumoconiosis and the screening of clinical therapeutic drugs.

[0032] Specifically, as Figure 1 shown, a method for constructing a coal-silicosis mouse model includes:

[0033] S1. Thoroughly grind coal dust and silica dust, and prepare a coal-silica mixed dust suspension with different free silica contents;

[0034] Prepare coal-silica mixed dust suspensions containing 15%, 25% and 50% free silica;

[0035] S2. Hang the anesthetized healthy adult male mice on a fixing device, and use the oropharyngeal inhalation method to instill a quantitative coal-silica mixed dust suspension. The control group is given an equal volume of normal saline to establish a coal-silicosis mouse model;

[0036] Hang the anesthetized healthy adult (6-8 weeks old) C57BL / 6N male mice on a fixing device, and use the oropharyngeal inhalation method to instill 50 μl of a quantitative coal-silica mixed dust suspension once a week for 4 consecutive weeks;

[0037] S3. Take the lung tissues of mice for pathological analysis and detection of fibrosis indexes to evaluate the modeling effect of the coal-silicosis mouse model.

[0038] Observe for 28 days, 56 days and 112 days respectively, and take the lung tissues of mice for pathological analysis and detection of fibrosis indexes.

[0039] In step S1, the coal-silica mixed dust suspension is a mixed dust suspension containing 15%, 25%, and 50% silica, and the concentration is 50 mg / mL for all; preferably, the mixed dust suspension containing 15% silica is used.

[0040] In step S2, use the oropharyngeal inhalation method to drip 50 μL of the mixed dust suspension respectively once a week for four consecutive weeks.

[0041] In step S3, take the lung tissues of mice for pathological analysis and detection of lung fibrosis indexes on the 28th day, 56th day, and 112th day after the first modeling respectively; preferably, the 56-day model is used. The described mouse model shows typical pathological features of coal-silicosis, and the lung fibrosis indexes increase.

[0042] It can be seen from the above embodiments that compared with the conventional tracheal exposure method and the non-tracheal exposure intratracheal instillation method, this method is a non-invasive modeling method, which reduces the artificial injury of the tracheal exposure method and also reduces the experimental interference that may be brought by the intratracheal instillation of the non-tracheal exposure method; the present invention can successfully establish an efficient and stable coal-silicosis mouse model by dripping a 15% SiO2 suspension through the oropharyngeal self-inhalation method, and has relatively high repeatability and safety; this dust exposure method conforms to the inhalation route of coal mine workers' occupational exposure to coal dust and better meets the requirements of medical models; the recommended dripping dose is a relatively safe dose for mice and can be appropriately adjusted according to the experimental purpose; this technology is simple and easy to operate, greatly reduces the risk of animal death, shortens the operation time, saves manpower, and one-time modeling can meet the basic requirements of the animal experiment sample size, reducing the biological clock experimental error caused by modeling in different batches and at different times; compared with the previous two methods, it reduces the experimental cost and the risk coefficient of the operator, and achieves the modeling effect expected by the experiment. In summary, the method for preparing the coal-silicosis mouse model greatly improves the scientific research efficiency in this field, accelerates people's understanding of the occurrence and development of coal-silicosis, provides a powerful tool for conquering coal worker's pneumoconiosis at an early date, and protects the health of coal industry workers.

[0043] In the above embodiments, the descriptions of each embodiment have their own emphases. For the parts not detailed or recorded in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0044] To further illustrate the relevant effects of the embodiments of the present invention, the following experiments are carried out.

[0045] (1) Preparation before the experiment: Healthy C57BL / 6N male mice were purchased from SciBest Biotechnology Co., Ltd. The mice were acclimated and housed in an SPF animal room for one week.

[0046] (2) Anesthetized mice were suspended on a fixed board and orally instilled with 50 μL of a 50 mg / mL coal silica dust suspension by spontaneous inhalation once a week for 4 weeks. The coal silica dust suspensions contained 15%, 25%, and 50% free silica, respectively. A suspension containing 15% free SiO2 was preferred.

[0047] (3) Lung tissues of mice were collected for pathological analysis (HE and Masson staining) and detection of pulmonary fibrosis indicators (type I and III collagen, α-SMA) on the 28th, 56th, and 112th day after the initial modeling. Day 56 was preferred.

[0048] Materials and methods

[0049] 1. Experimental animals.

[0050] One hundred and twenty healthy, 5-week-old C57BL / 6N male mice (18 ± 2.0 g) were purchased from SciBest Biotechnology Co., Ltd. Mice were housed in an SPF-grade animal room at the School of Public Health, Xinxiang Medical University, at an ambient temperature of 22 ± 1°C and a relative humidity of approximately 65%, with natural light and half-day and half-night intervals. They were fed purified water and standard chow daily. Animal husbandry and experiments were approved by the Experimental Animal Ethics Committee of Xinxiang Medical University.

[0051] 2. Instruments and reagents.

[0052] Pathology slicer (Shanghai Leica Instrument Co., Ltd.), optical microscope (Shanghai Leica Instrument Co., Ltd.), coal mine dust (from an underground coal mine in Pingdingshan, Henan Province), silica dust (SILICA, USA), hematoxylin-eosin staining reagent, Masson staining reagent (Wuhan Sewell Biotechnology Co., Ltd.), α-SMA antibody, type I collagen (collagen I), β-actin antibody, and horseradish peroxidase-labeled goat anti-rabbit secondary antibody (Affinity, USA).

[0053] 3. Preparation of coal-silicon mixed dust suspension.

[0054] The coal mine dust and standard silica dust were each sufficiently ground for 2 h and baked in an oven at 180 °C for 6 h to remove endotoxins. An appropriate amount of coal mine dust and silica were weighed and respectively prepared into coal dust and silica suspensions at a concentration of 50 mg / mL, and autoclaved at 120 °C for 15 min. According to the free silica content in the coal mine dust (about 5%), silica suspension was added to make the silica content in the coal-silica mixed dust suspension 15%, 25%, and 50% respectively.

[0055] 4. Establishment of animal models.

[0056] 120 mice were randomly divided into a control group, a mixed dust group containing 15% silica (15% S-mixed dust group), a mixed dust group containing 25% silica (25% S-mixed dust group), and a mixed dust group containing 50% silica (50% S-mixed dust group), with 30 mice in each group. Mice in the control group were instilled with 50 μL of sterile PBS by oropharyngeal inhalation (once a week for a total of four times), and mice in the 15%, 25%, and 50% S-mixed dust groups were each instilled with 50 μL of 50 mg / mL mixed dust containing different free silica by oropharyngeal inhalation (once a week for a total of four times). On the 28th, 56th, and 112th days after the first modeling, 10 mice were taken from each group, blood was collected after anesthesia with isoflurane, and lung tissues were taken for model evaluation.

[0057] 5. The method for measuring the degree of lung tissue injury in the modeled mice is as follows:

[0058] (1) Histopathological observation of lung tissue.

[0059] Lung tissues fixed with 4% paraformaldehyde solution were dehydrated with absolute ethanol, cleared, and then embedded in paraffin. 5-μm-thick sections were made with a microtome, deparaffinized, hydrated, and then subjected to HE staining and Masson staining, and finally sealed. Observation was carried out using an optical microscope. The degree of pathological injury of lung tissue was referenced to the Ashcroft scoring standard.

[0060] (2) Immunofluorescence detection of the expression of type I collagen and α-SMA.

[0061] Paraffin sections were deparaffinized, hydrated, rinsed, and then antigen repaired with citrate buffer. After washing three times with PBST, an inactivated endogenous HRP blocker was added. After washing, it was blocked, incubated with type I collagen and α-SMA primary antibodies (1:100) overnight. After washing again, secondary antibodies were incubated and DAB was used for color development. Then hematoxylin was used for counterstaining, dehydration, and clearing, and finally sealed with neutral gum. Observation and photography were carried out under a microscope, and the positive staining area of each group was analyzed using Image-Pro Plus 6.1 software.

[0062] 6. Statistical analysis.

[0063] Statistical analysis was performed using SPSS 22.0. Measurement data were expressed as mean ± standard deviation. One-way ANOVA was used to compare the differences between groups at the same time point and the differences between groups at different time points under the same silica proportion. A P value < 0.05 was considered statistically significant.

[0064] Results.

[0065] (1) Effects of exposure to coal-silica mixed dust with different free silica contents for different times on the pathological changes of mouse lung tissue.

[0066] The results showed that in the control group of mice at 28 d, 56 d, and 112 d, the lung tissue structure was normal, the alveolar wall was intact, and no inflammation occurred. In the 15% S-mixed dust group of mice at 28 d and 56 d, coal spots and coal-silica nodules appeared in the lung tissue, mainly concentrated around the bronchi, accompanied by perifocal emphysema. In the 25% S-mixed dust group of mice at 28 d and 56 d, the volume of coal spots and coal-silica nodules around the trachea in the lung tissue increased. In the 50% S-mixed dust group of mice at 28 d, silicotic nodules appeared on the basis of coal spots in the lung tissue, and the degree of pulmonary fibrosis increased. In the 50% S-mixed dust group of mice at 56 d, reticular fibrosis appeared in the lung tissue, and the fibrotic area increased. In the 15% S-mixed dust group of mice at 112 d, the number of coal-silica nodules in the lung tissue further increased; in the 25% S-mixed dust group and 50% S-mixed dust group of mice at 112 d, the coal-silica nodules in the lung tissue further fused to form large areas of fibrosis. See Figure 2 .

[0067] (2) Effects of exposure to coal-silica mixed dust with different free silica contents for different times on the collagen content of mouse lung tissue. As Figure 3 is the atlas of the effects of coal-silica mixed dust with different silica contents and exposure for different times on the collagen content of mouse lung tissue;

[0068] The results of Masson staining showed that a small amount of collagen appeared around the coal spots in the mixed dust group of mice at 28 d and 56 d, and the collagen content in the lung tissue of the 50% S-mixed dust group of mice at 56 d further increased. The collagen content in the lung tissue of the mixed dust group of mice at 112 d was higher than that at 28 d and 56 d, and the collagen content in the lung tissue of the 50% S-mixed dust group of mice at 112 d was higher than that in the 15% S-mixed dust group and 25% S-mixed dust group. A small amount of collagen appeared on the coal spots in the lung tissue of the 15% S-mixed dust and 25% S-mixed dust groups of mice at different time points, and the pathological characteristics were mainly coal-silica nodules; in the lung tissue of the 50% S-mixed dust group of mice, in addition to coal-silica nodules, typical silicotic nodules appeared, and the collagen content in the central part of the nodules increased, showing a concentric circle. The results of Ashcroft scoring showed ( Figure 4 ) that the score of the mixed dust group was higher than that of the control group, and the difference was statistically significant (P < 0.05). Figure 5The results showed that compared with the control group, the collagen content in the lung tissue of mice in the mixed dust group increased, and the difference was statistically significant (P<0.05).

[0069] (3) Effects of exposure to coal-silica mixed dust with different free silica contents for different times on the contents of α-SMA and type I collagen in the lung tissue of mice.

[0070] The results showed that compared with the control group, the contents of type I collagen and α-SMA in the lung tissue of mice in the mixed dust group increased, and the difference was statistically significant (P<0.05). The results of α-SMA showed that there was no significant difference in the expression of α-SMA among the three groups of 15% S-mixed dust, 25% S-mixed dust and 50% S-mixed dust at the same time point. On the contrary, the content of type I collagen in the lung tissue of mice in the 50% S-mixed dust group was higher than that in the 15% S-mixed dust group and the 25% S-mixed dust group, which was also consistent with the results of Masson staining. Among them Figure 6 It is a map of the effects of exposure to coal-silica mixed dust with different silica contents for different times on the contents of α-SMA and type I collagen in the lung tissue of mice.

[0071] In the present invention, a coal-silicosis mouse model was established using a mixed dust suspension prepared from coal dust and free silica dust to simulate coal-silicosis caused by coal dust and silica dust contacted by coal miners at different working faces. During the process of constructing the model, different degrees of pulmonary fibrosis were caused by different proportions of silica content in the mixed dust. As the proportion of silica increased, the degree of pulmonary fibrosis gradually increased, and it gradually changed from coal macule lesions to silicotic nodules. After 28 days of exposure to 15% S-mixed dust, coal macule lesions and a small amount of collagen appeared in the lung tissue; after 56 days of exposure to 15% S-mixed dust, the pathological changes in the lung tissue of mice were mainly coal macule lesions, and the collagen content increased compared with that at 28 days, showing a coal-silicotic nodule manifestation mainly composed of coal macule lesions. When the free silica content in the mixed dust increased to 25% and 50%, the pathological manifestations of the lung tissue were mainly silicotic nodules, accompanied by coal macule lesions, showing a coal-silicotic nodule manifestation mainly composed of silicotic nodules. In the present invention, a coal-silicosis model was constructed by exposing mice to 15% S-mixed dust for 56 days, and the pathological manifestations of its lung tissue were consistent with those of coal-silicosis, providing technical support for the research on the pathogenesis of coal-silicosis and the screening of therapeutic drugs.

[0072] As mentioned above, it is only a relatively preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any modification, equivalent replacement and improvement made by those skilled in the art within the technical scope disclosed by the present invention, as long as they are made within the spirit and principle of the present invention, shall be covered by the protection scope of the present invention.

Claims

1. A method for constructing a coal worker's pneumoconiosis mouse model, characterized in that, The method includes: S1. Sufficiently grind coal dust and silica dust, and prepare coal-silica mixed dust suspensions with different free silica contents; S2. Hang anesthetized healthy adult male mice on a fixing device, and use the method of oropharyngeal inhalation to instill a quantitative coal-silica mixed dust suspension. The control group is given an equal volume of normal saline to establish a coal-silicosis mouse model; S3. Take mouse lung tissues for pathological analysis and detection of fibrosis indexes to evaluate the modeling effect of the coal-silicosis mouse model.

2. The method for constructing a coal worker's pneumoconiosis mouse model according to claim 1, wherein In step S1, the coal-silica mixed dust suspension is a mixed dust suspension containing 15% silica, with a concentration of 50 mg / mL.

3. The method for constructing a coal worker's pneumoconiosis mouse model according to claim 1, wherein In step S1, the coal-silica mixed dust suspension is a mixed dust suspension containing 25% silica, with a concentration of 50 mg / mL.

4. The method for constructing a coal worker's pneumoconiosis mouse model according to claim 1, characterized in that, In step S1, the coal-silica mixed dust suspension is a mixed dust suspension containing 50% silica, with a concentration of 50 mg / mL.

5. The method for constructing a coal worker's pneumoconiosis mouse model according to claim 1, wherein In step S2, 50 μl of the mixed dust suspension is instilled respectively by the method of oropharyngeal inhalation once a week for four consecutive weeks.

6. The method for constructing a coal worker's pneumoconiosis mouse model according to claim 1, wherein, In step S3, mouse lung tissues are taken on the 28th day after the first modeling for pathological analysis and detection of lung fibrosis indexes.

7. The method for constructing a coal worker's pneumoconiosis mouse model according to claim 1, wherein In step S3, mouse lung tissues are taken on the 56th day after the first modeling for pathological analysis and detection of lung fibrosis indexes.

8. The method for constructing a coal workers' pneumoconiosis mouse model according to claim 1, characterized in that, In step S3, mouse lung tissues are taken on the 112th day after the first modeling for pathological analysis and detection of lung fibrosis indexes.

9. Use of a coal-silicosis mouse model in verifying pathological characteristic information of coal worker's pneumoconiosis, characterized in that, This application uses the coal-silicosis mouse model constructed by the method for constructing a coal-silicosis mouse model described in claim 1.

10. Use of a coal-silicosis mouse model in the screening of drugs for treating coal workers' pneumoconiosis, characterized in that, This application uses the coal-silicosis mouse model constructed by the method for constructing a coal-silicosis mouse model described in claim 1.

11. Use of a coal workers' pneumoconiosis mouse model for detecting pulmonary fibrosis indexes in mouse lung tissue, characterized in that, This application uses the coal-silicosis mouse model constructed by the method for constructing a coal-silicosis mouse model described in claim 1.

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