Application of isovitexin in treatment of pulmonary fibrosis

Abstract

The invention discloses application of isovitexin in treatment of pulmonary fibrosis, and belongs to the technical field of medicines. The isovitexin with different concentrations is used for treating a pulmonary fibrosis model, detecting pulmonary fibrosis markers and various ferroptosis related markers, and exploring the influence of the isovitexin on the pulmonary fibrosis animal model and how the isovitexin affects the growth of pulmonary fibrosis cells. Finally, a conclusion is drawn: (1) the pulmonary fibrosis is treated by the isovitexin, so that ferroptosis of cells is promoted, and collagen deposition is reduced; and (2) isovitexin is used, so that the progress of pulmonary fibrosis can be inhibited.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to application of isovitexin in the treatment of pulmonary fibrosis. Background Art

[0002] Isovitexin (ISO), also known as flavonoids, is a flavonoid obtained from passion fruit, cannabis and palm. It has multiple effects such as antioxidant, antibacterial, antiviral, anti-inflammatory, and tumor growth inhibition.

[0003] Pulmonary fibrosis is an interstitial lung disease, and its pathological characteristics are alveolar epithelial cell damage and proliferation, basement membrane erosion, alveolar consolidation and fibroblastic lesions. Finally, patients experience symptoms such as progressive dyspnea, cough, respiratory physiological function limitation and gas exchange disorder. Inflammatory cells, cytokines, myofibroblasts, epithelial-mesenchymal transition (EMT), protease system, lipid peroxidation and signal transduction mechanism play a key role in the occurrence and development of fibrosis. Its etiology is complex and the pathogenesis is still unclear.

[0004] Ferroptosis is an iron-dependent programmed cell death caused by unrestricted phospholipid peroxidation. The occurrence of this process mainly depends on the increase of metabolites reactive oxygen species (ROS), phospholipids containing polyunsaturated fatty acid chains (PUFA-PL) and iron accumulation. In addition, intracellular and intercellular signals and environmental stress can also indirectly affect ferroptosis by regulating cell metabolism and ROS levels. The triggering pathways of ferroptosis are: ① the classic Cystine / GSH / GPX4 regulation pathway; ② the ferroptosis suppressor protein 1 (FSP1) / CoQ10 pathway; ③ the dihydroorotate dehydrogenase (DHODH) pathway; ④ the GTP cyclohydrolase 1 (GCH1) / tetrahydrobiopterin (BH4) pathway; ⑤ the lipid metabolism pathway; ⑥ the iron metabolism pathway, etc. In addition, ferroptosis is also affected by cell metabolism and AMPK pathway. Summary of the invention

[0005] The invention provides application of isovitexin in preparing a product for treating pulmonary fibrosis. The CAS number of the isovitexin is 38953-85-4.

[0006] Furthermore, in the product, isovitexin is the only active ingredient.

[0007] Furthermore, the product is a medicine.

[0008] Furthermore, the medicine also contains a medically acceptable carrier.

[0009] Furthermore, the dosage form of the drug is any one of powder, tablet, granule, capsule, solution, emulsion and suspension.

[0010] The present invention also provides a traditional Chinese medicine composition for treating pulmonary fibrosis, which is composed of the following components in parts by weight: 20-30 parts of isovitexin, 1-5 parts of baicalin, 1-5 parts of nobiletin, 1-5 parts of isoliquiritigenin, 1-5 parts of tangeretin, 1-5 parts of acacetin, 1-5 parts of polydatin, and 1-5 parts of morusin.

[0011] Furthermore, the Chinese medicine composition is composed of the following Chinese medicine components in parts by weight: 25 parts of isovitexin, 1 part of baicalin, 1 part of nobiletin, 1 part of isoliquiritigenin, 1 part of tangeretin, 1 part of acacetin, 2 parts of polydatin, and 1 part of morusin.

[0012] The present invention also provides a method for preparing a Chinese medicine preparation for treating pulmonary fibrosis, the preparation method comprising the following steps:

[0013] The above Chinese medicine composition is crushed, and then water is added, ultrasonicated at 45-70°C for 30-60 minutes, filtered to obtain a filtrate, and the filtrate is vacuum concentrated and sprayed to obtain a preparation, or directly spray-dried to obtain a powder.

[0014] Furthermore, the amount of water added is twice the weight of the traditional Chinese medicine composition.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] (1) The present invention promotes cell ferroptosis and reduces collagen deposition by treating pulmonary fibrosis with isovitexin;

[0017] (2) The present invention uses isovitexin to inhibit the progression of pulmonary fibrosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Detection of markers related to the pulmonary fibrosis cell model constructed in Example 3.

[0019] Figure 2 The activity level of MRC-5 cells after TGF-β induction treated with ISO was detected by CCK8 in Example 4.

[0020] Figure 3The IC50 value of CCK8 in Example 4 for detecting MRC-5 cells induced by TGF-β after ISO treatment for 24 hours.

[0021] Figure 4 This is the detection of apoptosis level of MRC-5 cells induced by TGF-β after ISO treatment in Example 5.

[0022] Figure 5 This is the apoptosis (fluorescence) level of MRC-5 cells induced by TGF-β after ISO treatment in Example 6.

[0023] Figure 6 This is a scratch test in Example 7 to detect the effect of ISO on the migration ability of MRC-5 cells induced by TGF-β.

[0024] Figure 7 This is the detection of activity levels of MRC-5 cells induced by TGF-β after being treated with different death inhibitors in Example 8.

[0025] Figure 8 This is the detection of ferroptosis-related indicators in MRC-5 cells induced by TGF-β after ISO treatment in Example 9.

[0026] Fig. 9 This is the protein level detection of ferroptosis-related markers before and after ISO treatment of MRC-5 cells in Example 10.

[0027] Fig.10 The flow cytometry method in Example 11 was used to detect the ROS levels in MRC-5 cells before and after treatment with TGF-β.

[0028] Fig.11 This is a bar graph analysis of ROS levels before and after treatment of MRC-5 cells induced by TGF-β in Example 11.

[0029] Fig.12 This is the ROS (fluorescence) level before and after treatment of MRC-5 cells induced by TGF-β in Example 12.

[0030] Fig.13 The lipid peroxidation levels of MRC-5 cells induced by TGF-β and treated with different concentrations of ISO in Example 13 were detected by flow cytometry.

[0031] Fig.14 This is a quantitative analysis of the lipid peroxidation level of MRC-5 cells induced by TGF-β after treatment with different concentrations of ISO in Example 13.

[0032] Fig.15 HE staining of the animal model of pulmonary fibrosis constructed in Example 2.

[0033] Fig.16 Masson staining of the pulmonary fibrosis animal model constructed in Example 2.

[0034] Fig.17 Detection of markers related to the animal model of pulmonary fibrosis constructed in Example 2.

[0035] Fig.18 HE staining of the animal model of pulmonary fibrosis treated with ISO in Example 2.

[0036] Fig.19 This is the detection of relevant markers after the animal model of pulmonary fibrosis was treated with ISO in Example 2. DETAILED DESCRIPTION

[0037] Example 1 Construction of animal model of pulmonary fibrosis

[0038] In this experiment, the drug bleomycin (BLM) was used to induce C57BL / 6 mice to construct an animal model of pulmonary fibrosis. The specific treatment process is as follows:

[0039] 1.1 Grouping of mice

[0040] Fifteen 6-8 week old male C57BL / 6 mice were selected and housed in a barrier environment and divided into a control group and experimental groups (BLM group and BLM+ISO group).

[0041] 1.2 Construction of animal model

[0042] On day 0, the control group was not treated, and the experimental group was intratracheally administered (3 mg / kg) once. After BLM entered the animal's trachea, it was immediately rotated upright to evenly distribute the drug in the lung tissue; 21 days after modeling, two mice were randomly selected from the control group and the experimental group for euthanasia, and the lung tissues were collected for staining to observe whether the modeling was successful (HE staining was used to observe the degree of alveolitis in the lung tissue; Masson staining was used to observe the degree of pulmonary fibrosis in the lung tissue. The interstitial capillaries in the lung tissue of the model mice showed congestion and expansion, the alveolar wall became thicker and even broke, and inflammatory cell infiltration, mainly neutrophils, appeared in the alveolar cavity). Then, the BLM+ISO group was intraperitoneally injected with 50 mg / kg every 2 days, and the control group and BLM group were treated with an equal volume of normal saline. The mice were euthanized 7 days later, and the lung tissues of the mice were collected for detection and analysis.

[0043] Example 2 Detection of pulmonary fibrosis markers in mouse lung tissue

[0044] Lung tissues were immersed in 4% paraformaldehyde overnight, dehydrated according to standard procedures, and the tissues were paraffin-embedded.

[0045] 2.1 Staining of paraffin sections

[0046] The tissue was cut into 3 μm sections, and after routine dewaxing and hydration, it was stained with hematoxylin and eosin for 7 min each. The staining was performed according to the Masson staining kit. HE staining and Masson staining images were collected for analysis.

[0047] 2.2 Immunohistochemistry on paraffin sections

[0048] After conventional dewaxing and hydration, an antigen retrieval step was performed in citrate buffer (pH 6.0) at about 115°C for 2 min, and then the sections were blocked with goat serum for 1 h at room temperature. Subsequently, the sections were incubated with primary antibodies (anti-α-SMA, anti-Collagen I, anti-TGF-β) at 4°C overnight. After washing, the sections were incubated with HRP-labeled goat anti-mouse / rabbit IgG at room temperature for 1 hour and with streptavidin / HRP at room temperature for 20 min. Finally, the DAB color development kit was used for color development.

[0049] Example 3 Construction of pulmonary fibrosis cell model

[0050] In this experiment, human embryonic lung fibroblast MRC-5 cells were selected, and the specific processing process was as follows.

[0051] 1.1 Cell culture and plating

[0052] (1) MRC-5 cells were cultured in complete medium containing MEM non-essential amino acid solution (NEAA) containing 10% FBS at 37°C and 5% CO 2 .

[0053] (2) Cell passaging: When the cell density reaches 80%-90%, the cells are passaged and cultured continuously.

[0054] 1.2 Establishment of pulmonary fibrosis cell model

[0055] Transforming growth factor-β (5 ng / mL) was added to the culture medium and treated for 24 h to induce the cells to transform into myofibroblasts.

[0056] 1.3 Drug treatment of cells

[0057] (1) Myofibroblasts induced by TGF-β were plated in a 6-well plate. When the cell density reached about 70%, different concentrations of isovitexin (0, 1, 5, 10, 20 and 30 μM) were added to each well and cultured for 24 h.

[0058] (3) Treat the cells with trypsin and detect changes in the levels of related markers.

[0059] Example 4 Detection of cell activity levels in MRC-5 cells induced by TGF-β before and after ISO treatment

[0060] In this part of the experiment, CCK-8 reagent was used to detect the changes in cell activity over time in MRC-5 cells induced by TGF-β before and after treatment with different concentrations of ISO. The concentrations of ISO treatment were 0, 1, 5, 10, 20 and 30 μM, and the detection time was 24h, 48h, 72h, 96h and 120h, respectively.

[0061] Example 5 Flow cytometry detection of apoptosis levels in TGF-β-induced MRC-5 cells before and after ISO treatment

[0062] The MRC-5 cells induced by TGF-β were treated with different concentrations of ISO for 24 h, and then the Annexin V-PE / 7-AADApoptosis Detection Kit (Vazyme) was used to measure the changes in the levels of MRC-5 cells induced by TGF-β after treatment with different concentrations of ISO.

[0063] Example 6 Immunofluorescence detection of apoptosis levels in MRC-5 cells induced by TGF-β before and after ISO treatment

[0064] MRC-5 cells induced by TGF-β were treated with different concentrations of ISO (0 and 30 μM) for 24 h, and then the cells were stained with Propidium Iodide (PI) dye, and the fluorescence was detected by photographing.

[0065] Example 7 Detection of migration ability of TGF-β-induced MRC-5 cells before and after ISO treatment by scratch test

[0066] ISO was used to treat MRC-5 cells induced by TGF-β at different concentration gradients (0, 1, 5, 10, 20, 30 μM). When the cells grew to about 90%, a line was drawn in the middle and on both sides of the well with a 10 μL pipette tip. After washing the cells with PBS, the cells were cultured with 3% FBS culture medium and photographed to observe the migration of the cells.

[0067] Example 8 Combination of multiple death pathway inhibitors and ISO to treat MRC-5 cells induced by TGF-β

[0068] Different types of cell death inhibitors were used to treat MRC-5 cells induced by TGF-β, including five cell death inhibitors: ferroptosis inhibitors Fer-1 and Lip-1, apoptosis inhibitor Z-VAD-FMK, autophagy inhibitor 3-methyladenine (3-ME), pyroptosis inhibitor Bay 11-7821 (BAY), and necrosis inhibitor Necrosulfonamide (NSA). The groups were set as follows: CTRL (no drug treatment group), ISO (30μM) treatment group, ISO (30μM) combined with Fer-1 (10μM) treatment group, ISO (30μM) combined with Lip-1 (10μM) treatment group, ISO (30μM) combined with Z-VAD-FMK (10μM) treatment group, ISO (30μM) combined with 3-ME (5mM) treatment group, ISO (30μM) combined with BAY (10μM) treatment group, and ISO (30μM) combined with NSA (5μM) treatment group. After 24 h of ISO treatment, CCK8 reagent was used to detect cell viability.

[0069] Example 9 Detection of ferroptosis products in pulmonary fibrosis cell model before and after ISO treatment

[0070] ISO and ferroptosis inhibitors Fer-1 and Lip-1 were used to treat lung fibroblasts induced by TGF-β, and the groups were set as follows: CTRL (no drug treatment group), ISO (30μM) treatment group, ISO (30μM) combined with Fer-1 (10μM) treatment group, ISO (30μM) combined with Lip-1 (10μM) treatment group. The levels of ferroptosis-related products MDA and GSH were detected by MDA and GSH detection kits.

[0071] Example 10 Detection of ferroptosis-related proteins in pulmonary fibrosis cells before and after ISO treatment

[0072] MRC-5 cells induced by TGF-β were treated with different concentrations of ISO at 0, 1, 5, 10, 20 and 30 μM, and the cells were collected after 24 hours of treatment. The cells were collected and proteins were extracted with RIPA buffer and a mixture of protease inhibitors (mixing ratio 100:1). The proteins detected mainly included: GPX4, SLC7A11, ACSL4, ACSL3, CD71, FSP1 and FTH1.

[0073] Example 11 Detection of ROS levels in pulmonary fibrosis cells before and after ISO treatment by flow cytometry

[0074] MRC-5 cells induced by TGF-β were treated with different concentrations of ISO, 0, 1, 5, 10, 20 and 30 μM, and the cells were collected after 24 hours of treatment. C11-BODIPY was used to detect the level of ROS in each group of cells.

[0075] Example 12 Immunofluorescence detection of ROS levels in pulmonary fibrosis cells before and after ISO treatment

[0076] MRC-5 cells induced by TGF-β were treated with different concentrations of ISO, with the treatment concentrations being 0 and 30 μM, respectively. After 24 h of treatment, the cells were incubated with H2DCFDA dye in the dark for 30 min, and then photographed to detect the intracellular ROS level.

[0077] Example 13 Flow cytometry detection of lipid peroxidation levels in pulmonary fibrosis cells before and after ISO treatment

[0078] Different concentrations of ISO were used to treat MRC-5 cells induced by TGF-β, with the concentrations of 0, 1, 5, 10, 20 and 30 μM, and the cells were collected after 24 hours of treatment. DCFH-DA was then used to detect the level of lipid peroxidation in each group of cells.

[0079] In this study, we first detected fibrosis-related indicators in TGF-β-induced human embryonic lung cells MRC-5 cells. It can be seen that the levels of α-SMA, Vimentin and snail increased significantly, indicating that the pulmonary fibrosis cell model was successfully constructed ( Figure 1 ). Next, different concentrations of isovitexin ISO were used to treat TGF-β-induced lung fibroblasts to explore the effect of ISO on the activity of lung fibrosis cells. The results showed that ISO significantly affected the proliferation of lung fibroblasts after fibrosis, and the higher the concentration, the stronger the inhibitory effect ( Figure 2 ). We further calculated that the half-inhibitory concentration of ISO on the activity of pulmonary fibrosis cells was 38.78 μM ( Figure 3 ). Subsequently, we further detected the effect of ISO on the apoptosis of lung fibroblasts by flow cytometry and immunofluorescence. The results showed that ISO could promote the apoptosis of lung fibroblasts induced by TGF-β, and the degree of apoptosis increased in a concentration-dependent manner ( Figure 4 and Figure 5 ). Furthermore, we found that ISO can affect the migration ability of pulmonary fibrosis cells in a concentration-dependent manner when acting on them. Figure 6). The above experimental results show that ISO significantly affects the activity of lung fibroblasts induced by TGF-β. Next, we further explored the death pathway through which ISO affects lung fibrosis cells. Using different types of death inhibitors combined with ISO to treat TGF-β-induced lung fibroblasts, we found that ISO significantly induced cell death, but only after treatment with ferroptosis inhibitors (Fer-1 and Lip-1) could cell viability be properly restored, while other types of death inhibitors combined with ISO could only slightly restore cell viability ( Figure 7 ). Therefore, we concluded that ISO inhibits the activity of lung fibroblasts mainly through the pathway of ferroptosis, and can act as an inducer of ferroptosis to alleviate pulmonary fibrosis. Then, we verified this conclusion by detecting ferroptosis-related markers. For example, the levels of MDA and GSH. The ferroptosis metabolite MDA increased significantly after ISO treatment, while the antioxidant GSH decreased significantly after ISO treatment. At the same time, the levels of these markers were restored after combined treatment with ISO and ferroptosis inhibitors ( Figure 8 Then, we detected the changes in the levels of ferroptosis-related proteins ( Fig. 9 ), and in protein detection, it was found that the FTH1 level of lung fibroblasts decreased with the increase of ISO treatment concentration, while the levels of other related proteins did not change significantly. These results indicate that ISO-induced ferroptosis of lung fibrosis cells is carried out by reducing the expression of FTH1. Then, we used flow cytometry to detect ROS in ISO-treated TGF-β-induced lung fibroblasts ( Fig.10 and Fig.11 ) and lipid peroxidation levels ( Fig.13 and Fig.14 ), we can see that the levels of ROS and lipid peroxidation increased in an ISO concentration-dependent manner, further indicating that ISO can induce lung fibroblast death through ferroptosis. Secondly, we used H2DCFDA to stain ISO-treated lung fibrosis cells and detected the generated ROS by immunofluorescence ( Fig.12), and found that the proportion of green fluorescence increased significantly in cells treated with ISO, indicating that the level of ROS produced increased, which is consistent with the above-mentioned flow cytometry test results. After the in vitro cell model was verified, we constructed an animal model for further verification. We chose bleomycin-induced C57BL / 6 mice to construct an animal model of pulmonary fibrosis. After treatment, the lungs of pulmonary fibrosis mice and control group mice were compared by HE staining and Masson staining. It can be seen that the alveolar spaces of pulmonary fibrosis mice were widened, the fibrosis exudation in the alveolar cavity was severe, and the inflammatory cells around the blood vessels were severely infiltrated. The alveolar structure was partially destroyed, and the levels of related markers increased, indicating that the pulmonary fibrosis model was successfully constructed ( Fig.15 , Fig.16 and Fig.17 ). Subsequently, we divided the mice in the experimental group into an induction-only group and an ISO-treated group. Through HE staining experiments, we found that after ISO treatment, the degree of pulmonary fibrosis in mice was significantly alleviated ( Fig.18 ), further through immunohistochemistry experiments we can see that after ISO drug treatment, compared with the model group (BLM treatment group), the levels of lung fibrosis-related markers in the ISO treatment group were significantly reduced, which shows that ISO can reduce the degree of lung fibrosis in mice and has a certain therapeutic effect on lung fibrosis ( Fig.19 ).

[0080] In summary, isovitexin can not only directly inhibit the growth of induced lung fibroblasts through the apoptosis pathway, but also promote the death of pulmonary fibrosis cells by inducing ferroptosis, and the efficacy of isovitexin on animal models is also significant. Therefore, the use of isovitexin to prepare drugs for the treatment of pulmonary fibrosis is expected to have good application prospects.

[0081] Example 14 Preparation of a Chinese medicinal compound for preventing and treating pulmonary fibrosis

[0082] 1. Take the following Chinese medicinal components by weight: 25 parts of isovitexin; 1 part of baicalin; 1 part of nobiletin; 1 part of isoliquiritigenin; 1 part of tangeretin; 1 part of acacetin; 2 parts of polydatin; 1 part of sanshin. Grind the above Chinese medicinal components in a mortar and mix them well, filter through a 100-mesh sieve, and collect the mixed powder.

[0083] 2. Add 2 times the weight of water to the above powder, dissolve it at 60°C under ultrasound for 30 minutes, collect the filtrate after filtering, spray dry to obtain solid powder, make it into granular preparation, and finally prepare a Chinese medicine combination for the treatment of pulmonary fibrosis.

[0084] Example 15 Preparation of a Chinese medicinal compound for preventing and treating pulmonary fibrosis

[0085] 1. Take the following Chinese medicinal components by weight: 25 parts of isovitexin; 1 part of baicalin; 1 part of nobiletin; 1 part of isoliquiritigenin; 1 part of tangeretin; 1 part of acacetin; 2 parts of polydatin; 1 part of sanshin. Grind the above Chinese medicinal components in a mortar and mix them well, filter through a 100-mesh sieve, and collect the mixed powder.

[0086] 2. Add 2 times the weight of water to the above powder, dissolve it at 45°C under ultrasound for 60 minutes, collect the filtrate after filtering, spray dry to obtain solid powder, make it into granular preparation, and finally prepare a Chinese medicine combination for the treatment of pulmonary fibrosis.

[0087] Example 16 Preparation of a Chinese medicinal compound for preventing and treating pulmonary fibrosis

[0088] 1. Take the following Chinese medicinal components by weight: 25 parts of isovitexin; 1 part of baicalin; 1 part of nobiletin; 1 part of isoliquiritigenin; 1 part of tangeretin; 1 part of acacetin; 2 parts of polydatin; 1 part of sanshin. Grind the above Chinese medicinal components in a mortar and mix them well, filter through a 100-mesh sieve, and collect the mixed powder.

[0089] 2. Add 2 times the weight of water of the mixed raw materials to the above powder, dissolve it at 70°C under ultrasound for 30 minutes, collect the filtrate after filtration, and vacuum concentrate it to the same weight volume of the mixed raw materials to prepare a spray preparation, and finally prepare a Chinese medicine combination for the treatment of pulmonary fibrosis.

[0090] Example 17 Preparation of a Chinese medicinal compound for preventing and treating pulmonary fibrosis

[0091] 1. Take the following Chinese medicinal components by weight: 25 parts of isovitexin; 1 part of baicalin; 1 part of nobiletin; 1 part of isoliquiritigenin; 1 part of tangeretin; 1 part of acacetin; 2 parts of polydatin; 1 part of sanshin. Grind the above Chinese medicinal components in a mortar and mix them well, filter through a 100-mesh sieve, and collect the mixed powder.

[0092] 2. Add 2 times the weight of water of the mixed raw materials to the above powder, dissolve it at 45°C under ultrasound for 60 minutes, collect the filtrate after filtration, and vacuum concentrate it to the same weight volume of the mixed raw materials to prepare a spray preparation, and finally prepare a Chinese medicine combination for the treatment of pulmonary fibrosis.

[0093] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. The use of isovitexin in the preparation of a product for treating pulmonary fibrosis, characterized in that: The CAS number of the isovitexin is 38953-85-4.

2. The use according to claim 1, characterized in that: Isovitexanthin is the only active ingredient in the product.

3. The use according to claim 1 or 2, characterized in that: The product described is a drug.

4. The use according to claim 3, characterized in that: The medicine also contains a medically acceptable carrier.

5. The use according to claim 4, characterized in that: The dosage form of the drug is any one of powder, tablet, granule, capsule, solution, emulsion and suspension.

6. A Chinese medicine composition for treating pulmonary fibrosis, characterized in that: The traditional Chinese medicine composition is composed of the following components in parts by weight: 20-30 parts of isovitexin, 1-5 parts of baicalin, 1-5 parts of nobiletin, 1-5 parts of isoliquiritigenin, 1-5 parts of hesperidin, 1-5 parts of acacetin, 1-5 parts of polydatin and 1-5 parts of morusin.

7. The Chinese medicine composition according to claim 6, characterized in that: The traditional Chinese medicine composition consists of the following components in parts by weight: 25 parts of isovitexin, 1 part of baicalin, 1 part of nobiletin, 1 part of isoliquiritigenin, 1 part of tangeretin, 1 part of acacetin, 2 parts of polydatin and 1 part of moranin.

8. A method for preparing a Chinese medicine preparation for treating pulmonary fibrosis, characterized in that: The preparation method comprises the following steps: The Chinese medicine composition according to claim 6 or 7 is crushed, and then water is added, ultrasonicated at 45-70° C. for 30-60 minutes, filtered to obtain a filtrate, and the filtrate is vacuum concentrated and sprayed to obtain a preparation, or directly spray-dried to obtain a powder.

9. The preparation method according to claim 8, characterized in that: The amount of water added is twice the weight of the traditional Chinese medicine composition.

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