Method for analyzing action mechanism of aleppo avens for treating idiopathic pulmonary fibrosis based on network pharmacology

Abstract

The invention relates to the technical field of traditional Chinese medicine research, in particular to a method for analyzing an action mechanism of aleppo avens for treating idiopathic pulmonary fibrosis based on network pharmacology. According to the present invention, the UHPLC-Q-Orbitrap HRMS technology is adopted to qualitatively analyze the basic peak chromatogram (BPC) of the chemical components of the aleppo avens positive aqueous extract under the positive and negative ion mode for the first time; 27 chemical components are identified according to information such as relative retention time, accurate molecular mass, secondary fragment ions and the like by retrieving a Compound Discovery 2.1 database and looking up literatures, and structures, compound information and mass spectrum data of the 27 chemical components are determined. The pharmacological action mechanism of aleppo avens for improving the IPF is discussed by utilizing network pharmacology, finally, a'component-target-pathway 'network is constructed by utilizing Cytoscape 3.9. 1 software, and the result shows that the prevention and treatment of the IPF by the aleppo avens may be closely related to inflammation-related pathways.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine research, and specifically to a method for analyzing the mechanism of action of Lanbuzheng in treating idiopathic pulmonary fibrosis based on network pharmacology. Background Art

[0002] Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, cicatricial interstitial lung disease characterized by abnormal fibroblast proliferation, excessive deposition of extracellular matrix (ECM), inflammatory damage, and destruction of lung tissue function. Clinical manifestations include distorted lung structure and decreased lung function, which eventually lead to respiratory failure or even death. The average survival period is 3-5 years, and the incidence rate is increasing worldwide. Although a lot of research has been done on IPF, its pathogenesis is complex and the cause is still unclear. The main pathological changes are caused by inflammation, immune mechanism, epithelial-mesenchymal transition (EMT), and oxidative stress. At present, the treatment options for IPF are very limited. Lung transplantation is the only effective method for patients with advanced IPF and respiratory failure. The 2015 International Guidelines for Idiopathic Pulmonary Fibrosis recommend the use of pirfenidone and nintedanib as first-line clinical drugs, but they are expensive and have poor prognosis. Therefore, it is particularly urgent to find a safe and effective treatment for IPF.

[0003] In recent years, traditional Chinese medicine (TCM) has made significant progress in the prevention or treatment of pulmonary fibrosis through multi-target and systemic regulation. Through the synergistic regulation of multiple components, multiple targets and multiple pathways, it plays an important role in the prevention, treatment and rehabilitation of pulmonary fibrosis. Blue Buzheng is the dried whole herb of Geum japonicum Thunb. var. chinense Bolle and Geum aleppicum Jacq. of the Rosaceae family. It has the effects of invigorating qi and blood, nourishing yin, dispelling wind and relieving exterior symptoms, promoting blood circulation and detoxifying, moistening the lungs and resolving phlegm. It is used for symptoms such as consumptive tuberculosis and weak cough. It is recorded in classics such as "Guizhou Miao Medicine Research and Development", "Guizhou Chinese Medicine Resources", "Hunan Medicine Records", "Guizhou Herbs", and "Yunnan Chinese Herbs". It is a medicine used by ethnic minorities in our province. Modern pharmacological studies have shown that blue Buzheng has pharmacological activities such as anti-inflammatory and anti-tumor. In clinical practice, blue Buzheng is one of the main ingredients of the listed products "Fenghan Cold Capsules" and "Shengning Cold Liquid", and has a certain therapeutic effect on respiratory inflammatory diseases. However, there are no reports on Lanbu's research on improving and treating IPF.

[0004] Therefore, the current key research direction is to use network pharmacology and serum metabolomics methods to analyze the key components, potential targets, biological processes and metabolic pathways of Lanbuzheng and further explore the potential mechanism of Lanbuzheng in preventing and treating IPF. Summary of the invention

[0005] In order to solve the above technical problems existing in the prior art, the present invention provides a method for analyzing the chemical components of Lanbuzheng and a method for studying its mechanism of action for treating idiopathic pulmonary fibrosis, including the following contents:

[0006] A method for simultaneously analyzing multiple components in blue cloth, wherein the blue cloth is analyzed by UHPLC-Q-Orbitrap HRMS technology;

[0007] The specific chromatographic conditions are as follows:

[0008] WatersAcquity uplc hss T3 chromatographic column was used; mobile phase A: acetonitrile, mobile phase B: 0.1% formic acid water; column temperature: 40°C; flow rate: 0.3mL / min; injection volume: 10μL. Gradient elution conditions: 0-2min, 5%-5%A; 2-42min, 5%-95%A; 42-47min, 95%-95%A; 47-47.1min, 95%-5%A; 47.1-50min, 5%-5%A;

[0009] The specific mass spectrometry conditions are as follows:

[0010] The mass spectrometer system was Thermo Scientific Q Exactive Focus, using full scan

[0011] / dd-ms2, positive and negative ion scanning simultaneously, capillary voltage was 3.0 kV in positive ion mode, 2.5 kV in negative ion mode, heated electrospray ion source (HESI) temperature was 320 °C, capillary temperature was 350 °C, sheath gas flow rate was 35 arb, auxiliary gas flow rate was 10 arb. Scanning range was m / z 100-1500, MS1 resolution was 70000, MS / MS resolution was 17500, collision cell energy was

[0012] (NCE) is 20, 40 and 60 eV.

[0013] Xcalibur 3.0 software (Thermo Fisher Scientific) was used to process the mass spectrometry and chromatographic results. The elemental composition and molecular formula of the chromatographic peaks were accurately predicted based on the high-resolution mass spectrometry information, with an allowable error of no more than 5 × 10 -6 , and using Compound Discoverer 2.0 software (equipped with and database) for auxiliary analysis.

[0014] Furthermore, the blue cloth is the dried whole herb of Geum japonicum Thunb. var. chinense Bolle and Geum aleppicum Jacq. of the Rosaceae family.

[0015] Furthermore, the indigofera is an indigofera extract, which may be a water extract or an alcohol extract.

[0016] Furthermore, the Lanbuzheng extract is prepared by the following scheme: weigh the dried whole herb of Lanbuzheng, add distilled water to soak and boil, keep it slightly boiling, filter it with gauze while hot to obtain a filtrate; boil the residue again in the same way, filter it while hot, and combine the two filtrates; concentrate it under reduced pressure to form a clear paste, freeze-dry it, and obtain a powdery solid.

[0017] Furthermore, the test solution is prepared by the following method: accurately weigh the blue cloth water extract, put it in a volumetric flask, add methanol to dissolve it by ultrasonic, make the volume of methanol to the scale, centrifuge, and take the supernatant to obtain the test solution.

[0018] A method for studying the mechanism of action of Lanbuzheng in treating idiopathic pulmonary fibrosis comprises the following steps:

[0019] (1) Target prediction of chemical components of Lanbuzheng: Determine the target information of each component of the identified Lanbuzheng chemical components;

[0020] (2) Idiopathic pulmonary fibrosis (IPF) target prediction: search for “IPF” to obtain IPF disease targets;

[0021] (3) Acquisition of key targets: Based on the above target information, a Venn diagram of components and disease-related targets is drawn to obtain drug and disease intersection targets;

[0022] (4) Protein-protein interaction (PPI) analysis: import the above-mentioned intersection targets into the database and construct a PPI network diagram;

[0023] (5) GO and KEGG enrichment analysis: Using the database, GO and KEGG enrichment analysis was performed on common targets. The p-values ​​were ranked from small to large, the top 20 pathways were screened, and the results were visualized. On this basis, a “component-target-pathway” network was constructed.

[0024] Furthermore, the target prediction of the chemical components of LBZ is specifically to collect the structural information of each component using the Chemspider (www.chemspider.com) and Pubchem (pubchem.ncbi.nlm.nih.gov) databases for the identified chemical components, and use ChemDraw software to draw the chemical structure of each component, and save the results in sdf and mol file formats; SwissTargetPrediction (www.swisstargetprediction.ch / ) and PharmMapper (http: / / lilab-ecust.cn / pharmmapper / index.html) platforms are used to predict the targets related to the compounds, and the predicted target information is input according to the UniProKBt search function in the UniProt database (https: / / www.uniprot.org), and the prediction condition is limited to the target "Homo sapiens", and all target names are corrected to the official name (Officialgenesymbol) to obtain the target information of each component of LBZ.

[0025] Furthermore, the idiopathic pulmonary fibrosis IPF target prediction is specifically to use the OMIM database (www.omim.org) and the Genecard database (www.genecards.org) to enter the disease name "IPF" for retrieval to obtain IPF disease targets; in order to ensure the accuracy of the targets, the GeneCards database retrieval results need to use the score value as the screening condition, discard the targets below the median, and summarize the retrieval results of each database and remove duplicate genes to obtain IPF-related targets.

[0026] Furthermore, the key target acquisition is specifically as follows: according to the above target information, it is imported into Venny2.1.0 (https: / / bioinfogp.cnb.csic.es / tools / venny / index.html), a Venn diagram of components and disease-related targets is drawn, and the intersection target of drugs and diseases is obtained.

[0027] Furthermore, the protein-protein interaction (PPI) analysis was specifically as follows: the above-mentioned intersection targets were imported into the STRING (https: / / string-db.org / ) database, the species was selected as "Homo spanies", the confidence data was set to a network relationship higher than 0.9, and after removing free proteins, the results were imported into Cytoscape 3.7.1 software to construct a PPI network diagram.

[0028] Furthermore, the GO and KEGG enrichment analysis is specifically as follows: using the DAVID database (https: / / david.ncifcrf.gov / ), the species is limited to (Homo sapiens), GO and KEGG enrichment analysis is performed on the common targets (P<0.01), ranking them from small to large according to the p value, screening the top 20 pathways, and visualizing the results on the Micro-Information website (www.bioinforma tics.com.cn), on this basis, constructing a "component-target-pathway" network.

[0029] Compared with the prior art, the technical effects created by the present invention are embodied in:

[0030] (1) This application first uses UHPLC-Q-Orbitrap HRMS technology to qualitatively analyze the chemical components of blue cloth. The base peak chromatogram (BPC) under positive and negative ion modes is shown in ( Figure 1 A). By searching the Compound Discoverer 2.1 database and consulting the literature, a total of 27 chemical components were identified based on relative retention time, accurate molecular mass, secondary fragment ions and other information. The structures are shown in Figure 1 B. Its compound information and mass spectrum data are shown in Table 1.

[0031] (2) This application uses network pharmacology to explore the pharmacological mechanism of Lanbuzheng in improving IPF, and collects 734 targets related to 27 components identified in Lanbuzheng in the SwissTargetPrediction and PharmMapper databases. Subsequently, 1270 disease targets related to IPF were collected from the Genecards, OMIM and Drugbank databases. After taking the intersection of the component targets and the disease targets, 153 common targets were obtained, which may be potential targets for Lanbuzheng to improve IPF. The 153 key targets were analyzed using the String database and visualized as a protein-protein interaction (PPI) diagram using Cytoscape 3.9.1 software. The number of node connections represented the strength of the target interaction. The target Degree value was calculated using the plug-in CytoNCA in Cytoscape3.9.1 software. The change in node color from dark to light represented the corresponding target Degree value from large to small. The network diagram included 122 nodes and 368 edges, suggesting that the 122 targets may be potential targets for Lanbuzheng to prevent and treat IPF. Further, the 122 PPI targets were subjected to Go and KEGG enrichment analysis. The targets were related to 672 biological processes (biological processes, BP), 76 cellular components (cellular components, CC), and 133 molecular functions (molecular functions, MF). The top 10 results were visualized (see Figure 2), GO analysis results showed that the potential targets of Lanbuzheng were mainly enriched in positive regulation of protein kinase B signaling, positive regulation of MAPK cascade and inflammatory response for biological process (BP); cell composition (CC) was mainly enriched in extracellular region, cytoplasm and cytosol; molecular function (MF) was mainly enriched in identical protein binding, enzyme binding and protein kinase activity. KEGG was mainly enriched in HIF-1 signaling pathway, PI3K-Akt signaling pathway and arachidonic acid metabolism. Finally, the "component-target-pathway" network was constructed using Cytoscape 3.9.1 software, and the results suggested that the prevention and treatment of IPF by LBZ may be closely related to inflammation-related pathways. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 (A, B) are BPC diagrams of LBZ in negative ion and positive ion modes; (C) is the structure of the identified compound in LBZ.

[0033] Figure 2 This is the network pharmacology analysis of LBZ improving IPF, including (A) Venn diagram of the intersection of IPF targets and LBZ chemical component targets; (B) protein interaction network diagram; (C) component-target-pathway diagram; (D) GO enrichment analysis; (E) KEGG pathway analysis. DETAILED DESCRIPTION

[0034] The technical solution of the present invention is further defined below in conjunction with specific implementation methods, but the scope of protection required is not limited to the description.

[0035] Example:

[0036] Study on the mechanism of action of Lanbuzheng in improving idiopathic pulmonary fibrosis based on network pharmacology analysis

[0037] 1. Materials

[0038] 1.1 Drugs and reagents

[0039] LBZ sample (Weigh 500 g of dried whole herb of Lanbuzheng, add 10 times the amount of distilled water, soak for 30 min, boil, keep slightly boiling for 30 min, filter with gauze while hot to obtain the filtrate; decoct the residue once more in the same way, filter while hot, and combine the two filtrates. Concentrate under reduced pressure to 1 g·mL -1 The clear paste was placed in a freeze dryer and freeze-dried to obtain 139.5 g of powdered solid, with an extraction rate of 27.9%. The powder was stored in a -20°C refrigerator and was recorded as LBZ), acetonitrile, methanol (chromatographic grade, Honeywell, USA), chromatographic grade formic acid (Fisher, USA), Watsons water (Guangzhou Watsons Water Food and Beverage Co., Ltd.).

[0040] 1.2 Main instruments

[0041] Thermo Scientific Q Exactive Focus high-resolution mass spectrometer (Thermo Fisher Technology Co., Ltd., USA); DW-86L578J -80℃ refrigerator (Qingdao Haier Biomedical Co., Ltd.); DK-98-ⅡA electric hot water bath (Tianjin Test Instrument Co., Ltd.); MS105DC electronic balance (Mettler, Switzerland); KQ-100E ultrasonic extractor (Shanghai Kedao Ultrasonic Instrument Co., Ltd.); TGL16M high-speed centrifuge (Hunan Kaida Scientific Instrument Co., Ltd.)

[0042] 2. Methods

[0043] 2.1 Analysis of chemical composition of LBZ

[0044] 2.1.1 Preparation of test solution

[0045] Accurately weigh 0.1 g of LBZ and place it in a 10 mL volumetric flask. Add a small amount of 70% methanol and ultrasonically dissolve it for 10 min. Make up to the volume with methanol. Centrifuge at 12000 r / min and 4 °C for 10 min. Take the supernatant to obtain the test solution for LC-MS analysis.

[0046] 2.1.2 Chromatographic conditions

[0047] A Waters Acquity uplc hss T3 column (2.1×100 mm, 1.8 μm) was used; mobile phase A: acetonitrile, mobile phase B: 0.1% formic acid water; column temperature: 40°C; flow rate: 0.3 mL / min; injection volume: 10 μL. Gradient elution conditions: 0-2 min, 5%-5% A; 2-42 min, 5%-95% A; 42-47 min, 95%-95% A; 47-47.1 min, 95%-5% A; 47.1-50 min, 5%-5% A.

[0048] 2.1.3 Mass spectrometry conditions

[0049] The mass spectrometer system was Thermo Scientific Q Exactive Focus, using full scan / dd-ms2, positive and negative ion scanning simultaneously, capillary voltage was 3.0 kV in positive ion mode, 2.5 kV in negative ion mode, heated electrospray ion source (HESI) temperature was 320 °C, capillary temperature was 350 °C, sheath gas flow rate was 35 arb, auxiliary gas flow rate was 10 arb. The scanning range was m / z 100-1500, MS1 resolution was 70000, MS / MS resolution was 17500, and collision cell energy (NCE) was 20, 40 and 60 eV.

[0050] 2.1.4 LBZ composition analysis

[0051] Xcalibur 3.0 software (Thermo Fisher Scientific) was used for data processing. The elemental composition and molecular formula of the chromatographic peaks were accurately predicted based on the high-resolution mass spectrometry information, with an allowable error of no more than 5 × 10 -6 , and use Compound Discoverer2.0 software (equipped with and database) for auxiliary analysis.

[0052] 2.2 Network pharmacology analysis

[0053] 2.2.1 LBZ chemical composition target prediction

[0054] The chemical components identified under 2.1 were used to collect the structural information of each component using the Chemspider (www.chemspider.com) and Pubchem (pubchem.ncbi.nlm.nih.gov) databases, and the chemical structures of each component were drawn using ChemDraw software, and the results were saved in sdf and mol file formats. The SwissTargetPrediction (www.swisstargetprediction.ch / ) and PharmMapper (http: / / lilab-ecust.cn / pharmmapper / index.html) platforms were used to predict the targets related to the compounds. According to the UniProKBt search function in the UniProt database (https: / / www.uniprot.org), the predicted target information was entered, and the prediction condition was limited to the target "Homo sapiens". All target names were corrected to official names (Officialgene symbol), and the target information of each component of LBZ was obtained.

[0055] 2.2.2 IPF target prediction

[0056] The disease name "IPF" was searched using the OMIM database (www.omim.org) and the Genecard database (www.genecards.org) to obtain IPF disease targets. In order to ensure the accuracy of the targets, the GeneCards database search results need to be filtered using the score value, and targets below the median are discarded. The search results of each database are summarized and duplicate genes are removed to obtain IPF-related targets.

[0057] 2.2.3 Acquisition of key targets

[0058] According to the above target information, it was imported into Venny2.1.0 (https: / / bioinfogp.cnb.csic.es / tools / venny / index.html), and the Venn diagram of components and disease-related targets was drawn to obtain the intersection targets of drugs and diseases.

[0059] 2.2.4 Protein-protein interaction (PPI) analysis

[0060] The above intersection targets were imported into the STRING (https: / / string-db.org / ) database, the species was selected as “Homo spanies”, the confidence data was set to a network relationship higher than 0.9, and after removing free proteins, the results were imported into Cytoscape 3.7.1 software to construct a PPI network diagram.

[0061] 2.2.5GO and KEGG enrichment analysis

[0062] The DAVID database (https: / / david.ncifcrf.gov / ) was used to limit the species to Homo sapiens. GO and KEGG enrichment analysis was performed on the common targets (P<0.01). The pathways were ranked from small to large according to the p-value, and the top 20 pathways were screened. The results were visualized on the Microinformation website (www.bioinforma tics.com.cn), and a "component-target-pathway" network was constructed on this basis.

[0063] 3. Results

[0064] 3.1 Preliminary identification of chemical components in LBZ

[0065] In this study, UHPLC-Q-Orbitrap HRMS technology was used for the first time to qualitatively analyze the chemical components of the aqueous extract of blue cloth. The base peak chromatogram (BPC) under positive and negative ion modes was shown in Figure 1 (A, B). By searching the Compound Discoverer 2.1 database and consulting the literature, a total of 27 chemical components were identified based on relative retention time, accurate molecular mass, secondary fragment ions and other information. The structures are shown in Figure 1 C. Its compound information and mass spectrum data are shown in Table 1.

[0066] Table 1. Identification of chemical components of LBZ

[0067]

[0068]

[0069] 3.2GO and KEGG enrichment and network analysis of LBZ on IPF-related targets

[0070] Network pharmacology was used to explore the pharmacological mechanism of LBZ in improving IPF. 734 targets related to 27 components identified in LBZ were collected in the SwissTargetPrediction and PharmMapper databases. Subsequently, 1270 disease targets related to IPF were collected from the Genecards, OMIM, and Drugbank databases. After the intersection of the component targets and the disease targets was taken, 153 common targets were obtained ( Figure 2A). 153 key targets were analyzed using the String database and visualized as a protein-protein interaction (PPI) diagram using Cytoscape 3.9.1 software. The number of node connections represented the strength of the target interaction. The target Degree value was calculated using the plug-in CytoNCA in Cytoscape 3.9.1 software. The change of node color from dark to light represented the process of the corresponding target Degree value from large to small. The network diagram included 122 nodes and 368 edges, suggesting that the 122 targets may be potential targets for LBZ to prevent and treat IPF ( Figure 2 B). Further, 122 PPI targets were subjected to Go and KEGG enrichment analysis, and the targets were related to 672 biological processes (BP), 76 cellular components (CC), and 133 molecular functions (MF). The top 10 results were visualized ( Figure 2 D), GO analysis results showed that the potential targets of LBZ were mainly enriched in the positive regulation of protein kinase B signaling, positive regulation of MAPK cascade and inflammatory response for biological processes (BP); the cell composition (CC) was mainly enriched in the extracellular region, cytoplasm and cytosol; the molecular function (MF) was mainly enriched in the binding of the same protein, enzyme binding and protein kinase activity. KEGG was mainly enriched in the HIF-1 signaling pathway, PI3K-Akt signaling pathway and arachidonic acid metabolism (Arachidonic acid metabolism). Figure 2 E). Finally, the “component-target-pathway” network was constructed using Cytoscape 3.9.1 software ( Figure 2 C), the results suggest that the prevention and treatment of IPF by LBZ may be closely related to inflammation-related pathways.

[0071] Based on UHPLC-Q-Orbitrap HRMS technology, a total of 27 compounds were analyzed, mainly including flavonoids, tannins, lignins and terpenoids. Through the identification of 27 potential active ingredients of LBZ and network pharmacology analysis, PI3K / Akt, TNF and arachidonic acid pathways may be the key pathways of LBZ-mediated IPF. Activation of PI3K-Akt signaling pathway and TNF signaling pathway can release inflammatory factors, such as TNF-α and IL-6, which are involved in the formation of IPF. Arachidonic acid can be converted into lipid mediators under the action of lipoxygenase (LOX), such as arachidonic acid can be converted into lipid mediators such as leukotrienes (LTs) and lipotoxins (LXs) through the lipoxygenase (LOX) pathway, and converted into prostaglandins (PGs) and thromboxanes (TXs) through the cyclooxygenase (COX) pathway. These products may cause inflammation, fibrosis and other adverse changes. In summary, LBZ may regulate the HIF-1 signaling pathway, PI3K-Akt signaling pathway and arachidonic acid metabolism through targets such as TP53 and AKT, suggesting that LBZ intervention in IPF may be closely related to inflammation-related pathways.

[0072] Finally, it should be pointed out that the above embodiments are only representative examples of the present invention. Obviously, the technical solution of the present invention is not limited to the above embodiments, and there are many variations. All variations that can be directly derived or associated with the content disclosed by ordinary technicians in this field should be considered as the protection scope of the present invention.

Claims

1. A method for analyzing multiple components in blue cloth, characterized in that: The blue cloth was analyzed using UHPLC-Q-Orbitrap HRMS technology; The specific chromatographic conditions are as follows: A Waters Acquity uplc hss T3 column was used, with specifications of 2.1×100 mm, 1.8 μm; mobile phase A: acetonitrile, mobile phase B: 0.1% formic acid water; column temperature: 40°C; flow rate: 0.3 mL / min; injection volume: 10 μL. Gradient elution conditions: 0-2 min, 5%-5% A; 2-42 min, 5%-95% A; 42-47 min, 95%-95% A; 47-47.1 min, 95%-5% A; 47.1-50 min, 5%-5% A; The specific mass spectrometry conditions are as follows: The mass spectrometer system was Thermo Scientific Q Exactive Focus, using full scan / dd-ms2, positive and negative ion scanning simultaneously, capillary voltage was 3.0 kV in positive ion mode, 2.5 kV in negative ion mode, heated electrospray ion source (HESI) temperature was 320 °C, capillary temperature was 350 °C, sheath gas flow rate was 35 arb, auxiliary gas flow rate was 10 arb. The scanning range was m / z 100-1500, MS1 resolution was 70000, MS / MS resolution was 17500, and collision cell energy (NCE) was 20, 40 and 60 eV.

2. The method for analyzing multiple components in blue cloth according to claim 1, characterized in that: The blue cloth is the whole herb of the Rosaceae plant, the pubescent green grass or the green grass.

3. The method for analyzing multiple components in blue cloth according to claim 2, characterized in that: The indigofera is an indigofera extract.

4. The method for analyzing multiple components in a blue cloth according to claim 2, characterized in that: The Lanbuzheng extract is prepared by the following scheme: weighing the dried whole herb of Lanbuzheng, soaking it in distilled water and then boiling it, keeping it slightly boiling, filtering it with gauze while it is hot to obtain a filtrate; decocting the residue once more in the same way, filtering it while it is hot, and combining the two filtrates; concentrating it under reduced pressure into a clear paste, freeze-drying it, and obtaining a powdery solid.

5. The method for analyzing multiple components in blue cloth according to claim 1, characterized in that: The test solution is prepared by the following method: accurately weigh the blue cloth extract, put it in a volumetric flask, add methanol to dissolve it by ultrasonic, make the volume of methanol to the scale, centrifuge, and take the supernatant to obtain the test solution.

6. The method for analyzing multiple components in a blue cloth according to claim 1, characterized in that: Xcalibur 3.0 software was used to process the mass spectrometry and chromatographic analysis results. The elemental composition and molecular formula of the chromatographic peaks were accurately predicted based on the high-resolution mass spectrometry information, with an allowable error of no more than 5 × 10 -6 , and Compound Discoverer 2.0 software was used for auxiliary analysis.

7. A method for studying the mechanism of action of Lanbuzheng in treating idiopathic pulmonary fibrosis based on network pharmacology analysis, characterized in that: The steps include: (1) Target prediction of chemical components of Lanbuzheng: Determine the target information of each component of the identified Lanbuzheng chemical components; (2) Idiopathic pulmonary fibrosis (IPF) target prediction: search for "IPF" to obtain IPF disease targets; (3) Acquisition of key targets: Based on the above target information, a Venn diagram of components and disease-related targets is drawn to obtain drug and disease intersection targets; (4) Protein-protein interaction (PPI) analysis: import the above-mentioned intersection targets into the database and construct a PPI network diagram; (5) GO and KEGG enrichment analysis: Using the database, GO and KEGG enrichment analysis was performed on common targets. The p-values ​​were ranked from small to large, the top 20 pathways were screened, and the results were visualized. On this basis, a "component-target-pathway" network was constructed.

8. The method for studying the mechanism of action of Lanbuzheng in treating idiopathic pulmonary fibrosis according to claim 7, characterized in that: The target prediction of the chemical components of Lanbuzheng is specifically to collect the structural information of each component of the identified chemical components using the Chemspider and Pubchem databases, and use the ChemDraw software to draw the chemical structure of each component, and save the results in sdf and mol file formats; use the SwissTargetPrediction and PharmMapper platforms to predict the targets related to the compounds, and input the predicted target information according to the UniProKBt search function in the UniProt database. The prediction condition is limited to the target "Homo sapiens", and all target names are corrected to official names to obtain the target information of each component of Lanbuzheng.

9. The method for studying the mechanism of action of Lanbuzheng in treating idiopathic pulmonary fibrosis according to claim 7, characterized in that: The idiopathic pulmonary fibrosis IPF target prediction specifically uses the OMIM database and the Genecard database to input the disease name "IPF" for retrieval to obtain IPF disease targets; in order to ensure the accuracy of the targets, the GeneCards database retrieval results need to use the score value as the screening condition, discard targets below the median, and summarize the retrieval results of each database and remove duplicate genes to obtain IPF-related targets.

10. The method for studying the mechanism of action of Lanbuzheng in treating idiopathic pulmonary fibrosis according to claim 7, characterized in that: The key target acquisition is specifically as follows: according to the above target information, it is imported into Venny2.1.0, a Venn diagram of components and disease-related targets is drawn, and the intersection target of drugs and diseases is obtained.

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