A method for analyzing the active components of Radix Panacis radix in the treatment of acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology

Abstract

The present invention discloses a method and application thereof for analyzing the active ingredients of Radix Trichosanthis for treating acute pancreatitis based on HPLC-Q-TOF-MS, network pharmacology, and molecular docking. The analysis method uses HPLC-Q-TOF-MS / MS analysis technology to generate a molecular network of chemical components through accurate mass number, secondary mass spectrometry data, mass spectrometry fragmentation rules, and a database. Then, using a network pharmacology-related database, acute pancreatitis disease-related targets are obtained, a "component-disease-target" network diagram is constructed, a protein interaction network is constructed, and GO function and KEGG pathway enrichment analysis are performed. The 10 key chemical components obtained are: quercetin, coumarin glucoside, gentiopicroside, eriodictyol-7-O-glucoside, protocatechuic acid, 3-O-caffeoylquinic acid, astragalin, isoorientin, isovitexin, and mangiferin.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to a method for analyzing the effective components of Radix Panacis Tiliraceae for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology. Background Art

[0002] Acute pancreatitis is a digestive disease in which pancreatic enzymes are activated within the pancreas, triggering the release of inflammatory factors and leading to a local and systemic inflammatory response. Key clinical symptoms include pancreatic edema, secondary infection, hemorrhage and necrosis, peritonitis, and multiple organ failure. 20% to 25% of patients present with severe disease, developing local or systemic complications, multiple organ failure, and even life-threatening conditions. In acute pancreatitis, excessive white blood cell activation triggers a respiratory burst, producing oxygen free radicals. Excessive oxygen free radicals can also damage acinar cells, triggering both extracellular and intracellular activation of pancreatic enzymes, leading to a vicious cycle of pancreatic damage. Currently, drugs used clinically to treat acute pancreatitis include the synthetic octapeptide cyclic compound octreotide, the protease inhibitor ulinastatin, and omeprazole injection. Octreotide can mitigate pancreatic tissue damage, while ulinastatin not only inhibits trypsin and glycolytic enzymes but also suppresses the production of inflammatory factors and endogenous shock factor. In addition, the combination of Chinese and Western medicine, such as Dachengqi Decoction or Xuebijing combined with alanine-glutamine, also has certain therapeutic effects.

[0003] With the widespread promotion of traditional Chinese medicine (TCM) in clinical applications, it has achieved remarkable therapeutic effects in the treatment of acute pancreatitis. One or more chemical component groups contained in TCM can treat acute pancreatitis from multiple targets, multiple angles, and a comprehensive perspective. Different active ingredient groups have different focuses on the effects on acute pancreatitis, including blocking inflammatory reactions, improving pancreatic secretory function, regulating immune inflammatory reactions, effectively scavenging oxygen free radicals, and alleviating oxidative stress reactions. Therefore, it is very necessary to discover chemical components from TCM that have the potential to treat acute pancreatitis. Hypericum japonicum Thunb.ex Murray is the whole herb of the Hypericum japonicum plant of the genus Hypericum in the family Garciniaaceae. It is an annual herb. It has a sweet, bitter, and cool nature and a slightly fragrant odor. It enters the lung, liver, and stomach meridians. Its main functions include clearing away heat and detoxifying, promoting blood circulation and reducing swelling, promoting dampness and jaundice, and dispersing blood stasis and relieving pain.

[0004] Network pharmacology provides a holistic perspective on the relationship between drugs and diseases by constructing drug-disease-target networks. Because it offers a holistic and systematic visualization of the multi-component, multi-target, and multi-pathway characteristics of Traditional Chinese Medicine (TCM), it has been widely used to predict potential active ingredients in TCM and study their mechanisms of action on specific disease targets.

[0005] However, there is no literature report that Radix Panacis can treat acute pancreatitis, and a method for studying the effective ingredients of Radix Panacis for treating acute pancreatitis using network pharmacology methods is used. The present invention adopts high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS / MS) to analyze and identify the chemical components of Radix Panacis, uses network pharmacology to explore the mechanism of action of Radix Panacis in treating acute pancreatitis, and preliminarily verifies the network pharmacology results using an acute pancreatitis rat model, aiming to clarify the pharmacological material basis of Radix Panacis in treating acute pancreatitis and provide a theoretical basis for new drug research and development. It also provides certain reference and reference significance for the subsequent development and clinical application of Radix Panacis medicinal materials. Summary of the Invention

[0006] The purpose of the present invention is to determine a method for analyzing the effective components of Tianjihuang for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology, and to obtain the effective components, key targets and pathways with high enrichment significance of Tianjihuang for treating acute pancreatitis.

[0007] The technical solution of the present invention is a method for analyzing the effective components of Radix Panacis Nigrum for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology. The analysis method adopts HPLC-Q-TOF-MS / MS analysis technology to generate a molecular network of chemical components through accurate mass number, secondary mass spectrometry data, mass spectrometry fragmentation rules and database, and then uses the network pharmacology related database to obtain acute pancreatitis disease-related targets, construct a "component-disease-target" network diagram, construct a protein interaction network, and then perform GO function and KEGG pathway enrichment analysis.

[0008] The aforementioned method for analyzing the active ingredients of Radix Panacis radix for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology is carried out according to the following steps:

[0009] (1) Preparation of Rhizoma Cibotii extract: Weigh 350-400 g of Rhizoma Cibotii medicinal material, crush it, add 1.8-2.5 L of 70-90% ethanol, heat and reflux and extract 3-5 times, each time for 1.5-2.5 hours, combine the extracts, and recover the solvent under reduced pressure to obtain the Rhizoma Cibotii extract, which is stored in a refrigerator at -20°C for later use;

[0010] (2) HPLC-Q-TOF-MS / MS analysis: The retention time of the chromatographic peaks, high-resolution mass spectrometry data, and multi-stage mass spectrometry data were compared with the GNPS spectral library data. The molecular network of chemical components was generated by accurate mass, secondary mass spectrometry data, mass spectrometry fragmentation patterns, and the database to obtain the structural information of the chemical components of the Rhizoma Cibotii extract;

[0011] HPLC conditions: Waters XSelect CSH-C18 column, 150 × 4.6 mm, 2.5 μm; mobile phase A: 0.1% formic acid in water, mobile phase B: acetonitrile; gradient elution; flow rate: 0.4 mL / min, column temperature: 35°C, injection volume: 2 μL, split ratio: 1:4;

[0012] Mass spectrometry conditions: ESI ion source positive and negative ion modes; drying gas temperature 350°C; drying gas flow rate 10 L / min; nebulizer gas 45 psig; sheath gas temperature 350°C; sheath gas flow rate 10 L / min; capillary voltage 4000 V; fragmentor voltage 180 V; cone voltage 65 V; mass scan range: 100–2000 Da;

[0013] (3) Screening of active chemical component targets of Rhizoma Cibotii: The structural information of the chemical components of Rhizoma Cibotii extract identified by HPLC-Q-TOF-MS / MS analysis was input into the TCMSP database, SwissTargetPrediction database, and UniProt database to calibrate the combined targets, eliminate non-human genes, and obtain the targets of the chemical components of Rhizoma Cibotii;

[0014] (4) Screening of disease targets related to acute pancreatitis: Acute pancreatitis disease targets were obtained by using the three databases of OMIM, DrugBank and GeneCards with the keyword of Acute pancreatitis. The acute pancreatitis disease targets were intersected with the targets of the chemical components of the Radix Glehniae in step (3) using the Venny2.1 online analysis tool to obtain potential targets of the Radix Glehniae extract for the treatment of acute pancreatitis, i.e., potential targets.

[0015] (5) Constructing a “component-disease-target” network diagram: The potential targets in step (4) were predicted using the SwissTargetPrediction database, and after deleting duplicate data, the data were imported into the Cytoscape software to construct a “compound-disease-target” network, obtaining 10 key chemical components;

[0016] (6) Construction of protein interaction network: The potential target genes in step (4) were imported into the STRING database, “multiple protein” was selected, and the species was limited to “Homo sapiens” to obtain the protein interaction relationship. After exporting the data, it was imported into Cytoscape software, and the network parameters were calculated using the Network Analyze plug-in to screen out 10 key targets;

[0017] (7) GO function and KEGG pathway enrichment analysis: Using the Metascape data platform, we performed enrichment analysis on the main biological processes and metabolic pathways of chemical component-disease intersection targets to obtain pathways with high enrichment significance.

[0018] In the above step (1), the preparation of the Rhizoma Cibotii extract is as follows: 380 g of Rhizoma Cibotii medicinal material is weighed, crushed, 2 L of 80% ethanol is added, and the mixture is heated under reflux and extracted 4 times, each time for 2 hours. The extracts are combined, and the solvent is recovered under reduced pressure to obtain the Rhizoma Cibotii extract, which is then stored in a refrigerator at -20°C for later use.

[0019] In the aforementioned step (2), the gradient elution program of the HPLC chromatographic conditions is: 0-30 min, 5-65% B; 30-32 min, 65-90% B; 32-40 min, 90% B; 40-40.01 min, 90-5% B; 40.01-45.01 min, 5% B.

[0020] In the aforementioned step (5), the 10 key chemical components are: quercetin, coumarin glucoside, gentiopicroside, eriodictyol-7-O-glucoside, protocatechuic acid, 3-O-caffeoylquinic acid, astragaloside, isoorientin, isovitexin, and mangiferin.

[0021] In the aforementioned step (6), the key targets are albumin, glyceraldehyde-3-phosphate dehydrogenase, HRAS protein encoding gene, tyrosine protein kinase, heat shock protein 90α encoding gene, myeloid cell leukemia sequence 1, vascular endothelial growth factor receptor 2 encoding gene, phosphatidylinositol 3-kinase regulatory subunit 1, hepatocyte growth factor receptor HGFR, and PTK2 protein tyrosine kinase 2.

[0022] In the aforementioned step (7), the pathways with high enrichment significance are the calcium signaling pathway and the HIF-1 signaling pathway.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] 1. The present invention uses HPLC-Q-TOF-MS to comprehensively analyze the chemical components of Radix Glehniae and finds that Radix Glehniae has the effect of treating acute pancreatitis.

[0025] 2. A "component-disease-target" network diagram was constructed, and 10 chemical components with therapeutic effects on acute pancreatitis were obtained: quercetin, coumarin glucoside, gentiopicroside, eriodictyol-7-O-glucoside, protocatechuic acid, 3-O-caffeoylquinic acid, astragaloside, isoorientin, isovitexin, and mangiferin.

[0026] 3. A protein interaction network was constructed and 10 targets related to acute pancreatitis were screened out: albumin, glyceraldehyde-3-phosphate dehydrogenase, HRAS protein encoding gene, tyrosine protein kinase, heat shock protein 90α encoding gene, myeloid cell leukemia sequence 1, vascular endothelial growth factor receptor 2 encoding gene, phosphatidylinositol 3-kinase regulatory subunit 1, hepatocyte growth factor receptor HGFR, and PTK2 protein tyrosine kinase 2.

[0027] 3. Through GO function and KEGG pathway enrichment analysis: the pathways with high enrichment significance were determined to be calcium signaling pathway and HIF-1 signaling pathway.

[0028] 4. Tianjihuang can inhibit the secretion of digestive enzymes in rats with acute pancreatitis, reduce the degree of oxidative stress in the body, and have antioxidant effects, which was confirmed by the results of network pharmacology prediction. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Venn diagram of the active ingredients of Tianjihuang and the targets of acute pancreatitis;

[0030] Figure 2 The "chemical component-disease-target" network of the active ingredients of Radix Panacis radiata and the targets of acute pancreatitis;

[0031] Figure 3 Protein interaction PPI network diagram of core targets;

[0032] Figure 4 GO analysis of core targets;

[0033] Figure 5 KEGG enrichment analysis of core targets;

[0034] Figure 6 Effects of Tianjihuang on the pathological morphology of pancreatic tissue in rats with acute pancreatitis;

[0035] Figure 7 The levels of AMS, LPS, MDA, H2O2, GSH-PX and NO in the serum of rats in each group at 6h and 12h. DETAILED DESCRIPTION

[0036] The present invention will be described in detail below with reference to the examples and accompanying drawings. However, the following examples should not be considered as limiting the scope of the present invention.

[0037] Example 1:

[0038] (1) Preparation of Rhizoma Cibotii extract: Weigh 380 g of Rhizoma Cibotii medicinal material, crush it, add 2 L of 80% ethanol, heat and reflux and extract it 4 times, each time for 2 hours, combine the extracts, and recover the solvent under reduced pressure to obtain Rhizoma Cibotii extract, which is stored in a refrigerator at -20°C for later use.

[0039] (2) HPLC-Q-TOF-MS / MS analysis: The retention time of the chromatographic peaks, high-resolution mass spectrometry data, and multi-stage mass spectrometry data were compared with the GNPS spectral library data. The molecular network of chemical components was generated by accurate mass, secondary mass spectrometry data, mass spectrometry fragmentation patterns, and the database to obtain the structural information of the chemical components of the Rhizoma Cibotii extract;

[0040] HPLC conditions: Waters XSelect CSH-C18 column, 150 × 4.6 mm, 2.5 μm; mobile phase A: 0.1% formic acid in water, mobile phase B: acetonitrile; gradient elution; flow rate: 0.4 mL / min, column temperature: 35°C, injection volume: 2 μL, split ratio: 1:4;

[0041] The gradient elution program of HPLC chromatography conditions was as follows: 0-30 min, 5-65% B; 30-32 min, 65-90% B; 32-40 min, 90% B; 40-40.01 min, 90-5% B; 40.01-45.01 min, 5% B;

[0042] Mass spectrometry conditions: ESI ion source positive and negative ion modes; drying gas temperature 350°C; drying gas flow rate 10 L / min; nebulizer gas 45 psig; sheath gas temperature 350°C; sheath gas flow rate 10 L / min; capillary voltage 4000 V; fragmentor voltage 180 V; cone voltage 65 V; mass scan range: 100-2000 Da.

[0043] (3) Screening of active chemical component targets of Rhizoma Cibotii: The structural information of the chemical components of Rhizoma Cibotii extract identified by HPLC-Q-TOF-MS / MS analysis was input into the TCMSP database, SwissTargetPrediction database and UniProt database to calibrate the combined targets, eliminate non-human genes, and obtain the target of the chemical components of Rhizoma Cibotii.

[0044] (4) Screening of disease targets related to acute pancreatitis: Acute pancreatitis disease targets were obtained by using the three databases of OMIM, DrugBank, and GeneCards with the keyword "Acute pancreatitis". The acute pancreatitis disease targets were intersected with the targets of the chemical components of Rhizoma Cibotii using the Venny2.1 online analysis tool to obtain potential targets of Rhizoma Cibotii extract for the treatment of acute pancreatitis.

[0045] (5) Constructing a “component-disease-target” network diagram: The target information of potential targets was predicted using the SwissTargetPrediction database, and after deleting duplicate data, the data were imported into the Cytoscape software to construct a “compound-disease-target” network. Ten key chemical components were obtained: quercetin, coumarin glucoside, gentiopicroside, eriocarbamate-7-O-glucoside, protocatechuic acid, 3-O-caffeoylquinic acid, astragaloside, isoorientin, isovitexin, and mangiferin.

[0046] (6) Construction of protein interaction network: Potential target genes were imported into the STRING database, “multiple protein” was selected, and the species was limited to “Homo sapiens” to obtain the protein interaction relationship. After exporting the data, it was imported into Cytoscape software, and the network parameters were calculated using the Network Analyze plug-in. Ten key targets were screened out: albumin, glyceraldehyde-3-phosphate dehydrogenase, HRAS protein encoding gene, tyrosine protein kinase, heat shock protein 90α encoding gene, myeloid cell leukemia sequence 1, vascular endothelial growth factor receptor 2 encoding gene, phosphatidylinositol 3-kinase regulatory subunit 1, hepatocyte growth factor receptor HGFR, and PTK2 protein tyrosine kinase 2.

[0047] (7) GO function and KEGG pathway enrichment analysis: Using the Metascape data platform, we performed enrichment analysis on the main biological processes and metabolic pathways of chemical component-disease intersection targets, and found that the pathways with high enrichment significance were the calcium signaling pathway and the HIF-1 signaling pathway.

[0048] Example 2:

[0049] (1) Preparation of Rhizoma Cibotii extract: Weigh 350 g of Rhizoma Cibotii medicinal material, crush it, add 1.8 L of 70% ethanol, heat and reflux and extract it three times, each time for 1.5 h, combine the extracts, and recover the solvent under reduced pressure to obtain the Rhizoma Cibotii extract, which is stored in a refrigerator at -20°C for later use;

[0050] (2) HPLC-Q-TOF-MS / MS analysis: The retention time of the chromatographic peaks, high-resolution mass spectrometry data, and multi-stage mass spectrometry data were compared with the GNPS spectral library data. The molecular network of chemical components was generated by accurate mass, secondary mass spectrometry data, mass spectrometry fragmentation patterns, and the database to obtain the structural information of the chemical components of the Rhizoma Cibotii extract;

[0051] HPLC conditions: Waters XSelect CSH-C18 column, 150 × 4.6 mm, 2.5 μm; mobile phase A: 0.1% formic acid in water, mobile phase B: acetonitrile; gradient elution; flow rate: 0.4 mL / min, column temperature: 35°C, injection volume: 2 μL, split ratio: 1:4;

[0052] The gradient elution program of HPLC chromatography conditions was as follows: 0-30 min, 5-65% B; 30-32 min, 65-90% B; 32-40 min, 90% B; 40-40.01 min, 90-5% B; 40.01-45.01 min, 5% B;

[0053] Mass spectrometry conditions: ESI ion source positive and negative ion modes; drying gas temperature 350°C; drying gas flow rate 10 L / min; nebulizer gas 45 psig; sheath gas temperature 350°C; sheath gas flow rate 10 L / min; capillary voltage 4000 V; fragmentor voltage 180 V; cone voltage 65 V; mass scan range: 100-2000 Da.

[0054] (3) Screening of active chemical component targets of Rhizoma Cibotii: The structural information of the chemical components of Rhizoma Cibotii extract identified by HPLC-Q-TOF-MS / MS analysis was input into the TCMSP database, SwissTargetPrediction database and UniProt database to calibrate the combined targets, eliminate non-human genes, and obtain the target of the chemical components of Rhizoma Cibotii.

[0055] (4) Screening of disease targets related to acute pancreatitis: Acute pancreatitis disease targets were obtained by using the three databases of OMIM, DrugBank, and GeneCards with the keyword "Acute pancreatitis". The acute pancreatitis disease targets were intersected with the targets of the chemical components of Rhizoma Cibotii using the Venny2.1 online analysis tool to obtain potential targets of Rhizoma Cibotii extract for the treatment of acute pancreatitis.

[0056] (5) Constructing a “component-disease-target” network diagram: The target information of potential targets was predicted using the SwissTargetPrediction database, and after deleting duplicate data, the data were imported into the Cytoscape software to construct a “compound-disease-target” network. Ten key chemical components were obtained: quercetin, coumarin glucoside, gentiopicroside, eriocarbamate-7-O-glucoside, protocatechuic acid, 3-O-caffeoylquinic acid, astragaloside, isoorientin, isovitexin, and mangiferin.

[0057] (6) Construction of protein interaction network: Potential target genes were imported into the STRING database, “multiple protein” was selected, and the species was limited to “Homo sapiens” to obtain the protein interaction relationship. After exporting the data, it was imported into Cytoscape software, and the network parameters were calculated using the Network Analyze plug-in. Ten key targets were screened out: albumin, glyceraldehyde-3-phosphate dehydrogenase, HRAS protein encoding gene, tyrosine protein kinase, heat shock protein 90α encoding gene, myeloid cell leukemia sequence 1, vascular endothelial growth factor receptor 2 encoding gene, phosphatidylinositol 3-kinase regulatory subunit 1, hepatocyte growth factor receptor HGFR, and PTK2 protein tyrosine kinase 2.

[0058] (7) GO function and KEGG pathway enrichment analysis: Using the Metascape data platform, we performed enrichment analysis on the main biological processes and metabolic pathways of chemical component-disease intersection targets, and found that the pathways with high enrichment significance were the calcium signaling pathway and the HIF-1 signaling pathway.

[0059] Example 3:

[0060] (1) Preparation of Rhizoma Cibotii extract: Weigh 400 g of Rhizoma Cibotii medicinal material, crush it, add 2.5 L of 90% ethanol, heat and reflux and extract it 5 times, each time for 2.5 h, combine the extracts, and recover the solvent under reduced pressure to obtain the Rhizoma Cibotii extract, which is stored in a refrigerator at -20°C for later use;

[0061] (2) HPLC-Q-TOF-MS / MS analysis: The retention time of the chromatographic peaks, high-resolution mass spectrometry data, and multi-stage mass spectrometry data were compared with the GNPS spectral library data. The molecular network of chemical components was generated by accurate mass, secondary mass spectrometry data, mass spectrometry fragmentation patterns, and the database to obtain the structural information of the chemical components of the Rhizoma Cibotii extract;

[0062] HPLC conditions: Waters XSelect CSH-C18 column, 150 × 4.6 mm, 2.5 μm; mobile phase A: 0.1% formic acid in water, mobile phase B: acetonitrile; gradient elution; flow rate: 0.4 mL / min, column temperature: 35°C, injection volume: 2 μL, split ratio: 1:4;

[0063] The gradient elution program of HPLC chromatography conditions was as follows: 0-30 min, 5-65% B; 30-32 min, 65-90% B; 32-40 min, 90% B; 40-40.01 min, 90-5% B; 40.01-45.01 min, 5% B;

[0064] Mass spectrometry conditions: ESI ion source positive and negative ion modes; drying gas temperature 350°C; drying gas flow rate 10 L / min; nebulizer gas 45 psig; sheath gas temperature 350°C; sheath gas flow rate 10 L / min; capillary voltage 4000 V; fragmentor voltage 180 V; cone voltage 65 V; mass scan range: 100-2000 Da.

[0065] (3) Screening of active chemical component targets of Rhizoma Cibotii: The structural information of the chemical components of Rhizoma Cibotii extract identified by HPLC-Q-TOF-MS / MS analysis was input into the TCMSP database, SwissTargetPrediction database and UniProt database to calibrate the combined targets, eliminate non-human genes, and obtain the target of the chemical components of Rhizoma Cibotii.

[0066] (4) Screening of disease targets related to acute pancreatitis: Acute pancreatitis disease targets were obtained by using the three databases of OMIM, DrugBank, and GeneCards with the keyword "Acute pancreatitis". The acute pancreatitis disease targets were intersected with the targets of the chemical components of Rhizoma Cibotii using the Venny2.1 online analysis tool to obtain potential targets of Rhizoma Cibotii extract for the treatment of acute pancreatitis.

[0067] (5) Constructing a “component-disease-target” network diagram: The target information of potential targets was predicted using the SwissTargetPrediction database, and after deleting duplicate data, the data were imported into the Cytoscape software to construct a “compound-disease-target” network. Ten key chemical components were obtained: quercetin, coumarin glucoside, gentiopicroside, eriocarbamate-7-O-glucoside, protocatechuic acid, 3-O-caffeoylquinic acid, astragaloside, isoorientin, isovitexin, and mangiferin.

[0068] (6) Construction of protein interaction network: Potential target genes were imported into the STRING database, “multiple protein” was selected, and the species was limited to “Homo sapiens” to obtain the protein interaction relationship. After exporting the data, it was imported into Cytoscape software, and the network parameters were calculated using the Network Analyze plug-in. Ten key targets were screened out: albumin, glyceraldehyde-3-phosphate dehydrogenase, HRAS protein encoding gene, tyrosine protein kinase, heat shock protein 90α encoding gene, myeloid cell leukemia sequence 1, vascular endothelial growth factor receptor 2 encoding gene, phosphatidylinositol 3-kinase regulatory subunit 1, hepatocyte growth factor receptor HGFR, and PTK2 protein tyrosine kinase 2.

[0069] (7) GO function and KEGG pathway enrichment analysis: Using the Metascape data platform, we performed enrichment analysis on the main biological processes and metabolic pathways of chemical component-disease intersection targets, and found that the pathways with high enrichment significance were the calcium signaling pathway and the HIF-1 signaling pathway.

[0070] The present invention has done a lot of analysis and verification experiments, and the following are the results of the experimental research of the present invention:

[0071] 1. Materials

[0072] 1.1 Medicinal materials and reagents

[0073] Tianjihuang (Gynostemma pentaphyllum) was purchased from Kangmei Pharmaceutical Co., Ltd., originating in Jiangxi Province. Tianjihuang (Gynostemma pentaphyllum) was authenticated by Professor Wang Jianan of the Department of Traditional Chinese Medicine, Jining Medical College, and is stored in the Drug Analysis Laboratory of the Biomedical Platform of Jining Medical College. Sodium taurocholate (Shanghai MacLean Biochemical Technology Co., Ltd., Cat. No. S817405-1g); acetonitrile (Mallinckrodt Baker, USA, pesticide-free grade); acetic acid (Mallinckrodt Baker, USA, analytical grade); and purified water (Hangzhou Wahaha Company). α-amylase (AMS) and malondialdehyde (MDA) assay kits were obtained from Nanjing Jiancheng Bioengineering Research Institute, Cat. Nos. C016-1-1 and A003-1-2; and lipase (LPS), nitric oxide (NO), and hydrogen peroxide (H2O2) assay kits were obtained from Beijing Solebao Technology Co., Ltd., Cat. Nos. BC2340, BC1470, and BC3590.

[0074] 1.2 Instruments

[0075] Agilent 1290 ultra-high performance liquid chromatography system; Agilent 6530c Q / TOF quadrupole time-of-flight mass spectrometer (Agilent Technologies, USA); Synergy H1 multifunctional microplate reader (Bio-Tek Instrument Co., Ltd., USA); Sorvall ST 8R high-speed refrigerated centrifuge (Thermo Fisher Scientific Inc.); BS110S 1 / 10,000 electronic balance (Mettler Toledo, Switzerland); MIX-1500 microplate shaker (Hangzhou Miou Instrument Co., Ltd.); KQ-500DB ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.).

[0076] 1.3 Experimental animals

[0077] SPF male Sprague-Dawley rats, weighing 150 ± 20 g, were purchased from Qingdao Daren Fucheng Animal Husbandry Co., Ltd., license number: SCXK(Lu)20190003. This animal experiment was approved by the Animal Ethics Committee of Jining Medical College, ethics number 2019-YX-011. The animal housing environment was: relative humidity 23 ± 2°C, relative humidity 60 ± 10%. Rats were acclimated for 1 week before the experiment.

[0078] 2. Methods

[0079] 2.1 Preparation of Rhizoma Cibotii Extract

[0080] Weigh 380 g of Radix Rhizoma Cyperi, crush it, add 2 L of 80% ethanol, and heat and reflux and extract it 4 times, each time for 2 hours. Combine the extracts, and recover the solvent under reduced pressure to obtain the Radix Rhizoma Cyperi extract. Store it in a refrigerator at -20°C until use.

[0081] 2.2 Preparation of test solution

[0082] Weigh 1.0 g of Radix Rhizoma Cyperi, crush it, add 10 mL of 75% methanol, and cold-soak for 30 minutes, followed by ultrasonic extraction for 30 minutes. Centrifuge the extract at 12,000 rpm / min for 10 minutes, remove the supernatant, and filter it through a 0.22 μm microporous membrane to obtain the test solution.

[0083] 2.3 HPLC-Q-TOF-MS / MS Chromatographic Analysis and Mass Spectrometry Conditions

[0084] Chromatographic column: Waters XSelect CSH-C18 (150 × 4.6 mm, 2.5 μm); Mobile phase: 0.1% formic acid in water (A) and acetonitrile (B); Gradient elution: 0-30 min, 5-65% B; 30-32 min, 65-90% B; 32-40 min, 90% B; 40-40.01 min, 90-5% B; 40.01-45.01 min, 5% B. Flow rate: 0.4 mL / min; Column temperature: 35°C; Injection volume: 2 μL; Split ratio: 1:4. Mass spectrometry conditions: ESI ion source positive and negative ion modes; drying gas temperature 350°C; drying gas flow rate 10 L / min; nebulizer gas 45 psig; sheath gas temperature 350°C; sheath gas flow rate 10 L / min; capillary voltage 4000 V; fragmentor voltage 180 V; cone voltage 65 V; mass scan range: 100-2000 Da.

[0085] 2.4 Network Pharmacology

[0086] 2.4.1 Screening of active chemical component targets in Rhizoma Cibotii

[0087] The chemical component information of the Fructus Amomi extract identified by HPLC-Q-TOF-MS / MS analysis was entered into the TCMSP database (https: / / old.tcmsp-e.com / tcmsp.php) with oral bioavailability (OB) ≥ 10% and drug-likeness (DL) ≥ 0.1 as screening conditions. After screening, the SwissTargetPrediction database (http: / / www.swisstargetprediction.ch / ) was used to predict targets. The combined targets were then corrected using the UniProt database (https: / / www.uniprot.org / ), and non-human genes were eliminated to obtain the potential targets of the chemical components of Fructus Amomi.

[0088] 2.4.2 Screening of disease targets related to acute pancreatitis and construction of a “component-disease-target” network diagram

[0089] Disease targets for acute pancreatitis were obtained using the keyword "Acute pancreatitis" from the OMIM (https: / / www.omim.org / ), DrugBank (https: / / www.drugbank.ca / ), and GeneCards (https: / / www.genecards.org / ). Targets with a relevance score ≥ 5 were retrieved from GeneCards, and duplicate targets were removed. The Venny2.1 online analysis tool was used to intersect disease-related targets with active ingredient targets to identify potential targets for the treatment of acute pancreatitis with Rhizoma Cibotii extract. The chemical components and gene targets of Rhizoma Cibotii were imported into Cytoscape to construct a "component-disease-target" network diagram.

[0090] 2.4.3 Construction of protein-protein interaction (PPI) network

[0091] To illustrate the systemic effects of the potential target proteins of Rhizoma Cibotii extract in the treatment of acute pancreatitis, the above-mentioned potential target genes were imported into the STRING database (https: / / www.string-db.org / ), and the "multiple protein" was selected. The species was limited to "Homo sapiens" to obtain the protein interaction relationship. The relevant data were exported and imported into Cytoscape software. The network topology parameters were calculated using the Network Analyze plug-in, and the key targets and interacting chemical components were screened.

[0092] 2.4.4GO function and KEGG pathway enrichment analysis

[0093] The Metascape (http: / / metascape.org) data platform was used to perform enrichment analysis on the main biological processes and metabolic pathways of chemical component-disease intersection targets, including molecular functions, biological processes, cellular components, and KEGG. "Homosapiens" was selected with P < 0.01, and the results were visualized using the Microbiome Information website.

[0094] 2.5 Animal experiments

[0095] 2.5.1 Grouping, Modeling, and Drug Administration

[0096] Healthy male Sprague-Dawley rats were randomly divided into a blank group, a model group, and a group receiving 0.5 g / kg of Rhizoma Cibotii extract, with 6 rats in each group, and the drug was administered orally. The rats were fasted for 24 hours before modeling and deprived of water until 2 hours before modeling. After anesthesia, the rats were opened layer by layer along the midline of the abdomen below the xiphoid process, approximately 1-1.5 cm in depth, to fully expose the abdominal cavity. The duodenum and pancreas were located along the pylorus of the stomach and fully exposed. A needle was inserted through the major duodenal papilla into the common bile duct to a depth of approximately 0.5-1 cm, and the opening of the pancreaticobiliary duct at the duodenal ampulla was clamped with an artery clamp. An acute pancreatitis model was induced in rats by slowly and evenly injecting 0.2 mL of 2.5% sodium taurocholate solution using an infusion pump.

[0097] Blood was collected from the medial canthal vein at 6 and 12 hours, centrifuged at 4000 rpm for 10 minutes at 4°C, and the supernatant was collected and stored at -80°C. Twelve hours after blood collection, the rats were killed by cervical dislocation, and the pancreas was fixed in 4% paraformaldehyde and stained with HE for histological observation.

[0098] 2.5.2 Biochemical index detection

[0099] The operation was carried out according to the kit instructions to determine the contents of AMS, LPS, MDA, H2O2, GSH-Px and NO, and the results were statistically analyzed.

[0100] 2.5.3 Statistical analysis

[0101] Graphpad Prism 8.0.2 software was used for statistical analysis of all experimental data. The experimental results are expressed as (±s). One-way analysis of variance was used for comparison between groups, and P < 0.05 was considered statistically significant.

[0102] 3. Results

[0103] 3.1 Analysis of chemical composition of Tianjihuang

[0104] Chemical composition analysis of Rhizoma Cibotii was performed using HPLC-Q-TOF-MS in both positive and negative ion modes. Key identification information, including compound retention times, high-resolution mass spectrometry (HRMS), and secondary mass spectrometry (MS / MS) data, was obtained using conventional identification methods. Chromatographic peak retention times, HRMS, and multi-stage MS data were compared with reference materials and literature data. Molecular formulas were calculated based on accurate mass values ​​(the error between the measured and theoretical values ​​was less than 5 ppm). The fragmentation patterns of the MS / MS data and reference materials were analyzed, and potential compound structures were inferred based on reference literature. GNPS (http: / / gnps.ucsd.edu) was used to generate a molecular network of Rhizoma Cibotii components. First, the raw MS data were converted to the mzXML file format using ProteoWizard software (Los Angeles, CA, USA). The mass errors for both parent ions and MS / MS fragment ions were less than 0.02 Da. The high-resolution mass spectrometry data were then compared with the GNPS library data. A total of 42 chemical components were identified, as shown in Table 1.

[0105] Table 1 Retention time, molecular formula, high-resolution mass spectrometry data and secondary mass spectrometry fragment information of the chemical components in Rhizoma Cibotii

[0106]

[0107]

[0108]

[0109] *Indicates core ingredients

[0110] 3.2 Network pharmacology research

[0111] 3.2.1 Target screening for acute pancreatitis-related diseases

[0112] Based on the OMIM, DrugBank and GeneCard databases, 1153 acute pancreatitis-related targets were obtained. The 130 potential targets of the chemical components of Rhizoma Cibotii were screened and intersected with the above 1153 acute pancreatitis targets. Venny2.1 online analysis obtained a total of 51 intersection targets, such as Figure 1 shown

[0113] 3.2.2 Construction of “Compound-Disease-Target” Network

[0114] According to the set screening conditions, a total of 10 effective chemical components were screened out. The target information was predicted using the SwissTargetPrediction database, and after deleting duplicate data, it was imported into the Cytoscape software to construct a "compound-disease-target" network. Figure 2 As shown, there are 10 chemical components: quercetin, coumarin glucoside, gentiopicroside, eriodictyol-7-O-glucoside, protocatechuic acid, 3-O-caffeoylquinic acid, astragaloside, isoorientin, isovitexin, and mangiferin. There are 51 target nodes, indicating that 51 targets are associated with the main components and diseases. There are 100 edges between the targets. The greater the number of edges, the greater the likelihood that the target plays a key role in the mechanism. Quercetin, with 7 edges, is the most likely component of Tianjihuang for treating acute pancreatitis. A higher degree indicates that the node is most closely connected in the network. CA2 has the highest degree of 8, suggesting that CA2 is a key target for Tianjihuang for treating acute pancreatitis.

[0115] 3.2.3 Construction of PPI Network

[0116] The target protein PPI network of Radix Achyranthis Bidentatae extract in the treatment of acute pancreatitis Figure 3 As shown, there are 43 nodes and 249 edges. Node degree is used as an evaluation parameter. Higher values ​​indicate greater importance in the network, larger nodes, darker colors, and more connected targets. The top 10 core targets by degree include: albumin (ALB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), HRAS protein-encoding gene (HRAS), tyrosine protein kinase (SRC), heat shock protein 90α-encoding gene (HSP90AA1), myeloid cell leukemia sequence 1 (MCL1), vascular endothelial growth factor receptor 2-encoding gene (KDR), phosphatidylinositol 3-kinase regulatory subunit 1 (PIK3R1), hepatocyte growth factor receptor HGFR (MET), and PTK2 protein tyrosine kinase 2 (PTK2). Darker colors, larger diameters, and closer proximity to the center indicate higher degrees. Dark green nodes represent core targets.

[0117] 3.2.4 GO functional analysis and KEGG pathway enrichment

[0118] The Metascape data platform was used to perform enrichment analysis on the main biological processes and metabolic pathways of chemical component-disease intersection targets. The results of GO enrichment analysis showed 20 cellular biological processes (BP), mainly involving the regulation of kinase activity, protein phosphorylation and response to hormones, etc.; 12 cellular components (CC), mainly including the cytoplasmic perinuclear region, cytoplasmic vesicle cavity and cell apex, etc.; 16 molecular functions (MF), mainly including protein tyrosine kinase activity, protein domain-specific binding and protein serine / threonine kinase activity, etc. The top 10 with the largest counts in the three groups were selected for drawing. The results of KEGG pathway enrichment analysis showed that 51 cross-target proteins were enriched in 16 pathways (P<0.05), such as Figure 4 、 Figure 5 shown.

[0119] Pathways with significant enrichment included calcium signaling and the HIF-1 signaling pathway. Hypoxia-inducible factor (HIF-1α) in the HIF-1 signaling pathway has been shown to play a key role in the pathogenesis of acute pancreatitis. HIF-1α is an indirect response to oxidative stress. By integrating network pharmacology analysis results with related research and conducting animal experiments using oxidative stress as an indicator, we explored the therapeutic effects of Rhizoma Cibotii extract on acute pancreatitis.

[0120] 3.2 Animal experiments

[0121] 3.3.1 Observation of pancreatic histopathological changes by HE staining

[0122] The pancreatic tissue was fixed with paraformaldehyde, embedded in paraffin, sliced, waxed, and stained to observe the pathological changes of the rat pancreatic tissue. Figure 6 As shown, the pancreatic cell structure of the control group was intact and no abnormalities were found; in the acute pancreatic model group, the acinar edema, some cells were obviously necrotic, the tissue gap was enlarged, and there was inflammatory cell infiltration; in the Tianjihuang treatment group, the pathological changes were greatly improved compared with the model group and were similar to those of the control group.

[0123] 3.3.2 Effects of Tianjihuang on related indicators in the serum of rats in each group

[0124] AMS and lipase LPS are the most commonly used test indicators for clinical diagnosis of acute pancreatitis. [9]. The levels of AMS and LPS in the serum of rats in the model group were increased, which was significantly different from those in the control group (P<0.01). Compared with the rats in the model group, the levels of AMS and LPS in the serum of the group administered with the Radix Panacis chinensis extract were decreased, and the difference was statistically significant (P<0.01). The difference in AMS levels between 6h and 12h was very small, while the difference in LPS levels between 6h and 12h was large. Compared with the model group, the serum LPS activity of rats in the Radix Panacis chinensis administration group was different at 6h (P<0.05), but there was no significant difference at 12h (P>0.05). This shows that the total extract of Radix Panacis chinensis can inhibit the secretion of amylase and lipase and reduce the autodigestive effect of the pancreas.

[0125] Compared with the control group, the levels of MDA, H2O2 and NO in the serum of the rats in the model group were significantly increased (P<0.01), and GSH-P X Compared with the model group, the contents of MDA, H2O2 and NO in the serum of the rats in the group receiving Rhizoma Cibotii extract were significantly decreased (P<0.01), and GSH- X The content of H2O2, GSH-P X The difference between the content of NO and the content of MDA in the model group at 6h and 12h was not significant; the content of MDA in the model group decreased compared with that at 6h and 12h. Figure 7 shown.

[0126] 4. Discussion

[0127] The pathogenesis of acute pancreatitis primarily involves obstruction of the pancreatic duct, which blocks pancreatic secretion and hinders the exocytosis of zymogen granules, prompting their activation within cells. Subsequently, a cascade of digestive enzymes is activated, leading to autodigestion. This leads to inflammation, oxidative stress, organ damage, and severe complications. Oxidative stress is a key mechanism in the pathophysiology of acute pancreatitis. When acute pancreatitis occurs, the inflammatory response triggers oxidative stress. Inflammatory cells produce excessive oxygen free radicals, which exceed the clearance capacity of the cellular antioxidant defense system. This leads to excessive intracellular oxygen free radical production, causing cell and tissue damage. Oxidative stress is particularly significant in vulnerable tissues, such as the pancreas. HIF-1α plays a key role in the pathogenesis of acute pancreatitis. It is considered an indirect response to oxidative stress, which refers to the excessive production of reactive oxygen species (ROS) in the body, which damages the oxidative state of proteins, lipids, and DNA. Superoxide dismutase converts ROS into hydrogen peroxide (H2O2), which is then cleared by the enzyme MDA. High concentrations of NO can aggravate oxidative stress damage. GSH-Px can catalyze the reduction of toxic peroxides into non-toxic hydroxyl compounds and promote the decomposition of H2O2.

[0128] The present invention identified 42 chemical components from the total extract of Radix Panacis chinensis and conducted network pharmacology research, obtaining 130 chemical component targets, 1,153 targets for acute pancreatitis, and 51 targets obtained by the intersection of chemical components and acute pancreatitis. The STRING database was used to perform PPI analysis on the 51 targets related to the treatment of acute pancreatitis by Radix Panacis chinensis, and the GO and KEGG enrichment analysis of the intersection targets was performed using the Metascape database. It was found that its key proteins and enriched pathways were closely related to inflammation and oxidative stress. Animal experimental studies have shown that the total extract of Radix Panacis chinensis can alleviate the pathological changes in rats with acute pancreatitis induced by sodium taurocholate. The levels of AMS, LPS, MDA, H2O2 and NO in rat serum were significantly reduced, and the level of GSH-Px in serum was significantly increased. Comparing the results of LPS at 6h and 12h, the content at 12h was significantly lower. LPS may be related to the modeling method. After the injection of sodium taurocholate, the obstruction of the pancreatic bile duct is opened, causing sodium taurocholate to flow into the duodenal ampulla, thereby performing normal digestion and secretion, and its stimulation to the pancreas is then weakened. Since LPS is more sensitive than AMS, the content of LPS decreases at 12h, while AMS still maintains a high level.

[18] The total extract of Rhizoma Cibotii can inhibit the secretion of digestive enzymes in rats with acute pancreatitis, reduce the degree of oxidative stress in the body, and have antioxidant effects, which was confirmed by the results of network pharmacology prediction.

[0129] In summary, this study identified the chemical components of Rhizoma Cibotii using HPLC-Q-TOF-MS analysis, combined with network pharmacology analysis and rats with acute pancreatitis. The results showed that the total extract of Rhizoma Cibotii could inhibit the secretion of digestive enzymes and reduce the degree of oxidative stress in rats with acute pancreatitis, demonstrating a certain antioxidant effect. This finding is consistent with the results of network pharmacology predictions. This study provides valuable insights and references for the subsequent development and clinical application of Rhizoma Cibotii medicinal materials.

Claims

1. A method for analyzing the active ingredients of Radix Panacis radix for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology, characterized by: The analysis method uses HPLC-Q-TOF-MS / MS analysis technology to generate a mass spectrometry molecular network of chemical components through accurate mass, secondary mass spectrometry data, mass spectrometry fragmentation patterns and databases. Then, using network pharmacology-related databases, it obtains disease-related targets of acute pancreatitis, constructs a "component-disease-target" network diagram, constructs a protein interaction network, and then performs GO function and KEGG pathway enrichment analysis. The preparation method of the Rhizoma Cibotii extract is as follows: 350-400 g of Rhizoma Cibotii medicinal material is weighed, crushed, 1.8-2.5 L of 70-90% ethanol is added, and the mixture is heated under reflux and extracted 3-5 times, each time for 1.5-2.5 hours. The extracts are combined, and the solvent is recovered under reduced pressure to obtain the Rhizoma Cibotii extract, which is then stored in a -20°C refrigerator for later use; HPLC conditions: Waters XSelect CSH-C18 column, 150 × 4.6 mm, 2.5 μm; mobile phase A: 0.1% formic acid in water; mobile phase B: acetonitrile; gradient elution; flow rate: 0.4 mL / min; column temperature: 35°C; injection volume: 2 μL; split ratio: 1:4; Mass spectrometry conditions: ESI ion source positive and negative ion modes; drying gas temperature 350°C; drying gas flow rate 10 L / min; nebulizer gas 45 psig; sheath gas temperature 350°C; sheath gas flow rate 10 L / min; capillary voltage 4000 V; fragmentor voltage 180 V; cone voltage 65 V; mass scan range: 100–2000 Da; The "compound-disease-target" network was constructed, and 10 key chemical components were obtained: quercetin, coumarin glucoside, gentiopicroside, eriodictyol-7- O -Glucoside, protocatechuic acid, 3- O - Caffeoylquinic acid, astragalin, isoorientin, isovitexin and mangiferin.

2. The method for analyzing the active ingredients of Radix Panacis radix for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology as claimed in claim 1, characterized in that: The method is carried out according to the following steps: (1) Preparation of Radix Panacis chinensis extract: Weigh 350-400 g of Radix Panacis chinensis medicinal material, crush it, add 1.8-2.5 L of 70-90% ethanol, heat and reflux and extract 3-5 times, each time for 1.5-2.5 hours, combine the extracts, recover the solvent under reduced pressure, and obtain Radix Panacis chinensis extract, which is stored in a -20°C refrigerator for future use; (2) HPLC-Q-TOF-MS / MS analysis: The retention time of the chromatographic peak, high-resolution mass spectrometry data, and multi-stage mass spectrometry data were compared with the GNPS spectral library data. The molecular network of chemical components was generated by accurate mass number, secondary mass spectrometry data, mass spectrometry fragmentation rules, and database to obtain the structural information of the chemical components of the extract of Rhizoma Cibotii. HPLC conditions: Waters XSelect CSH-C18 column, 150 × 4.6 mm, 2.5 μm; mobile phase A: 0.1% formic acid in water; mobile phase B: acetonitrile; gradient elution; flow rate: 0.4 mL / min; column temperature: 35°C; injection volume: 2 μL; split ratio: 1:4; Mass spectrometry conditions: ESI ion source positive and negative ion modes; drying gas temperature 350°C; drying gas flow rate 10 L / min; nebulizer gas 45 psig; sheath gas temperature 350°C; sheath gas flow rate 10 L / min; capillary voltage 4000 V; fragmentor voltage 180 V; cone voltage 65 V; mass scan range: 100–2000 Da; (3) Screening of active chemical component targets of Rhizoma Cibotii: The structural information of the chemical components of Rhizoma Cibotii extract identified by HPLC-Q-TOF-MS / MS analysis was input into the TCMSP database, SwissTargetPrediction database, and UniProt database to calibrate the combined targets, eliminate non-human genes, and obtain the target of the chemical components of Rhizoma Cibotii; (4) Screening of disease targets related to acute pancreatitis: Acute pancreatitis disease targets were obtained by using the three databases of OMIM, DrugBank and GeneCards with the keyword of Acute pancreatitis. The acute pancreatitis disease targets were intersected with the targets of the chemical components of Rhizoma Cibotii in step (3) using the Venny2.1 online analysis tool to obtain potential targets of Rhizoma Cibotii extract for the treatment of acute pancreatitis, i.e., potential targets. (5) Constructing a "component-disease-target" network diagram: The potential targets in step (4) were predicted using the SwissTargetPrediction database, and after deleting duplicate data, the data were imported into the Cytoscape software to construct a "compound-disease-target" network, and 10 key chemical components were obtained; (6) Construction of protein interaction network: The targets of the chemical components of Rhizoma Cibotii and the potential target genes in step (4) were imported into the STRING database, "multiple protein" was selected, and the species was limited to "Homo sapiens" to obtain the protein interaction relationship. After exporting the data, it was imported into Cytoscape software, and the network parameters were calculated using the Network Analyze plug-in to screen out 10 key targets; (7) GO function and KEGG pathway enrichment analysis: Using the Metascape data platform, we performed enrichment analysis on the main biological processes and metabolic pathways of chemical component-disease intersection targets to obtain pathways with high enrichment significance.

3. The method for analyzing the medicinal components of Radix Panacis radix for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology as claimed in claim 2, characterized in that: In the step (1), the preparation of the Rhizoma Cibotii extract is as follows: 380 g of Rhizoma Cibotii medicinal material is weighed, crushed, added with 2 L of 80% ethanol, heated and refluxed for extraction 4 times, each time for 2 h, the extracts are combined, the solvent is recovered under reduced pressure to obtain the Rhizoma Cibotii extract, and stored in a refrigerator at -20°C for later use.

4. The method for analyzing the active ingredients of Radix Panacis radix for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology as claimed in claim 2, characterized in that: In step (2), the gradient elution program of the HPLC chromatographic conditions is: 0-30 min, 5-65% B; 30-32 min, 65-90% B; 32-40 min, 90% B; 40-40.01 min, 90-5% B; 40.01-45.01 min, 5% B.

5. The method for analyzing the active ingredients of Radix Panacis radix for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology as claimed in claim 2, characterized in that: In the step (5), the 10 key chemical components are: quercetin, coumarin glucoside, gentiopicroside, eriodictyol-7- O -Glucoside, protocatechuic acid, 3- O - Caffeoylquinic acid, astragalin, isoorientin, isovitexin and mangiferin.

6. The method for analyzing the active ingredients of Radix Panacis radix for treating acute pancreatitis based on HPLC-Q-TOF-MS and network pharmacology as claimed in claim 2, characterized in that: In step (6), the key targets are albumin, glyceraldehyde-3-phosphate dehydrogenase, HRAS protein encoding gene, tyrosine protein kinase, heat shock protein 90α encoding gene, myeloid cell leukemia sequence 1, vascular endothelial growth factor receptor 2 encoding gene, phosphatidylinositol 3-kinase regulatory subunit 1, hepatocyte growth factor receptor HGFR and PTK2 protein tyrosine kinase 2.

7. The method for analyzing the active ingredients of Radix Panacis radiata for treating acute pancreatitis based on HPLC-Q-TOF-MS, network pharmacology, and molecular docking as claimed in claim 2, characterized in that: In the step (7), the pathways with high enrichment significance are the calcium signaling pathway and the HIF-1 signaling pathway.

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