Application of polyphenol extract from Phellodendron chinense fruit in the preparation of drugs with hepatoprotective effects

Abstract

The present invention relates to the field of pharmaceutical chemistry technology, and specifically to the application of a polyphenol extract of abercrombie & Fitch fruit in the preparation of a drug with a liver-protecting effect. The polyphenol extract of abercrombie & Fitch fruit includes neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, arbutin, 7-hydroxycoumarin, phellodendron glycosides, isoquercetin, hyperoside, salicylic acid, dianthin, caffeic acid, D-(-)-quinic acid, citric acid, trifoliate glycosides, kaempferol, protocatechualdehyde, cynarin, 3,5-dimethoxybenzoic acid, daidzein, malic acid, jasmonic acid, and quercetin polyphenol components. The present invention applies the non-medicinal part of the fruit of the medicinal plant abercrombie & Fitch to the treatment of liver protection for the first time, and predicts its possible mechanism of action through network pharmacology, laying a foundation for further experimental verification of the action target and pathway mechanism of the liver protection of the fruit of abercrombie & Fitch fruit.

Description

Technical Field

[0001] The present invention relates to the technical field of medicinal chemistry, and in particular to application of a polyphenol extract of abercrombie & Fitch fruit in preparing a medicine with a liver-protecting effect. Background Art

[0002] In today's society, increasingly competitive pressures, coupled with unhealthy diets, alcohol consumption, and excessive fat intake, pose a serious threat to the health of the human liver. Without timely medication and effective control, liver damage can gradually worsen into cirrhosis and even liver cancer. The occurrence and progression of liver disease poses a significant threat to human health. Therefore, elucidating the pathogenesis of liver disease and exploring effective methods to protect the liver from its root causes are crucial to human health.

[0003] Phellodendron amurense is a perennial woody plant of the genus Phellodendron in the Rutaceae family. Its bark, the main medicinal part, is known for its heat-clearing and dampness-drying properties, purging internal heat and eliminating phlegm, and detoxifying and treating sores. It is used for damp-heat diarrhea, jaundice, dark urine, vaginal discharge, itchy vaginal discharge, painful stranguria caused by heat, beriberi, paralysis, bone steaming, fever, night sweats, nocturnal emission, sores, swelling, and eczema. However, limited basic research on its fruit has yet to be effectively utilized. Summary of the Invention

[0004] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a use of a polyphenol extract of Phellodendron amurense fruit in the preparation of a medicament having a hepatoprotective effect, and to develop the medicinal value of Phellodendron amurense fruit.

[0005] In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention is:

[0006] Provided is the use of a polyphenol extract from the fruit of Phellodendron amurense in the preparation of a medicine with a hepatoprotective effect.

[0007] Furthermore, the polyphenol extract of the fruit of Phellodendron amurense includes neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, arbutin, 7-hydroxycoumarin, phellodendron glycoside, isoquercetin, hyperoside, salicylic acid, dianthusin, caffeic acid, D-(-)-quinic acid, citric acid, trifolioside, kaempferol, protocatechualdehyde, cynarin, 3,5-dimethoxybenzoic acid, daidzein, malic acid, jasmonic acid, and quercetin polyphenol components.

[0008] Furthermore, the preparation method of the polyphenol extract of Phellodendron amurense fruit comprises the following steps:

[0009] The powder of Phellodendron amurense fruit is added with ethanol, heated and ultrasonically extracted, and then filtered to obtain a crude material of Phellodendron amurense fruit; the crude material of Phellodendron amurense fruit is concentrated, passed through a macroporous adsorption resin, and eluted with pure water and ethanol in sequence, the ethanol eluate is collected, most of the solvent is removed on a rotary evaporator, and then freeze-dried on a freeze dryer to obtain a polyphenol extract of Phellodendron amurense fruit.

[0010] Furthermore, ethanol is added to the Phellodendron amurense fruit powder to make the alcohol content reach 30-90%, and then the powder is heated and ultrasonically extracted.

[0011] Furthermore, ethanol is added to the Phellodendron amurense fruit powder to make the alcohol content reach 50%, and then the powder is subjected to heating and ultrasonic extraction.

[0012] Furthermore, the temperature of the heated ultrasonic extraction is 40-80° C., and the time is 30-90 min.

[0013] Furthermore, the temperature of the heated ultrasonic extraction was 60° C. and the time was 75 min.

[0014] The beneficial effects of the present invention are:

[0015] This invention is the first to apply the non-medicinal parts of the fruit of the medicinal plant Phellodendron amurense to liver protection treatment, and predicts its possible mechanism of action through network pharmacology, laying the foundation for further experimental verification of the target and pathway mechanism of the liver protection of Phellodendron amurense fruit.

[0016] The present invention develops the pharmacological activity of the fruit of the non-medicinal part of Phellodendron amurense, which can not only provide a new idea for the sustainable development and utilization of Phellodendron amurense, but also expand the application space of traditional Chinese medicine resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Figure 1 shows the pathological observation of liver tissue in mice with acetaminophen-induced liver damage after different doses of polyphenol extract from Phellodendron amurense fruit (×400), where A is the blank group; B is the model group; C is the positive drug group; D is the PSF-p low-dose group; E is the PSF-p medium-dose group; and F is the PSF-p high-dose group.

[0018] Figure 2 This is a Venn diagram of chemical components of Phellodendron amurense fruit and liver damage targets;

[0019] Figure 3 This is the "ingredient-target-disease" network diagram of the chemical components of Phellodendron truncatum fruit, where the prisms represent 96 intersection targets, the triangles represent 20 active ingredients, and the circles represent diseases;

[0020] Figure 4 This is the protein interaction (PPI) network diagram of the hepatoprotective targets of the chemical components of Phellodendron amurense fruit;

[0021] Figure 5 Three-in-one histogram for GO functional enrichment analysis;

[0022] Figure 6 KEGG pathway enrichment analysis diagram for key targets;

[0023] Figure 7This is a diagram of chemical components, targets and signaling pathways in the fruit of Phellodendron truncatum;

[0024] Figure 8 This is the total ion chromatogram of the polyphenol extract from the fruit of Phellodendron amurense by UPLC-Q-Orbitrap HRMS (positive ion mode);

[0025] Figure 9 This is the total ion chromatogram of the polyphenol extract from the fruit of Phellodendron amurense using UPLC-Q-Orbitrap HRMS (negative ion mode). DETAILED DESCRIPTION

[0026] The specific embodiments of the present invention are described below to facilitate understanding of the present invention by those skilled in the art. However, it should be clear that the present invention is not limited to the scope of the specific embodiments. For those skilled in the art, as long as various changes are within the spirit and scope of the present invention as defined and determined by the appended claims, these changes are obvious, and all inventions and creations utilizing the concepts of the present invention are protected.

[0027] Example 1

[0028] This embodiment discloses a method for preparing the polyphenol extract of the fruit of Phellodendron amurense.

[0029] To 500 g of Phellodendron amurense fruit powder, 40-fold the amount of 50% ethanol was added in portions. Ultrasonic extraction was performed at 60°C for 75 min, and the filtrates were filtered and combined. The filtrate was concentrated under reduced pressure on a rotary evaporator to remove most of the solvent and then freeze-dried in a freeze-dryer to obtain a crude Phellodendron amurense fruit extract. A glass chromatography column was fixed with absorbent cotton at the bottom end and loaded with an appropriate amount of ethanol-activated macroporous adsorption resin D101. The column was washed with distilled water until no ethanol and no bubbles were present. Degreased cotton was added to the top end. An aqueous solution of the crude Phellodendron amurense fruit extract with a polyphenol concentration of 1.0 mg / mL was slowly poured into the chromatography column. The column was eluted sequentially with pure water and 50% ethanol. The 50% ethanol eluate was collected, the solvent was removed on a rotary evaporator, and the column was freeze-dried in a freeze-dryer to obtain approximately 6 g of Phellodendron amurense fruit polyphenol extract, designated PSF-p.

[0030] Test Example 1

[0031] This Test Example 1 discloses a pharmacodynamic study of a polyphenol extract from the fruit of Phellodendron amurense.

[0032] 1. Preparation of Pharmacodynamic Sample Solutions

[0033] (1) Preparation of polyphenol extract from Phellodendron amurense fruit: According to the method of Example 1, the 50% ethanol eluate of the macroporous resin was collected, concentrated and dried, and 0.5% CMC-Na solution was added to prepare high, medium and low dose groups of suspension.

[0034] (2) Preparation of positive drugs: Grind the bifendate tablets into powder, weigh an appropriate amount of powder, and add 0.5% CMC_Na solution to make a suspension.

[0035] (3) Preparation of APAP solution: Grind the bifendate tablets into powder, weigh an appropriate amount of powder, and add 0.5% CMC-Na solution to prepare a suspension.

[0036] 2. Animal Grouping, Modeling, and Drug Administration

[0037] Forty-eight male Kunming mice, weighing 20±2 g, were randomly divided into six groups (n=8): control, model, bifendate (150 mg / kg), and high-, medium-, and low-dose groups (400, 200, and 100 mg / kg) of polyphenol extract from the fruit of Phellodendron amurense. Each treatment group received the corresponding drug (10 mL / kg) intragastrically. The normal and model groups received an equal volume of 0.9% sodium chloride solution intragastrically once daily for 7 consecutive days. The mice were fasted for 12 hours before the last dose. Two hours later, all groups, except the model group, were orally administered with APAP at a dose of 500 mg / kg to induce acute liver injury. The mice were fasted for 16 hours with or without water deprivation, and serum and liver samples were collected.

[0038] 3. Sample Collection and Processing

[0039] (1) Serum sample: Blood was collected from the mouse eyeball and allowed to stand for 2 h. The blood was centrifuged at 3500 r / min for 10 min to separate the serum and stored at 4°C.

[0040] (2) Liver tissue samples: After the mice were killed, the livers were quickly dissected and removed. Blood and other residues on the surface of the livers were repeatedly rinsed with physiological saline. After absorbing water with filter paper, the livers were weighed and the liver tissues at the same location were cut. Physiological saline was added at a weight to volume ratio of 1:9. Liver tissue homogenates were prepared using a mechanical homogenizer in an ice-water bath. The livers were then centrifuged at 2000 r / min for 15 min. The supernatant was aspirated to obtain a 10% liver tissue homogenate sample. The 10% liver tissue homogenate sample was taken and physiological saline was added at a volume ratio of 1:9 to obtain a 1% liver tissue homogenate sample.

[0041] (3) Liver pathological tissue sections: Liver tissues were cut from the same location in each group and fixed in 4% paraformaldehyde solution. The samples were then routinely paraffin-embedded, sectioned, and stained with HE for histopathological examination.

[0042] (4) The determination of serum biochemical indicators AST and ALT and liver tissue biochemical indicators MDA and SOD were performed according to the detailed steps of the kit and measured using an enzyme-labeled instrument.

[0043] 4. Data Processing

[0044] The data were expressed as mean ± S, and SPSS 26.0 software was used for statistical analysis. One-way analysis of variance was used to compare the sample means among multiple groups.

[0045] 5. Results

[0046] 5.1 Biochemical index results

[0047] According to the specific requirements of the kit, the biochemical parameters to be tested in mice were measured and analyzed using SPSS software. The results are shown in Table 1.

[0048] Table 1 Effects of polyphenol extracts from Phellodendron amurense fruit on biochemical parameters in mice with liver injury

[0049]

[0050] Note: Compared with the blank group, * P <0.05,** P <0.01; compared with the model group, # P <0.05, ## P <0.01.

[0051] The results showed that compared with the blank group, the serum ALT and AST concentrations of the model group mice were significantly increased ( P <0.01); the MDA content in liver tissue homogenate increased significantly (P<0.01), and the SOD activity decreased ( P <0.01).

[0052] Compared with the model group, the ALT concentrations in the serum of the positive drug and extract high-dose, medium-dose and low-dose groups were significantly reduced ( P <0.01), which was statistically significant. Compared with the model group, the serum AST concentrations in the high-dose and medium-dose groups of the positive drug and extract were significantly reduced ( P <0.01), the serum AST concentration in the low-dose extract group was significantly reduced ( P <0.05), which was statistically significant. Compared with the model group, the MDA concentration in the liver tissue homogenate of the positive drug and extract high-dose, medium-dose and low-dose groups was significantly reduced ( P <0.01), which was statistically significant. Compared with the model group, the SOD concentration in the liver tissue homogenate of the high-dose extract group was significantly reduced ( P <0.01), the SOD concentration in liver tissue homogenate of the positive drug and extract medium dose groups was significantly decreased ( P <0.05), which is statistically significant.

[0053] 5.2 Pathological section results

[0054] Pathological section results Figure 1 As shown, the liver tissue capsule of the control group was intact, the hepatic lobules were not clearly divided, and the hepatic cords were arranged relatively neatly. The central vein endothelium was intact, and the hepatocytes were arranged radially around the central vein with normal morphology, without obvious degeneration and necrosis. The hepatic sinusoidal structure was normal, without obvious congestion, dilation, or inflammatory infiltration. The interlobular arteries, interlobular veins, and interlobular bile ducts in the portal area were relatively intact, without obvious fibrous tissue proliferation or inflammatory cell infiltration around them. No other obvious pathological changes were observed. In the model group, a large number of hepatocytes around the central veins of the hepatic lobules of the mice were degenerated and necrotic, with blurred hepatocyte structure, pyknosis, fragmentation and lysis of the nuclei, obstruction of the hepatic sinusoids in the necrotic area, dilation of the hepatic sinusoids in the periphery, and a large number of red blood cells were observed, without obvious inflammatory cell infiltration. Compared with the model group, the pathological changes in the liver tissue of the positive, high, medium, and low-dose drug groups were alleviated to varying degrees.

[0055] Test Example 2

[0056] This experimental example discloses research on the material basis of the polyphenol extract of Phellodendron amurense fruit.

[0057] 1. Preparation of Sample Solution

[0058] 10 mg of the polyphenol extract PSF-p from the fruit of Phellodendron amurense obtained according to the method of Example 1 was accurately weighed and placed in a 10 mL volumetric flask. After dissolving in methanol, the mixture was fixed to volume and filtered through a microporous membrane to obtain the product.

[0059] 2. Chromatographic and mass spectrometry conditions

[0060] Chromatographic conditions: chromatographic column: Agilent SB-C18 (3.0 mm×100 mm, 1.8 μm); mobile phase: 0.1% formic acid aqueous solution (A)-acetonitrile (B), gradient elution: 0-10 min, 93%-88% A; 10-15 min, 88% A; 15-25 min, 88%-85% A; 25-30 min, 85% A; 30-33 min, 85%-82% A; 33-38 min, 82% A; 38-55 min, 82%-60% A; 55-60 min, 60% A); flow rate: 0.3 mL / min; injection volume: 5 μL.

[0061] Mass spectrometry conditions: An electrospray ionization (ESI) source was used in both positive and negative ion modes. The spray voltage was 3.5 kV, the auxiliary gas temperature was 350°C, the auxiliary gas flow rate was 10 arb, the sheath gas flow rate was 35 arb, and the ion transfer tube temperature was 320°C. Scan mode was Full MS / dd-MS2, with a primary resolution of 70,000 and a secondary resolution of 17,500. The scan range was from mass-to-charge ratio (m / z) 100 to 1500, and step collision energies were 20, 40, and 60 eV.

[0062] 3. Preliminary exploration of hepatoprotective active ingredients, targets, and pathways of polyphenol extracts from Phellodendron amurense fruit based on network pharmacology

[0063] 3.1 Screening of PSF-p chemical components and collection of corresponding targets

[0064] The chemical components corresponding to the polyphenol extracts from the fruits of Phellodendron amurense were obtained by searching the TCMSP database (http: / / ibts.hkbu.edu.hk / LSP / tcmsp.php).

[0065] 3.2 Targets for acute liver injury

[0066] The GeneCards database (https: / / www.genecards.org / ) was searched using the keyword “acute liver injury” to obtain targets corresponding to diseases related to liver injury, and the retrieved disease targets were further screened using a score value > 5.0.

[0067] 3.3 Screening of common drug-disease targets

[0068] Chemical component targets and disease targets were input into the Venny 2.1 online software drawing tool platform, common targets were screened, and Venn diagrams were drawn.

[0069] 3.4 Construction of component-target-disease network

[0070] The chemical components of the polyphenol extract of Phellodendron amurense fruit screened under "3.3" and the common targets of liver damage were imported into Cytoscape 3.7.2 analysis software to construct the "component-target-disease" network of the effective liver-protective parts of Phellodendron amurense fruit.

[0071] 3.5 PPI Network Analysis

[0072] Import the common targets obtained under "3.4" into the STRING database (https: / / string-db.org /

[0073] cgi / input.pl), a protein-target interaction network was constructed, with a comprehensive score greater than 0.4 considered significant. A PPT network diagram and related network information were generated. The network data was imported into Cytoscape software, and core targets were screened by calculating the degree value of each target and adjusting the node size and color based on the degree value.

[0074] 3.6 GO bioinformatics and KEGG pathway enrichment analysis

[0075] The common targets identified under "3.4" were imported into the David database (https: / / david.ncifcrf.gov / ) for GO analysis (including biological process (BP), cellular component (CC), and molecular function (MF)) and KEGG pathway enrichment analysis. The data were exported and the top 10 BP, CC, and MF data and the top 20 KEGG pathway enrichment data were selected using P-values ​​and gene counts. The data were then imported into the Bioinformatics website (http: / / www.bioinformatics.com.cn / ) to plot a three-in-one bar chart for GO functional enrichment analysis and a bubble chart for KEGG pathway enrichment analysis.

[0076] 3.7 Active ingredient-target-signaling pathway network

[0077] The top 10 signal pathways ranked by KEGG pathway analysis results were combined with the core components and core targets of the polyphenol extract of Phellodendron amurense fruit to construct a "component-target-pathway" network diagram, and the targets that did not intersect with the components and pathways were deleted from the diagram.

[0078] 4. Results

[0079] 4.1 Chemical composition analysis results of PSF-p

[0080] The UPLC-Q-Orbitrap HRMS system was used to collect the relevant mass spectrometry data of each chemical component in PSF-p in the order of retention time. The total ion current of PSF-p was obtained by analyzing the test solution. The mass spectrometry information was collected in the positive and negative ion modes based on the total ion current and relevant literature information. Figure 8-9 (Deviation within ±10). Compound Discoverer 3.1 was used for retention time correction, peak identification, and peak extraction. Substance identification was performed based on secondary mass spectrometry information using the ThermomzCloud online database and the ThermomzValut local database. Chemical components of Phellodendron truncatum fruit were searched in positive and negative ion modes. The results are shown in Table 2. A total of 28 chemical components were identified in the PSF-p, predominantly polyphenols.

[0081] Table 2 Identification and analysis of chemical components in PSF-p positive and negative ion modes

[0082]

[0083] 4.2 Preliminary results of network pharmacology

[0084] 4.2.1 Chemical component screening and target collection results

[0085] Through searching and screening of traditional Chinese medicine database, 20 components of Phellodendron amurense fruit were finally identified, involving 131 targets.

[0086] Table 3 Screening of active ingredients from Phellodendron amurense fruit

[0087]

[0088] 4.2.2 Results of Targets Corresponding to Acute Liver Injury

[0089] The search results of the disease gene database involved 3,781 disease targets.

[0090] 4.2.3 Screening of Common Drug-Disease Targets

[0091] Through Venny 2.1 online software analysis, there are 96 intersection targets between liver damage and Phellodendron truncatum fruit, see Figure 2 .

[0092] 4.2.4 Construction and topological analysis of component-target-disease networks

[0093] The 20 active ingredients of Phellodendron amurense fruit screened under "4.2.1" and the 96 intersection targets obtained under 4.2.3 were input into Cytoscape 3.7.2 analysis software to draw the "ingredient-target-disease" interaction network diagram, as shown in Figure 2. Figure 3 The topological analysis of the 20 components was performed using Network Analyzer, as shown in Table 4. In Table 4, 12 core components can be screened out with the degree value being greater than or equal to the median of 6 as the limit.

[0094] Table 4 Topological analysis of the active ingredients-key targets-disease network of Phellodendron amurense fruit

[0095]

[0096] 4.2.5 PPI Network Analysis

[0097] The 96 intersection targets were imported into the STRING database, and the protein interaction PPI network diagram was obtained by analysis with a score greater than 0.4 as the screening condition, as shown in Figure 4The PPI network diagram data were exported and imported into Cytoscape 3.7.2 analysis software. Thirty core targets were further analyzed and screened in the MCODE module of the analysis software, including IL6, TNF, PTGS2, TP53, BCL2, CASP3, SRC, JUN, PPARG, NFKB1, HSP90AB1, HMOX1, NOS3, IL4, RELA, VCAM1, MAPK14, NOS2, PRKACA, SOD1, XDH, CDK2, SELE, PLAU, CDK1, MMP3, AR, E2F1, MMP1, and IFNB1.

[0098] 4.2.6 GO bioinformatics and KEGG pathway enrichment analysis

[0099] GO analysis was performed on 30 key targets for the treatment of liver injury using the DAVID database, with a P value less than 0.05 as the screening condition, and 283 valid entries were screened out (including 214 biological process entries, 25 cell composition entries, and 44 molecular function entries). Figure 5 Shown are the lowest filtered P10 entries of values ​​(in ascending order). The results showed that the biological processes mainly involved response to xenobiotic stimulus, response to hypoxia, positive regulation of apoptotic process, response to lipopolysaccharide, negative regulation of gene expression, positive regulation of transcription, DNA-templated, inflammatory response, positive regulation of gene expression, cellular response to hypoxia, and negative regulation of apoptotic process. The main factors affecting cellular composition included caveolae, extracellular space, extracellular region, macromolecular complex, ficolin-1-rich granule lumen, cell surface, lysosome, secretory granule lumen, cytosol, and plasma membrane.Molecular functions mainly include enzyme binding, heme binding, identical protein binding, protein homodimerization activity, oxidoreductase activity, beta-amyloid binding, primary amine oxidase activity, protein binding, flavin adenine dinucleotide binding, and transcription regulatory region sequence-specific DNA binding.

[0100] Then, the KEGG signaling pathway analysis was performed on these 96 key targets and P < 0.05 was used as the screening condition, resulting in 126 signaling pathways. P The values ​​are sorted in descending order, and the top 20 signaling pathways are screened out (see Figure 6 ), such as Lipid and atherosclerosis, Fluidshear stress and atherosclerosis, Tumor necrosis factor signaling pathway (TNF signaling pathway), IL-17 signaling pathway, AGE-RAGE signaling pathway in diabetic complications, Hepatitis B, PI3K-Akt signaling pathway, etc.

[0101] 4.2.7 Active ingredient-target-signaling pathway network

[0102] The active ingredient-target-signaling pathway diagram of Phellodendron truncatum fruit is shown in Figure 7In the graph, the higher the degree value, the darker the node color. Analysis of the graph reveals that 18 active ingredients in Phellodendron truncatum fruit are involved in the treatment of liver injury: citric acid, neochlorogenic acid, fraxin, salicylic acid, cyperoside, chlorogenic acid, caffeic acid, 7-hydroxycoumarin, scopoletin, hyperoside, (dihydro)cortex glycoside, daidzein, trifolioside, kaempferol, jatrorrhizine, berberine, dianthin, and limonin. Key ingredients include daidzein, kaempferol, and salicylic acid. Core targets include PTGS2, NFKB1, RELA, HSP90AB1, JUN, and BCL2. Possible mechanisms of Phellodendron truncatum fruit's hepatoprotective effects include lipid and inflammatory pathways such as atherosclerosis, tumor necrosis factor signaling, IL-17 signaling, and hepatitis B.

[0103] This invention is the first to apply the non-medicinal parts of the fruit of the medicinal plant Phellodendron amurense to liver protection treatment, and predicts its possible mechanism of action through network pharmacology, laying the foundation for further experimental verification of the target and pathway mechanism of the liver protection of Phellodendron amurense fruit.

[0104] The invention develops the pharmacological activity of the non-medicinal part of the fruit of Phellodendron amurense, which can provide a new idea for the sustainable development and utilization of Phellodendron amurense and expand the application space of traditional Chinese medicine resources.

[0105] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the present invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the present invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

[0106] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. Use of polyphenol extract of Phellodendron truncatum fruit in the preparation of a drug having a liver-protecting effect, characterized in that: The preparation method of the polyphenol extract of Phellodendron truncatum fruit comprises the following steps: Take the fruit powder of Phellodendron truncatum, add 40 times the amount of 50% ethanol in several times, ultrasonically extract at 60℃ for 75min, filter and combine the filtrate; concentrate the filtrate under reduced pressure on a rotary evaporator to remove the solvent, and then freeze-dry it in a freeze dryer to obtain the crude extract of Phellodendron truncatum fruit; fix the glass chromatography column, add absorbent cotton at the lower end, load the macroporous adsorption resin D101 activated by ethanol, wash with distilled water until there is no ethanol in the column and ensure that there are no bubbles in the column, and add absorbent cotton at the upper end; take the crude extract of Phellodendron truncatum fruit with a polyphenol concentration of 1.0 mg / mL and slowly pour it into the chromatography column; elute with pure water and 50% ethanol in turn and collect the 50% ethanol eluate, remove the solvent on a rotary evaporator, and then freeze-dry it in a freeze dryer to obtain the polyphenol extract of Phellodendron truncatum fruit.

2. The use according to claim 1, characterized in that: The polyphenol extract of Phellodendron chinense fruit includes neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, arbutin, 7-hydroxycoumarin, phellodendron glycoside, isoquercetin, hyperoside, salicylic acid, dianthin, caffeic acid, D-(-)-quinic acid, citric acid, trifolioside, kaempferol, protocatechualdehyde, cynarin, 3,5-dimethoxybenzoic acid, daidzein, malic acid, jasmonic acid, and quercetin polyphenol components.

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