Penthorum chinense pursh compound as well as preparation method and application thereof

Abstract

The invention relates to the field of penthorum chinense pursh compounds, and provides a penthorum chinense pursh compound, a preparation method and application thereof, and the preparation method comprises the following steps: crushing a penthorum chinense pursh medicinal material, adding a 70% methanol-water mixture which is 10 times of the amount of penthorum chinense pursh, heating and extracting for three times at 50 DEG C, heating for 1.5 hours each time, filtering, concentrating the filtrate to 10 L, adding one time of water, diluting and filtering; c18 is used for preparing crude components, the detection wavelength is 280 nm, a mobile phase is an acetonitrile-water system of 0.1% formic acid, and the acetonitrile-water system comprises three sections, namely a first acetonitrile crude section, a second acetonitrile crude section and a third acetonitrile crude section. Research finds that the novel penthorum chinense pursh compound penthorum chinense pursh glycoside E can have a good treatment effect on ethanol-induced hepatocellular diseases, and liver injury related complications are reduced.

Description

Technical Field

[0001] The present invention relates to the field of compounds of Penthorum chinense Pursh, and more specifically, to a compound of Penthorum chinense Pursh, a preparation method and an application thereof. Background Art

[0002] Penthorum chinense Pursh, scientific name Penthorum chinense Pursh, was first recorded in "Herbal for Relief of Famine" in the Ming Dynasty and is a local traditional Chinese medicine in Gulin County. It is mainly distributed in the Wumeng Mountain area at an altitude of about 1000 meters and has the effects of relieving jaundice and clearing away damp-heat.

[0003] As a common raw material for beverages, ethanol often causes damage to liver cells and affects physical health after the beverages prepared with it are consumed.

[0004] Therefore, we propose a compound of Penthorum chinense Pursh, a preparation method and an application thereof and its preparation method to solve the above problems. Summary of the Invention

[0005] Technical Problems to be Solved Aiming at the problems existing in the prior art, the purpose of the present invention is to provide a compound of Penthorum chinense Pursh, a preparation method and an application, which solves the problem of ethanol-induced damage to liver cells.

[0006] Technical Solutions

[0007] To solve the above problems, the present invention adopts the following technical solutions.

[0008] A compound of Penthorum chinense Pursh, including Figure 1 the structure shown.

[0009] A preparation method of the above-mentioned compound of Penthorum chinense Pursh includes crushing Penthorum chinense Pursh medicinal materials, adding a 70% methanol-water mixture 10 times the amount of Penthorum chinense Pursh, heating and extracting 3 times at 50°C for 1.5 hours each time, filtering, concentrating the filtrate to 10 liters, and then adding one-fold water for dilution and filtering; Performing C18 preparative rough fractionation, with a detection wavelength of 280 nm and a mobile phase of an acetonitrile-water system with 0.1% formic acid, including three segments: acetonitrile rough segment one, acetonitrile rough segment two, and acetonitrile rough segment three; Concentrating acetonitrile rough segment two to 1 liter at 50°C, filtering, and performing C18 preparation again. The mobile phase is a 5% acetonitrile-water system containing 0.1% formic acid for preparation, with a detection wavelength of 280 nm. Collect the target F4 target segment, concentrate it to a small volume at 50°C, and repeatedly prepare and separate it with a 14% methanol-water system and a 5% acetonitrile-water system. Collect the qualified F4 and F4-2 solutions, and after concentrating under reduced pressure at 50°C and freeze-drying, obtain F4 solid and F4-2 solid, where F4 is casuarine, and F4-2 is the above-mentioned compound of Penthorum chinense Pursh.

[0010] Crude acetonitrile fraction 1: Concentrated to 2 L at 50 °C, filtered, and separated again by C18 preparation. The mobile phase was a 1% acetonitrile - water system containing 0.1% formic acid. The target fractions F1 and F3 containing the target substance were collected. The collected solutions of the two fractions were concentrated to 5% of the original volume at 50 °C, and then refined in an acetonitrile system. The mobile phase was 1% acetonitrile - water. The qualified solution was collected, concentrated to an appropriate volume at 50 °C, and then freeze - dried to obtain 1.67 g of solid F1 and 4.24 g of solid F3. Through structural analysis, F1 was determined to be 1 - galloylglucose, and F3 was phyllembin F.

[0011] Use of a swertia compound as described above in the preparation of a food, health product, or drug for preventing and / or treating ethanol - induced hepatocyte injury.

[0012] Beneficial effects: Compared with the prior art, the advantages of the present invention are as follows: Using Gulin swertia as raw material, extracting by methanol reflux method, and then obtaining 4 active substances through multi - stage separation and directional collection and purification by preparative liquid chromatography. Through nuclear magnetic resonance and mass spectrometry data analysis, they were finally identified as 1 - galloylglucose, phyllembin F, casuarinin, and a new swertia compound. The new swertia compound is tentatively named swertiamarin E in Chinese.

[0013] Through experiments, it was found that the new swertia compound swertiamarin E can have a good therapeutic effect on ethanol - induced hepatocyte diseases, potentially treating alcoholic liver diseases. For example, alcoholic liver diseases (such as fatty liver, hepatitis, cirrhosis) are common liver diseases globally. If swertiamarin can pass the safety and effectiveness verification, it can become a new choice for clinical treatment, providing a more effective intervention means for alcoholics or patients with alcoholic liver diseases, and reducing the incidence of serious complications such as liver fibrosis and liver failure.

[0014] Reducing liver injury - related complications, ethanol - induced hepatocyte injury may trigger systemic inflammatory responses, abnormal liver function, etc., and then affect other organs (such as the kidneys, nervous system). Protecting hepatocytes can indirectly reduce the risk of multiple organ dysfunction and improve the overall prognosis of patients.

[0015] Possibility of combination therapy. This swertia compound can be used in combination with existing hepatoprotective drugs (such as polyene phosphatidylcholine, silymarin, etc.) to enhance the therapeutic effect, or reduce the dosage and side effects of existing drugs, providing more choices for individualized treatment plans. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is a schematic diagram of the molecular structure of the swertia compound proposed by the present invention; Figure 2 It is a schematic diagram of the HMBC data of the swertia compound proposed by the present invention; Figure 3 Schematic diagram of 1H-1H COSY data of the compound of Penthorum chinense Pursh in the present invention; Figure 4 Schematic diagram of the molecular structure of 1-O-galloylglucopyranose of the compound of Penthorum chinense Pursh in the present invention; Figure 5 Schematic diagram of the molecular structure of phyllemblin F of the compound of Penthorum chinense Pursh in the present invention; Figure 6 Schematic diagram of the molecular structure of casuarinin of the compound of Penthorum chinense Pursh in the present invention. Detailed implementation manners

[0017] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention; obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0018] Embodiment 1

[0019] Referring to Figure 1 In this embodiment of the present application, a compound of Penthorum chinense Pursh is provided, including Figure 1 the structure shown.

[0020] Embodiment 2

[0021] A preparation method of a compound of Penthorum chinense Pursh, using Gulin Penthorum chinense Pursh as the raw material, extracting by methanol reflux method, and then obtaining 4 active substances through multi-stage separation and directional collection and purification by preparative liquid chromatography. Through nuclear magnetic resonance and mass spectrometry data analysis, they are finally identified as 1-galloylglucose, phyllemblin F, casuarinin, and 1 new compound of Penthorum chinense Pursh. The temporary Chinese name of the new compound of Penthorum chinense Pursh is named as Penthorum chinense glycoside E.

[0022] 10 kg of Penthorum chinense Pursh herbs were crushed, extracted with 10 times the amount of 70% methanol-water at 50 °C for 3 times, 1.5 hours each time. After filtration, the filtrate was concentrated to 10 L at 50 °C, then diluted with 1 time the amount of water and filtered. C18 preparative rough fractionation was carried out. Using a preparative liquid chromatography system filled with C18 packing material, the raw materials were roughly separated. The preparation process included column equilibration, sample injection, mobile phase elution, collection of eluate. The detection wavelength was 280 nm, and the mobile phase was a 5% acetonitrile-water system containing 0.1% formic acid, which was divided into 3 segments, namely acetonitrile rough segment 1, acetonitrile rough segment 2, and acetonitrile rough segment 3. According to the typical chromatogram of gallic acid-glucoside compounds, the elution order was sorted by time and recorded as acetonitrile rough segment one, acetonitrile rough segment two, and acetonitrile rough segment three. The collected solution was detected and judged by an analytical liquid chromatography system, and the qualified solution that met the quality requirements was selected and judged as a qualified solution.

[0023] Acetonitrile rough segment 1: Concentrated to 2 L at 50 °C, filtered, and separated again by C18 preparation. The mobile phase was a 1% acetonitrile-water system containing 0.1% formic acid. The target segments F1 and F3 containing the target substances were collected. The two collected solutions were concentrated to 5% of the original volume at 50 °C, and then refined in an acetonitrile system. The mobile phase was 1% acetonitrile-water. The qualified solution was collected, concentrated to an appropriate volume at 50 °C, and then freeze-dried to obtain 1.67 g of solid F1 and 4.24 g of solid F3. After structural analysis, F1 was determined to be 1-galloylglucose, and F3 was determined to be phyllanthusi F.

[0024] Acetonitrile rough segment 2: Concentrated to 1 L at 50 °C, filtered, and prepared again by C18. The mobile phase was a 5% acetonitrile-water system containing 0.1% formic acid for preparation. The detection wavelength was 280 nm. The target segment F4 containing the target was collected. It was concentrated to a small volume at 50 °C, and then repeatedly prepared and separated using a 14% methanol-water system and a 5% acetonitrile-water system. The qualified F4 and F4-2 solutions were collected, concentrated under reduced pressure at 50 °C and then freeze-dried to obtain 9.8 g of solid F4 and 0.369 g of solid F4-2. After structural analysis, F4 was determined to be casuarinin, and F4-2 was determined to be a new Penthorum chinense Pursh compound, named penthorin E.

[0025] Structural identification results of the active substances in Penthorum chinense Pursh: F1: 1-O-Galloyl-β-D-glucopyranose.

[0026] White powder, the quasi-molecular ion peak m / z = 331.34 [M-H]- by LC-MS, and the molecular formula is C13H16O10.

[0027] 1H-NMR (400 MHz, Methonal-d4): δ 7.12 (2H, s, H-2', H-6'), 5.65 (1H, d, 7.6, H-1), 3.85 (1H, dd, 1.5, 12.1, H-6), 3.70 (1H, dd, J = 4.5, 12.1 Hz, H-6), 3.40 - 3.50 (4H, m, H-2, H-3, H-4, H-5). 13C-NMR (100 MHz, Methonal-d4) δ: 95.9 (C-1), 74.1 (C-2), 78.2 (C-3), 71.1 (C-4), 78.8 (C-5), 62.3 (C-6), 120.7 (C-1'), 110.5 (C-2'), 146.5 (C-3'), 140.3 (C-4'), 146.5 (C-5'), 110.5 (C-6'), 167.0 (C-7). The above 1H NMR and 13C NMR data are basically consistent with those reported in the literature for 1-O-galloylglucopyranose, so it was identified as 1-O-galloylglucopyranose. Its molecular formula is as Figure 4 shown.

[0028] F3: Phyllanemblinin F.

[0029] White powder, with the quasi-molecular ion peak m / z = 669.40 [M-H]- by LC-MS, and the molecular formula C27H26O20.

[0030] 1H NMR (400 MHz, Methanol-d4) δ: 7.18 (2H, s, galloyl H-2’,6’), 7.05 (1H, s, neochebuloyl H-3’’), 5.65 (1H, d, 8.0, glc H-1), 5.29 (1H, d, 1.3, neochebuloyl H-2’’’), 4.50 (1H, dd, 2.3, 12.1, H-6a), 4.35 (1H, dd, 5.2, 12.0, H-6b), 3.91 (1H, dd, 1.3, 8.8, H-3’’’), 3.20 (1H, m, H-4’’’), 2.91 (1H, m, H-5’’’a), 2.42 (1H, dd, 4.9, 17.2, H-5’’’b), 13C NMR (100 MHz, Methanol-d4) δ: 175.2, 174.6, 172.3, 166.8, 164.4, (neochebuloyl C-1’’’,6’’’,7’’’,7’’, galloyl C-7’), 146.8 (C-6’’), 146.4 (C-3’,5’), 144.6 (C-4’’), 140.6 (C-5’’), 140.3 (C-4’), 120.7 (C-1’), 117.7 (C-2’’), 116.2 (C-1’’), 110.7 (C-2’,6’), 109.3 (C-3’’), 95.9 (C-1), 78.6 (C-2’’’), 78.0 (C-5), 76.0 (C-3), 74.0 (C-2), 71.1 (C-4), 64.9 (C-6), 45.6 (C-3’’’), 37.2 (C-4’’’), 35.0 (C-5’’’). The above 1H NMR and 13C NMR data are basically consistent with those reported in the literature. Therefore, this compound was identified as Phyllanemblinin F. Its molecular formula is as Figure 5 shown. F4: Strictinin, also known as Phyllanemblinin A, Phyllanemblinin B, and Pu-erh Fat-reducing Element (Strictinin).

[0031] White powder. The quasi-molecular ion peak of LC-MS is m / z = 634.01 [M-H]-, and its molecular formula is C27H22O18.

[0032] 1H NMR (400 MHz, Methanol-d4) δ: 7.15 (2H, s, H-2″,6″), 6.70- (1H, s,H-2'), 6.56 (1H, s, H-6'), 5.68 (1H, d, J = 8.1 Hz, H-1), 5.24 (1H, dd, J =13.2, 6.3 Hz, H-6), 4.87 (1H, overlap), 4.05 (1H,dd, 9.9, 5.9, H-5), 3.82(1H, d, J = 13.0 Hz, H-3), 3.73 (1H, t, 9.3, H-6), 3.63 (1H, m, H-2); 13C NMR(100 MHz, Methanol-d4) δ 169.9 (C-7'), 169.6 (C-7'''), 166.8 (C-7''), 146.5(C-3', 5'), 145.8 (C-6'', 6'''), 144.8(C-4''), 144.8(C- 4'''), 140.4 (C-4'),137.6 (C-5''),137.3 (C-5'''), 126.5 (C-1''), 126.3 (C- 1'''), 120.5 (C-1'),116.8 (C-2''), 116.6 (C- 2'''), 110.5(C-2', 6'), 108.6 (C-3''), 108.6 (C-3'''), 96.2 (C-1), 76.0 (C-2), 74.7 (C-3), 73.2 (C-4), 73.6 (C-5), 64.2 (C-6). The above 1H NMR and 13C NMR data are basically consistent with those reported in the literature. Therefore, the compound was identified as penchinenin. Its molecular formula is as Figure 6 shown.

[0033] F4-2: Penchinensin A.

[0034] This penchinenin compound was isolated from the leaves of Penthorum chinense Pursh (Penchinensis Herba), a plant of the genus Penthorum in the Saxifragaceae family. After a preliminary literature search, it was found to be a new penchinenin compound. For the sake of convenience in description, it was temporarily named penchinenin E in Chinese and Penchinensin A in English. As Figure 1 shown.

[0035] White powder, with the quasi-molecular ion peak of LC-MS at m / z = 669.40 [M-H]-, and the molecular formula of C27H26O20.

[0036] 1H NMR (Methanol-d4, 400 MHz) shows three benzene ring hydrogen proton signals, δ 7.14 (2H, s, H-2''',6''') and 7.02 (1H, s, H-7), one β-configured glucose anomeric hydrogen proton signal δ 5.75 (1H, d, 8.2, H-1') and other glucose hydrogen proton signals δ 5.06 (1H, t, 9.6, H-4'), δ 3.83 (1H,t, 9.3, H-3'), δ 3.55 - 3.78 (4H, m, H-2', H-5' and H-6'), one oxygen-linked methine H signal δ 5.27 (1H, d, 8.3, H-3), two methine H signals δ 4.81 (1H, overlap, H-4) and 3.72 (1H, m, H-11) and one methylene H signal δ 2.73 (1H, dd, 17.2, 5.0, H-12a), 2.66 (1H, dd, 17.1, 6.4, H-12b).

[0037] 13C NMR (Methanol-d4, 100 MHz) showed signals of 27 carbon atoms, including 5 carbonyl carbon signals at δ 179.8, 174.7, 173.8, 168.7, 166.9; 12 benzene ring carbon atom signals at δ 146.5×2, 146.4, 145.6, 140.4, 138.9, 122.9, 120.6, 118.4, 111.1, 110.5×2; 7 oxygenated methylene or methine carbon atom signals at δ 95.8, 79.4, 76.6, 75.8, 74.3, 72.4, 62.0 and 3 methylene or methine carbon atom signals at δ 44.3, 42.6, 35.0. Combining the above data, it can be determined that the compound contains 1 β-glucosyl fragment [δH 5.75 (1H, d, 8.2, H-1’), 3.83 (1H, t, 9.3, H-3’), 3.55 - 3.78 (4H, m, H-2’, H-5’ and H-6’) and δC 95.8, 76.6, 75.8, 74.3, 72.4, 62.0], 1 1,3,4,5-tetrasubstituted galloyl fragment [δH 7.14 (2H, s, H-2’’’,6’’’) and δC 166.9, 146.5×2, 140.4, 120.6, 110.5×2], 1 1,2,3,4,5-pentasubstituted galloyl fragment [δH 7.02 (1H, s, H-7) and δC 168.7, 146.4,145.6, 138.9, 122.9, 118.4, 111.1]. According to HNMR and combined with the HSQC spectrum, it can be determined that C-3 [δH 5.27 and δC 79.4], C-4 [δH 4.81 and δC 44.3] and C-11 [δH 3.72 and δC 42.6] are methine carbon signals, and C-12 [δH 2.73, 2.66 and δC 35.0] is a methylene carbon signal.

[0038] In the 1H-1H COSY spectrum, it can be clearly seen that there are cross-correlations between H-12 / H-11, H-11 / H-4, and H-4 / H-3. Combining with H-12 / C-13, H-12 / C-14, H-12 / C-4, H-11 / H-13, H-11 / H-12, H-4 / 12, H-4 / C-12, H-4 / C-2, H-4 / C-11, H-3 / C-4 in HMBC, the connection order of C-3, C-4, C-11, and C-12 can be determined.

[0039] The correlation of H-1’ / C-7’’ in the HMBC spectrum can determine that the 1,3,4,5-tetrasubstituted galloyl group is acylated at the C-1’ position of the glucosyl group; the correlation of H-4’ / C-15 can determine that the 1,2,3,4,5-pentasubstituted galloyl group is acylated at the C-4’ position of the glucosyl group; the correlations of H-3 / C-5, H-4 / C-5, and H-11 / C-5 can determine that C-4 and C-5 are directly connected. The configurations of C-3, C-4, and C-11 in this Penthorum chinense Pursh compound cannot be determined. This Penthorum chinense Pursh compound is a new compound, named Penthorum chinense Pursh glycoside E. The main HMBC and 1H-1H COSY data are as Figure 2 and Figure 3 shown.

[0040] Example 3

[0041] Use of a Penthorum chinense Pursh compound as described above in the preparation of a food, health product, or drug for preventing and / or treating ethanol-induced hepatocyte injury.

[0042] The above Penthorum chinense Pursh compound was used in experiments on ethanol-induced hepatocytes, and the experimental data are as follows: Statistical analysis was performed using IBM SPSS Statistics 22.0 software, and one-way ANOVA was used for comparative analysis: Table 1 Changes in ALT in cells of each group ( ±SD) Group Number of samples (example) ALT (U / L) Blank group 3 10.411±1.358 Model group 3 <![CDATA[20.760±1.840 *** > Model + F1 group 3 <![CDATA[15.365±0.343 ### > Model + F3 group 3 <![CDATA[14.615±1.886 ### > Model + F4 group 3 18.912±1.411 Model + F4-2 group 3 <![CDATA[15.515±2.282 ### > Note: Compared with the blank group, in the model group, * P<0.05, ** P<0.01, *** P<0.001; Compared with the model group for the remaining groups, # P<0.05, ## P<0.01, ### P<0.001; Table 1 statistical results show that compared with the blank group, the alanine aminotransferase (ALT) in the cells of the model group increased (P<0.001). Among them, the model group was the ethanol-induced hepatocyte injury group. Human hepatoma cells: code HepG2, product number icell-h092, icell.

[0043] Compared with the model group, the alanine ammonia ALT in the cells of the model + F1 group, model + F3 group, model + F4 group, and model + F4-2 group decreased (P<0.001).

[0044] Table 2 Changes in AST in cells of each group ( ±SD) Group Number of samples (example) AST (U / L) Blank group 3 35.009±4.620 Model group 3 <![CDATA[60.594±4.143 *** > Model + F1 group 3 <![CDATA[48.754±3.995 # > Model + F3 group 3 <![CDATA[41.670±7.452 ## > Model + F4 group 3 <![CDATA[46.331±2.593 ## > Model + F4-2 group 3 <![CDATA[45.170±4.624 ## > Note: Compared with the blank group, * P < 0.05, ** P < 0.01, *** P < 0.001; Compared with the model group, # P < 0.05, ## P < 0.01, ### P < 0.001; Table 2 Statistical results show that compared with the blank group, the aspartate aminotransferase (AST) in the cells of the model group increased (P < 0.001).

[0045] Compared with the model group, AST in the cells of the model + F1 group decreased (P < 0.05), and AST in the cells of the model + F3 group, model + F4 group, and model + F4-2 group decreased (P < 0.01).

[0046] Table 3 Changes in LDH in the cells of each group ( ±SD) Group Number of samples (example) LDH (U / gprot) Blank group 3 3527.104±28.847 Model group 3 <![CDATA[5434.782±382.677 *** > Model + F1 group 3 <![CDATA[4421.666±347.759 ### > Model + F3 group 3 <![CDATA[4397.538±335.541 ### > Model + F4 group 3 5155.494±267.119 Model + F4-2 group 3 <![CDATA[4406.431±179.905 ### > Note: Compared with the blank group, * P < 0.05, ** P < 0.01, *** P < 0.001; Compared with the model group, # P < 0.05, ## P < 0.01, ### P < 0.001; Table 3 Statistical results show that compared with the blank group, the lactate dehydrogenase (LDH) in the cells of the model group increased (P < 0.001).

[0047] Compared with the model group, LDH in the cells of the model + F1 group, model + F3 group, and F4-2 group decreased (P < 0.001), and there was no significant difference in LDH in the cells of the model + F4 group (P > 0.05).

[0048] Statistical analysis was performed using IBM SPSS Statistics 22.0 software, and one-way ANOVA was used for comparative analysis.

[0049] Table 4 Changes in SOD in the cells of each group ( ±SD) Group Number of samples (example) SOD (U / mgprot) Blank group 3 39.452±1.060 Model group 3 <![CDATA[28.794±2.712 *** > Model + F1 group 3 <![CDATA[33.243±1.024 # > Model + F3 group 3 <![CDATA[36.706±3.151 ### > Model + F4 group 3 <![CDATA[32.733±1.323 # > Model + F4-2 group 3 <![CDATA[34.817±2.030 ## > Note: Compared with the blank group, * P < 0.05, ** P < 0.01, *** P < 0.001; Compared with the model group,# P < 0.05, ## P < 0.01, ### P < 0.001; Table 4 statistical results showed that compared with the blank group, SOD in the cells of the model group decreased (P < 0.001).

[0050] Compared with the model group, SOD increased in the cells of the model + F1 group, model + F4 group, and model + F4-2 group (P < 0.05), and SOD increased in the cells of the model + F3 group (P < 0.001).

[0051] Table 5 Changes in MDA in cells of each group ( ±SD) Group Number of samples (example) MDA (nmol / mgprot) Blank group 3 1.837±0.307 Model group 3 <![CDATA[3.406±0.225 *** > Model + F1 group 3 <![CDATA[2.095±0.414 ## <!-- 6 -->]]> Model + F3 group 3 <![CDATA[2.459±0.605 # > Model + F4 group 3 2.966±0.444 Model + F4-2 group 3 <![CDATA[2.459±0.508 # > Note: Compared with the blank group, * P < 0.05, ** P < 0.01, *** P < 0.001; Compared with the model group for the remaining groups, # P < 0.05, ## P < 0.01, ### P < 0.001; Table 5 statistical results showed that compared with the blank group, malondialdehyde (MDA) in the cells of the model group increased (P < 0.001).

[0052] Compared with the model group, MDA decreased significantly in the cells of the model + F1 group (P < 0.01), MDA decreased in the cells of the model + F3 group and model + F4-2 group (P < 0.05), and there was no significant difference in MDA in the cells of the model + F4 group (P > 0.05).

[0053] Table 6 Changes in GSH in cells of each group ( ±SD) Group Number of samples (example) GSH (μmol / gprot) Blank group 3 126.433±6.870 Model group 3 <![CDATA[87.305±6.979 *** > Model + F1 group 3 <![CDATA[99.661±5.216 # > Model + F3 group 3 <![CDATA[103.301±5.487 ## > Model + F4 group 3 <![CDATA[100.669±4.852 # > Model + F4-2 group 3 <![CDATA[105.993±7.296 ## > Note: Compared with the blank group, * P < 0.05, ** P < 0.01, *** P < 0.001; Compared with the model group for the remaining groups, # P < 0.05, ## P < 0.01, ### P < 0.001; Table 6 statistical results showed that compared with the blank group, glutathione (GSH) in the cells of the model group decreased (P < 0.001).

[0054] Compared with the model group, the GSH levels in the cells of the model + F1 group and the model + F4 group increased (P < 0.05), and the GSH levels in the cells of the model + F3 group and the model + F4-2 group increased (P < 0.01).

[0055] Table 7 Changes in CAT in cells of each group ( ±SD) Group Number of samples (example) CAT (U / mgprot) Blank group 3 6.511±0.484 Model group 3 <![CDATA[3.833±0.208 *** > Model + F1 group 3 <![CDATA[4.920±0.516 # > Model + F3 group 3 <![CDATA[5.294±0.551 ## > Model + F4 group 3 4.405±0.331 Model + F4-2 group 3 <![CDATA[4.959±0.329 # > Note: Compared with the blank group, * P < 0.05, ** P < 0.01, *** P < 0.001; Compared with the model group, # P < 0.05, ## P < 0.01, ### P < 0.001; The statistical results in Table 7 showed that compared with the blank group, the catalase (CAT) in the cells of the model group decreased (P < 0.001).

[0056] Compared with the model group, the CAT in the cells of the model + F1 group and the model + F4-2 group increased (P < 0.05), the CAT in the cells of the model + F3 group increased (P < 0.01), and the difference in CAT in the cells of the model + F4 group was not significant (P > 0.05).

[0057] In practical applications, it has the following advantages.

[0058] I. Significance for the study of liver diseases Revealing the mechanism of liver injury: The protective effect of this Phyllanthus amarus compound may help scientists further clarify the specific molecular mechanisms of ethanol-induced hepatocyte injury (such as oxidative stress, inflammatory response, apoptosis pathways, etc.). By studying the interaction between the Phyllanthus amarus compound and its targets, the pathological process of liver injury can be deeply understood, providing a new perspective for subsequent research.

[0059] Expanding the research direction of liver protection drugs: Currently, there are limited specific drugs for alcoholic liver injury. This Phyllanthus amarus compound may become a lead compound for new liver protection drugs. Its target or pathway (such as activation of antioxidant enzymes, inhibition of inflammatory factors, etc.) can be used as a key basis for drug design, promoting the research and development process of related drugs.

[0060] II. Significance for clinical treatment Potential drugs for the treatment of alcoholic liver disease: Alcoholic liver disease (such as fatty liver, hepatitis, cirrhosis) is a common liver disease globally. If the swertia compound can pass the safety and efficacy verification, it can become a new option for clinical treatment, providing more effective intervention means for alcoholics or patients with alcoholic liver disease, and reducing the incidence of serious complications such as liver fibrosis and liver failure.

[0061] Reducing liver injury-related complications: Ethanol-induced hepatocyte injury may trigger systemic inflammatory responses, abnormal liver function, etc., which in turn affect other organs (such as the kidneys, nervous system). Protecting hepatocytes can indirectly reduce the risk of multiple organ dysfunction and improve the overall prognosis of patients.

[0062] Possibility of combination therapy: The swertia compound can be used in combination with existing hepatoprotective drugs (such as polyene phosphatidylcholine, silymarin, etc.) to enhance the therapeutic effect, or reduce the dosage and side effects of existing drugs, providing more options for individualized treatment plans.

[0063] III. Significance for public health Reducing the social burden of alcohol-related liver diseases: Liver injury caused by alcohol consumption is one of the major global health problems. According to World Health Organization data, alcoholic liver disease accounts for 30% - 50% of the causes of cirrhosis. If the swertia compound can be translated into practical applications, it can effectively reduce the incidence and mortality of alcoholic liver disease, reduce the consumption of medical resources, and alleviate the burden on families and society.

[0064] Potential for preventive application: For people who need to be exposed to alcohol for a long time (such as alcoholics, alcohol industry practitioners), the swertia compound may be used as a preventive drug to reduce the risk of liver injury, achieve the goal of "preventing diseases before they occur", and improve the public health level.

[0065] IV. Significance for drug development and industry Promoting the industrialization of innovative drugs: If the swertia compound has good drug-forming properties (such as high stability, high bioavailability, and low side effects), it can attract pharmaceutical companies to invest in research and development, form new pharmaceutical products, promote innovation in the pharmaceutical industry, and create economic value.

[0066] Expanding the application of natural products or synthetic swertia compounds: If the swertia compound is derived from natural products (such as plants, microorganisms), it can promote the development and utilization of natural drugs; if it is a synthetic swertia compound, the synthetic route can be optimized, the production cost can be reduced, and the foundation for large-scale production can be laid.

[0067] V. Significance for basic science and interdisciplinary research Enrich the theory of cell protection mechanisms: This research may discover new cell protection pathways or targets, providing new theories for fields such as cell biology and pharmacology, and promoting the development of basic science. For example, if the compounds in Phyllanthus niruri L. play a role by regulating the gut microbiota-liver axis, it can expand the research on the "gut-liver axis" in liver injury.

[0068] Promote interdisciplinary cooperation: From the screening of compounds in Phyllanthus niruri L., mechanism research to clinical translation, multidisciplinary collaboration is required (such as medicinal chemistry, molecular biology, clinical medicine, epidemiology, etc.). This research can promote the cross-integration of different fields and accelerate the transformation of scientific research results.

[0069] VI. Limitations and Future Directions Although this discovery is of great significance, further verification is still needed: Safety assessment: Animal toxicology experiments and clinical trials are required to confirm the safety and tolerance of the compounds in Phyllanthus niruri L. in vivo.

[0070] In-depth study of the mechanism of action: Identify key targets and signaling pathways to avoid the problem of unclear mechanisms caused by "pleiotropy".

[0071] Explore clinical applicability: Stratified studies are needed for the efficacy differences in different populations (such as different alcohol consumption levels and stages of liver injury).

[0072] The above are only specific implementation manners of the present invention. It should be noted that for those of ordinary skill in the art of this technology, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A kind of Penthorum chinense Pursh compound, characterized in that, It has the structure described in Formula 1: Formula 1.

2. A preparation method of the compound of Penthorum chinense Pursh as described in claim 1, characterized in that, Including: The Herba Penthori was crushed, and a 70% methanol-water mixture 10 times the amount of Herba Penthori was added, heated and extracted 3 times at 50 °C, each heating for 1.5 hours, filtered, the filtrate was concentrated to 10 L, and then diluted with one-fold water and filtered; Coarse separation by C18 preparation. Using a preparative liquid phase system filled with C18 packing, the raw materials were coarsely separated. The preparation process included column equilibration, sample injection, mobile phase elution, collection of eluate, detection wavelength was 280 nm, and the mobile phase was an acetonitrile-water system with 0.1% formic acid, including three segments: acetonitrile coarse segment 1, acetonitrile coarse segment 2, and acetonitrile coarse segment 3. According to the typical chromatogram of gallic acid-glucoside compounds, the elution peak order was sorted by time and recorded as acetonitrile coarse segment 1, acetonitrile coarse segment 2, and acetonitrile coarse segment 3. The collected solution was detected and discriminated by an analytical liquid phase system, and the solution meeting the quality requirements was screened out and determined as a qualified solution; The acetonitrile coarse segment 2 was concentrated to 1 L at 50 °C, filtered, and C18 was prepared again. The mobile phase was prepared with a 5% acetonitrile-water system containing 0.1% formic acid, the detection wavelength was 280 nm, the target F4 segment was collected, concentrated to a small volume at 50 °C, and repeatedly prepared and separated with a 14% methanol-water system and a 5% acetonitrile-water system. The qualified F4 and F4-2 solutions were collected, concentrated under reduced pressure at 50 °C and then freeze-dried to obtain solid F4 and solid F4-2, where F4 is casuarine, and F4-2 is the Penthorum chinense Pursh compound described in claim 1.

3. Use of the Penthorum chinense Pursh compound described in claim 1 in the preparation of a medicament for preventing and / or treating ethanol-induced hepatocyte injury.

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