Application of rhizoma polygonati extract in preparation of medicine for treating liver injury

Abstract

The invention relates to the technical field of biological pharmacy, in particular to application of a rhizoma polygonati extract in preparation of a medicine for treating liver injury, and the rhizoma polygonati extract is prepared from the following main active ingredients: hexadecanamide, L-valine, 2-pyrrolidine carboxylic acid, 4-oxoproline, manninotriose, succinic acid, choline, D-(-)-ribose, stachyose and L-threonine. The invention proves that the prophylactic administration of the rhizoma polygonati extract can improve the acetaminophen-induced liver injury, so that the rhizoma polygonati extract has important clinical significance on pain patients who need to take acetaminophen for a long time, and can be developed into an effective liver protection auxiliary medicine.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical technology, specifically to the application of Polygonatum extract in the preparation of drugs for treating liver injury. Background Technology

[0002] Drug-induced liver injury (DAI) is one of the leading causes of acute liver failure worldwide, accounting for approximately 50% of all ATI cases. Acetaminophen (APAP), a commonly used over-the-counter antipyretic and analgesic, has become the most common cause of DAI due to its narrow therapeutic window. When ingested in excess of safe doses, excessive APAP is metabolized in the liver to produce N-acetyl-p-benzoquinone imine, which depletes hepatic glutathione, promotes the accumulation of reactive oxygen species, and triggers severe oxidative stress, DNA damage, and inflammatory responses, ultimately leading to hepatocyte necrosis and apoptosis. Currently, N-acetylcysteine ​​remains the gold standard clinical treatment for APAP overdose; however, its therapeutic window is extremely narrow. Even with timely administration, some patients may still experience liver damage progression or treatment failure. Furthermore, N-acetylcysteine ​​administration may cause adverse reactions such as nausea, vomiting, and allergic reactions, affecting patient tolerance and treatment completion. Therefore, there is an urgent clinical need to develop novel therapeutic drugs or adjunctive treatment strategies for APAP-induced liver injury. Summary of the Invention

[0003] To address the above problems, this invention provides the application of Polygonatum extract in the preparation of drugs for treating liver injury.

[0004] This invention is achieved through the following technical solution: The application of Polygonatum extract in the preparation of drugs for treating liver injury, wherein the main active ingredients in Polygonatum extract include hexadecamide, L-valine, 2-pyrrolidinecarboxylic acid, 4-oxoproline, mannitol, succinic acid, choline, D-(-)-ribose, stachyose and L-threonine.

[0005] Preferably, the liver injury is drug-induced liver injury caused by acetaminophen.

[0006] Preferably, the Polygonatum extract is the only effective ingredient in the preparation of the drug for treating liver injury.

[0007] Preferably, the drug further includes pharmaceutically acceptable pharmaceutical excipients.

[0008] Preferably, the drug further includes pharmaceutically acceptable pharmaceutical excipients, which are one or more of the following: diluents, disintegrants, precipitation inhibitors, flow aids, binders, dispersants, suspending agents, isotonic agents, thickeners, emulsifiers, preservatives, and stabilizers.

[0009] Preferably, the Polygonatum extract is obtained by the following method: Polygonatum slices are dried, pulverized and sieved to obtain a dry powder; water is added to the dry powder for ultrasonic extraction to obtain the Polygonatum extract; the mass-volume ratio of the dry powder to water is 1g:20mL~40mL; the ultrasonic extraction time is 30min~60min; and the ultrasonic extraction frequency is 20kHz~40kHz.

[0010] Preferably, the thickness of the Polygonatum slices is 0.1cm to 0.5cm.

[0011] Preferably, the drying temperature is 40℃~60℃.

[0012] Preferably, the particle size of the dried powder is 60 mesh to 120 mesh.

[0013] Compared with the prior art, the present invention has the following beneficial effects: This invention provides the application of Polygonatum sibiricum extract in the preparation of drugs for treating liver injury. The main active components of the Polygonatum sibiricum extract include hexadecylamide, L-valine, 2-pyrrolidone carboxylic acid, 4-oxoproline, mannotriose, succinic acid, choline, D-(-)-ribose, stachyose, and L-threonine. This invention systematically elucidates for the first time that the protective effect of this Polygonatum sibiricum extract with specific active components against APAP-induced liver injury is achieved synergistically through a "triple inhibition mechanism": simultaneously inhibiting oxidative stress, significantly reducing MDA levels and restoring SOD activity; inhibiting apoptosis, reducing TUNEL-positive cells, and downregulating Caspase-3 expression; and inhibiting inflammatory responses, downregulating key inflammatory factors such as TNF-α and IL-17 and related signaling pathways. This multi-component, multi-target integrated regulatory mechanism overcomes the limitations of most existing hepatoprotective drugs with their single pathway of action, fully demonstrating the unique advantages of traditional Chinese medicine's multi-component, multi-pathway holistic regulation. This invention demonstrates that prophylactic administration of Polygonatum extract can improve APAP-induced liver damage, which has important clinical significance for pain patients, such as those with arthritis, who need to take APAP long-term, and can be developed into an effective adjuvant drug for liver protection. Attached Figure Description

[0014] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0015] Figure 1 This is a graph showing the component analysis results of the Polygonatum extract of this invention; Figure 1In the diagram, A is the TIC diagram of Polygonatum extract under positive ion mode; B is the TIC diagram of Polygonatum extract under negative ion mode.

[0016] Figure 2 This is a graph showing the changes in mouse body weight and liver index results of the present invention; Figure 2 In the graph, A represents the change in mouse body weight; B represents the liver index.

[0017] Figure 3 This is a graph showing the serum biochemical results of the present invention; Figure 3 In the table, A represents the level of alanine aminotransferase; B represents the level of aspartate aminotransferase; C represents the level of alkaline phosphatase; and D represents the level of total bilirubin.

[0018] Figure 4 This is a diagram showing the results of liver H&E staining according to the present invention.

[0019] Figure 5 This is a graph showing the results of malondialdehyde content and superoxide dismutase activity in this invention. Figure 5 In the table, A represents malondialdehyde content; B represents superoxide dismutase activity.

[0020] Figure 6 This image shows the results of liver TUNEL staining and Caspase 3 immunohistochemistry in this invention.

[0021] Figure 7 This is a diagram showing the KEGG enrichment pathway results of liver transcriptomics in this invention.

[0022] Figure 8 The present invention relates to the liver TNF-α and IL-17 mRNA levels; Figure 8 In the table, A represents the relative mRNA level of TNF-α; B represents the relative mRNA level of IL-17. Detailed Implementation

[0023] To facilitate understanding of the present invention, a more comprehensive description is provided below, along with preferred embodiments. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0024] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this invention and in its specification is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0025] The beneficial effects of the present invention will be illustrated below through specific embodiments.

[0026] The Polygonatum used in this invention is a common commercially available product.

[0027] Example 1 Extraction method of Polygonatum extract.

[0028] Fresh Polygonatum rhizome was sliced ​​into 0.3 cm thin slices and dried in a 50℃ forced-air drying oven. The dried powder was then pulverized and passed through an 80-mesh sieve. The dried powder was accurately weighed into an Erlenmeyer flask, and distilled water was added at a ratio of 1 g:30 mL. The mixture was then ultrasonically extracted for 30 min at a frequency of 30 kHz. After extraction, the extract was filtered to obtain the Polygonatum rhizome extract, which was stored at 4℃.

[0029] Example 2 Extraction method of Polygonatum extract.

[0030] Fresh Polygonatum sibiricum was sliced ​​and dried in a 40℃ forced-air drying oven, then pulverized and passed through a 60-mesh sieve. The dried powder was accurately weighed into an Erlenmeyer flask, and distilled water was added at a ratio of 1g:20mL. The mixture was then ultrasonically extracted for 45 minutes at a frequency of 20kHz. After extraction, the extract was filtered to obtain the Polygonatum sibiricum extract, which was stored at 4℃.

[0031] Example 3 Extraction method of Polygonatum extract.

[0032] Fresh Polygonatum sibiricum was sliced ​​and dried in a 60℃ forced-air drying oven, then pulverized and passed through a 120-mesh sieve. The dried powder was accurately weighed into an Erlenmeyer flask, and distilled water was added at a ratio of 1g:40mL. The mixture was then ultrasonically extracted for 60 minutes at a frequency of 40kHz. After extraction, the extract was filtered to obtain the Polygonatum sibiricum extract, which was stored at 4℃.

[0033] The components of the Polygonatum sibiricum extract obtained in Example 1 were analyzed using ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap mass spectrometry. The main active components identified in the extract included hexadecylamide (52.807%), L-valine (8.514%), 2-pyrrolidinecarboxylic acid (6.754%), 4-oxoproline (3.773%), mannotriose (3.603%), succinic acid (3.574%), choline (3.116%), D-(-)-ribose (2.404%), stachyose (1.986%), and L-threonine (1.946%). Figure 1 As shown.

[0034] It should be noted that the present invention also performs component analysis on the Polygonatum extracts obtained in Examples 2 and 3, and the main active ingredients identified are the same as those in Example 1, which will not be described again.

[0035] Experimental Example 1 1. Polygonatum extract improves body weight and liver index in mice with APAP-induced liver injury.

[0036] A mouse model of drug-induced liver injury was established by intraperitoneal injection of APAP. Three groups were set up: a blank control group, an APAP model group, and a Polygonatum sibiricum extract group. Mice in the Polygonatum sibiricum extract group were administered 10 mL / kg body weight of the Polygonatum sibiricum extract prepared in Example 1 by gavage daily for 7 consecutive days. Mice in the blank control group and the APAP model group were administered an equal volume of sodium carboxymethyl cellulose by gavage daily for 7 consecutive days. On the 8th day of the experiment, mice in the APAP model group and the Polygonatum sibiricum extract group were injected intraperitoneally with 300 mg / kg APAP, while mice in the blank control group were injected intraperitoneally with an equal volume of physiological saline. Food and water were allowed, and blood was collected 12 hours later by enucleation. The liver was harvested, washed with cold physiological saline, and stored. Changes in mouse body weight during the experiment are shown below. Figure 2 As shown in Figure A, during the first 7 days after administration, the body weight of mice in all groups increased slowly over time. After administration of APAP, the body weight of mice in the model group decreased significantly, while Polygonatum extract effectively alleviated the decrease in body weight.

[0037] Liver index = liver weight / body weight. Polygonatum extract can significantly reduce the liver index in APAP mice.

[0038] 2. Polygonatum extract improves serum biochemistry in mice with APAP-induced liver injury.

[0039] Blood samples were collected in 1.5 mL centrifuge tubes without anticoagulants and allowed to stand at room temperature for 2 hours to allow for natural coagulation. Subsequently, the samples were centrifuged at 4°C and 3000 rpm / min for 15 minutes. The supernatant was transferred to new centrifuge tubes and stored at -80°C for subsequent analysis. Serum biochemical parameters in mice were detected using an automated biochemical analyzer. The results are as follows: Figure 3 As shown, Polygonatum extract was found to inhibit the increase of ALT, AST, ALP and TBIL.

[0040] 3. Polygonatum extract improves liver tissue pathology in mice with APAP-induced liver injury.

[0041] Immediately after euthanizing the mouse, the liver tissue was removed and cut into pieces approximately 1 cm in size. 3Tissue blocks were quickly immersed in 4% paraformaldehyde fixative and fixed at 4°C for 24 hours. After fixation, the tissue blocks were dehydrated with a gradient of ethanol, cleared with xylene, and embedded in paraffin to form paraffin blocks. Serial sections of 4 μm thickness were prepared using a paraffin microtome. The sections were then placed on glass slides and immersed in 40°C warm water to allow for full tissue expansion. The tissue samples were then immersed in xylene for 10 minutes, followed by another 10 minutes of immersion in fresh xylene. Next, the samples were immersed in anhydrous ethanol for 5 minutes to wash away the xylene used for dewaxing, allowing water to enter the tissue. They were then immersed in 95%, 85%, and 70% ethanol solutions for 5 minutes each to achieve full hydration. The tissue samples were washed three times with PBS solution for 5 minutes each time. The sections were then immersed in hematoxylin staining solution for 10 minutes. Excess hematoxylin staining solution was washed away with distilled water, and the sections were then immersed in 1% hydrochloric acid-ethanol solution for color separation. Rinse with tap water, immerse in dilute ammonia solution to blue the cell nuclei for 5 minutes. Rinse with distilled water. Immerse the sections in eosin staining solution for 10 minutes, then rinse with distilled water. Dehydrate once each with 70%, 80%, and 90% ethanol, twice with 95% ethanol, and three times with 100% ethanol, 1 minute each time. Immerse the dehydrated tissue sections twice with xylene, 3 minutes each time, then dry the tissue sections, mount with neutral resin, and observe the pathological morphological changes of the liver tissue under an optical microscope. H&E staining results are as follows. Figure 4 As shown, compared with the blank control group, the livers of the APAP group exhibited uneven coloring, necrosis, loosely arranged spots, blood extravasation, and vacuolar degeneration, indicating that APAP caused significant liver damage. Treatment with Polygonatum extract reduced liver structural damage and decreased the area of ​​necrosis.

[0042] 4. Polygonatum extract reduces oxidative stress in mice with APAP-induced liver damage.

[0043] Approximately 100 mg of liver tissue was harvested and placed sequentially with 1 mL of pre-cooled physiological saline in a tissue homogenizer. After homogenization, the tissue was transferred to a 2 mL centrifuge tube. The centrifuge tube was set to a speed of 4000 rpm. After 10 minutes, 0.1 mL of the supernatant was pipetted into a test tube, labeled, and 3 mL of Coomassie Brilliant Blue G250 reagent was added. After mixing and incubation for 5 minutes, the absorbance was measured at 595 nm using standard tube 1 as a blank control. The protein concentration was determined based on the standard curve. Malondialdehyde and superoxide dismutase were measured according to the kit instructions. Results are as follows: Figure 5 As shown, compared with the blank control group, the MDA level in the model group was significantly increased. P <0.01; SOD activity decreased significantly. P<0.01 indicates that APAP induced hepatocyte damage. Polygonatum extract reduced malondialdehyde levels in APAP-affected mice and restored superoxide dismutase activity.

[0044] 5. Polygonatum extract inhibits apoptosis in APAP-induced liver injury mice.

[0045] Liver tissue fixed in paraformaldehyde solution was embedded in paraffin and cut into 50 μm thick sections. The sections were then placed on glass slides and immersed in 40°C warm water to allow for full tissue expansion. The tissue samples were then immersed in xylene for 10 min, followed by another 10 min immersion in xylene. The tissue samples were then first immersed in anhydrous ethanol for 5 min to wash away the xylene used for dewaxing, allowing water to enter the tissue; subsequently, they were immersed in 95%, 85%, and 70% ethanol for 5 min each to achieve full hydration. The tissue samples were then washed three times with PBS solution for 5 min each time. 100 μL of diluted proteinase K was added to each sample, and the samples were incubated at room temperature for 20 min to promote cell permeability. The sections were washed three times with PBS for 5 min each time to remove residual proteinase K. 100 μL of equilibration buffer was added, and the samples were equilibrated at room temperature for 20 min. The TUNEL reaction mixture was prepared by adding 1 μL of rTdT... ＋ 1 μL of biotin-labeled dUTP ＋Mix 98 μL of equilibration buffer thoroughly. Add 100 μL of TUNEL reaction mixture to the specimen, seal with film, and react in a dark and humidified chamber at 37°C for 1 h. Immerse tissue sections in PBS solution three times for 5 min each time. Then add 100 μL of DAB mixture; when a light brown background appears under the microscope after approximately 10 min, rinse with deionized water. Counterstain with hematoxylin, rinsing immediately with tap water after approximately 3 seconds. Dehydrate with gradient alcohols: 70%, 85%, and 95% ethanol for 1 min each. Immerse the dehydrated tissue sections twice in xylene for 3 min each time, then dry the sections and mount with neutral resin. After dewaxing and hydration, tissue sections were placed in an autoclave containing 0.01M citric acid at pH 6.0 and heated at 120°C for 2 minutes. After natural cooling, the sections were rinsed three times with PBS buffer, 2 minutes each time. The sections were then incubated in a 3% H₂O₂ solution at room temperature for 10 minutes, followed by rinsing three times with PBS buffer, 2 minutes each time. Finally, the sections were rinsed with a 5% H₂O₂ solution. BSA-covered samples were sealed in a humidified chamber at room temperature for 30 min. Caspase 3 was diluted 1:100 with primary antibody diluent to prepare the primary antibody working solution. The prepared primary antibody working solution was added dropwise to each group of mouse liver tissue sections and incubated overnight at 4°C in a humidified chamber. The next day, excess primary antibody working solution was washed away with PBS buffer. HRP-labeled secondary antibody working solution was added dropwise and covered each group of mouse liver tissue sections, and incubated at 37°C for 30 min in a humidified chamber. The sections were then washed three times with PBS for 5 min each time. During secondary antibody incubation, DAB chromogenic solution was added and allowed to stand for 2 min for color development, followed by rinsing with tap water. Hematoxylin was counterstained for 3 min, followed by rinsing with tap water. The sections were then dehydrated with a gradient of alcohols, i.e., 70%, 85%, and 95% ethanol for 1 min each. The dehydrated tissue sections were soaked twice in xylene for 3 min each time. The tissue sections were then dried and mounted with neutral resin. The sections were observed and photographed under a microscope. The results are as follows: Figure 6 As shown, compared with the control group, a large number of TUNEL-positive apoptotic cells were observed concentrated in the damaged area of ​​the liver in the APAP group. These cells exhibited chromatin atrophy and marginalization, nuclear membrane rupture, and significantly increased Caspase 3 expression levels, indicating that APAP induced severe liver cell apoptosis. In the Polygonatum extract group, hepatocytes showed clear morphology and distinct intercellular spaces, with only a small amount of positive expression. Caspase 3 expression levels were decreased, suggesting that Polygonatum extract can inhibit apoptosis and alleviate APAP-induced liver damage.

[0046] 6. Polygonatum extract inhibits inflammation-related pathways in mice with APAP-induced liver injury.

[0047] Total RNA was extracted using the Trizol method and quality-checked using an Agilent 2100 bioanalyzer to ensure a RNA integrity RIN ≥ 7, meeting library construction requirements. Subsequently, mRNA libraries were constructed from qualified samples, and paired-end sequencing was performed on an Illumina high-throughput sequencing platform. The raw sequencing data were first processed using Cutadapt software to remove adapters and low-quality sequences, followed by quality assessment using FastQC. The high-quality data after quality control were compared with a reference genome, gene expression levels were normalized using FPKM values, and differential expression analysis was performed using DESeq2 software. The selection criteria were |log2FC| > 1 and adjusted for high quality. P Values ​​<0.05 were used to obtain a list of differentially expressed genes. To further elucidate the biological functions of differentially expressed genes, KEGG pathway enrichment analysis was performed using the clusterProfiler R package, setting... P <0.05 was used as the significance threshold, and the key enriched pathways were visualized using the Pathview package. The results are as follows: Figure 7 As shown, it was found that Polygonatum extract mainly participates in regulating the interaction of inflammatory factor receptors, the TNF signaling pathway, and the IL-17 signaling pathway.

[0048] 6. Polygonatum extract inhibits the TNF and IL17 pathways in APAP mice.

[0049] Take 30 mg of mouse liver tissue and place it in an RNase-free grinding tube. Add 1 mL of TransZol Up and two enzyme-free zirconia grinding beads to each tube, and grind at low temperature for 120 s. Add 1 mL of TransZol Up to each cell sample and pipette repeatedly for 30 s. Transfer the ground tissue or lysed cell sample to a new enzyme-free EP tube and incubate at room temperature for 5 min. Add 200 μL of chloroform to each EP tube, shake vigorously for 30 s, and incubate at room temperature for 3 min. Centrifuge at 10000 g, 4 °C for 15 min. At this point, the sample shows obvious stratification. Aspirate 300 μL of the colorless aqueous phase and transfer it to a new enzyme-free EP tube. Add 300 μL of anhydrous ethanol and mix by inversion. Transfer the solution in the EP tube to a centrifuge column and centrifuge at 12000 g for 30 s, discarding the eluent. Add 500 μL of CB9, centrifuge at 12000 g for 30 s, and discard the eluent. Repeat once. Add 500 μL of WB9 containing anhydrous ethanol, centrifuge at 12000g for 30s, and discard the eluent. Repeat once. Centrifuge at 12000g at room temperature for 2min to completely remove residual ethanol. Place the centrifuge column in an RNase-free centrifuge tube, add 50 μL of RNase-free water to the center of the column, and incubate at room temperature for 1min. Centrifuge at 12000g at room temperature for 1min to elute RNA. Measure the RNA concentration using a micro spectrophotometer and store the RNA at -80℃ for later use. Add 3 μg of RNA template, 1 μL of anchored Oligo(dT)18 primer, and RNase-free water, mix well, incubate at 65℃ for 5min, and then incubate on ice for 2min. Add 2×TS reaction mixture, TransScript RT / RI enzyme mixture, and gDNA removal agent, mix well, and incubate at 42℃ for 30min. Heat at 85℃ for 5s to inactivate TransScript RT / RI and gDNA removal agent. The mouse target gene-specific primers and the reverse-transcribed RNA were diluted and added to an octet tube. Then, 2×TransStart Tip Green quantitative PCR premix, passive reference dye, template, and RNase-free water were added, mixed thoroughly, and amplified in a PCR instrument. Using 2... -ΔΔCt The method was used to calculate relative gene expression. The results are as follows: Figure 8 As shown, compared with the blank control group, the mRNA expression levels of TNF-α and IL-1β in the model group were significantly increased; while Polygonatum extract could significantly inhibit the expression of TNF-α and IL-1β mRNA, suggesting that Polygonatum extract can improve APAP-induced hepatocyte apoptosis.

[0050] It should be noted that the Polygonatum extracts obtained in Examples 2 and 3 were used in the experiments described in Example 1. The experimental results showed that the Polygonatum extracts prepared in Examples 2 and 3 had similar protective effects against APAP-induced liver injury in mice as the extract in Example 1, significantly improving serum biochemical indicators, alleviating liver pathological damage, inhibiting oxidative stress and apoptosis, and downregulating the expression of inflammatory pathway-related factors. This confirms that within the extraction method parameters defined in claim 1 of this invention, the obtained Polygonatum extracts all possess stable and significant hepatoprotective effects.

[0051] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0052] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the invention. Those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the scope of protection of the present invention. Therefore, the scope of protection of this invention should be determined by the appended claims.

Claims

1. The application of Polygonatum extract in the preparation of drugs for treating liver injury, characterized in that, The main active ingredients in the Polygonatum extract include hexadecamide, L-valine, 2-pyrrolidinecarboxylic acid, 4-oxoproline, mannitol, succinic acid, choline, D-(-)-ribose, stachyose and L-threonine.

2. The application according to claim 1, characterized in that, The liver injury was drug-induced liver injury caused by acetaminophen.

3. The application according to claim 1, characterized in that, The Polygonatum extract is the only effective ingredient in the preparation of drugs for treating liver damage.

4. The application according to claim 3, characterized in that, The drug also includes pharmaceutically acceptable excipients.

5. The application according to claim 4, characterized in that, The drug also includes pharmaceutically acceptable pharmaceutical excipients, which are one or more of the following: diluents, disintegrants, precipitation inhibitors, flow aids, binders, dispersants, suspending agents, isotonic agents, thickeners, emulsifiers, preservatives, and stabilizers.

6. The application according to claim 1, characterized in that, The Polygonatum extract was obtained by the following method: Polygonatum slices were dried, pulverized and sieved to obtain a dry powder; water was added to the dry powder and ultrasonic extraction was performed to obtain the Polygonatum extract; the mass-volume ratio of the dry powder to water was 1g:20mL~40mL; the ultrasonic extraction time was 30min~60min; and the ultrasonic extraction frequency was 20kHz~40kHz.

7. The application according to claim 6, characterized in that, The thickness of the Polygonatum slices is 0.1cm to 0.5cm.

8. The application according to claim 6, characterized in that, The drying temperature is 40℃~60℃.

9. The application according to claim 6, characterized in that, The particle size of the dried powder is 60 mesh to 120 mesh.

Images