An anti-alcoholic liver-protecting beverage, a preparation method and application thereof

Abstract

The application discloses an alcoholism-relieving and liver-protecting beverage as well as a preparation method and application thereof, and belongs to the technical field of biotechnology.The alcoholism-relieving and liver-protecting beverage disclosed by the application comprises the following raw materials: Chinese jujube seeds, radix puerariae, dendrobium, rhizoma imperatae, chrysanthemum, raw rehmannia, raw licorice, corn silk and lemon slices.The alcoholism-relieving and liver-protecting beverage disclosed by the application can effectively relieve acute alcohol-induced liver injury by improving liver function, regulating lipid metabolism, reducing liver tissue pathological injury and regulating core metabolic pathways, and provides an experimental basis for the development and application of the alcoholism-relieving and liver-protecting functional beverage.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and more specifically to a hangover relief and liver protection beverage, its preparation method, and its application. Background Technology

[0002] Currently, alcoholic liver disease has become a major liver health problem in my country. Data from the 2024 "Epidemiology and Disease Burden of Liver Disease in China" shows that there are over 400 million people with chronic liver disease in my country, of which approximately 60 million have alcoholic liver disease, with a prevalence rate as high as 11.6% in the 40-49 age group. After alcohol enters the body, about 90% is metabolized in the liver. Ethanol is oxidized to acetaldehyde by alcohol dehydrogenase. Acetaldehyde has extremely high reactivity and can directly form adducts with proteins and DNA, damaging hepatocytes. Simultaneously, chronic excessive alcohol consumption induces abnormal activation of CYP2E1, producing large amounts of reactive oxygen species and depleting glutathione, inducing severe oxidative stress, and activating the NF-κB inflammatory pathway, releasing pro-inflammatory factors such as TNF-α and IL-6, exacerbating hepatocyte necrosis and apoptosis. Recent studies have also confirmed that ferroptosis is involved in alcohol-induced hepatocyte damage. This multi-faceted pathogenesis means that single-target intervention is insufficient for effective protection.

[0003] However, existing hangover remedies and liver protection products have significant shortcomings. At the pharmaceutical level, while metadoxine can promote alcohol metabolism, it is not a specific hangover remedy, and long-term use may cause peripheral neuropathy. Its excipient, sodium metabisulfite, can induce allergic asthma. Naltrexone has limited safety evidence in patients with decompensated cirrhosis. The 2024 American College of Gastroenterology guidelines also do not recommend disulfiram for alcoholic liver disease complicated by alcohol use disorder. At the health supplement level, the 2023 "Chinese Guidelines for the Diagnosis and Treatment of Drug-Induced Liver Injury" lists health supplements and dietary supplements as potential causes of liver injury. Blindly taking them not only fails to protect the liver but may also lead to drug-induced liver injury or even liver failure. In some individuals with specific constitutions, it may even trigger drug-induced autoimmune liver disease. Although there are products on the market containing traditional hangover remedies such as kudzu root and Japanese raisin tree fruit, related studies indicate that the hangover remedy and liver protection efficacy of the kudzu root-Japanese raisin tree fruit pair is highly dependent on the scientific compatibility ratio and preparation process. If the compatibility is unreasonable or the process parameters are not properly controlled, the product quality and efficacy will vary considerably. Meanwhile, some such products do not fully consider the differences in the applicable population: kudzu root is cooling in nature, and excessive consumption by those with weak spleen and stomach can cause gastrointestinal discomfort such as abdominal distension and diarrhea; excessive consumption of Japanese raisin tree fruit or improper combination can also cause adverse reactions such as nausea and diarrhea. Therefore, providing a hangover relief and liver protection beverage with scientific formulation, clearly defined active ingredient content, safety and effectiveness, and a wide range of applicable populations, as well as its preparation method and application, is a problem that urgently needs to be solved by those skilled in the art.

[0004] Therefore, providing a hangover-relieving and liver-protecting beverage, its preparation method, and its application is a problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0005] In view of this, the present invention provides a hangover relief and liver protection beverage, its preparation method and application.

[0006] This invention standardizes the traditional decoction preparation process of hangover relief and liver protection drinks, ensuring the repeatability of the operation and the consistency of the samples, and providing standardized samples for subsequent quality control, efficacy research or daily application.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: A hangover relief and liver protection beverage, by weight, includes the following ingredients: 15 parts of Japanese raisin tree fruit, 10 parts of kudzu root, 10 parts of dendrobium, 20 parts of imperata root, 5 parts of chrysanthemum, 6 parts of rehmannia root, 6 parts of licorice root, 10 parts of corn silk, and 5 parts of lemon slices.

[0008] Furthermore, the preparation method of the above-mentioned hangover relief and liver protection beverage includes the following steps: (1) Weighing Weigh each herb separately and mix them thoroughly. (2) Soaking Place the mixed medicinal materials in a decoction container, add ultrapure water, with a mass-to-volume ratio of mixed medicinal materials to ultrapure water of 87:720 g / ml, so that the medicinal materials are completely submerged; let it stand and soak at room temperature for 40 minutes. (3) First decoction and filtration Heat over high heat until boiling, then immediately reduce to low heat and simmer for 30 minutes. After decoction, hot filtration was performed immediately; the decoction and dregs were separated by atmospheric pressure filtration using pre-weighed filter paper; the container and dregs were washed several times with 30 mL of boiling ultrapure water, and the washing liquid was combined with the filtrate to obtain the first decoction. The first decoction was collected in a clean container. The dregs, along with the filter paper, were retained in the original container.

[0009] (4) Second decoction and filtration Add boiling ultrapure water to the dregs, wherein the volume ratio of the boiling ultrapure water to the ultrapure water used in the first soaking is 650:720; heat again over a low flame, maintaining a gentle boil, and simmer precisely for 40 minutes. Repeat the filtration process (using new pre-weighed filter paper) to obtain the second decoction; (5) Combining, concentrating and adjusting volume Combine the first decoction with the second decoction to obtain a combined liquid; place the combined liquid in a decoction container and heat it over a low flame to concentrate it, stirring occasionally, until the volume of the liquid is reduced to 1 / 6 of the volume of the ultrapure water used in the first soaking. (6) Filtration and dispensing Filter the concentrated drug solution while it is still hot using filter paper to remove any precipitates that may form. After the filtered drug solution has cooled to room temperature, dispense it into sterile sample vials and seal them. Labeling: Mark the sample name, batch number, concentration, preparation date, and operator; Storage: For long-term storage (1 year), freeze at -20℃ or -80℃; for short-term storage (2 months), refrigerate at 2-8℃.

[0010] All containers and utensils should be thoroughly cleaned before use to avoid contamination; during decoction, care should be taken to prevent the liquid from overflowing, such as by using a splash guard or adjusting the heat; if using reflux condensation, ensure that the cooling water is unobstructed; during concentration, constant stirring is required to prevent scorching at the bottom; record all original data to ensure traceability.

[0011] Furthermore, the application of the aforementioned hangover-relieving and liver-protecting beverage in the preparation of a beverage for treating acute alcoholic liver injury.

[0012] Furthermore, the application of the aforementioned hangover-relieving and liver-protecting beverage in the preparation of liver-protecting drinks.

[0013] As can be seen from the above technical solution, compared with the prior art, this invention discloses a hangover-relieving and liver-protecting beverage, its preparation method, and its application. By constructing a mouse model of acute alcoholic liver injury, the hepatoprotective effect and potential molecular mechanism of the hangover-relieving and liver-protecting beverage were systematically evaluated. Results showed that compared with the NC group, Mod mice had significantly increased serum ALT and AST levels, increased LDL-C and TG levels, and decreased HDL-C levels. Inflammatory infiltration and a large number of lipid droplet deposits were observed in the liver tissue, indicating that the acute alcoholic liver injury model was successfully constructed. After intervention with the hangover-relieving and liver-protecting beverage, GG group mice showed significantly reduced serum ALT and AST levels, effectively improved lipid metabolism disorders, and significantly reduced pathological damage and fatty degeneration in liver tissue, with an overall condition close to the blank control group. Transcriptome analysis revealed that acute alcohol exposure significantly altered the gene expression profile of liver tissue. Differentially expressed genes were primarily enriched in fatty acid metabolism, alcohol metabolism, and the PPAR signaling pathway, manifested as upregulation of lipid synthesis-related genes and downregulation of genes related to antioxidant and lipid breakdown. Intervention with a hangover-relieving and liver-protecting beverage reversed the gene expression profile of the model group back to that of the control group, with significant downregulation of lipid synthesis-related genes. Simultaneously, differentially expressed genes were enriched in monooxygenase activity, defense responses, and the IL-17 signaling pathway, suggesting that the beverage restores liver metabolic homeostasis by inhibiting lipid synthesis, promoting lipid breakdown, and enhancing the liver's antioxidant defense and inflammatory regulation capabilities, thereby protecting against acute alcoholic liver injury. In conclusion, a hangover-relieving and liver-protecting beverage can effectively alleviate acute alcohol-induced liver injury by improving liver function, regulating lipid metabolism, reducing pathological damage to liver tissue, and regulating core metabolic pathways, providing experimental evidence for its development and application as a functional hangover-relieving and liver-protecting beverage. 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 embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0015] Figure 1 The changes in initial body weight (A), final body weight (B), and liver index (C) of mice are shown.

[0016] Figure 2 The changes in serum biochemical parameters in mice are shown. A: serum alanine aminotransferase (ALT) level; B: serum aspartate aminotransferase (AST) level; C: serum low-density lipoprotein cholesterol (LDL-C) level; D: serum high-density lipoprotein cholesterol (HDL-C) level; E: serum triglyceride (TG) level; F: serum total cholesterol (TC) level. * indicates statistically significant differences. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

[0017] Figure 3 HE staining results of liver tissue from mice in each group (left: 200×; right: 400×).

[0018] Figure 4 The results of Oil Red O staining for fatty degeneration in mouse liver tissue (left: 200×; right: 400×).

[0019] Figure 5 This section presents differential gene expression analysis in mouse liver tissue. A: Principal component analysis (PCA) plot; B: Venn diagram of gene co-expression; C: Bar chart of differentially expressed genes, with red representing total differentially expressed genes, blue representing upregulated genes, and green representing downregulated genes. The differential gene screening thresholds were padj < 0.05 and |log2FoldChange| ≥ 2.

[0020] Figure 6 Differential gene expression analysis in mouse liver tissue; A: Volcano plot of differential genes in Mod vs NC group; B: Volcano plot of differential genes in GG vs Mod group.

[0021] Figure 7 Bubble plot of GO function enrichment in mouse liver tissue Mod vs NC group.

[0022] Figure 8 Bubble plot of GO function enrichment in mouse liver tissue (GG vs Mod group).

[0023] Figure 9 Bubble plot of KEGG pathway enrichment in mouse liver tissue Mod vs NC group.

[0024] Figure 10 Bubble plot of KEGG pathway enrichment in mouse liver tissue (GG vs Mod group).

[0025] Figures 7-10 In this context, padj < 0.05 is the enrichment significance threshold, the size of the bubble represents the number of enriched genes, and the color intensity represents the enrichment significance (padj value). Detailed Implementation

[0026] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0027] Precision electronic balance (accuracy 0.001 g); glass decoction apparatus with reflux condenser (e.g., round-bottom flask and condenser) or ceramic casserole (for non-laboratory use); graduated cylinder (100 mL), beaker, glass rod vacuum filtration system (including Buchner funnel, filtration flask, slow quantitative filter paper) or conventional filter screen (pore size ≤ 75 μm); heating mantle or adjustable temperature furnace; electric heating drying oven (optional, for drying medicinal residue); desiccator, sample vials, cryovials; timer, label paper; thermometer, hygrometer, barometer (for environmental recording). Example 1

[0028] A method for preparing a hangover relief and liver-protecting beverage includes the following steps: (1) Weighing Weigh each of the following herbs separately: 15 g of Japanese raisin tree fruit, 10 g of kudzu root, 10 g of dendrobium, 20 g of imperata root, 5 g of chrysanthemum, 6 g of rehmannia root, 6 g of licorice root, 10 g of corn silk, and 5 g of lemon slices; mix the herbs evenly. (2) Soaking Place the mixed medicinal materials in a decoction container, add ultrapure water, with a mass-to-volume ratio of mixed medicinal materials to ultrapure water of 87:720 g / mL, so that the medicinal materials are completely submerged; let it stand and soak at room temperature for 40 minutes. (3) First decoction and filtration Heat over high heat until boiling, then immediately reduce to low heat and simmer for 30 minutes. After decoction, hot filtration was performed immediately; the decoction and dregs were separated by atmospheric pressure filtration using pre-weighed filter paper; the container and dregs were washed several times with 30 mL of boiling ultrapure water, and the washing liquid was combined with the filtrate to obtain the first decoction. (4) Second decoction and filtration Add boiling ultrapure water to the dregs, with a volume ratio of boiling ultrapure water to the ultrapure water used in the first soaking of 650:720; heat again over low heat, maintaining a gentle boil, and simmer precisely for 40 minutes. Repeat the filtration process to obtain the second decoction; (5) Combining, concentrating and adjusting volume Combine the first decoction with the second decoction to obtain a combined liquid; place the combined liquid in a decoction container and heat it over a low flame to concentrate it, stirring occasionally, until the volume of the liquid is reduced to 1 / 6 of the volume of the ultrapure water used in the first soaking. (6) Filtration and dispensing Filter the concentrated drug solution while it is still hot using filter paper; after the filtered drug solution has cooled to room temperature, dispense it into sterile sample bottles and seal them; refrigerate at 4°C.

[0029] Example 2: Evaluation of the hepatoprotective effect of a hangover-relieving and liver-protecting beverage on mice with acute alcoholic liver injury. Test drug: A hangover relief and liver protection beverage, 40 mL / bottle, stored at 4℃ for 7 days.

[0030] Mouse type: 4-week-old KM mice.

[0031] Other main reagents: 52% (v / v) Hongxing Erguotou Baijiu.

[0032] Experimental equipment: optical microscope (Jiangnan, model: BM2000), centrifuge (Hunan Xiangyi, model: TG16-W), rotary microtome (Shenzhen Ruiwode, model: S700), 96-well plate shaker (Haimen Qilinbeier, model: MLI-2), cryostat (Thermo Fisher Scientific, model: Thermo CryoStar™ NX50), ELISA reader (BioTeK, model: Epoch), vortex mixer (Servicebio, model: MV-100), fully automated biochemical analyzer (Shenzhen Mindray Medical, model: BS360S).

[0033] (1) Animal experiments Twenty-four male KM mice were randomly divided into three groups (n=8): a blank control group (NC), an acute alcoholic liver injury model group (Mod), and a liver-protecting beverage intervention group (GG). The blank control group and the model group were administered an equal volume of distilled water by gavage daily, while the intervention group was administered the liver-protecting beverage by gavage daily, with a gavage volume of 0.2 mL / mouse. This intervention lasted for 6 consecutive days, during which the general physiological status of the mice was observed daily. On day 7, except for the blank control group, the other two groups were administered 52% vol Hongxing Erguotou liquor by gavage at a dose of 12 mL / kg·BW to establish an acute alcoholic liver injury model. The blank control group was administered an equal volume of distilled water by gavage. Four hours after model establishment, the model group and the intervention group were administered distilled water and the liver-protecting beverage (0.2 mL / mouse), respectively, while the blank control group was administered an equal volume of distilled water by gavage. After the last gavage, the mice were kept on a fasting for 12 hours, but water was allowed. Blood and tissue samples were then collected. After the collected blood was centrifuged, the serum was separated and stored at -80°C for later use. The liver tissue was fixed with 4% paraformaldehyde for subsequent histopathological analysis, and the remaining tissue was quickly stored at -80°C for transcriptomics sequencing.

[0034] (2) Weight monitoring and organ index calculation Before the experiment, the initial body weight of the mice was weighed and recorded. During the intervention period, the general growth status of the mice was observed daily. After the last gavage, the final body weight of the mice was weighed and recorded. Before sample collection, the fasting body weight of the mice was weighed. After the mice were sacrificed, the liver was quickly separated, the surface bloodstains were washed with physiological saline, the moisture was absorbed with filter paper, and the wet weight of the organ was accurately weighed. The organ index was calculated using the following formula: Organ Index (%) = Wet weight of organ (g) / Fasting body weight of mouse (g) × 100%.

[0035] Before the experiment began, the initial body weight of mice in each group was assessed at baseline, and the results showed that ( Figure 1 A), there were no significant differences in initial body weight among mice in the NC, Mod, and GG groups. p > 0.05), indicating balanced grouping and consistent baseline. Final body weight was measured after the last intervention. Figure 1 B), there were no significant differences in the terminal body remodeling of the three groups of mice. p > 0.05). Liver index results showed ( Figure 1 C), there was no significant difference in liver index among the three groups of mice. p > 0.05); Compared with the NC group, the liver index in the Mod group showed a slight upward trend, but did not reach the level of statistical difference. There was also no significant difference between the GG group and the Mod and NC groups.

[0036] (3) Serum biochemical tests Blood was collected from the eyes of mice, and the serum was separated by centrifugation after standing. A fully automated biochemical analyzer was used to detect the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in the mouse serum. The simplified steps are as follows: After centrifugation to separate the serum from the collected blood samples, the samples were loaded into the instrument according to the predetermined parameters, and the system automatically completed reagent dispensing, reaction, and concentration calculation.

[0037] Serum liver function and lipid-related indicators test results are as follows Figure 2 As shown, the experiment measured six core indicators: ALT, AST, LDL-C, HDL-C, TG, and TC, and compared the differences among the NC, Mod, and GG groups. Compared with the NC group, the serum ALT level in the Mod group mice was significantly increased ( p < 0.05), AST levels increased significantly ( p < 0.01); Among the lipid metabolism-related indicators, the levels of LDL-C and TG were significantly increased in the Mod group ( p < 0.05), HDL-C levels were significantly reduced ( p <0.05, while TC levels showed no statistically significant difference between the NC and Mod groups ( p > 0.05). After intervention with the hangover-relieving and liver-protecting beverage, the serum ALT and AST levels in the GG group mice showed a significantly lower trend compared to the Mod group, and the LDL-C and TG levels were also significantly lower than those in the Mod group. The HDL-C level, however, rebounded compared to the Mod group. Furthermore, there were no significant differences in ALT, AST, LDL-C, HDL-C, and TG levels between the GG group and the NC group. p >0.05); no statistically significant difference was found in TC levels among the three groups. p > 0.05).

[0038] (4) HE staining HE staining of mouse liver tissue was performed strictly according to standard operating procedures. First, 4 μm thick paraffin sections were baked in a 65°C oven for at least 60 min to ensure complete dewaxing. The sections were then dewaxed sequentially with xylene I and II (12 min each), followed by a gradient rehydration process with anhydrous ethanol I and II (3 min each), 90% ethanol (3 min), and 70% ethanol (3 min), and finally washed with tap water for 5 min. For staining, hematoxylin was first applied for 6 min to stain the cell nuclei. After staining, the sections were thoroughly rinsed with running water and then immersed in PBS blue solution for 1 min to turn the hematoxylin blue. Cytoplasmic staining was performed using a self-prepared eosin Y alcohol solution, immersing for approximately 35 s, followed by rapid rinsing with tap water to remove excess stain. All staining steps were performed at room temperature. After staining, the sections were drained and briefly baked in a 65°C oven, then cleared with xylene for 5 min. Finally, the sections were mounted with neutral resin. The prepared slides were observed and images were acquired under an optical microscope.

[0039] HE staining results ( Figure 3 The results showed that in the NC group, the hepatocytes were neatly arranged, the lobular structure was intact and clear, the hepatocyte morphology was regular, the cytoplasm was uniform, and no obvious cell degeneration, necrosis, or inflammatory infiltration was observed; the liver tissue morphology was normal. In the Mod group, the hepatocyte cytoplasm was loose and lightly stained, the hepatocyte cords were disordered, and a small number of inflammatory cell infiltration foci and scattered hepatocyte necrosis were also observed, suggesting that acute alcohol exposure induced pathological damage to the mouse liver tissue. In the GG group, the lobular structure was clear, the hepatocyte cords were more neatly arranged, and a small number of inflammatory foci were still present in the tissue; the overall pathological morphology was similar to that of the NC group.

[0040] (5) Oil Red O staining Freshly prepared frozen sections of mouse liver tissue (approximately 8-10 μm thick) were removed from a -80°C freezer and allowed to air dry at room temperature for 10 min. The sections were then washed with 60% isopropanol for approximately 1 min to enhance the lipid affinity for the dye. After draining, the sections were completely immersed in Oil Red O working solution and stained in the dark for 10-15 min. Immediately after staining, rapid differentiation was initiated with 60% isopropanol to remove background staining. This step should be controlled under a microscope until the tissue background is nearly colorless and the lipid droplets are bright red. After differentiation, the reaction was gently terminated by rinsing with running distilled water. To visualize the cell nuclei, the sections were counterstained with Myersin for approximately 1-2 min, followed by bluing with running water. Finally, the sections were mounted with an aqueous mounting medium (such as glycerol gelatin) to prevent the lipids from being dissolved by organic solvents. The prepared sections were immediately observed and imaged under an optical microscope.

[0041] Oil Red O staining results ( Figure 4The results showed that in the NC group, no obvious red lipid droplet deposition was observed in the liver tissue of mice, and the hepatocyte morphology was normal, with no signs of fatty degeneration. In the Mod group, a large number of densely distributed red lipid droplets were observed in the liver tissue, widely deposited in the hepatocyte cytoplasm, suggesting that acute alcohol exposure induced significant hepatocyte fatty degeneration and severe lipid accumulation. In the GG group, the deposition of red lipid droplets in the liver tissue was significantly reduced compared to the Mod group, with only a few scattered lipid droplets observed, and the degree of lipid accumulation was significantly reduced. This indicates that the hangover-relieving and liver-protecting beverage can effectively improve alcohol-induced hepatocyte fatty degeneration and reduce intrahepatic lipid accumulation.

[0042] (6) Liver transcriptomics sequencing and bioinformatics analysis Total RNA was extracted from mouse liver tissue using TRIzol reagent and treated with DNase I to remove genomic DNA contamination. RNA purity, concentration, and integrity were measured using a 2100 bioanalyzer and an ND-2000 ultra-micro spectrophotometer, respectively. Qualified samples were used to construct transcriptome libraries using the Illumina TruSeq™ RNA kit. After quantification and standardization, the libraries were subjected to paired-end sequencing on the BGI Genomics DNBSEQ-T7 sequencing platform (performed by Tianjin Novogene Technology Co., Ltd.).

[0043] Raw sequencing data were first processed using Cutadapt software to remove adapter sequences and low-quality reads, and then quality controlled using FastQC. Bowtie2 and TopHat2 were then used to align the sequencing reads to a reference genome, and StringTie was used for transcript assembly and gene expression level estimation. The selection criteria for differentially expressed genes were set as |log2FC|>2 and padj < 0.05. GO functional annotation and KEGG pathway enrichment analysis were performed on the selected differentially expressed genes.

[0044] Transcriptome sequencing and differential gene analysis results showed that ( Figure 5 Principal component analysis (PCA) Figure 5 A) indicates that the Mod group showed significant separation from the NC and GG groups in both PC1 and PC2 dimensions. While the GG and NC groups showed some differentiation in the PC1 direction, they clustered closely in the PC2 direction, suggesting that acute alcohol exposure significantly altered the gene expression characteristics of liver tissue. Intervention with the hangover-relieving and liver-protecting beverage could cause the gene expression profile of the model group to revert towards that of the control group. Venn diagram ( Figure 5 B) shows a total of 10,377 co-expressed genes across the three groups, 212 genes specifically expressed in the Mod group, and 272 genes specifically expressed in the GG group. Statistical results of differentially expressed genes ( Figure 5C) shows that, using padj < 0.05 |log2FoldChange|≥2 as the screening criterion, a total of 235 differentially expressed genes were identified in the Mod vs NC group (170 upregulated and 65 downregulated), and a total of 270 differentially expressed genes were identified in the GG vs Mod group (142 upregulated and 128 downregulated).

[0045] Volcano Map Analysis ( Figure 6 A and B) further revealed changes in the expression of key differentially expressed genes: In the Mod vs NC group, genes closely related to lipid synthesis and metabolism (such as ELOVL3) were significantly upregulated, while genes involved in antioxidant stress (GSTT2) and lipolysis (ACOT1) were significantly downregulated, suggesting that alcohol exposure exacerbates lipid accumulation and oxidative damage in liver tissue by promoting lipid synthesis, inhibiting lipolysis, and reducing antioxidant defense. In the GG vs Mod group, the aforementioned lipid synthesis-related genes (such as ELOVL3) were significantly downregulated, while genes involved in metabolic regulation (FGF21) and inflammation regulation were significantly upregulated, indicating that the hangover relief and liver-protecting beverage can exert a liver-protective effect by inhibiting lipid synthesis and restoring metabolic homeostasis.

[0046] GO functional enrichment analysis results show that ( Figure 7 , Figure 8 The differentially expressed genes in the Mod vs NC group were significantly enriched in biological functions such as fatty acid metabolism, adipocyte differentiation, and alcohol metabolism (padj < 0.05), suggesting that acute alcohol exposure mainly induces hepatocyte steatosis and liver damage by perturbing lipid metabolism and alcohol breakdown-related pathways. The differentially expressed genes in the GG vs Mod group were significantly enriched in functional items such as monooxygenase activity, lipid droplets, fatty acid metabolism, and positive regulation of the defense response, indicating that intervention with hangover-relieving and liver-protecting beverages can improve alcohol-induced liver dysfunction by regulating oxidase activity, inhibiting lipid droplet formation, regulating fatty acid metabolism, and enhancing the liver's defense response. KEGG pathway enrichment analysis showed (… Figure 9 , Figure 10 The differentially expressed genes in the Mod vs NC group were significantly enriched in pathways such as PPAR signaling, retinol metabolism, steroid biosynthesis, arachidonic acid metabolism, and fatty acid degradation (padj < 0.05). The enrichment of the PPAR signaling pathway, a core regulatory pathway of lipid metabolism, further suggests that alcohol exposure exacerbates hepatic lipid accumulation by interfering with the balance between lipid synthesis and breakdown. The differentially expressed genes in the GG vs Mod group were also significantly enriched in the PPAR signaling pathway, as well as in pathways such as steroid hormone biosynthesis, retinol metabolism, IL-17 signaling, and arachidonic acid metabolism. This indicates that the hangover-relieving and liver-protecting beverage can alleviate alcohol-induced hepatic lipid metabolism disorders and liver damage by regulating the PPAR signaling pathway and pathways related to lipid metabolism and inflammatory responses.

[0047] (7) Statistical analysis Experimental data are expressed as mean ± standard deviation (mean ± SD). Unpaired two-tailed Student's t-test was used for univariate comparisons between two groups; one-way ANOVA was used for univariate comparisons between three or more groups; all statistical analyses were performed using GraphPad Prism 9.5 software. Statistical significance was defined as: * p < 0.05, ** p <0.01, *** p < 0.001, **** p < 0.0001.

[0048] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A hangover relief and liver-protecting beverage, characterized in that, By weight, it includes the following ingredients: 15 parts of Japanese raisin tree fruit, 10 parts of kudzu root, 10 parts of dendrobium, 20 parts of imperata root, 5 parts of chrysanthemum, 6 parts of rehmannia root, 6 parts of licorice root, 10 parts of corn silk, and 5 parts of lemon slices.

2. The method for preparing a hangover-relieving and liver-protecting beverage according to claim 1, characterized in that, Includes the following steps: (1) Weighing Weigh each herb separately and mix them thoroughly. (2) Soaking Place the mixed medicinal materials in a decoction container, add ultrapure water, and ensure that the mass-to-volume ratio of the mixed medicinal materials to ultrapure water is 87:720g / mL, so that the medicinal materials are completely submerged; let it stand and soak at room temperature for 40 minutes. (3) First decoction and filtration Heat over high heat until boiling, then immediately reduce to low heat and simmer for 30 minutes. After decoction, hot filtration was performed immediately; the decoction and dregs were separated by atmospheric pressure filtration using pre-weighed filter paper; the container and dregs were washed several times with 30 mL of boiling ultrapure water, and the washing liquid was combined with the filtrate to obtain the first decoction. (4) Second decoction and filtration Add boiling ultrapure water to the dregs, wherein the volume ratio of the boiling ultrapure water to the ultrapure water used in the first soaking is 650:720; heat again over a low flame, maintaining a gentle boil, and simmer precisely for 40 minutes. Repeat the filtration process to obtain the second decoction; (5) Combining, concentrating and adjusting volume The first decoction and the second decoction were combined to obtain the combined liquid; Place the combined liquid in a decoction container and heat it over a low flame to concentrate it, stirring occasionally, until the volume of the liquid is reduced to 1 / 6 of the volume of the ultrapure water used in the first soaking. (6) Filtration and dispensing Filter the concentrated drug solution while it is still hot using filter paper; after the filtered drug solution has cooled to room temperature, dispense it into sterile sample bottles and seal them. Storage: For long-term storage, freeze at -20℃ or -80℃; for short-term storage, refrigerate at 2-8℃.

3. The application of the hangover relief and liver protection beverage according to claim 1 in the preparation of a beverage for treating acute alcoholic liver injury.

4. The application of the hangover relief and liver protection beverage according to claim 1 in the preparation of liver protection drinks.

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