Use of guangming salt four decoction in preparation of drugs for preventing and treating acute liver injury

Abstract

This invention discloses the application of Guangming Salt Four-Flavor Decoction Powder in the preparation of drugs for the prevention and treatment of acute liver injury, belonging to the field of biomedical technology. The study found that Guangming Salt Four-Flavor Decoction Powder has a significant hepatoprotective effect on carbon tetrachloride-induced acute liver injury, and its mechanism of action exhibits a time-dependent biphasic pattern: early on, it is mainly characterized by inflammation suppression centered on nuclear factor-κB; later, it is characterized by cytochrome P450-mediated detoxification and restoration of antioxidant metabolic homeostasis. This provides systematic multi-omics scientific evidence for the modernization research and clinical translation of this classic Mongolian medicine compound.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the application of Guangming Salt Four-Flavor Decoction Powder in the preparation of drugs for the prevention and treatment of acute liver injury. Background Technology

[0002] The liver is a core organ for metabolism and detoxification in the human body, and it is highly susceptible to damage from external toxins, unhealthy lifestyle habits, and physical and chemical stimuli. The incidence of various liver injury diseases is rising year by year, seriously threatening human health. Clinically, there are many types of liver injury, such as acute liver injury, alcoholic liver injury, and non-alcoholic fatty liver disease. Different types of liver injury differ significantly in their pathogenesis, pathological progression, clinical manifestations, and intervention needs, leading to significant differences in clinical diagnosis, treatment, and drug development directions, requiring clear differentiation.

[0003] Acute liver injury is a sudden, rapidly progressing acute pathological injury characterized by hepatocyte necrosis, inflammatory infiltration, and oxidative stress. The key pathogenic mechanism is the excessive activation of pro-inflammatory cytokines and comprehensive disruption of liver metabolic function. It is often induced by sudden factors such as drugs, toxins, infections, physical and chemical stimuli, and stress, without a long-term chronic lesion accumulation process. The onset is rapid, causing extensive hepatocyte degeneration and necrosis, and acute liver failure within a short period. Clinical manifestations include a sudden increase in transaminase levels, jaundice, and severe pain in the liver area. In severe cases, it can induce acute liver failure with a high mortality rate. Clinically, there is an urgent need for highly effective drugs that can quickly and promptly block acute hepatocyte necrosis and alleviate acute inflammatory damage to the liver. This approach differs significantly from the therapeutic targets and intervention logic of anti-alcohol toxicity in alcoholic liver injury and metabolic regulation in non-alcoholic fatty liver disease.

[0004] Currently, there is no research or technology disclosure regarding its use in the prevention and treatment of acute liver injury. Because the pathogenesis and pathological characteristics of acute liver injury are distinctly different from other chronic liver injuries, its protective effect against chronic liver injury cannot be used to infer its preventive and therapeutic effects on acute liver injury. Therefore, there is an urgent need to develop drugs that have a clear effect on the sudden, rapid, and severe hepatocellular necrosis and inflammatory damage characteristics of acute liver injury. Summary of the Invention

[0005] The purpose of this invention is to provide the application of Guangming Salt Four-Flavor Decoction Powder in the preparation of drugs for the prevention and treatment of acute liver injury, thereby solving the problems existing in the prior art. This invention has found that Guangming Salt Four-Flavor Decoction Powder has a significant hepatoprotective effect on carbon tetrachloride-induced acute liver injury, and its mechanism of action exhibits a time-dependent biphasic pattern: early on, it is mainly characterized by inflammation suppression centered on nuclear factor-κB; later, it is characterized by cytochrome P450-mediated detoxification and restoration of antioxidant metabolic homeostasis. This provides a systematic multi-omics scientific basis for the modernization research and clinical translation of this classic Mongolian medicine compound.

[0006] To achieve the above objectives, the present invention provides the following solution: This invention provides the application of Guangming Salt Four-Flavor Decoction Powder in the preparation of drugs for the prevention and treatment of acute liver injury, including acute liver injury induced by carbon tetrachloride.

[0007] Furthermore, the Bright Salt Four-Flavor Soup Powder is made from Bright Salt, dried ginger, long pepper, and Terminalia chebula.

[0008] Furthermore, the drug has the effect of inhibiting abnormal increases in tumor necrosis factor-α, interleukin-6, and interleukin-1β.

[0009] Furthermore, the drug has the effect of preventing and treating liver tissue structure destruction, inflammatory infiltration, hepatocyte necrosis, and early liver fibrosis.

[0010] The present invention discloses the following technical effects: This invention reveals that Guangming Salt Four-Flavor Decoction, a classic Mongolian medicine compound, exhibits significant preventive and therapeutic effects against acute liver injury. Guangming Salt Four-Flavor Decoction exerts a significant protective effect against carbon tetrachloride-induced acute liver injury through a time-dependent biphasic mechanism: in the acute phase (day 2), it primarily inhibits the NF-κB / cytokine inflammatory signaling network in a dose-dependent manner; in the repair phase (day 5), its action shifts to restoring cytochrome P450-mediated detoxification function, glutathione-dependent antioxidant defense, and overall metabolic homeostasis. This biphasic regulatory mechanism has been consistently validated at both the transcriptomic and metabolomic levels. This invention provides a systematic multi-omics scientific basis for the modern research and clinical translation of Guangming Salt Four-Flavor Decoction. Attached Figure Description

[0011] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments 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.

[0012] Figure 1 This study investigated the effects of Guangming Salt Four-Flavor Decoction Powder (MD-4) on the levels of pro-inflammatory cytokines. A represents the serum interleukin-6 (IL-6) level in each group of mice on days 2, 5, and 7; B represents the serum interleukin-1β (IL-1β) level in each group of mice on days 2, 5, and 7; and C represents the serum tumor necrosis factor-α (TNF-α) level in each group of mice on days 2, 5, and 7. Data are expressed as mean ± standard error. p < 0.05 p < 0.01, p < 0.001, p < 0.0001; Figure 2 The study investigated the effects of Guangming Salt Four-Flavor Decoction Powder (MD-4) on hepatocyte necrosis and early fibrotic remodeling. A represents representative sections of mouse livers stained with hematoxylin and eosin (HE) on days 2, 5, and 7; B represents representative sections of mouse livers stained with masson trichrome on days 2, 5, and 7. Figure 3 The transcriptomic analysis results for day 2 are shown in the differential gene volcano plot; where A is the differential gene volcano plot of the control group-model group; and B is the differential gene volcano plot of the model group-Guangming Salt Four-Flavor Decoction group. Figure 4 The transcriptomic analysis results for day 2 are shown below - KEGG enrichment analysis. A represents the KEGG enrichment analysis of upregulated differentially expressed genes in the control group-model group; B represents the KEGG enrichment analysis of downregulated differentially expressed genes in the control group-model group; C represents the KEGG enrichment analysis of upregulated differentially expressed genes in the model group-Guangming Salt Four-Flavor Decoction group; and D represents the KEGG enrichment analysis of downregulated differentially expressed genes in the model group-Guangming Salt Four-Flavor Decoction group. Figure 5 The transcriptomic analysis results for day 2 are shown below - GO functional enrichment analysis. A represents the GO functional enrichment analysis of upregulated differentially expressed genes in the control group and model group; B represents the GO functional enrichment analysis of downregulated differentially expressed genes in the control group and model group; C represents the GO functional enrichment analysis of downregulated differentially expressed genes in the model group and Guangming Yansiwei Tangsan group; and D represents the GO functional enrichment analysis of upregulated differentially expressed genes in the model group and Guangming Yansiwei Tangsan group. Figure 6 The transcriptomic analysis results on day 5 are presented as differential gene volcano plots; where A is the differential gene volcano plot of the control group-model group; and B is the differential gene volcano plot of the model group-Guangming Salt Four-Flavor Decoction group. Figure 7 The transcriptomic analysis results on day 5 are shown in the KEGG enrichment analysis. A represents the KEGG enrichment analysis of upregulated differentially expressed genes in the control group-model group; B represents the KEGG enrichment analysis of downregulated differentially expressed genes in the control group-model group; C represents the KEGG enrichment analysis of upregulated differentially expressed genes in the model group-Guangming Salt Four-Flavor Decoction group; and D represents the KEGG enrichment analysis of downregulated differentially expressed genes in the model group-Guangming Salt Four-Flavor Decoction group. Figure 8The transcriptome analysis results on day 5 are shown below - GO functional enrichment analysis. Among them, A is the representative GO functional enrichment analysis of differentially expressed genes upregulated in the control group and model group; B is the representative GO functional enrichment analysis of differentially expressed genes downregulated in the control group and model group; C is the representative GO functional enrichment analysis of differentially expressed genes downregulated in the model group and Guangming Salt Four-Flavor Decoction group; and D is the representative GO functional enrichment analysis of differentially expressed genes upregulated in the model group and Guangming Salt Four-Flavor Decoction group. Figure 9 The results of serum metabolomics analysis on day 2 are shown below; A represents the chemical classification and distribution of serum metabolites under negative ion mode; B represents the chemical classification and distribution of serum metabolites under positive ion mode; C represents the score plot of unsupervised principal component analysis under negative ion mode; and D represents the score plot of unsupervised principal component analysis under positive ion mode. Figure 10 The differential metabolite profiles are shown on day 2. A represents the volcano plot of differential metabolites between the control and model groups under negative ion mode (VIP > 1, P < 0.05); B represents the volcano plot of differential metabolites between the control and model groups under positive ion mode (VIP > 1, P < 0.05); C represents the volcano plot of differential metabolites between the model group and the Guangming Salt Four-Flavor Decoction Powder group under negative ion mode; and D represents the volcano plot of differential metabolites between the model group and the Guangming Salt Four-Flavor Decoction Powder group under positive ion mode. Figure 11 The results of hierarchical cluster analysis of differential metabolites between the control group and the model group on day 2 are shown. Among them, A is the hierarchical cluster heatmap of differential metabolites between the control group and the model group under negative ion mode; B is the hierarchical cluster heatmap of differential metabolites between the control group and the model group under positive ion mode. Figure 12 The results of hierarchical cluster analysis of differential metabolites between the Guangming Salt Four-Flavor Decoction group and the model group on day 2 are shown. Among them, A is the hierarchical cluster heatmap of differential metabolites between the Guangming Salt Four-Flavor Decoction group and the model group under negative ion mode; B is the hierarchical cluster heatmap of differential metabolites between the Guangming Salt Four-Flavor Decoction group and the model group under positive ion mode. Figure 13 The results of KEGG pathway enrichment analysis of differentially metabolites in serum on day 2 are shown below. A represents the KEGG enrichment bubble diagram of differentially metabolites in the control group and model group under negative ion mode; B represents the KEGG enrichment bubble diagram of differentially metabolites in the control group and model group under positive ion mode; C represents the KEGG enrichment bubble diagram of differentially metabolites in the Guangming Salt Four-Flavor Decoction Powder group and model group under negative ion mode; and D represents the KEGG enrichment bubble diagram of differentially metabolites in the Guangming Salt Four-Flavor Decoction Powder group and model group under positive ion mode. Figure 14 The results of serum metabolomics analysis on day 5 are shown below; A represents the chemical classification and distribution of serum metabolites under negative ion mode; B represents the chemical classification and distribution of serum metabolites under positive ion mode; C represents the principal component analysis score plot under negative ion mode; and D represents the principal component analysis score plot under positive ion mode. Figure 15 The differential metabolite profiles are shown on day 5. A represents the volcano plot of differential metabolites between the control and model groups under negative ion mode (VIP > 1, |log2FC| > 1, P < 0.05); B represents the volcano plot of differential metabolites between the control and model groups under positive ion mode (VIP > 1, |log2FC| > 1, P < 0.05); C represents the volcano plot of differential metabolites between the model group and the Guangming Salt Four-Flavor Decoction Powder group under negative ion mode; and D represents the volcano plot of differential metabolites between the model group and the Guangming Salt Four-Flavor Decoction Powder group under positive ion mode. Figure 16 The results of hierarchical cluster analysis of differential metabolites between the control group and the model group on day 5 are shown. Among them, A is the hierarchical cluster heatmap of differential metabolites between the control group and the model group under negative ion mode; B is the hierarchical cluster heatmap of differential metabolites between the control group and the model group under positive ion mode. Figure 17 The results of hierarchical cluster analysis of differential metabolites between the Guangming Salt Four-Flavor Decoction Group and the model group on day 5 are shown. Among them, A is the hierarchical cluster heatmap of differential metabolites between the Guangming Salt Four-Flavor Decoction Group and the model group under negative ion mode; B is the hierarchical cluster heatmap of differential metabolites between the Guangming Salt Four-Flavor Decoction Group and the model group under positive ion mode. Figure 18 The results of KEGG pathway enrichment analysis of serum metabolites on day 5 are shown. Among them, A is the KEGG enrichment bubble diagram of differential metabolites between the control group and the model group under negative ion mode; B is the KEGG enrichment bubble diagram of differential metabolites between the control group and the model group under positive ion mode; C is the KEGG enrichment bubble diagram of differential metabolites between the Guangming Salt Four Flavors Decoction Powder group and the model group under negative ion mode. Figure 19 The results are as follows: transcriptome-metabolome association analysis; A is the Spearman correlation heatmap of core genes and metabolites in the Guangming Salt Four-Flavor Decoction Powder intervention group on day 2; B is the Spearman correlation heatmap of core genes and metabolites in the Guangming Salt Four-Flavor Decoction Powder intervention group on day 5. In the image, GMYSWS or MD-4 are both abbreviations for Guangming Salt Four Flavors Soup Powder. Detailed Implementation

[0013] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0014] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0015] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0016] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0017] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0018] Example 1 I. Drugs, Chemical Reagents and Consumables Guangming Salt Four-Flavor Decoction Powder (hereinafter referred to as MD-4; made from Guangming salt, dried ginger, long pepper, and chebula; batch number conforms to the 1998 edition of the "Drug Standards of the Ministry of Health of the People's Republic of China - Mongolian Medicine Volume", standard number ZZ8329) was provided by Inner Mongolia Mongolian Medicine Co., Ltd. Carbon tetrachloride (CCl4) and olive oil were purchased from Sigma-Aldrich, Inc., USA. The enzyme-linked immunosorbent assay (ELISA) kits for tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) were purchased from Jiangsu Jingmei Biotechnology Co., Ltd., China.

[0019] II. Laboratory Animals and Experimental Protocols Male C57BL / 6 mice (6-8 weeks old, weighing 20-22g) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. All mice were housed in a specific pathogen-free environment with a 12h / 12h circadian rhythm, at a controlled temperature of 22±2℃, and with free access to food and water. After one week of acclimatization, the mice were randomly divided into 5 groups of 6 mice each. Control group: single intraperitoneal injection of olive oil (solvent), daily gavage with normal saline; Model group: single intraperitoneal injection of carbon tetrachloride-olive oil mixture, daily gavage with normal saline; Low-dose group of Guangming Salt Four-Flavor Decoction Powder: carbon tetrachloride modeling + daily gavage administration of 0.308g / kg MD-4; Medium-dose group of Guangming Salt Four-Flavor Decoction Powder: carbon tetrachloride modeling + daily gavage administration of 0.617g / kg MD-4; High-dose group of Guangming Salt Four-Flavor Decoction Powder: carbon tetrachloride modeling + daily gavage administration of 1.233g / kg MD-4.

[0020] Establishment of a carbon tetrachloride-induced acute liver injury model: Carbon tetrachloride was dissolved in olive oil (volume ratio 1:4, final concentration 20%) and injected intraperitoneally at a dose of 2 mL / kg body weight to establish an acute liver injury model (actual carbon tetrachloride dose: 0.4 mL / kg); the control group received an equal volume of olive oil. MD-4 medicinal material was ground into a uniform powder and prepared into a suspension of the appropriate concentration with sterile physiological saline before use. Based on the body surface area conversion formula, the human clinical dose (3g per dose) was converted to an equivalent mouse dose of 0.617g / kg, which was designated as the medium dose; the low and high doses were 0.5 times and 2 times the medium dose, respectively. Mice were administered the medication by gavage once daily at a fixed time, with the gavage volume adjusted according to the mouse's body weight. Mice were sacrificed on day 2 (peak inflammation), day 5 (damage-repair transition period), and day 7 (repair period) after model establishment, and the following tests were then performed: (1) Collection of serum and liver tissue samples At each time point, mice were briefly anesthetized, and blood was collected from the orbital venous plexus. After the blood was allowed to stand for 30 minutes, it was centrifuged at 3500 rpm for 10 minutes at room temperature to obtain serum, which was then stored at -80°C for analysis. Immediately after sacrifice, liver tissue was harvested: a portion was fixed with 4% paraformaldehyde for histopathological examination; the remaining tissue was flash-frozen in liquid nitrogen and stored at -80°C for molecular analysis.

[0021] (2) Quantitative detection of serum pro-inflammatory cytokines Serum tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) concentrations were detected using a commercially available double-antibody sandwich ELISA kit from Jiangsu Jingmei Biotechnology Co., Ltd., according to the manufacturer's instructions. Recombinant cytokines were serially diluted twofold (15.625-1000 pg / mL) to plot standard curves. 100 μL of sample was added to antibody-coated 96-well plates and incubated at 37°C for 180 min. After washing three times with phosphate-buffered saline (PBST) containing 0.05% Tween-20, 100 μL of biotin-labeled secondary antibody was added, and the plates were incubated at room temperature for 60 min. After another wash, 100 μL of tetramethylbenzidine (TMB) substrate was added, and the plates were incubated in the dark for 20 min. The reaction was terminated with stop solution, and the absorbance was read at 450 nm. Cytokine concentrations were calculated using a four-parameter logistic regression analysis.

[0022] (3) Histopathological examination Liver tissue fixed in 4% paraformaldehyde was dehydrated with graded ethanol, embedded in paraffin, and sectioned to 4 μm. Hematoxylin and eosin (H&E) staining was used to observe hepatocyte necrosis, lobular structure destruction, and inflammatory cell infiltration. Masson's trichrome staining was used to stain adjacent sections to assess collagen deposition and early fibrosis: sections were stained with Masson's trichrome for 3 min, differentiated twice with 0.2% acetic acid (1-5 s each time), treated with 1% phosphomolybdic acid for 5-8 min, counterstained with 1% aniline blue for 2-4 min, and rinsed again with 0.2% acetic acid. All sections were dehydrated, cleared with xylene, mounted with neutral resin, and representative images were acquired under an optical microscope.

[0023] III. Experimental Results 1. Effects of Guangming Salt Four-Flavor Decoction Powder on Serum Pro-inflammatory Cytokine Levels The results are as follows Figure 1 As shown in the figure, compared with the blank control group, the serum IL-6 level in the model group mice was significantly increased throughout the observation period (p < 0.01 or p < 0.05), indicating that the inflammatory response persisted. Figure 1 (A). IL-1β levels significantly increased on day 2 after modeling (p < 0.001) and remained highly expressed on days 5 and 7 (p < 0.001; p < 0.0001). Figure 1 (B). The inflammatory mediator TNF-α, with the highest absolute concentration, remained significantly high throughout the 7-day observation period (p < 0.05 or p < 0.001). Figure 1 (C). All three cytokines peaked on day 2 and then slowly declined, but remained significantly higher than baseline levels on day 7, indicating that the acute systemic inflammatory response induced by carbon tetrachloride has a certain degree of persistence.

[0024] The intervention with Guangming Salt Four-Flavor Decoction Powder significantly and dose-dependently inhibited the expression of the aforementioned cytokines. Based on body surface area, the clinically equivalent dose (medium-dose group) consistently and stably inhibited all three cytokines at all time points, showing a statistically significant reduction from day 2. This broad-spectrum inhibitory effect persisted throughout the 7-day observation period, demonstrating a balanced effect consistent with clinical application. The high-dose group showed a slightly stronger overall inhibitory effect, with IL-1β levels approaching those of the control group by day 7; however, this enhancement required twice the clinically equivalent dose, and the improvement was limited compared to the medium-dose group. The low-dose group showed a weaker intervention effect: only on day 7 did IL-6 levels show a statistically significant difference (p < 0.05), while IL-1β and TNF-α showed no significant improvement at any time point. In summary, both medium and high doses of Guangming Salt Four-Flavor Decoction Powder can sustainably and significantly alleviate carbon tetrachloride-induced systemic inflammatory responses. The medium-dose group achieved a good balance between efficacy and clinical application, while the low-dose group showed a delayed onset of anti-inflammatory effect and limited overall efficacy.

[0025] 2. The effect of Guangming Salt Four-Flavor Decoction Powder on improving pathological damage to liver tissue. The results are as follows Figure 2 As shown. HE staining results showed that in the blank control group, the liver lobule structure was intact, the hepatocytes were arranged regularly, the portal morphology was normal, and there was no obvious inflammatory cell infiltration at each time point. Figure 2 (A) The model group showed severe pathological damage: on day 2 of modeling, disordered liver lobule structure, portal dilation with extensive inflammatory cell infiltration, significant hepatocyte ballooning degeneration, focal necrosis, and sinusoidal congestion were observed; these pathological changes persisted on day 5; although some spontaneous repair occurred on day 7, necrotic lesions and inflammatory infiltration remained clearly visible. Intervention with Guangming Salt Four-Flavor Decoction could dose-dependently alleviate the aforementioned pathological damage. Specifically, the medium-dose group showed significant and progressive histological improvement at all time points, with a gradual reduction in liver lobule destruction, inflammatory infiltration, and hepatocyte necrosis, indicating that the drug has a stable and repeatable protective effect at clinically equivalent doses. The high-dose group showed a slightly stronger effect, especially with basic recovery of liver structure on day 7; however, compared to the medium-dose group, its additional histological benefit was limited, suggesting that the medium-dose group had reached an effective therapeutic dose and was more suitable for clinical application. The low-dose group showed only mild and delayed signs of repair, and a clear dose-response relationship was observed among the three groups.

[0026] Masson's trichrome staining results showed that the liver tissue of the blank control group had almost no collagen deposition ( Figure 2(B) In the model group, collagen accumulation was detectable around the portal area from day 2, and progressively worsened over time, reaching its most significant level on day 7, suggesting that the fibrotic remodeling process was initiated early in acute liver injury. Both the medium- and high-dose groups showed significant anti-fibrotic effects with consistent temporal patterns: no significant inhibitory effect on collagen deposition was observed on day 2, but significant inhibition of collagen accumulation was observed from day 5 onwards. The anti-fibrotic effect of the medium-dose group continued to increase throughout the observation period, and by day 7, it significantly reduced collagen deposition at the clinically equivalent dose. The high-dose group showed a slightly better anti-fibrotic effect on day 7, with collagen levels approaching the normal physiological range, but the benefit was limited due to the dosage being twice that of the medium-dose group. The low-dose group showed no significant improvement.

[0027] The above histological results were highly consistent with the serum cytokine detection results, jointly confirming that Guangming Salt Four-Flavor Decoction Powder can effectively alleviate carbon tetrachloride-induced hepatocellular necrosis and early fibrosis. Considering all detection indicators, the medium-dose group showed the most balanced overall efficacy and can be considered the preferred dose for subsequent translational and clinical studies.

[0028] Example 2 I. Untargeted Serum Metabolomics Analysis by Ultra-High Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) 1. Sample pretreatment Take 100 μL of serum sample, add 400 μL of ice bath extraction buffer (methanol:acetonitrile = 1:1, v / v, containing isotope-labeled internal standard), vortex for 3 min, and let stand at -20℃ for 1 h; centrifuge at 14000×g for 15 min at 4℃, aspirate 450 μL of supernatant to a new centrifuge tube, and vacuum dry; reconstitute the residue with 100 μL of acetonitrile:water (1:1, v / v), vortex for 2 min, centrifuge, and then analyze.

[0029] 2. Chromatographic and Mass Spectrometric Conditions Detection was performed using a Waters Ultra High Performance Liquid Chromatography (UHPLC) system in tandem with a Thermo Fisher Q-Exactive HF-X high-resolution mass spectrometer. Chromatographic separation was performed using a Waters ACQUITY UHPLC HSS T3 column (2.1 mm × 100 mm, 1.8 μm) at 40 °C. Mobile phase A was an aqueous solution containing 0.1% formic acid, and mobile phase B was acetonitrile. Gradient elution program: 0–1 min, 1% B; 1–9 min, 1%–99% B; 9–11 min, 99% B; 11–11.1 min, 99%–1% B; 11.1–14 min, 1% B. Flow rate: 0.4 mL / min. Data were acquired using electrospray ionization in both positive and negative ion modes.

[0030] 3. Data Processing and Analysis The raw data was converted to mzML format using Proteo Wizard software. Peak identification, peak alignment, retention time correction, and peak area extraction were performed using XCMS software to generate a three-dimensional data matrix containing mass-to-charge ratio, retention time, and peak area. Metabolite identification was completed by comparing the data with a self-built secondary mass spectrometry database and public databases such as HMDB and KEGG. Multivariate statistical analysis was performed using SIMCA-P software, including unsupervised principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Differential metabolites were screened based on the criteria of variable projection importance > 1, |log2 fold change| > 1, and P < 0.05, and KEGG pathway enrichment analysis was performed on the differential metabolites.

[0031] II. Liver Transcriptome Sequencing and Analysis 1. Sample pretreatment Approximately 30 mg of liver tissue was flash-frozen in liquid nitrogen, and total RNA was extracted using TRIzol reagent. RNA purity and concentration (A) were determined using a NanoDrop 2000 spectrophotometer. 260 / A 280 RNA integrity was assessed using an Agilent 2100 Bioanalyzer (RNA integrity value > 7.0), with a ratio of 1.8–2.1.

[0032] 2. Sequencing and Analysis Qualified RNA samples were used to construct libraries using the NEBNext® Ultra™ II RNA library preparation kit. After quantification using Qubit 3.0 quantitative PCR and real-time quantitative PCR, paired-end 150 bp sequencing was performed on the Illumina NovaSeq 6000 platform. Low-quality bases and adapter sequences were removed using Trim Galore software. Effective reads were aligned to the mouse reference genome (GRCm38 / mm10) using HISAT2 software, and the number of gene reads was counted using featureCounts software. Differentially expressed genes were screened using the DESeq2 package in R, with a screening threshold of |log2 fold change| > 1 and a corrected false discovery rate (P < 0.05). Gene ontology (GO) functional annotation and KEGG pathway enrichment analysis were performed using the clusterProfiler R package.

[0033] III. Experimental Results 1. Transcriptomic analysis reveals the time-dependent hepatoprotective effect of Guangming Salt Four-Flavor Decoction Powder (1) Transcriptome changes on day 2 were characterized by inflammatory activation and early metabolic repair. The results are as follows Figures 3-5 As shown. On day 2, a total of 893 differentially expressed genes were screened between the control group and the model group, including 409 upregulated genes and 484 downregulated genes. Figure 3A); A total of 689 differentially expressed genes were screened between the model group and the Guangming Salt Four-Flavor Decoction Powder intervention group, of which 290 genes were upregulated and 399 genes were downregulated. Figure 3 B).

[0034] The differentially expressed genes upregulated in the model group were mainly enriched in glutathione metabolism, cytochrome P450-mediated heterologous metabolism, drug metabolism-cytochrome P450, chemical carcinogenic pathways, and DNA replication and repair pathways, suggesting that oxidative stress, toxic response, and proliferative stress occur in the body during acute liver injury. Figure 4 A). Downregulated differentially expressed genes were mainly enriched in hematopoietic cell lineages, B cell receptor signaling pathways, PI3K-Akt signaling pathways, and calcium signaling pathways, indicating that immune development and cell survival-related functions were suppressed. Figure 4 B).

[0035] After intervention with Guangming Salt Four-Flavor Decoction, the differentially expressed genes that were upregulated were mainly enriched in steroid hormone biosynthesis, retinol metabolism, pyruvate metabolism, fatty acid degradation, glycolysis / gluconeogenesis, and cytochrome P450-related metabolic pathways. Figure 4 The C-value suggests that liver metabolism and detoxification functions have been restored; the downregulated differentially expressed genes are mainly enriched in the nuclear factor-κB signaling pathway, cytokine-cytokine receptor interaction, chemokine signaling pathway, tumor necrosis factor signaling pathway, C-type lectin receptor signaling pathway, and NOD-like receptor signaling pathway. Figure 4 The D), indicating that the extensive inflammatory cascade response was suppressed.

[0036] Gene Ontology (GO) functional analysis further validated the above results. The upregulated genes in the model group mainly participate in molecular functions such as DNA replication and antioxidant activity. Figure 5 A), downregulated genes are enriched in B cell proliferation, lymphocyte activation, and monocyte proliferation ( Figure 5 B). After intervention with Guangming Salt Four-Flavor Decoction, the downregulated genes were mainly related to neutrophil chemotaxis, granulocyte migration, and chemokine-mediated signal transduction. Figure 5 C), upregulated genes are enriched in fatty acid metabolism, sterol metabolism, cholesterol biosynthesis and amino acid biosynthesis ( Figure 5 (D).

[0037] (2) Transcriptome changes on day 5 suggest that the function has shifted from inflammation regulation to metabolic repair. The results are as follows Figures 6-8 As shown. On day 5 of modeling, a total of 525 differentially expressed genes were screened between the control group and the model group, including 213 upregulated genes and 312 downregulated genes. Figure 6(A); Compared with the Guangming Salt Four Flavors Decoction Powder (MD-4) group, a total of 409 differentially expressed genes were screened in the model group, of which 194 genes were upregulated and 215 genes were downregulated. Figure 6 (B) Compared to day 2, the number of differentially expressed genes decreased, suggesting that transcriptional levels tended to partially stabilize during the repair process.

[0038] The differentially expressed genes upregulated in the model group were still enriched in glutathione metabolism, cytochrome P450-related drug metabolism, heterologous substance metabolism, and chemical oncogenic pathways. Figure 7 The A-values ​​suggest that oxidative stress and toxic stress persist; the downregulated differentially expressed genes are mainly enriched in drug metabolism, retinol metabolism, glutathione metabolism, bile secretion, porphyrin metabolism, and cofactor biosynthesis pathways. Figure 7 The result of B indicates that the core functions of the liver are still impaired.

[0039] After intervention with Guangming Salt Four-Flavor Decoction, the differentially expressed genes that were upregulated were significantly enriched in drug metabolism-cytochrome P450, glutathione metabolism, pyruvate metabolism, AMPK signaling pathway, calcium signaling pathway, insulin signaling pathway, and carbon metabolism pathway. Figure 7 The C-value indicates enhanced liver detoxification capacity, improved antioxidant defense capacity, and restored energy metabolism function; the downregulated differentially expressed genes are mainly enriched in complement and coagulation cascade, cytokine-cytokine receptor interaction, C-type lectin receptor signaling pathway, hematopoietic cell lineage and phagosome-related pathway (C-type lectin receptor signaling pathway, hematopoietic cell lineage and phagosome-related pathway). Figure 7 The D-value indicates that innate immune activation and inflammation-related signals are continuously suppressed.

[0040] Gene Ontology (GO) analysis showed that on day 5, the upregulated genes in the model group were significantly enriched in functional items such as cellular response to foreign stimuli, foreign substance metabolism, fatty acid metabolism, glutathione transferase activity, and antioxidant activity. Figure 8 A); Downregulated genes were significantly enriched in damage response, glutathione metabolism, skin development, skeletal morphogenesis, and functions related to odor binding and amide transmembrane transporter activity (A). Figure 8 B). After intervention with Guangming Salt Four-Flavor Decoction, downregulated genes were significantly enriched in neutrophil migration, granulocyte chemotaxis, myeloid leukocyte migration, and the regulation of chemokine and interleukin-1β production. Figure 8 C); Upregulated genes are enriched in heterologous metabolism, fatty acid metabolism, nucleotide metabolism, extracellular matrix-related structures, and glutathione-related antioxidant functions (C). Figure 8 (D). The above results indicate that the intervention effect of Guangming Salt Four-Flavor Decoction Powder gradually shifted from early inflammation suppression on day 2 to a more significant metabolic, detoxification, and repair effect on day 5.

[0041] 2. Dynamic intervention effect of Guangming Salt Four-Flavor Decoction Powder on serum metabolome in mice with acute liver injury (1) Effects of Guangming Salt Four-Flavor Decoction on Serum Metabolic Profile and KEGG Pathway Analysis on Day 2 of Modeling Untargeted metabolomics analysis was performed on serum samples from all groups of mice on day 2 of modeling using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS / MS). Total ion chromatograms obtained in both negative and positive ion modes showed extremely low baseline peak intensity in the blank samples; the peak shapes of the quality control samples were consistent with those of all biological samples, and retention time and peak area showed good repeatability. This indicates that the instrument was stable and the data quality met the requirements for subsequent analysis.

[0042] The statistical results of serum metabolite chemical classification on day 2 showed that lipids and lipid-like molecules were the most abundant components in both detection modes (negative ion mode: 35.19%, positive ion mode: 32.32%), followed by organic acids and their derivatives (negative ion mode: 18.10%, positive ion mode: 17.22%) and organic heterocyclic compounds (negative ion mode: 15.67%, positive ion mode: 19.11%). Figure 9 (AB). The above results indicate that the serum metabolome is mainly composed of lipids and organic acids, and the two detection modes are highly complementary.

[0043] Unsupervised principal component analysis (PCA) results showed that the cumulative contribution rate of principal component 1 (PC1) and principal component 2 (PC2) was 60.38% (40.31% + 20.07%) in negative ion mode and 57.44% (37.99% + 19.45%) in positive ion mode. Figure 9 (CD). The principal component score plots show a clear separation trend among the three sample groups, with a large gap between the model group and the control group, suggesting that the overall serum metabolic profile of the body changes after modeling.

[0044] Differential metabolites between groups were screened using variable projection importance > 1 and P < 0.05 as screening criteria. Compared with the control group, 28 differential metabolites were detected in the model group under negative ion mode (10 upregulated and 18 downregulated), and 92 differential metabolites were detected under positive ion mode (44 upregulated and 48 downregulated). Figure 10 (AB). Compared with the model group, the Guangming Salt Four-Flavor Decoction Powder group showed 46 differentially expressed metabolites (22 upregulated and 24 downregulated) in negative ion mode and 150 differentially expressed metabolites (93 upregulated and 57 downregulated) in positive ion mode. The number of differentially expressed metabolites was significantly higher in the Guangming Salt Four-Flavor Decoction Powder group than in the model group-control group, especially in the positive ion mode. Figure 10 (CD).

[0045] Hierarchical cluster analysis was performed on the differentially expressed metabolites between the model group and the control group: under negative ion mode, the two groups of samples formed clearly separated clusters ( Figure 11In the model group (A), metabolites such as toluenesulfonic acid and lysophosphatidylcholine 18:3 showed an upregulation trend, while lipid-related metabolites such as cyclic lysophosphatidylcholine (18:1) and Lehualide I showed a downregulation trend. A bidirectional regulatory pattern was also observed under the positive ion mode, suggesting that modeling induces widespread disturbances in lipid and organic acid metabolism. Figure 11 B).

[0046] Under positive ion mode, intervention with Guangming Salt Four-Flavor Decoction Powder induced significant changes in more metabolites. The intervention group and the model group formed independent metabolic expression profile clusters, showing good intra-group aggregation and significant inter-group differences. (Heatmap) Figure 12 B) showed that, compared to the model group, the intervention group had significantly upregulated levels of multiple metabolites, mainly including N-methylethanolamine phosphate, L-phenylalanyl-L-proline, L-carnitine, and various acylcarnitine compounds (such as 3,4-dimethylhexanoylcarnitine and 5-hydroxydecanoylcarnitine), suggesting that the intervention with Guangming Salt Four-Flavor Decoction Powder enhanced amino acid transport and fatty acid β-oxidation-related metabolism. In addition, adenosine, N... 6 O 2’ - Purine nucleoside metabolites such as dimethyladenosine were upregulated in the intervention group, suggesting that energy metabolism had recovered to some extent.

[0047] Meanwhile, some metabolites in the intervention group were downregulated compared to the model group, including various lysophosphatidylcholine (LysoPC(24:0 / 0:0), PC(20:1(11Z) / 18:2(9Z,12Z)) etc.) and lipid-related substances such as dodecanoic acid and palmitoylethanolamine, suggesting that Guangming Salt Four Flavors Decoction Powder can inhibit and regulate the abnormally elevated lipid metabolism in the model group.

[0048] At the bottom of the heatmap, metabolites such as 1-carboxyethyl tryptophan, lysyl-histyl-glycine, gonadotropin-releasing hormone, 5-methyl pelarginine, and flumazenil showed significant upregulation in the intervention group, further demonstrating that the intervention effect of Guangming Salt Four-Flavor Decoction involves multiple metabolic axes, including amino acid metabolism, neurotransmitter regulation, and hormone synthesis. In summary, Guangming Salt Four-Flavor Decoction can exert multi-target and multi-pathway regulatory effects on model-induced metabolic disorders.

[0049] KEGG pathway enrichment analysis was performed on differentially expressed metabolites between the model group and the control group: under negative ion mode, the significantly enriched pathways included choline metabolism in cancer (enrichment factor ≈ 0.45), valine / leucine / isoleucine biosynthesis, and glycerophospholipid metabolism. Figure 13 In the positive ion mode, it mainly enriches vascular smooth muscle contraction (enrichment factor ≈ 1.0), platelet activation, and unsaturated fatty acid biosynthesis pathways (A); Figure 13 (B) suggests that the metabolic disorders in the model group mainly involve abnormalities in lipid metabolism, amino acid metabolism, and inflammation-related pathways.

[0050] Pathway enrichment analysis was performed on differentially expressed metabolites between the Guangming Salt Four-Flavor Decoction group and the model group: Purine metabolism and cytochrome P450-mediated heterologous metabolism pathways were significantly enriched under negative ion mode (enrichment factor ≈ 0.14, -log 10 (P value is the highest) Figure 13 In the positive ion mode, ethanol metabolism (enrichment factor ≈ 0.75), arginine and proline metabolism, sphingolipid signaling pathway, and unsaturated fatty acid biosynthesis pathway were enriched. Figure 13 (D). The combined results of the two detection modes indicate that Guangming Salt Four-Flavor Decoction Powder mainly exerts its intervention effect by regulating sphingolipid and unsaturated fatty acid metabolism, amino acid metabolism, and heterologous detoxification pathways.

[0051] (2) Effects of Guangming Salt Four-Flavor Decoction on Serum Metabolic Profile and KEGG Pathway Analysis on Day 5 of Modeling Ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS / MS) was used to analyze serum samples from mice in each group on day 5 of modeling. The results showed that the total ion chromatogram peaks were well-shaped in both negative and positive ion modes, the peak intensities of the quality control samples were stable, and the repeatability met the requirements for metabolomics data analysis.

[0052] The chemical composition of serum metabolites on day 5 was highly consistent with that on day 2. In both detection modes, lipids and lipid-like molecules accounted for the highest proportion (negative ion mode: 35.60%, positive ion mode: 32.33%), followed by organic acids and their derivatives (negative ion mode: 18.97%, positive ion mode: 20.51%) and organic heterocyclic compounds (negative ion mode: 14.96%, positive ion mode: 17.41%). Figure 14 (AB), further confirming that lipid metabolism-related substances are the main components of the serum metabolome.

[0053] Principal component analysis (PCA) results showed that the cumulative contribution rate of principal component 1 (PC1) and principal component 2 (PC2) was 48.80% (32.05% + 16.75%) in negative ion mode and 43.00% (23.38% + 19.62%) in positive ion mode. Figure 14 (CD). Significant separation trends were observed among the three groups within the principal component space. The distribution of the Guangming Salt Four-Flavor Decoction group was between that of the model group and the control group, suggesting that Guangming Salt Four-Flavor Decoction can regulate metabolic disorders induced by modeling. The samples from each group showed good clustering and excellent intra-group repeatability.

[0054] Differential metabolites were screened based on the criteria of variable projection importance > 1, |log2 fold change| > 1, and P < 0.05. Compared with the control group, 44 differential metabolites were detected in the model group under negative ion mode (14 upregulated and 30 downregulated), and 54 differential metabolites were detected under positive ion mode (17 upregulated and 37 downregulated). Both modes showed that downregulated metabolites were predominant, suggesting that overall metabolic activity is inhibited in the body under pathological conditions. Figure 15 (AB). Compared with the model group, the Guangming Salt Four-Flavor Decoction group showed 28 differentially expressed metabolites (23 upregulated and 5 downregulated) under negative ion mode and 22 differentially expressed metabolites (10 upregulated and 12 downregulated) under positive ion mode. The differentially expressed metabolites under negative ion mode were mainly upregulated, suggesting that the intervention of Guangming Salt Four-Flavor Decoction Powder can effectively restore the levels of suppressed metabolites in the model group. Figure 15 (CD).

[0055] The model-control group differential metabolite clustering heatmap showed that under negative ion mode, metabolites such as ferulic acid-4-sulfate and imperatorin were significantly upregulated in the model group, while lipid metabolites such as lysophosphatidylethanolamine and various organic acid metabolites were significantly downregulated. Figure 16 In the positive ion mode, lysophosphatidylcholine and amino acid metabolism-related products also undergo significant changes. Figure 16 (B). Cluster analysis of differential metabolites between the Guangming Salt Four-Flavor Decoction group and the model group showed that under negative ion mode, multiple metabolites such as 4-vinylphenol sulfate and swazil lignan C in the treatment group were significantly upregulated compared with those in the model group. Figure 17 In the positive ion mode, metabolites such as ethyl adipic acid and sangrino V are upregulated, while metabolites such as phosphatidylserine and fenmetrazine are downregulated. Figure 17 The B indicates that Guangming Salt Four Flavors Decoction Powder has a certain selectivity and specificity in regulating metabolic disorders.

[0056] Pathway enrichment results for differentially metabolites between the model group and the control group showed that the negative ion mode enriched a total of 8 pathways, among which glycerol ester metabolism (enrichment factor ≈ 0.5, -log) was the most significant. 10 The most significant result was achieved at P≈1.4, with the remaining results including tryptophan metabolism, phenylalanine metabolism, the pentose phosphate pathway, and amino acid biosynthesis. Figure 18 (A); enriched three pathways in the positive ion mode, with folic acid biosynthesis (enrichment factor ≈ 1.0, -log) 10 P≈2.0) is the most prominent ( Figure 18 B).

[0057] The pathway enrichment results of differential metabolites between the Guangming Salt Four-Flavor Decoction group and the model group showed that the negative ion mode enriched a total of 5 pathways, among which drug metabolism-cytochrome P450 (enrichment factor ≈ 0.33) was the most significant, and the others included the pentose phosphate pathway, carbon metabolism and amino acid biosynthesis. Figure 18 (C); no significant enrichment pathway was observed in the positive ion mode.

[0058] Based on the metabolomics analysis results on days 2 and 5, the serum metabolic disorders in mice after modeling were characterized by abnormalities in lipid metabolism (glycerides, lysophospholipids), amino acid metabolism, and energy metabolism, and the extent of the disorders changed with the extension of modeling time. After intervention with Guangming Salt Four-Flavor Decoction, the levels of various differentially metabolites were restored to varying degrees. Its mechanism of action may involve multiple metabolic axes, including lipid metabolism remodeling, amino acid metabolism regulation, energy metabolism restoration (pentose phosphate pathway, carbon metabolism), and CYP450 enzyme activity regulation, suggesting that Guangming Salt Four-Flavor Decoction has a systemic, multi-target metabolic intervention effect on mice with acute liver injury.

[0059] 3. Transcriptome-metabolome association analysis validates the conclusions of multi-omics integration. To further validate the pathway conclusions obtained from multi-omics joint enrichment analysis at the molecular interaction level, this invention conducted Spearman correlation analysis on the core genes and metabolites in the co-enriched pathways of the Guangming Salt Four Flavors Decoction Powder intervention group at two time points.

[0060] The correlation heatmap for day 2 included 20 transcriptome genes and 6 metabolites, and the hierarchical clustering results showed a significantly polarized structure. Figure 19 (A). The genes were clearly divided into two modules with opposite functions: the left module was dominated by metabolic enzyme genes such as Ugt1a6a, Ugt2a3, Adh1, and Gstk1; the right module was dominated by inflammatory signaling genes such as Cxcl2, Src, Camk2d, Cxcl1, and Lbp. Lysophosphatidylcholine LysoPC (20:4) and LysoPC (24:0) were strongly positively correlated with the right inflammatory module (r=0.51~0.91) and strongly negatively correlated with the left metabolic enzyme module (r=-0.42~-0.93), suggesting that even in the Guangming Salt Four-Flavor Decoction intervention group, the accumulation of residual lysophosphatidylcholine on day 2 continued to drive the inflammatory signaling axis, consistent with the high enrichment of the glycerol metabolism pathway at this time point. Cholesterol ester CE (5:0) showed a strong positive correlation with Camk2d (r=0.96) and Src (r=0.91), providing specific metabolite-kinase pairing evidence for sustained receptor activation in chemical carcinogenic pathways, and confirming that inflammation suppression is the main therapeutic axis in the early stage of Guangming Salt Four Flavors Decoction Powder.

[0061] The heatmap associated with day 5 includes 9 transcriptome genes and 5 metabolites. Figure 19(B). 7,8-Dihydrobiopterin showed a significant negative correlation with all nine genes (r = -0.59 to -0.73). These genes included members of the CYP450-related oxidoreductase system such as Gsta1, Gsta2, Fmo3, and Aldh3b3. This suggests that in the Guangming Salt Four-Flavor Decoction intervention group, individuals with stronger CYP450-mediated detoxification function had lower levels of 7,8-dihydrobiopterin, reflecting biopterin pathway disorder and oxidative load. This confirms at the transcriptional and metabolic levels that the restoration of liver detoxification function is the core therapeutic effect of Guangming Salt Four-Flavor Decoction during the repair phase. 7-Pseoserotonose 7-phosphate showed a strong positive correlation with key enzymes in the pentose phosphate pathway, Tkfc, Rpia, and Pklr (r = 0.72 to 0.79), indicating that the pentose phosphate pathway (PPP) was significantly activated during the metabolic repair phase, providing support for the body's antioxidant defense and NADPH production. This corroborates the conclusion of pathway enrichment related to the restoration of cofactor biosynthesis.

[0062] By combining the temporal changes at both time points, a coherent mechanistic shift can be clearly presented: the heatmap on day 2 shows a predominantly bipolar antagonistic pattern, with lysophospholipid-driven inflammatory signals and metabolic enzyme inhibition coexisting, reflecting the continued presence of lipid toxicity, indicating that Guangming Salt Four-Flavor Decoction mainly exerts an anti-inflammatory effect in the early stages; the heatmap on day 5 shows a systematic negative correlation between CYP450 system genes and oxidative stress metabolites, and a positive correlation between pentose phosphate pathway enzymes and sedoheptulose-7-phosphate, suggesting a shift in action towards detoxification function reconstruction and antioxidant metabolic homeostasis. These temporal changes, in terms of regulatory direction and intensity, are highly consistent with the pathway conclusions of multi-omics joint enrichment analysis, providing complete molecular chain evidence from gene expression to metabolic phenotype for the dual-phase hepatoprotective mechanism of Guangming Salt Four-Flavor Decoction.

[0063] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. The application of Guangming Salt Four-Flavor Decoction Powder in the preparation of drugs for the prevention and treatment of acute liver injury, characterized in that, The acute liver injury mentioned includes carbon tetrachloride-induced acute liver injury.

2. The application according to claim 1, characterized in that, The Guangming Salt Four-Flavor Soup Powder is made from Guangming salt, dried ginger, long pepper, and Terminalia chebula.

3. The application according to claim 1, characterized in that, The drug has the effect of inhibiting the abnormal increase of tumor necrosis factor-α, interleukin-6 and interleukin-1β.

4. The application according to claim 1, characterized in that, The drug has the effect of preventing and treating liver tissue structure damage, inflammatory infiltration, hepatocyte necrosis and early liver fibrosis.

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