A composition for soothing and protecting liver and its preparation method and application

Abstract

The application provides a composition for soothing liver and protecting liver and a preparation method and application thereof, and belongs to the technical field of compositions. The application comprises the following raw materials: radix puerariae, dandelion, phyllanthus emblica juice, asparagus, hovenia dulcis thunb, eucommia ulmoides oliver flower, tricholoma matsutake sing, American ginseng, dried tangerine or orange peel, liquorice, double-petaled red rose, kale juice, rice, oat flour and mung bean. The composition of the application realizes overall regulation on the complex pathological mechanism of NAFLD through the composite effect of multiple components, multiple targets and multiple pathways, can not only directly improve the lipid metabolism state of the liver, but also can regulate the structure and function of intestinal flora, reduce the endotoxin load, block the abnormal transmission of inflammatory signals from the intestine to the liver, and thus systematically improve the function of the intestinal-liver axis. No obvious adverse reactions are found, the application has the significant advantages of comprehensive mechanism of action, high safety, great potential for long-term application, and can effectively make up for the deficiencies of existing treatment methods in terms of treatment effect sustainability and comprehensive intervention ability.

Description

Technical Field

[0001] This invention belongs to the field of composition technology, and particularly relates to a composition for soothing and protecting the liver, its preparation method and application. Background Technology

[0002] In recent years, the incidence of non-alcoholic fatty liver disease (NAFLD) has been rising globally, gradually evolving into a complex metabolic disease characterized by lipid metabolism disorders, accompanied by chronic inflammatory responses and gut microbiota homeostasis imbalance, posing a long-term challenge to population health and healthcare systems. However, due to the multi-factor and multi-pathway interactions involved in its pathogenesis, there is currently a lack of specific therapeutic drugs that can exert stable therapeutic effects against its overall pathological process.

[0003] Currently, clinical intervention for NAFLD is still based on lifestyle management, including dietary adjustments, exercise interventions, and weight control. Although these methods can improve metabolic status and slow disease progression to some extent, their effectiveness is highly dependent on patients' long-term self-management ability. In practice, problems such as insufficient compliance and difficulty in maintaining intervention intensity are common, leading to unstable efficacy and difficulty in achieving continuous and controllable disease management.

[0004] Meanwhile, existing drug treatment strategies often target single pathological aspects, such as lipid metabolism abnormalities or inflammatory responses. Their mechanisms of action are relatively limited, making it difficult to simultaneously address multiple interrelated pathogenic factors, including hepatic lipid deposition, chronic inflammation, and gut microbiota imbalance. In particular, gut microbiota dysbiosis is considered closely related to disease progression in the development of NAFLD, yet current treatments are insufficient in regulating the overall gut-liver axis function. Furthermore, existing interventions often focus on a specific stage of the disease, lacking a comprehensive intervention program that can be flexibly applied across different stages of disease development, catering to both early intervention in high-risk groups and treatment needs of confirmed patients.

[0005] Therefore, existing drugs have limitations in terms of comprehensive intervention capabilities, long-term safety, and adaptability to different stages of the disease, making it difficult to meet the actual needs of long-term management and early intervention for NAFLD. Developing a composition that can simultaneously improve lipid metabolism abnormalities, inhibit chronic inflammatory responses, and regulate gut microbiota imbalance through multi-pathway synergistic effects has become a crucial problem urgently needing to be solved in this field. Summary of the Invention

[0006] In view of this, one of the objectives of the present invention is to provide a composition that soothes and protects the liver.

[0007] A second objective of this invention is to provide a method for preparing the composition.

[0008] A third objective of this invention is to provide the application of the composition or the preparation method in the preparation of products for preventing non-alcoholic liver injury.

[0009] A fourth objective of this invention is to provide the application of the composition or the preparation method in the preparation of products that regulate intestinal flora.

[0010] The fifth objective of this invention is to provide a product for preventing non-alcoholic liver injury.

[0011] To achieve the above-mentioned objectives, the present invention provides the following technical solution: A composition for soothing and protecting the liver, comprising the following ingredients by weight: 10-20 parts kudzu root, 2-4 parts dandelion, 3-8 parts amla juice, 1-3 parts asparagus, 5-8 parts Japanese raisin tree fruit, 0.5-1.5 parts eucommia male flower, 0.5-1.5 parts matsutake mushroom, 0.5-1.5 parts American ginseng, 0.2-0.5 parts dried tangerine peel, 0.5-0.8 parts licorice root, 0.3-0.8 parts double-petaled red rose, 2-3 parts kale juice, 20-30 parts rice, 3-5 parts oat flour, and 1-2 parts mung beans.

[0012] Preferably, the ingredients, by weight, include the following: 15 parts kudzu root, 3 parts dandelion, 5 parts amla juice, 2 parts asparagus, 6 parts Japanese raisin tree fruit, 1 part eucommia male flower, 1 part matsutake mushroom, 1 part American ginseng, 0.3 parts dried tangerine peel, 0.6 parts licorice, 0.5 parts double-petaled red rose, 2.5 parts kale juice, 24 parts rice, 4 parts oat flour, and 1.5 parts mung beans.

[0013] The present invention also provides a method for preparing the composition, comprising the following steps: Pueraria root was extracted with water, and the extract was collected and concentrated to obtain concentrate 1; After mixing Hovenia dulcis, dandelion, asparagus, dried tangerine peel and licorice, extract with water, collect the extract, and concentrate to obtain concentrate 2; Rice, matsutake mushrooms, eucommia male flowers, American ginseng and double-petaled red roses were mixed and then ultra-finely pulverized to obtain ultra-fine powder 1. After stir-frying the mung beans, they were then ultra-finely ground to obtain ultra-fine powder 2. Concentrate 1, concentrate 2, ultrafine powder 1, ultrafine powder 2, oat flour, amla juice, kale juice and sweetener are mixed and ground using a colloid mill. The ground liquid is then dried and matured to obtain the composition.

[0014] Preferably, the steps of kudzu root water extraction include: mixing kudzu root and water at a mass ratio of 1:8, decocting for 60 minutes, and then collecting the extract; After mixing the jujube seed, dandelion, asparagus, dried tangerine peel and licorice, add water at a mass ratio of 1:8 and decoct for 60 minutes. Collect the first extract and the residue. Mix the residue with water at a mass ratio of 1:8 and decoct for 30 minutes. Collect the second extract. Combine the two extracts.

[0015] Preferably, the particle size of ultrafine powder 1 is 120~130μm; the particle size of ultrafine powder 2 is 120~130μm.

[0016] Preferably, the grinding slurry is dried at 100~110℃.

[0017] The present invention also provides the use of the composition or the preparation method thereof in the preparation of products for the prevention and / or treatment of non-alcoholic liver injury.

[0018] Preferably, the product regulates liver lipid metabolism and inhibits inflammation.

[0019] The present invention also provides the use of the composition or the preparation method in the preparation of products that regulate intestinal flora.

[0020] The present invention also provides a product for preventing non-alcoholic liver injury, the product comprising the composition described above.

[0021] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a liver-protecting composition that, through the synergistic effect of multiple ingredients, achieves multi-level intervention on key pathological aspects of NAFLD, including lipid metabolism abnormalities, chronic inflammatory responses, and gut-hepatic axis dysfunction. This invention not only directly improves hepatic lipid metabolism but also systemically improves gut-hepatic axis function by regulating gut microbiota structure and function, reducing endotoxin load, and blocking abnormal transmission of inflammatory signals from the gut to the liver. No significant adverse reactions have been observed.

[0022] The composition of this invention achieves holistic regulation of the complex pathological mechanisms of NAFLD through the combined effects of multiple components, multiple targets, and multiple pathways. It possesses significant advantages such as a comprehensive mechanism of action, high safety, and great potential for long-term application, effectively compensating for the shortcomings of existing treatments in terms of sustained efficacy and comprehensive intervention capabilities. It effectively addresses the problems of existing treatments primarily targeting single aspects of lipid metabolism or inflammatory responses, making it difficult to holistically regulate the complex metabolic-inflammatory-gut microecological imbalance in NAFLD; the unstable efficacy or potential adverse reaction risks associated with long-term use of existing drugs, making it difficult to meet the needs of long-term intervention and early prevention of NAFLD; and the lack of a comprehensive treatment plan that can function effectively at different stages of the disease (including early prevention in high-risk populations and treatment in diagnosed patients). Attached Figure Description

[0023] Figure 1 The production process of the composition of the present invention; Figure 2 The production process of the composition in Comparative Example 1; Figure 3 The sheet composition obtained in Example 1; Figure 4 Experimental design and dosing timelines for different compositions for the prevention of non-alcoholic liver injury; Figure 5 Experimental design and dosing timeline for the use of the composition to prevent NAFLD; Figure 6 The study investigated the effects of the combined intervention on body weight, tissue weight, serum biochemical parameters, and inflammatory markers in mice. A represents physiological parameters—body weight, liver weight, and white adipose tissue weight; B represents serum lipopolysaccharide; C represents serum biochemical parameters—total cholesterol, triglycerides, alanine aminotransferase (ALT), and aspartate aminotransferase (AST); and D represents serum inflammatory factors—tumor necrosis factor-α, interleukin-1β, interleukin-6, and interleukin-10. Different letters indicate statistically significant differences between groups (P < 0.05). Figure 7 The effects of the combined intervention on liver histological changes and CD86 and FABP4 expression in NAFLD mice were investigated. A represents liver H&E staining and Oil Red O staining; B represents NAS score; C represents liver immunohistochemistry; D represents quantitative expression of CD86 in the liver; and E represents quantitative expression of FABP4 in the liver. Different letters indicate statistically significant differences between groups (P<0.05). P<0.05; Figure 8 The effects of combined intervention on the structure and function of gut microbiota in NAFLD mice were investigated. A represents the Shannon index; B represents the Venn diagram; C represents PCOA analysis; D represents a bar chart of species composition at the phylum level; E represents a bar chart of species composition at the family level; and G represents microbial symbiotic network analysis. Different letters indicate statistically significant differences between groups (P < 0.05). Figure 9 LDA score for LEfse analysis; Figure 10 Prediction of KEGG functional pathways in gut microbiota; Figure 11The effects of the combined intervention on the liver transcriptome are shown below; where A is the MOD vs CON differential gene volcano plot; B is the PRE vs MOD differential gene volcano plot; C is the TX vs MOD differential gene volcano plot; D is the MOD vs CON GO enrichment analysis plot; E is the TX vs MOD GO enrichment analysis plot; F is the PRE vs MOD GO enrichment analysis plot; COD is the control group; MOD is the high-fat diet model group; PRE is the prevention group; and TX is the treatment group. Figure 12 KEGG enrichment analysis of the effects of combined intervention on the liver transcriptome; where G is the MOD vs CON KEGG enrichment analysis plot; H is the PRE vs MOD KEGG enrichment analysis plot; and I is the TX vs MOD KEGG enrichment analysis plot. Figure 13 The combined intervention had no significant effect on the weight of the colon, spleen, and kidney in mice; where A represents the weight of the colon; B represents the weight of the spleen; and C represents the weight of the kidney; different letters indicate statistically significant differences between groups (P<0.05). Figure 14 This study investigated the effects of a combination of interventions under conditions of gut microbiota depletion on body weight, metabolic indicators, inflammatory markers, liver histology, and the expression of CD86 and FABP4. In this study, A represents body weight; B represents aspartate aminotransferase (AST); C represents total cholesterol; D represents tumor necrosis factor-α; E represents liver H&E staining and Oil Red O staining; F represents NAS score; G represents liver immunohistochemistry; H represents quantified liver CD86 expression; and I represents quantified liver FABP4 expression. Different letters indicate statistically significant differences between groups (P < 0.05). Figure 15 A schematic diagram of experimental grouping and time flow for validating gut microbiota-related mechanisms; Figure 16 This study investigated the effects of fecal microbiota transplantation (FMT) on body weight, tissue weight, serum biochemical parameters, and inflammatory markers in NAFLD mice. In this study, A represents body weight; B represents white adipose tissue weight; C represents liver weight; D represents IL-1β content; E represents asthenospermia ulmoides (AS) content; F represents total thrombocytopenic purpura (TC) content; G represents triglyceride (TG) content; and H represents altrine thrombocytopenia (ALT) content. Different letters indicate statistically significant differences between groups (P < 0.05). Figure 17 The study investigated the effects of fecal microbiota transplantation (FMT) on liver histological changes and CD86 and FABP4 expression in NAFLD mice. In the figures, A represents H&E staining and Oil Red O staining; B represents the NAS score; C represents the immunohistochemical staining of liver CD86 and FABP4; D represents the quantitative results of liver CD86 immunohistochemical expression; and E represents the quantitative results of liver FABP4 immunohistochemical expression. Different letters indicate statistically significant differences between groups (P < 0.05). Detailed Implementation

[0024] This invention provides a composition for soothing and protecting the liver, comprising the following ingredients by weight: 10-20 parts kudzu root, 2-4 parts dandelion, 3-8 parts amla juice, 1-3 parts asparagus, 5-8 parts Japanese raisin tree fruit, 0.5-1.5 parts eucommia male flower, 0.5-1.5 parts matsutake mushroom, 0.5-1.5 parts American ginseng, 0.2-0.5 parts dried tangerine peel, 0.5-0.8 parts licorice root, 0.3-0.8 parts double-petaled red rose, 2-3 parts kale juice, 20-30 parts rice, 3-5 parts oat flour, and 1-2 parts mung bean. Preferably, the ingredients, by weight, include the following: 15 parts kudzu root, 3 parts dandelion, 5 parts amla juice, 2 parts asparagus, 6 parts Japanese raisin tree fruit, 1 part eucommia male flower, 1 part matsutake mushroom, 1 part American ginseng, 0.3 parts dried tangerine peel, 0.6 parts licorice root, 0.5 parts double-petaled red rose, 2.5 parts kale juice, 24 parts rice, 4 parts oat flour, and 1.5 parts mung beans. This invention does not specifically limit the source of the above ingredients; commercially available products commonly used in the field are acceptable.

[0025] In the composition of this invention, kudzu root and dandelion are the principal herbs. Kudzu root promotes the upward movement of clear yang, detoxifies alcohol, and promotes the production of body fluids and relaxes muscles. Alcohol toxicity damages the liver, and kudzu root can promote alcohol metabolism and relieve liver heat after drinking. Dandelion clears liver and stomach heat and disperses heat stagnation in the liver meridian. It has a direct clearing effect on liver fire rising (red eyes, irritability). Together, these two herbs, one promoting upward movement and the other clearing and draining, directly target the two core causes of liver damage: alcohol toxicity and liver heat.

[0026] In this invention, Phyllanthus emblica, Hovenia dulcis, Eucommia ulmoides male flowers, and Matsutake mushroom are used as assistant herbs. Phyllanthus emblica clears heat and cools the blood, promotes fluid production, enhances the liver's detoxification ability, and counteracts the excessive coldness of the principal herb. Hovenia dulcis is a traditional remedy for alcohol poisoning, promoting diuresis and eliminating dampness, accelerating the excretion of alcohol and metabolic waste. Eucommia ulmoides male flowers warm and tonify the liver and kidneys, preventing excessive purging from damaging liver yang, embodying the principle of "soothing while tonifying." Matsutake mushroom tonifies deficiency and strengthens the body, improving the liver's antioxidant and repair capabilities. The combined effects of Phyllanthus emblica, Hovenia dulcis, Eucommia ulmoides male flowers, and Matsutake mushroom assist the principal herb in strengthening alcohol detoxification and liver cleansing, while also protecting the liver (tonifying the liver and kidneys, and repairing), ensuring that detoxification does not harm the body's vital energy.

[0027] In this invention, the composition includes American ginseng, dried tangerine peel, double-petaled red rose, and kale as adjuvants. American ginseng replenishes qi and nourishes yin. The liver is yin in nature and yang in function; excessive liver-soothing can easily deplete qi and damage yin. American ginseng can nourish the yin and blood of the liver, preventing excessive liver-soothing. Dried tangerine peel regulates qi and strengthens the spleen. Liver stagnation easily weakens the spleen; dried tangerine peel not only helps soothe the liver (promote qi circulation) but also prevents liver disease from affecting the spleen. Double-petaled red rose directly soothes the liver, relieves stagnation, invigorates blood, and regulates menstruation. Its aroma is penetrating and can penetrate deep into the liver meridian, relieving emotional liver stagnation. Modern nutritional science has confirmed that kale is rich in glucosinolates and chlorophyll, which can induce phase II detoxification enzymes in the liver, enhancing the liver's clearance of toxins (including alcohol metabolites). American ginseng, dried tangerine peel, double-petaled red rose, and kale together serve as adjuvants to address concurrent symptoms: soothing the liver, strengthening the spleen, nourishing yin, enhancing detoxification, and comprehensively optimizing the liver-protecting effect.

[0028] In this invention, licorice is used as the guiding herb. Licorice harmonizes the various herbs, relieving urgency and harmonizing the middle jiao. It can both moderate the cold nature of kudzu root and dandelion, and tonify the spleen and replenish qi, ensuring that the entire formula works synergistically without conflict. This ensures the overall formula works smoothly and avoids stomach upset, making it suitable for long-term use.

[0029] This invention uses kudzu root and dandelion as the principal ingredients to clear away alcohol toxicity and liver heat, rose and tangerine peel as auxiliary ingredients to soothe liver stagnation, American ginseng and eucommia male flowers to nourish the liver, and jujube fruit and kale to accelerate detoxification. Licorice is used to harmonize the effects, achieving a liver-soothing and liver-protecting effect that is clearing without being cold, soothing without being depleting, and nourishing without being stagnant.

[0030] The composition of this invention is scientifically formulated, and the components complement and synergistically enhance each other in terms of medicinal properties, meridian tropism, and modern pharmacological effects, forming a stable and highly effective compound therapeutic system. It has the effects of soothing the liver and regulating qi, relieving emotional stress, replenishing qi and nourishing yin, and improving fatigue.

[0031] The present invention also provides a method for preparing the composition, comprising the following steps: extracting kudzu root with water, collecting the extract and concentrating it to obtain concentrate 1; mixing Japanese raisin tree fruit, dandelion, asparagus, dried tangerine peel and licorice root, extracting them with water, collecting the extract, and concentrating it to obtain concentrate 2; mixing rice, matsutake mushroom, Eucommia ulmoides male flower, American ginseng and double-petaled red rose and then pulverizing them into ultrafine powder 1; stir-frying mung beans and then pulverizing them into ultrafine powder 2; mixing concentrate 1, concentrate 2, ultrafine powder 1, ultrafine powder 2, oat flour, amla juice, kale juice and sweetener, grinding them with a colloid mill, drying and aging the grinding liquid to obtain the composition.

[0032] In the preparation method of this invention, the step of water extraction of kudzu root includes: mixing kudzu root with water at a mass ratio of 1:8, decocting for 60 minutes, and collecting the extract. The extract is preferably concentrated under reduced pressure at -0.08 to -0.04 MPa to a relative density of 1.05 to 1.10 (measured at 60°C) to obtain concentrate 1. Mixing jujube seed, dandelion, asparagus, tangerine peel, and licorice root, preferably with water at a mass ratio of 1:8, decocting for 60 minutes, and collecting the first extract and filter residue; mixing the filter residue with water at a mass ratio of 1:8, decocting for 30 minutes, and collecting the second extract; combining the two extracts, preferably under reduced pressure at -0.08 to -0.04 MPa to a relative density of 1.05 to 1.10 (measured at 60°C), and collecting concentrate 2. Kudzu root has a high starch content, and when mixed with other materials for extraction, it easily forms a starch gelatinization system, hindering the dissolution of other active ingredients. This invention extracts kudzu root separately and then mixes it with other ingredients, which can increase the content of active ingredients.

[0033] In this invention, the ultrafine pulverization is preferably carried out using an ultrafine pulverizer. The particle size of ultrafine powder 1 is preferably 120~130μm; the particle size of ultrafine powder 2 is preferably 120~130μm.

[0034] In a specific embodiment of the present invention, the preferred method for stir-frying mung beans is to wash and dry the mung beans, heat the pan, add the mung beans, stir-fry over low heat while continuously turning them, and stir-fry for 15-20 minutes until the surface of the mung beans turns light yellowish-brown, thus obtaining the stir-fried mung beans. Raw mung beans are cold in nature and are not suitable for people with weak spleen and stomach or indigestion. After stir-frying, they become mild in nature, strengthen the spleen and stomach, and reduce the burden on the gastrointestinal tract.

[0035] This invention utilizes a colloid mill to further refine the particle size of the concentrated liquid to approximately 10 μm, resulting in a more stable and uniform material system. This leads to a smoother, more delicate texture, better solubility, and reduced stratification and sedimentation after reconstitution. The grinding liquid is preferably dried and matured using a drum drying process, with a preferred drying temperature of 100-110°C and a preferred drum rotation speed of 15-20 Hz. After drying and maturation, the grinding liquid yields a flake composition, preferably with a flake thickness of 0.3-0.4 mm. Drum drying minimizes damage to heat-sensitive nutrients, preserving the natural value of the raw materials to a greater extent. Furthermore, drum-dried products exhibit better rehydration properties, resulting in a smoother, more delicate, and viscous texture after reconstitution.

[0036] The sweeteners used in this invention are preferably any one or more of mogrosides, xylitol, and fructooligosaccharides. When the sweeteners added in this invention consist of multiple components, each component is added in an equal mass ratio. The mass of the sweetener added in this invention is 1% to 7% of the mass of the composition.

[0037] The present invention also provides the use of the composition or the preparation method in the preparation of products for preventing non-alcoholic liver injury, the products having the effects of regulating liver lipid metabolism and inhibiting inflammation.

[0038] The present invention also provides the use of the composition or the preparation method in the preparation of products that regulate intestinal flora.

[0039] The present invention also provides a product for preventing non-alcoholic liver injury, the product comprising the composition described above.

[0040] The compositions of this invention can be extended to the treatment of other metabolic-related diseases. For example, steatohepatitis, insulin-resistant liver injury, and obesity are all closely related to hepatic lipid metabolism disorders, chronic inflammatory responses, and gut microbiota imbalances. Because their pathological mechanisms highly overlap with NAFLD, the compositions of this invention have strong cross-disease adaptability and can serve as adjunctive therapy for various metabolic liver diseases, especially providing new treatment approaches for complex pathological processes such as metabolic disorders and inflammatory responses that cannot be fully improved by existing treatments.

[0041] The composition of this invention not only has a clear therapeutic effect but also possesses dual application value in prevention. Its application is not limited to the treatment of patients diagnosed with NAFLD, but can also be used for early intervention and disease prevention in high-risk populations. For patients with existing hepatic lipid metabolism abnormalities or inflammatory responses, systemic treatment using conventional drugs can be administered; while for individuals without obvious clinical symptoms but with risk factors such as obesity, metabolic syndrome, or abnormal liver function indicators, preventative intervention can be achieved through long-term, low-dose, continuous use, thereby delaying or blocking disease progression. By implementing differentiated application strategies in different populations, this invention can not only function at different stages of disease development but also improve the foresight of interventions and the overall therapeutic effect.

[0042] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0043] Example 1 A liver-soothing and liver-protecting composition, comprising the following ingredients by weight: 15 parts kudzu root, 3 parts dandelion, 5 parts amla juice, 2 parts asparagus, 6 parts Japanese raisin tree fruit, 1 part eucommia male flower, 1 part matsutake mushroom, 1 part American ginseng, 0.3 parts dried tangerine peel, 0.6 parts licorice root, 0.5 parts double-petaled red rose, 2.5 parts kale juice, 24 parts rice, 4 parts oat flour, and 1.5 parts mung bean.

[0044] The preparation method is as follows: Kudzu root and water were mixed at a mass ratio of 1:8 and decocted for 60 minutes at 100℃. The extract was collected and concentrated under reduced pressure at -0.08 to -0.04 MPa to a relative density of 1.05 to 1.10 (measured at 60℃) to obtain concentrate 1.

[0045] The following ingredients were mixed: Hovenia dulcis seeds, asparagus, dandelion, dried tangerine peel, and licorice root. These were then mixed with water at a mass ratio of 1:8. The mixture was decocted twice at 100℃. The first extraction lasted 60 minutes, and the second extraction lasted 30 minutes. The extracts were collected and concentrated under reduced pressure (-0.08 to -0.04 MPa) to a relative density of 1.05–1.10 (measured at 60℃), yielding concentrate 2.

[0046] Rice, matsutake mushrooms, eucommia male flowers, American ginseng and double-petaled red roses were mixed and then ultra-finely pulverized to obtain ultra-fine powder 1 (particle size 120~130μm). After stir-frying the mung beans, they were ultra-finely pulverized to obtain ultra-fine powder 2 (particle size 120~130μm). Concentrate 1, Concentrate 2, Ultrafine Powder 1, Ultrafine Powder 2, Oat flour, Phyllanthus emblica juice, Kale juice, and sweetener (the sweetener is a mixture of xylitol and fructooligosaccharides in equal mass ratio, with the sweetener accounting for 6.3% of the total mass of the composition) are mixed and ground using a colloid mill. The ground liquid is then dried and matured using a drum dryer at a temperature of 105°C and a drum speed of 20 Hz to obtain a sheet-like composition (e.g., ...). Figure 3 (As shown).

[0047] Example 2 A composition for soothing and protecting the liver, comprising the following ingredients in parts by weight: 12 parts kudzu root, 4 parts dandelion, 4 parts amla juice, 3 parts asparagus, 7 parts Japanese raisin tree fruit, 0.8 parts eucommia male flower, 0.8 parts matsutake mushroom, 0.8 parts American ginseng, 0.5 parts dried tangerine peel, 0.8 parts licorice, 0.4 parts double-petaled red rose, 2 parts kale juice, 28 parts rice, 3 parts oat flour, and 2 parts mung beans.

[0048] The ingredients are: 10-20 parts kudzu root, 2-4 parts dandelion, 3-8 parts amla juice, 1-3 parts asparagus, 5-8 parts Japanese raisin tree fruit, 0.5-1.5 parts eucommia male flower, 0.5-1.5 parts matsutake mushroom, 0.5-1.5 parts American ginseng, 0.2-0.5 parts dried tangerine peel, 0.5-0.8 parts licorice root, 0.3-0.8 parts double-petaled red rose, 2-3 parts kale juice, 20-30 parts rice, 3-5 parts oat flour, and 1-2 parts mung beans. Preferably, the ingredients, by weight, include the following: The preparation method is the same as in Example 1.

[0049] Example 3 A composition for soothing and protecting the liver, comprising the following ingredients in parts by weight: 18 parts kudzu root, 2 parts dandelion, 7 parts amla juice, 1 part asparagus, 6 parts Japanese raisin tree fruit, 1.2 parts eucommia male flower, 1.2 parts matsutake mushroom, 1.2 parts American ginseng, 0.3 parts dried tangerine peel, 0.5 parts licorice, 0.7 parts double-petaled red rose, 3 parts kale juice, 22 parts rice, 5 parts oat flour, and 1 part mung bean.

[0050] The preparation method is the same as in Example 1.

[0051] Comparative Example 1 A composition for soothing and protecting the liver, comprising the following ingredients in parts by weight: 15 parts kudzu root, 3 parts dandelion, 5 parts amla juice, 2 parts asparagus, 6 parts Japanese raisin tree fruit, 1 part eucommia male flower, 1 part matsutake mushroom, 1 part ginseng powder, 24 parts rice and 4 parts mung beans.

[0052] Preparation process such as Figure 2 As shown, specifically: Mixture 1, which is a mixture of kudzu root, Japanese raisin tree fruit, dandelion, eucommia male flower, matsutake mushroom and asparagus, is mixed with water at a mass ratio of 1:8. The mixture is then boiled in water (100℃) twice for 1 hour each time. The extracts are combined and concentrated to 1 / 20 of the original volume to obtain a concentrated solution. Rinse the rice twice with water, mix it with 90℃ water at a mass ratio of 1:2, and soak for 4 hours to obtain wet rice. After stir-frying the mung beans, they are then ultra-finely pulverized to obtain ultra-fine powder (particle size 120~130μm). The concentrate, ultrafine powder, wet rice, ginseng powder, amla juice and sweetener (the sweetener is a mixture of erythritol and mogroside in equal mass ratio, and the mass of the sweetener is 1.6% of the total mass of the composition) are mixed and ground using a colloid mill. The grinding liquid is then dried and matured to obtain the composition.

[0053] Example 4 The effects of different compositions on the prevention of non-alcoholic liver injury.

[0054] The efficacy of the composition provided in Comparative Example 1 (denoted as X-WQT, dissolved in 0.9% sodium chloride solution, concentration of 62.5 mg / mL) and the composition provided in Comparative Example 1 (denoted as Y-WQT, dissolved in 0.9% sodium chloride solution, concentration of 62.5 mg / mL) in preventing non-alcoholic liver injury was compared.

[0055] 1. Experimental methods.

[0056] C57BL / 6J mice (purchased from Beijing Vital River Biotechnology Co., Ltd.) were randomly divided into 4 groups (n=6 per group) after one week of acclimatization: Control group (CON): fed standard feed throughout weeks 1-13; High-fat diet model group (MOD): fed a high-fat diet (HFD) from week 1 to week 9, and switched to a standard diet from week 10 to week 13 to maintain the same dietary conditions as the groups related to the combined food (WQT) intervention. Comparative Example 1: Composition Prevention Group (Y-PRE): The same dietary regimen as the MOD group was adopted, but Y-WQT was continuously administered via gavage (0.2 mL) throughout weeks 1 to 13. Example 1 Composition Prevention Group (X-PRE): The same diet regimen as the MOD group was adopted, but X-WQT was continuously administered by gavage (0.2 mL) throughout the 1st to 13th weeks.

[0057] The standard feed (CD) was purchased from Jiangsu Medison Biomedical Co., Ltd., and its main components are crude protein, crude fat, crude fiber, calcium, phosphorus, lysine, methionine and cystine. The high-fat feed (HFD) is based on the standard feed, with the addition of 27% lard, 8% sucrose, 10.8% whole milk powder, 13.5% casein and 2% calcium bicarbonate. The content of each component is the mass percentage of its total mass in the high-fat feed.

[0058] Experimental feeding protocol such as Figure 4 As shown, after 13 weeks of feeding, the mice were weighed, blood was collected from each group of mice, and they were euthanized by dislocation. The liver weight was measured, the white fat was separated and weighed, and the TC and TG content in the blood was measured. The data were then statistically analyzed.

[0059] 2. Experimental results.

[0060] Table 1. Experimental data on the prevention of non-alcoholic liver injury by different compositions ( - x±s)

[0061] To compare with COD, This means p < 0.01. # represents p < 0.05; # is for comparison with MOD, ## represents p < 0.01, and ## represents p < 0.05.

[0062] According to the results in Table 1, the composition provided in Example 1 can significantly alleviate HFD-induced lipid accumulation, reduce white fat in the body, and reduce liver fat deposition, thereby reducing liver weight. At the same time, the composition provided in Example 1, due to the addition of several traditional Chinese medicines such as tangerine peel, licorice, double-petaled red rose, and American ginseng, significantly reduced the total cholesterol and triglycerides in the serum of mice.

[0063] The composition provided by this invention, with the core effects of dried tangerine peel and American ginseng, combined with the auxiliary regulatory effects of other components, can form a synergistic effect of multiple targets and pathways in regulating glucose and lipid metabolism and improving metabolic disorders. The various raw materials in the composition of this invention each perform their specific functions and work synergistically to control lipids at the source, regulate metabolism through pathways, improve insulin resistance, exert anti-inflammatory and antioxidant effects, and regulate neuroendocrine homeostasis in a multi-dimensional synergistic manner, comprehensively improving lipid metabolism abnormalities, hepatic steatosis, and metabolic disorders caused by high-fat diets and metabolic syndrome. This allows for the observation of a decrease in serum total cholesterol and triglycerides in animal experiments, making it particularly suitable for mouse models of metabolic syndrome or fatty liver, and showing superior efficacy in improving non-alcoholic fatty liver disease in mice compared to the composition provided in Comparative Example 1.

[0064] Example 5 The effectiveness of the composition of the present invention was verified.

[0065] Using the composition provided in Example 1 as a sample, denoted as WQT, dissolved in 0.9% sodium chloride solution, the concentration was 62.5 mg / mL, to verify its effects on regulating liver lipid metabolism, inhibiting inflammatory response, and improving intestinal flora imbalance.

[0066] 1. The composition of the present invention has the effect of regulating liver lipid metabolism and inhibiting inflammatory response.

[0067] 1.1 Experimental methods.

[0068] C57BL / 6J mice (purchased from Beijing Vital River Biotechnology Co., Ltd.) were randomly divided into 5 groups (n=8 per group) after one week of acclimatization: Control group (CON): fed standard diet (CD) throughout weeks 1-13; High-fat diet model group (MOD): fed a high-fat diet (HFD) from week 1 to week 9, and switched to a standard diet from week 10 to week 13 to maintain the same dietary conditions as the groups related to the WQT intervention of the present invention.

[0069] Prevention group (PRE): The same dietary regimen as the MOD group was adopted, but WQT was continuously administered via gavage (0.2 mL) throughout weeks 1 to 13.

[0070] Treatment group (TX): The same diet as the MOD group was followed, but WQT was administered via gavage (0.2 mL) only during weeks 10-13.

[0071] To evaluate the role of gut microbiota in the prophylactic effect of WQT, an antibiotic clearance group (ABX) was set up on the basis of prophylactic intervention: recipient mice were treated with antifungal drugs and antibiotics to deplete gut microbiota, and then continued to be treated with the same diet and WQT administration method as the PRE group; except for microbiota treatment, all other conditions were kept the same.

[0072] The antifungal drug combined with antibiotic treatment consisted of two phases: Phase 1: Antifungal treatment, with amphotericin B (1 mg / kg) administered orally by gavage once daily for 3 consecutive days to reduce intestinal fungal load. Phase 2: Antibacterial treatment, with a mixture of antibiotics including ampicillin (100 mg / kg), gentamicin (100 mg / kg), metronidazole (100 mg / kg), and vancomycin (50 mg / kg) administered orally by gavage once daily for 7 consecutive days to inhibit intestinal bacteria. All drugs were dissolved in physiological saline and administered at a gavage volume of 10 mL / kg. All solutions were freshly prepared before use. Mouse weight and general condition were monitored daily during the treatment period. To reduce the risk of cross-contamination, mice in the antibiotic-treated group were housed in dedicated cages with individual bedding and water bottles.

[0073] The standard feed (CD) and high-fat feed (HFD) are the same as in Example 4.

[0074] The dosing timelines for each group are as follows: Figure 5 As shown, this study systematically evaluates the preventive and therapeutic effects of WQT and examines whether its preventive intervention effect changes under conditions of gut microbiota depletion, thereby analyzing the relationship between gut microbiota and the intervention effect of WQT.

[0075] After the 13th week of the intervention trial, the following indicators were measured: (1) Physiological and biochemical indicators: In the CON, MOD, PRE and TX groups, blood was collected from mice after an overnight fast. The blood was allowed to stand at room temperature for 30 min to coagulate, and then the serum was separated by centrifugation at 3,000×g for 10 min (4℃) and stored for testing. The indicators included: ① Serum lipopolysaccharide; ② Serum biochemical indicators: TC, TG, ALT, AST; ③ Serum inflammatory factors: TNF-α, IL-1β, IL-6, IL-10; Physiological index detection: After the experiment, the mice were euthanized and dissected, and the following indicators were measured: body weight, liver weight, and white fat weight.

[0076] (2) Detection of liver pathology and immunohistochemical indicators: CON, MOD, PRE and TX groups, liver H&E staining, Oil Red O staining, NAS staining, liver immunohistochemistry, liver CD86 expression quantification, liver FABP4 expression quantification.

[0077] (3) Fecal metabolomics analysis: Fecal samples from four groups (CON, MOD, PRE, and TX) were analyzed by 16S rRNA gene sequencing and bioinformatics analysis. The indicators included: Shannon index, Venn diagram, PCOA analysis, species composition at the phylum level, species group at the family level, LDA score of LEfse analysis, microbial symbiotic network analysis, and KEGG functional pathway prediction of gut microbiota.

[0078] (4) Liver transcriptomic analysis: Transcriptome sequencing analysis was performed on liver tissues from CON, MOD, PRE, and TX mice. This included differential expression analysis, GO enrichment analysis, and KEGG enrichment analysis.

[0079] (5) Weight detection of colon, spleen and kidney in mice: After the experiment, mice in the CON, MOD, PRE and TX groups were sacrificed and dissected, and the indicators were weighed, including colon weight, spleen weight and kidney weight.

[0080] (6) Physiological, metabolic, verification, liver histology and CD86 and FABP4 detection under the condition of intestinal flora depletion: PRE and ABX group mice were tested, and the indicators included: body weight, aspartate aminotransferase, total cholesterol, tumor necrosis factor α, liver H&E staining, Oil Red O staining, NAS score, liver immunohistochemistry, liver CD86 expression quantification, and liver FABP4 expression quantification.

[0081] 1.2 Experimental results.

[0082] 1.2.1 Effects of the composition on body weight and tissue weight, serum biochemistry and inflammatory markers in mice.

[0083] according to Figure 6 The results showed that, compared with the MOD group, the PRE group mice had significantly lower liver weight and white adipose tissue weight (P<0.05); significantly lower serum biochemical indicators (TC, TG, ALT, AST) (P<0.05); significantly lower levels of pro-inflammatory factors (TNF-α, IL-1β, IL-6); and significantly lower serum lipopolysaccharide (LPS) levels (P<0.05). Figure 6 (A~D in the original text). This indicates that the preventive intervention of the composition of the present invention can significantly alleviate HFD-induced lipid accumulation and inflammatory response, and reduce endotoxin load associated with intestinal barrier damage.

[0084] Compared with the MOD group, the TX group showed a significant decrease in white adipose tissue weight (P<0.05), a significant decrease in serum biochemical indicators TC, ALT and AST (P<0.05), and a significant decrease in serum LPS level (P<0.05). Figure 6 (A~C in the text). Regarding inflammatory markers, TNF-α and IL-6 were significantly reduced in the TX group ( Figure 6 (D in the original text). In summary, the therapeutic intervention of the composition of the present invention can improve HFD-induced metabolic disorders and inflammatory states to a certain extent.

[0085] 1.2.2 Effects of the composition on histological changes in liver tissue and expression of CD86 and FABP4 in NAFLD mice To evaluate the effects of the composition of this invention on the structure and pathological damage of mouse liver tissue, H&E staining and Oil Red O staining were performed on liver tissues of each group, and immunohistochemical (IHC) detection of CD86 and FABP4 was carried out; at the same time, the degree of liver lesions was quantified according to the NAFLD activity score (NAS, 0-8 points).

[0086] Compared with the CON group, the MOD group showed significant lipid droplet deposition, hepatocyte ballooning, and hepatocyte swelling / damage in the liver tissue, accompanied by mild inflammatory cell infiltration. Figure 7(A) The NAS score shows that the MOD group scored 4 points ( Figure 7 (B in the text). At the molecular phenotypic level, IHC results showed that the expression of CD86 and FABP4 in the liver tissue of the MOD group was significantly increased (P<0.05). Figure 7 The values ​​in C and D indicate that the HFD-induced NAFLD model is accompanied by enhanced lipid accumulation and upregulation of inflammation-related signals.

[0087] Compared with the MOD group, both the PRE and TX groups showed a reduction in lipid deposition and ballooning degeneration in liver tissue. Figure 7 In the A group, the NAS score dropped to 2 points, significantly lower than that of the MOD group ( Figure 7 (B in the text). IHC quantitative analysis showed that the CD86 expression level in the TX group was significantly lower than that in the MOD group (P<0.05), while there was no significant difference between the PRE group and the MOD group (P<0.05). Figure 7 (C, D in the original text). Regarding FABP4, the PRE group showed significantly lower expression than the MOD group (P<0.05), while the TX group only showed a decreasing trend and was not significantly different from the MOD group (P>0.05). Figure 7 (C, E in the text).

[0088] The above results indicate that the intervention of the composition of the present invention can alleviate HFD-related hepatic lipid deposition and histological damage to a certain extent, and shows a group-specific improvement pattern in inflammation and lipid metabolism-related markers. Overall, both the preventive and therapeutic interventions of the composition of the present invention are associated with the improvement of NAFLD phenotype. Under the experimental conditions, the preventive intervention showed more consistent improvement in lipid deposition-related indicators, while the therapeutic intervention was more prominent in inflammation-related markers (such as CD86).

[0089] 1.2.3 Effects of combined intervention on the structure and function of gut microbiota in NAFLD mice.

[0090] To further elucidate the microbiological basis of the composition of this invention in preventing NAFLD, 16S rRNA sequencing analysis was performed on fecal samples from the CON, MOD, PRE, and TX groups of mice.

[0091] The α-diversity results showed that the Shannon index of the MOD group was significantly lower than that of the CON group; while the Shannon index of the PRE and TX groups rebounded to a level that was not significantly different from that of the CON group, and was significantly higher than that of the MOD group (P<0.05). Figure 8 Figure A in the figure indicates that the intervention of the composition of the present invention can alleviate the decline in NAFLD-related microbial diversity to a certain extent. The Venn diagram further shows that, compared to the MOD group, the PRE and TX groups shared an increased number of ASVs with the control group, while exhibiting more intervention-specific ASVs (…). Figure 8 (B in the middle).

[0092] β-diversity analysis (PCoA) showed a clear separation of gut microbiota structures among the groups: the MOD group samples clustered independently, while the community structures of the PRE and TX groups were closer to those of the CON group. Figure 8 The presence of C indicates that the composition of this invention can significantly remodel the intestinal microecological imbalance induced by HFD. At the phylum level, the MOD group showed a trend of increased relative abundance of Firmicutes and decreased relative abundance of Bacteroidetes, resulting in an increased Firmicutes / Bacteroidetes ratio; the PRE and TX groups alleviated this imbalance to some extent, making their overall community structure closer to that of the CON group ( Figure 8 (D) At the family level, compared with the CON group, the MOD group Muribaculaceae and Akkermansiaceae The relative abundance decreased significantly, while Ruminococcaceae and Lactobacillaceae The relative abundance was significantly increased. Compared with the MOD group, the PRE group... Muribaculaceae and Akkermansiaceae Significantly increased, at the same time Ruminococcaceae and Lactobacillaceae Significant decrease; the TX group showed Muribaculaceae Significantly increased, and Ruminococcaceae and Lactobacillaceae Significantly reduced ( Figure 8 The E in the figure indicates that the corrective effect of preventive and therapeutic interventions on microbial imbalance is group-specific, showing differences in the adjustment range and combination characteristics of key dominant departments.

[0093] LEfSe analysis further screened and identified the landmark taxa with differential abundance in each group. Both PRE and TX groups were enriched with several characteristic bacterial groups with high LDA scores. Figure 9 The F in the figure reveals that the intervention of the composition of the present invention can drive the gut microbiota to produce identifiable group-specific changes and form relatively stable differential abundance characteristics.

[0094] To assess changes in gut microbiota interaction structure, we further conducted co-occurrence network analysis. The results showed that the MOD group exhibited a significantly simplified network structure with fewer nodes and connections, indicating that the NAFLD state may be accompanied by weakened microbiota interactions and decreased community stability. In contrast, the CON, PRE, and TX groups displayed more complex network structures. Figure 8 (G in Table 2) indicates that the intervention of the composition of the present invention is associated with an increase in the complexity of microbial community interactions.

[0095] Functional predictive analysis showed that, compared with the CON group, the proportion of functions related to "carbohydrate metabolism" was significantly increased in the MOD group (P<0.05). Compared with the MOD group, functions such as "amino acid metabolism," "endocrine system," and "terpenoid and polyketide metabolism" were significantly increased in the PRE group (P<0.05), while functions such as "amino acid metabolism," "endocrine system," and "cell growth and death" were also significantly enhanced in the TX group (P<0.05). Figure 10 (H in the text).

[0096] The above results indicate that the intervention of the composition of the present invention is related to the optimization of the complexity and functional potential of the gut microbiota to a certain extent, and may promote the adjustment of NAFLD-related microecological imbalance towards a more stable state.

[0097] Table 2. Topological characteristics of gut microbiota symbiotic networks in the CON, MOD, PRE, and TX groups.

[0098] In summary, the intervention with the composition of this invention is associated with a reduction in HFD-induced gut microbiota dysbiosis. Specifically, the intervention group showed improved α-diversity, a shift in community conformation in the β-diversity space towards the control group, and an increase in shared ASVs with the control group, accompanied by the appearance of intervention-specific ASVs. At the taxonomic level, the intervention alleviated MOD to some extent. Firmicutes / Bacteroidetes Imbalance, and exhibiting consistent but varying regulatory characteristics in key advantageous disciplines (e.g.) Muribaculaceae , Akkermansiaceae The relative abundance increased, and Ruminococcaceae , Lactobacillaceae The relative abundance decrease indicates group differences in the intensity and combination patterns of gut microbiota restoration between preventive and therapeutic interventions. Simultaneously, co-occurrence network analysis showed that the intervention group maintained / restored higher network complexity, supporting its consistency with richer potential interactions and higher ecological robustness. Functional prediction further revealed that the intervention of the present invention's composition was accompanied by a redistribution of metabolic-related functional potential, manifested as a significant enhancement of amino acid metabolism and endocrine system-related functions. In summary, the above structural and functional changes collectively suggest that the present invention's composition may be associated with improved NAFLD-related phenotypes by reshaping gut microbiota composition, interaction networks, and metabolic potential, providing a microbiological basis for its preventive and therapeutic effects.

[0099] 1.2.4 Effects of combined intervention on the liver transcriptome.

[0100] To further elucidate the molecular basis of the composition of the present invention in preventing and treating NAFLD, transcriptome sequencing analysis was performed on liver tissues of mice in the CON, MOD, PRE, and TX groups.

[0101] Volcano plot analysis of differential expression revealed a large number of differentially expressed genes (DEGs) in comparisons of MOD vs CON, PRE vs MOD, and TX vs MOD. Figure 11 The A~C in the figure indicate that HFD can induce extensive changes in the liver transcriptome profile; while the intervention of the composition of the present invention is associated with partial reversal of this abnormal transcriptional feature, and the magnitude of the reversal differs between preventive and therapeutic interventions.

[0102] GO enrichment analysis showed that, compared with the CON group, the DEGs in the MOD group were mainly enriched in biological processes such as fatty acid metabolism, small molecule decomposition, and purine nucleotide metabolism (P<0.05). Figure 11 The D in the figure reflects a significant remodeling of liver metabolic-related biological processes under HFD exposure. In the comparison between PRE and MOD, differentially expressed genes showed more significant enrichment in lipidolysis, fatty acid metabolism, and purine nucleotide metabolism (P<0.05). Figure 11 The F values ​​indicate that preventive intervention has a more concentrated transcriptional reversion effect on lipid-related metabolic processes. The differentially expressed genes between TX and MOD are mainly enriched in fatty acid metabolism, small molecule breakdown, and organic acid breakdown processes (P<0.05). Figure 11 The E in the data shows that therapeutic interventions are also associated with transcriptional remodeling of metabolic processes, but the composition of their enriched entries differs somewhat from that of preventive interventions.

[0103] KEGG enrichment analysis further showed that, compared with the CON group, the MOD group had significantly enriched pathways such as "transcriptional dysregulation in cancer", "AGE-RAGE signaling pathway (diabetic complications)" and "mTOR signaling pathway", while the basic energy metabolism-related pathways such as "carbon metabolism" and "pyruvate metabolism" were significantly decreased (P<0.05). Figure 12 The presence of H in the H indicates a shift in liver metabolic homeostasis and stress / inflammatory signals under HFD conditions. In contrast, both the PRE and TX groups showed upregulation and enrichment of pathways such as carbon metabolism, fatty acid degradation, and fatty acid metabolism, while insulin resistance, AGE-RAGE signaling pathway (a diabetic complication), and MAPK signaling pathway were significantly downregulated (P<0.05). Figure 12 The G~I in the figure indicates that the intervention of the composition of the present invention is consistent with the restoration of metabolic pathways and the inhibition of inflammation / stress-related signaling networks.

[0104] In summary, liver transcriptomics results indicate that the preventive and therapeutic effects of the composition of the present invention are closely related to the transcriptional reversion of fatty acid metabolism and energy metabolism-related pathways, and are accompanied by a reduction in metabolic inflammation and stress signaling pathways such as AGE-RAGE and mTOR / MAPK. The above-mentioned molecular-level changes are consistent with the direction of NAFLD phenotype improvement, providing transcriptomics evidence for the composition of the present invention to alleviate the progression of NAFLD.

[0105] 1.2.5 The combination intervention had no significant effect on the weight of the colon, spleen and kidneys in mice.

[0106] To assess the potential adverse reactions of the composition of the present invention, the general condition and behavior of mice were dynamically observed after each gavage during the experiment. No abnormal reactions such as lethargy, reduced activity or vomiting were observed.

[0107] Further organ index analysis showed that after the prevention and treatment intervention with the composition of the present invention, there were no significant differences in the weight of the colon, spleen and kidney of mice among the groups (P>0.05). Figure 13 (A~C in the original text). The above results indicate that, within the dosage and intervention period of this study, the composition of the present invention did not cause detectable overall toxicity or changes in the weight of major organs, and its safety profile is good.

[0108] 1.2.6 Effects of combined intervention under conditions of gut microbiota depletion on body weight, metabolic indicators, inflammatory markers, liver histology, and expression of CD86 and FABP4.

[0109] To clarify the role of the gut microbiota in the preventive effect of the compositions of this invention, we used antibiotics combined with antifungal drugs to deplete the gut microbiota in the antibiotic clearance prevention group (ABX). After the microbiota depletion was completed, the ABX group continued to receive preventive intervention with the compositions of this invention according to the same protocol as the PRE group.

[0110] Compared with the PRE group, the body weight, serum biochemical indicators (AST, TG) and pro-inflammatory factor (TNF-α) levels of mice in the ABX group were significantly increased (P<0.05); Figure 14 (A~D in the original text).

[0111] Histological observation showed that lipid deposition and hepatocyte ballooning were more pronounced in the ABX group than in the PRE group. Figure 14 (E, F in the text).

[0112] At the molecular phenotypic level, immunohistochemical quantitative results showed that the expression of CD86 and FABP4 in liver tissue of the ABX group was not significantly different from that of the PRE group (P>0.05). Figure 14 (G~I in the middle).

[0113] In summary, under the background of gut microbiota depletion, the preventive intervention of the composition of the present invention showed a reduced improvement in body weight, metabolic indicators and liver histological phenotype, indicating that its preventive effect depends to some extent on the integrity of the gut microbiota.

[0114] 2. The mediating role of gut microbiota in the preventive effect of the composition of the present invention.

[0115] 2.1 Experimental methods.

[0116] After one week of acclimatization, C57BL / 6J mice were randomly divided into 3 groups (n = 8 in each group): Antibiotic clearance group (ABX): Same as the ABX group in "1.1 Experimental Methods"; In the control donor microbiota transplantation group (AFC) after antibiotic clearance: recipient mice were treated with antifungal drugs and antibiotics to deplete their gut microbiota, and were fed a high-fat diet (HFD) for the next 9 weeks. From weeks 10 to 13, they were switched to a standard diet (CD), and were given prophylactic gavage intervention with the composition of the present invention (WQT) throughout weeks 1 to 13, at the same gavage dosage as in the PRE group method in "Example 5". From weeks 10 to 13, they received fecal microbiota transplantation (FMT) with fresh mixed fecal suspension from CON group donor mice once daily for 4 consecutive weeks. Following antibiotic clearance, mice received model donor microbiota transplantation (AFM): After treatment with antifungal drugs and antibiotics to deplete the gut microbiota, recipient mice were fed a high-fat diet (HFD) for the next 9 weeks, then switched to a standard diet (CD) from weeks 10 to 13. Throughout weeks 1-13, mice received prophylactic gavage intervention with the composition of this invention (WQT), at the same dosage as the PRE group method in "Example 5". From weeks 10 to 13, mice received fresh mixed fecal suspension from donor mice in the MOD group once daily for 4 consecutive weeks for fecal microbiota transplantation (FMT).

[0117] FMT Method: Fresh fecal particles from donor mice were collected under aseptic conditions and processed within 30 minutes of defecation. Fecal samples were collected and weighed according to donor source (CON or MOD), and a 100 mg / mL suspension was prepared by adding pre-cooled sterile saline. After thorough homogenization, the suspension was filtered through a 70 μm filter to remove large particles. The prepared suspension was stored on ice throughout the process and administered via gavage within 2 hours, with a daily gavage volume of 0.2 mL. To verify the effectiveness of microbial clearance and transplantation, quantitative detection could be performed by extracting fecal DNA, or periodic monitoring could be conducted using culture-based methods to confirm the effectiveness of microbial removal and subsequent reconstruction.

[0118] The experimental timelines for each group are as follows: Figure 15As shown, body weight and general condition were monitored weekly during the experiment, and metabolic-related indicators and liver endpoints were assessed at the end of week 13 of the HFD intervention. This experiment was used to compare phenotypic differences after microbiota reconstruction from different donor sources in the context of microbiota depletion, thereby evaluating the role of the gut microbiota in the preventive intervention effect of the composition of the present invention and its relationship with NAFLD-related phenotypic improvement.

[0119] 2.2 Experimental results.

[0120] 2.2.1 Effects of fecal microbiota transplantation on body weight, tissue weight, serum biochemistry, and inflammatory markers in NAFLD mice.

[0121] To further verify the mediating role of gut microbiota in the preventive effect of Wuqing Decoction, fecal microbiota transplantation (FMT) was performed on recipient mice using fresh fecal suspensions from donor mice in the CON and MOD groups, respectively, under antibiotic clearance.

[0122] Compared with the ABX group, the AFC group mice showed significantly reduced white adipose tissue weight and liver weight, and significantly decreased serum biochemical levels of TC, TG and ALT (P<0.05). Figure 16 (A~H in the group). The AFM group also showed improved metabolic phenotype, manifested by a significant decrease in body weight and white adipose tissue weight, and a significant decrease in the levels of inflammatory factor IL-1β and serum TC, TG and ALT (P<0.05; Figure 16 (A~H in the text).

[0123] In summary, FMT administered against a background of gut microbiota depletion can partially improve the metabolic and inflammation-related phenotypes of recipient mice, indicating that the replenishment / reconstruction of the gut microbiota and the metabolic benefits associated with the preventive intervention of the composition of this invention show a consistent trend.

[0124] 2.2.2 Effects of fecal microbiota transplantation on liver histological changes and CD86 and FABP4 expression in NAFLD mice.

[0125] At the liver histological level, H&E and Oil Red O staining results showed that FMT treatment reduced liver lipid deposition, decreased the degree of related histological damage, and significantly reduced NAS score (P<0.05). Figure 17 (A, B in the original text).

[0126] At the molecular phenotypic level, immunohistochemical quantification showed that, compared with the ABX group, the expression of CD86 and FABP4 in the liver tissue of the AFC group was significantly reduced (P<0.05); while there was no significant difference between the AFM group and the ABX group (P>0.05). Figure 17 (C, D in the original text).

[0127] It is evident that microbial transplantation from different donor sources has varying regulatory effects on liver inflammation and lipid metabolism-related markers. Microbial transplantation from the control donor (AFC) is consistent with more significant histological improvement and a decrease in molecular markers.

[0128] Based on the above results, it can be seen that by using the composition of the present invention as the core intervention method, and forming a synergistic regulatory pathway of "metabolism-inflammation-microecology" around the key pathological links of NAFLD, the following beneficial effects can be achieved: (1) Addressing the problem of insufficient coverage of single-target treatment for NAFLD due to multi-link coupling: The occurrence and development of NAFLD involves the interaction of multiple links such as hepatic lipid / energy metabolism disorders, activation of inflammatory and stress signals, and intestinal microecological imbalance. Intervention of a single target or a single link is often difficult to achieve systemic improvement. The composition of this invention is consistent with the reversion of energy metabolism pathways such as fatty acid metabolism and carbon metabolism at the liver level, and is accompanied by downregulation of metabolic inflammation / stress-related pathways such as AGE-RAGE, insulin resistance, and mTOR / MAPK; at the intestinal level, it is consistent with the improvement of microbial diversity, community structure, key microbiota and interaction network complexity, thereby achieving multi-dimensional intervention for the complex pathological process of NAFLD and breaking through the technical limitation of "only acting on a single link".

[0129] (2) Addressing the issue of insufficient safety and stability required for long-term management: NAFLD is characterized by chronic progression, and clinical management emphasizes the sustainability of long-term intervention. Some existing drugs have fluctuating efficacy or long-term medication risks, which limits their long-term use. The composition of this invention, under the experimental dosage and intervention period, showed no typical adverse reactions in mice after continuous gavage observation, and no significant changes were observed in the weight indicators of organs such as the colon, spleen, and kidneys. This supports the good tolerability and safety of the composition of this invention under the above-mentioned research conditions, providing an acceptable safety basis for long-term intervention and early prevention.

[0130] (3) Addressing the lack of a comprehensive intervention strategy across different disease stages: Existing interventions are mostly focused on the treatment stage after diagnosis, while a systematic approach is lacking for early prevention and disease progression control in high-risk populations. This invention simultaneously establishes two intervention pathways: prevention and treatment. In both intervention scenarios, trends consistent with improvements in metabolic phenotype, liver histological damage, and abnormal molecular pathways were observed. Furthermore, the verifiability of the "microecological link" was further supported by strategies such as antibiotic depletion and fecal microbiota transplantation, demonstrating that the integrity and reconstruction of the gut microbiota are consistently associated with the preventive benefits of the composition of this invention, providing a mechanistic basis for a comprehensive intervention across different stages.

[0131] The composition of this invention synergistically regulates metabolic and inflammatory networks, and incorporates the gut microbiota as a key regulated link into the overall framework, thereby forming a technical path and evidence support with translational value in improving metabolic disorders, reducing liver damage, and meeting the needs of long-term management and phased intervention.

[0132] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A composition for soothing and protecting the liver, characterized in that, The ingredients, by weight, include the following: 10-20 parts kudzu root, 2-4 parts dandelion, 3-8 parts amla juice, 1-3 parts asparagus, 5-8 parts Japanese raisin tree fruit, 0.5-1.5 parts eucommia male flower, 0.5-1.5 parts matsutake mushroom, 0.5-1.5 parts American ginseng, 0.2-0.5 parts dried tangerine peel, 0.5-0.8 parts licorice root, 0.3-0.8 parts double-petaled red rose, 2-3 parts kale juice, 20-30 parts rice, 3-5 parts oat flour, and 1-2 parts mung beans.

2. The composition according to claim 1, characterized in that, The ingredients, by weight, include the following: 15 parts kudzu root, 3 parts dandelion, 5 parts amla juice, 2 parts asparagus, 6 parts Japanese raisin tree fruit, 1 part eucommia male flower, 1 part matsutake mushroom, 1 part American ginseng, 0.3 parts dried tangerine peel, 0.6 parts licorice root, 0.5 parts double-petaled red rose, 2.5 parts kale juice, 24 parts rice, 4 parts oat flour, and 1.5 parts mung beans.

3. A method for preparing the composition according to claim 1 or 2, characterized in that, Includes the following steps: Pueraria root was extracted with water, and the extract was collected and concentrated to obtain concentrate 1; After mixing Hovenia dulcis, dandelion, asparagus, dried tangerine peel and licorice, extract with water, collect the extract, and concentrate to obtain concentrate 2; Rice, matsutake mushrooms, eucommia male flowers, American ginseng and double-petaled red roses were mixed and then ultra-finely pulverized to obtain ultra-fine powder 1. After stir-frying the mung beans, they were then ultra-finely ground to obtain ultra-fine powder 2. Concentrate 1, concentrate 2, ultrafine powder 1, ultrafine powder 2, oat flour, amla juice, kale juice and sweetener are mixed and ground using a colloid mill. The ground liquid is then dried and matured to obtain the composition.

4. The preparation method according to claim 3, characterized in that, The steps for water extraction of kudzu root include: mixing kudzu root and water at a mass ratio of 1:8, decocting for 60 minutes, and then collecting the extract. After mixing the jujube seed, dandelion, asparagus, dried tangerine peel and licorice, add water at a mass ratio of 1:8 and decoct for 60 minutes. Collect the first extract and the residue. Mix the residue with water at a mass ratio of 1:8 and decoct for 30 minutes. Collect the second extract. Combine the two extracts.

5. The preparation method according to claim 3, characterized in that, The particle size of ultrafine powder 1 is 120~130μm; the particle size of ultrafine powder 2 is 120~130μm.

6. The preparation method according to claim 3, characterized in that, The grinding slurry is dried at 100~110℃.

7. Use of the composition of claim 1 or 2 or the preparation method of any one of claims 3 to 6 in the preparation of products for the prevention and / or treatment of non-alcoholic liver injury.

8. The application according to claim 7, characterized in that, The product regulates liver lipid metabolism and inhibits inflammation.

9. The use of the composition of claim 1 or 2 or the preparation method of any one of claims 3 to 6 in the preparation of products that regulate intestinal flora.

10. A product for preventing non-alcoholic liver injury, characterized in that, The product includes the composition of claim 1 or 2.

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