A composition for assisting in the protection of chemical-induced liver injury, and a preparation method and use thereof

Abstract

The application provides a composition for assisting in protecting chemical liver injury and a preparation method and application thereof, and relates to the technical field of medicines.The composition for assisting in protecting chemical liver injury provided by the application has the following raw material components: radix puerariae, ganoderma lucidum, hovenia dulcis thunb, schisandra chinensis, curcuma longa, dioscorea, glycyrrhiza uralensis, fructus jujubae, taraxacum mongolicum, chrysanthemum, polygonatum and pericarpium citri reticulatae.The components are extracted by decocting with water, the extraction liquid is mixed and dried, and the obtained composition has the function of assisting in protecting chemical liver injury, and is applied to products with the functions of improving damaged liver cells, reducing transaminase, reducing blood lipids and improving the shape of the liver, and the effect is remarkable.The composition can effectively improve the chemical liver injury of model mice in the dosage range of the components, and has similar efficacy compared with silymarin which is a clinically applied drug.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a composition for adjuvant protection against chemically induced liver injury, its preparation method, and its application. Background Technology

[0002] The liver is a vital organ involved in metabolic processes. Studies have confirmed that long-term alcohol consumption, high-fat diets, exposure to carbon tetrachloride, and abnormal drug metabolism can lead to fatty liver, which in turn causes liver damage. Liver damage is mostly caused by excessive production of free radicals and lipid peroxidation. Excessive free radical production can damage membrane structure and function, causing severe toxic damage to hepatocytes. This persistent damage gradually develops into liver fibrosis, eventually progressing to decompensated cirrhosis and even inducing malignant lesions such as primary liver cancer, constituting a complete chain reaction of liver disease progression.

[0003] Early-stage liver disease is often difficult to detect due to the lack of obvious symptoms. By the time noticeable symptoms are discovered during examination, the disease has already progressed to a severe stage. Furthermore, apart from liver transplantation, there are no medications that can reverse the gradual deterioration of the patient's condition. Liver fibrosis, however, is an early pathological manifestation and is reversible. Therefore, it is necessary to develop products that can reverse liver fibrosis and effectively protect against chemically induced liver damage.

[0004] Currently, there are many types of drugs available for treating chemically induced liver injury, primarily Western medicine. However, the long-term use of Western medicine can cause liver and kidney damage, and the recurrence of symptoms upon discontinuation, thus failing to provide protection against liver damage. Modern pharmacological research indicates that the research and development of natural medicines driven by traditional Chinese medicine theory is a feasible method for treating liver injury. Therefore, developing compositions from medicinal and edible herbs that have an auxiliary protective effect against chemically induced liver injury is an effective approach.

[0005] Chinese invention patent CN118986917A discloses a milk thistle, turmeric, and schisandra capsule for adjuvant protection against chemically induced liver injury, composed of the following ingredients: turmeric extract, milk thistle extract, schisandra extract, and a fortifying composition, wherein the fortifying composition consists of dandelion extract, artichoke leaf extract, zinc, vitamin E, and green tea extract. This capsule can prevent and treat chemically induced liver injury, reducing the damage to the liver caused by drugs and chemicals. However, current prior art only demonstrates that the simultaneous use of multiple components has a better adjuvant protective effect than a single component, without creatively studying the synergistic effect between the components, and cannot prove the advantage of the synergistic combination of components in alleviating chemically induced liver injury.

[0006] Chinese invention patent CN114521648A discloses a food composition with auxiliary protective function against chemically induced liver damage and its preparation method. The food composition comprises the following components in parts by weight: 20-25 parts kudzu root, 12-20 parts Japanese raisin tree fruit, 10-15 parts polygonatum rhizome, 4-6 parts yam, 4-6 parts jujube, 2-3 parts mung bean, 2-4 parts schisandra fruit, 2-4 parts dried tangerine peel, 1-3 parts poria cocos, 1-3 parts alisma rhizome, 1-3 parts dandelion, and 1-2 parts licorice root. This food composition indicates that kudzu root, Japanese raisin tree fruit, polygonatum rhizome, yam, and jujube are essential main active ingredients and cannot be arbitrarily replaced. It can protect the liver, improve chemically induced liver damage, repair damaged liver cells, and enhance the body's antioxidant capacity. However, in this technical field, there are many components that have a protective effect against liver damage. Further in-depth research is needed on the synergistic effects between these components in order to obtain a new composition that can synergistically enhance the effects of each component and provide auxiliary protection against chemically induced liver damage. Summary of the Invention

[0007] This invention addresses the problems existing in the prior art by providing a composition for the adjuvant protection against chemically induced liver injury, its preparation method, and its application. Through the interaction of the various components, it delivers a superior liver-protective effect compared to compositions with other conventional components, effectively protecting the liver, improving chemically induced liver injury, repairing damaged liver cells, improving liver function, reducing inflammation, and significantly alleviating gas bubble-like lesions in liver tissue.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] First, the present invention provides a composition for adjuvant protection against chemically induced liver injury, the raw material components of which include: kudzu root, Ganoderma lucidum, Japanese raisin tree fruit, Schisandra chinensis, turmeric, Chinese yam, licorice, jujube, dandelion, chrysanthemum, Polygonatum odoratum and dried tangerine peel.

[0010] Preferably, the composition, by weight, comprises the following raw material components: 20-30 parts of kudzu root, 15-22 parts of Ganoderma lucidum, 15-22 parts of Hovenia dulcis, 6-10 parts of Schisandra chinensis, 6-10 parts of turmeric, 6-10 parts of Dioscorea opposita, 3-5 parts of licorice root, 3-5 parts of jujube, 3-5 parts of dandelion, 1-3 parts of chrysanthemum, 1-4 parts of Polygonatum odoratum, and 1-3 parts of dried tangerine peel.

[0011] More preferably, the composition, by weight, comprises the following raw material components: 25-28 parts of kudzu root, 18-20 parts of Ganoderma lucidum, 18-20 parts of Hovenia dulcis, 6-8 parts of Schisandra chinensis, 6-8 parts of turmeric, 6-8 parts of Dioscorea opposita, 3-4 parts of licorice root, 3-4 parts of jujube, 3-4 parts of dandelion, 2-3 parts of chrysanthemum, 2-3 parts of Polygonatum odoratum, and 2 parts of dried tangerine peel.

[0012] More preferably, the composition, by weight, comprises the following raw material components: 28 parts kudzu root, 20 parts Ganoderma lucidum, 20 parts Hovenia dulcis, 8 parts Schisandra chinensis, 8 parts turmeric, 8 parts Dioscorea opposita, 4 parts licorice, 4 parts jujube, 4 parts dandelion, 3 parts chrysanthemum, 3 parts Polygonatum odoratum, and 2 parts dried tangerine peel.

[0013] Then, the present invention provides a method for preparing the above composition, comprising the steps of: decocting and extracting kudzu root, Ganoderma lucidum, Hovenia dulcis, Schisandra chinensis, Curcuma longa, Dioscorea opposita, Glycyrrhiza uralensis, Jujube, Taraxacum mongolicum, Chrysanthemum morifolium, Polygonatum odoratum and Citrus reticulata peel, mixing the extract and drying to obtain the composition.

[0014] Preferably, before decoction and extraction, each raw material is cleaned, washed, and dried.

[0015] Preferably, the decoction extraction specifically involves: extracting 1-3 times, with each extraction lasting 1-3 hours; more preferably, the decoction extraction specifically involves: extracting 2 times, with each extraction lasting 2 hours.

[0016] Preferably, the decoction extraction uses 8-15 times the weight of the raw material in water; more preferably, the decoction extraction uses 10 times the weight of the raw material in water.

[0017] Furthermore, the present invention provides the use of the above composition in the preparation of a medicine having the functions of assisting in the protection of chemically induced liver injury, improving damaged hepatocytes, reducing transaminase, reducing blood lipids, and improving liver morphology.

[0018] Furthermore, the present invention provides a health food with the function of assisting in the protection against chemically induced liver damage, comprising the above-mentioned composition.

[0019] Preferably, the health food further includes components: excipients acceptable in the food industry.

[0020] Finally, the present invention provides a medicine with auxiliary protective function against chemically induced liver injury, comprising the above-described composition.

[0021] Preferably, the pharmaceutical product further includes components: excipients acceptable in the pharmaceutical field.

[0022] In the composition of this invention, kudzu root, a legume, is cool in nature, sweet and pungent in taste, and enters the spleen and stomach meridians. It contains isoflavone components such as puerarin, puerarin xylose, daidzein, daidzein glycoside, β-sitosterol, and arachidic acid. It can be used to relieve exterior syndromes and reduce fever, promote body fluid production, promote rash eruption, and raise yang to stop diarrhea. It is used for exogenous fever and headache, hypertension with stiff neck, thirst, diabetes, incomplete measles eruption, dysentery, and diarrhea. Studies have shown that kudzu root can increase cerebral and coronary blood flow, protect the liver, and lower blood sugar and lipids.

[0023] Reishi mushroom, also known as Lingzhi grass, divine mushroom, zhi grass, immortal herb, and auspicious herb, is recorded in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica) as having six varieties: purple, red, blue, yellow, white, and black. It refers to the whole plant of *Ganoderma lucidum* or *Ganoderma sinense*, belonging to the Polyporaceae family. It is neutral in nature and sweet in taste, and enters the heart, lung, liver, and kidney meridians. The main chemical components of reishi mushroom include polysaccharides, nucleosides, furans, sterols, alkaloids, triterpenoids, oils, various amino acids and proteins, enzymes, and various trace elements. It is used to treat weakness, cough, asthma, insomnia, indigestion, and malignant tumors. Animal pharmacological experiments show that reishi mushroom has an inhibitory effect on the nervous system, a hypotensive effect and strengthens cardiac contractility in the circulatory system, and an expectorant effect on the respiratory system. In addition, it also has hepatoprotective, antioxidant, anti-aging, antibacterial, anticoagulant, platelet aggregation inhibitory, and immune-enhancing effects.

[0024] Hovenia dulcis seeds, as described in the *Tang Materia Medica*, are the mature seeds of *Hovenia dulcis*, *Hovenia dulcis*, and *Hovenia hairyensis*, all belonging to the Rhamnaceae family. The fruit with its inflorescence axis is also sometimes used. It has a sweet and sour taste, and is neutral in nature; it enters the spleen meridian. The chemical components of Hovenia dulcis seeds include ryegrassine, hovenia glycoside, glucose, and potassium malate, among others. Hovenia dulcis seeds have a significant diuretic effect, hovenia dulcis saponins have a hypotensive effect, and the homogenate of Hovenia dulcis seeds has anti-lipid peroxidation effects and enhances cold and heat resistance.

[0025] Schisandra chinensis, also known as Xuanji, Huiji, Wumeizi, and Shanhuajiao, is warm in nature and sour in taste. It enters the lung and kidney meridians. Its main components include deoxyschisandrin, neoschisandrin, schisandrol, and schisandrin ester. Its main functions are astringency, replenishing qi and promoting body fluid production, tonifying the kidneys and calming the mind. It has excitatory effects on both the nervous and respiratory systems, influencing both excitation and inhibition processes in the cerebral cortex, thus balancing them. It has antitussive, expectorant, blood pressure lowering, transaminase lowering, and choleretic effects, and protects liver cells. It enhances the body's defense against non-specific stimuli, increases cellular immune function, and significantly enhances superoxide dismutase activity, thus possessing high immunomodulatory, antioxidant, and anti-aging effects. It also has inhibitory effects on Staphylococcus aureus, Klebsiella pneumoniae, Salmonella enterica, and Pseudomonas aeruginosa.

[0026] Turmeric is the dried rhizome of the turmeric plant (Zingiberaceae family). It is pungent, bitter, and warm in nature; it enters the spleen and liver meridians. It contains volatile oils, the main components of which are curcumin, aromatic curcumin, gingerene, phellandrene, sapindus mucilage, eucalyptol, curcumone, curcumol, caryophyllene, borneol, camphor, curcumin, and demethoxycurcumin. It has lipid-lowering, anti-tumor, anti-inflammatory, choleretic, antioxidant, and antihypertensive effects, while also protecting the gastric mucosa and liver cells.

[0027] Yam is neutral in nature and sweet in taste; it enters the spleen, lung, and kidney meridians. It contains diosgenin, mucilage, choline, starch, glycoprotein, free amino acids, vitamin C, amylase, etc. It is mainly used for spleen deficiency with poor appetite, fatigue, loose stools and diarrhea, lung deficiency with cough and asthma, kidney deficiency with seminal emission, leukorrhea and frequent urination, and internal heat and thirst.

[0028] Jujubes are warm in nature and sweet in taste; they enter the spleen, stomach, and heart meridians. They contain organic acids, triterpenoid glycosides, alkaloids, flavonoids, sugars, vitamins, amino acids, volatile oils, and trace elements. They are mainly used to tonify the spleen and stomach, replenish qi and promote body fluid production, and detoxify. They treat poor appetite due to stomach deficiency, loose stools due to spleen weakness, insufficient qi, blood, and body fluids, disharmony between the nutritive and defensive qi, and palpitations.

[0029] Dandelion is the dried whole herb of *Taraxacum officinale*, *Taraxacum mongolicum*, or several other species in the same genus, belonging to the Asteraceae family. It is cold in nature and bitter and sweet in taste; it enters the liver and stomach meridians. It contains taraxasterol, taraxacin, taraxacin, inositol, lactucin, taraxerol, caffeic acid, and resins, etc. It is mainly used for clearing heat and detoxifying, reducing swelling and dissipating nodules, promoting urination and relieving strangury, treating boils and carbuncles, mastitis, scrofula, red eyes, sore throat, lung abscess, intestinal abscess, damp-heat jaundice, and painful urination due to heat. In addition, dandelion has the effect of clearing the liver and improving eyesight, and is used to treat red and swollen eyes caused by liver fire.

[0030] Licorice is the dried root and rhizome of *Glycyrrhiza uralensis*, *Glycyrrhiza inflata*, or *Glycyrrhiza glabra*, all belonging to the genus *Glycyrrhiza* of the legume family. It is neutral in nature and sweet in taste; it enters the heart, lung, spleen, and stomach meridians. Licorice contains many chemical components, primarily glycyrrhizin and glycyrrhetinic acid. It tonifies the spleen and replenishes qi, clears heat and detoxifies, resolves phlegm and relieves cough, alleviates spasms and pain, and harmonizes the properties of other herbs. It is used for spleen and stomach weakness, fatigue, palpitations, shortness of breath, cough with excessive phlegm, abdominal and limb spasms and pain, carbuncles and boils, and to alleviate the toxicity and harshness of other medications. It has the effects of tonifying qi and replenishing the middle jiao, moistening the lungs and relieving cough, clearing heat and detoxifying, relieving spasms and pain, and harmonizing the properties of other herbs.

[0031] Chrysanthemum, also known as sweet chrysanthemum, white chrysanthemum, yellow chrysanthemum, and medicinal chrysanthemum, is cool in nature and has a sweet and bitter taste; it enters the lung and liver meridians. Its chemical composition includes volatile oil, the main components of which are borneol, camphor, and chrysanthemum cyclic ketones. It also contains chrysanthemin, adenine, choline, flavonoids, stachydrine, vitamin A, vitamin B1, vitamin E, amino acids, and robinin, among other components. It is used to dispel wind and clear heat, calm the liver, and improve eyesight. It is used for wind-heat colds, headaches, dizziness, red and swollen eyes, and blurred vision.

[0032] Polygonatum odoratum, the rhizome of the plant Polygonatum odoratum in the Liliaceae family, is sweet in taste and neutral in nature; it enters the lung and stomach meridians. Its chemical components include steroidal saponins, flavonoids, and their glycosides. It nourishes yin and moistens dryness, promotes body fluid production and quenches thirst. It is used for lung and stomach yin deficiency, dry cough due to heat, dry throat and thirst, and internal heat with thirst.

[0033] Dried tangerine peel (Chenpi) is the dried, mature peel of the citrus fruit (Citrus reticulata) and its cultivated varieties, belonging to the Rutaceae family. It is warm in nature and has a pungent and bitter taste; it enters the spleen and lung meridians. Dried tangerine peel contains hesperidin, hesperidin, neohesperidin, p-hydroxyphosphonic acid, flavonoids, and other components. It has the functions of regulating qi and strengthening the spleen, drying dampness and resolving phlegm. It is used for chest and abdominal distension, poor appetite, vomiting and diarrhea, and cough with excessive phlegm.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] 1. In the composition of the present invention, kudzu root, Ganoderma lucidum, Japanese raisin tree fruit, Schisandra chinensis, turmeric, Chinese yam, licorice, jujube, dandelion, chrysanthemum, Polygonatum odoratum and dried tangerine peel are used in combination. Through scientific and specific mixing and matching, the components interact with each other, bringing about a synergistic effect to help protect against chemical liver damage. It significantly reduces the serum alanine aminotransferase and aspartate aminotransferase levels in mice with liver damage, reduces the serum triglyceride, cholesterol and low-density lipoprotein levels, and increases the high-density lipoprotein level.

[0036] 2. The composition of the present invention, through the interaction of its components, provides a superior liver-protective effect compared to compositions with other conventional components, effectively protecting the liver, improving chemically induced liver damage, repairing damaged liver cells, improving liver function, reducing inflammation, and significantly alleviating gas bubble-like lesions in liver tissue.

[0037] 3. The composition of the present invention addresses the root cause of liver stagnation by clearing and nourishing the liver, and addresses the symptoms by tonifying the kidneys and replenishing essence, regulating qi and strengthening the spleen, thus treating both the root cause and the symptoms. The composition of the present invention is derived from medicinal and edible herbs, with a wide range of raw material sources, low cost, and safety and non-toxicity. Moreover, the formula is concise and reasonable, the production process is simple, suitable for industrialization, long-term use will not cause a decrease in patient tolerance, the effect is mild and long-lasting, and it is safer and suitable for daily use, providing new ideas for the further development of products with liver-protective effects. Attached Figure Description

[0038] Figure 1 This is a diagram showing the effect of the composition of the present invention on the pathological morphology of mouse liver tissue.

[0039] Figure 2 The figure shows the effect of different dosages of the composition in Example 1 on the pathological morphology of mouse liver tissue. Detailed Implementation

[0040] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0041] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0042] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention are obtained through conventional commercial channels. Products from different manufacturers do not have a significant impact on the effectiveness.

[0043] Example 1

[0044] A composition for adjuvant protection against chemically induced liver injury, comprising, by weight, the following ingredients: 28 parts kudzu root, 20 parts Ganoderma lucidum, 20 parts Hovenia dulcis, 8 parts Schisandra chinensis, 8 parts turmeric, 8 parts Dioscorea opposita, 4 parts licorice root, 4 parts jujube, 4 parts dandelion, 3 parts chrysanthemum, 3 parts Polygonatum odoratum, and 2 parts dried tangerine peel.

[0045] Preparation method of the composition: Mix the above raw materials, then add water in 10 times the mass of the raw materials, decoct and extract twice, each time for 2 hours, to obtain the extract; mix the extracts and freeze dry to obtain the composition.

[0046] Example 2

[0047] A composition for adjuvant protection against chemically induced liver injury, comprising, by weight, the following ingredients: 25 parts kudzu root, 18 parts Ganoderma lucidum, 18 parts Hovenia dulcis, 6 parts Schisandra chinensis, 6 parts turmeric, 6 parts Dioscorea opposita, 3 parts licorice root, 3 parts jujube, 3 parts dandelion, 2 parts chrysanthemum, 2 parts Polygonatum odoratum, and 2 parts dried tangerine peel.

[0048] The composition was prepared using the same method as in Example 1.

[0049] Example 3

[0050] A composition for adjuvant protection against chemically induced liver injury, comprising, by weight, the following ingredients: 20 parts kudzu root, 15 parts Ganoderma lucidum, 15 parts Hovenia dulcis, 6 parts Schisandra chinensis, 6 parts turmeric, 6 parts Dioscorea opposita, 3 parts licorice root, 3 parts jujube, 3 parts dandelion, 1 part chrysanthemum, 1 part Polygonatum odoratum, and 1 part dried tangerine peel.

[0051] The composition was prepared using the same method as in Example 1.

[0052] Example 4

[0053] A composition for adjuvant protection against chemically induced liver injury, comprising, by weight, the following ingredients: 30 parts kudzu root, 22 parts Ganoderma lucidum, 22 parts Hovenia dulcis, 10 parts Schisandra chinensis, 10 parts turmeric, 10 parts Dioscorea opposita, 5 parts licorice root, 5 parts jujube, 5 parts dandelion, 3 parts chrysanthemum, 4 parts Polygonatum odoratum, and 3 parts dried tangerine peel.

[0054] The composition was prepared using the same method as in Example 1.

[0055] Comparative Example 1

[0056] Unlike Example 1, Ganoderma lucidum was replaced with Polygonatum sibiricum.

[0057] A composition for adjuvant protection against chemically induced liver injury, comprising, by weight, the following ingredients: 28 parts kudzu root, 20 parts polygonatum rhizome, 20 parts jujube fruit, 8 parts schisandra fruit, 8 parts turmeric, 8 parts yam, 4 parts licorice root, 4 parts jujube, 4 parts dandelion, 3 parts chrysanthemum, 3 parts polygonatum rhizome, and 2 parts dried tangerine peel.

[0058] Preparation method of the composition: Mix the above raw materials, then add water in 10 times the mass of the raw materials, decoct and extract twice, each time for 2 hours, to obtain the extract; mix the extracts and freeze dry to obtain the composition.

[0059] Comparative Example 2

[0060] Unlike Example 1, Solomon's Seal is replaced with mung bean.

[0061] A composition for adjuvant protection against chemically induced liver injury, comprising, by weight, the following ingredients: 28 parts kudzu root, 20 parts Ganoderma lucidum, 20 parts Hovenia dulcis, 8 parts Schisandra chinensis, 8 parts turmeric, 8 parts Dioscorea opposita, 4 parts licorice root, 4 parts jujube, 4 parts dandelion, 3 parts chrysanthemum, 3 parts mung bean, and 2 parts dried tangerine peel.

[0062] Preparation method of the composition: Mix the above raw materials, then add water in 10 times the mass of the raw materials, decoct and extract twice, each time for 2 hours, to obtain the extract; mix the extracts and freeze dry to obtain the composition.

[0063] Comparative Example 3

[0064] A Polygonatum extract: Polygonatum is mixed with 10 times its weight of water, decocted and extracted twice, each time for 2 hours, to obtain an extract; the extracts are mixed and freeze-dried to obtain the Polygonatum extract.

[0065] Comparative Example 4

[0066] A Ganoderma lucidum extract: Ganoderma lucidum is mixed with 10 times its weight of water, decocted and extracted twice, each time for 2 hours, to obtain an extract; the extracts are mixed and freeze-dried to obtain the Ganoderma lucidum extract.

[0067] Comparative Example 5

[0068] Unlike Example 1, the weight ratio of the raw materials in the composition is different:

[0069] The composition is made from the following raw materials in parts by weight: 28 parts kudzu root, 10 parts Ganoderma lucidum, 20 parts Japanese raisin tree fruit, 4 parts Schisandra chinensis, 12 parts turmeric, 8 parts Chinese yam, 4 parts licorice, 4 parts jujube, 4 parts dandelion, 8 parts chrysanthemum, 8 parts Polygonatum odoratum and 2 parts dried tangerine peel.

[0070] Everything else is the same as in Example 1.

[0071] Experiment 1: Animal Experiment

[0072] Carbon tetrachloride liver injury model

[0073] 1. Principle

[0074] Carbon tetrachloride (CCl4) is activated by hepatic microsomal enzymes to become chloroform free radicals (CCl3•), which covalently bind to proteins, leading to impaired protein synthesis and disordered lipid metabolism, resulting in the accumulation of triglycerides (TG) in hepatocytes. Chloroform free radicals can rapidly combine with O2 to transform into chloroform peroxide free radicals (CCl3O2•), causing lipid peroxidation, which in turn causes cell membrane degeneration and damage, leading to enzyme leakage and various types of cytopathic effects, even necrosis.

[0075] 2. Laboratory animals

[0076] C57BL / 6J mice (weighing 18-22 grams), male, purchased from Beijing Huafukang Biotechnology Co., Ltd., SPF grade, all mice were approved by the Laboratory Animal Management and Use Committee of the Affiliated Hospital of Inner Mongolia Medical University (Y.2025014).

[0077] 2.1 Experimental methods and procedures

[0078] 2.1.1 Dosage grouping and modeling method

[0079] The experiment consisted of 11 groups: blank group, model group, Example 1-Example 4 groups, and Comparative Example 1-Comparative Example 5 groups, with 10 mice in each group. Except for the blank group, a liver injury model was established using CCl4 (analytical grade) via gavage. The concentration of CCl4 administered to mice via gavage was 1%, diluted with edible vegetable oil, at a volume of 5 mL / kg BW (equivalent to a CCl4 dose of 80 mg / kg BW).

[0080] 2.1.2 Experimental Procedure

[0081] After successful modeling, the example and comparative groups were administered the corresponding compositions orally at a dose of 0.5 g / kg daily via gavage; the blank and model groups were administered the same dose of distilled water for 30 consecutive days. Animals were weighed twice weekly to adjust the gavage dose. On day 30 of the experiment, all animals were fasted overnight for 16 hours. The model group, example and comparative groups were administered CCl4 (5 mL / kg of 1% CCl4 in edible vegetable oil solution) via gavage once, while the blank group was administered the same amount of edible vegetable oil via gavage. The example and comparative groups continued to receive the compositions until the end of the experiment (with an interval of at least 4 hours between CCl4 gavage).

[0082] Finally, after administering CCl4, the animals were euthanized after 48 hours, blood was collected to separate serum, and the liver was placed in a 4% paraformaldehyde solution.

[0083] Data Analysis

[0084] All experimental data were analyzed by t-test using GraphPad Prism5 software, and the data are expressed as mean ± standard deviation (SD).

[0085] 2.2 Measurement Indicators

[0086] Liver function and blood lipid level tests: The serum separated after the experiment was tested to detect the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), cholesterol (TC), high-density lipoprotein (HDLC) and low-density lipoprotein (LDLC).

[0087] Liver index detection: Weigh the mice and measure their liver weight, and calculate the liver index.

[0088] Liver histopathological examination: Fresh liver tissue was fixed in 4% paraformaldehyde solution, embedded in the tissue, and stained with hematoxylin and eosin (HE).

[0089] 2.3 Experimental Results

[0090] 2.3.1 Effects on serum AST, ALT, TG, TC, HDLC and LDLC in mice

[0091] The results of AST and ALT levels in different groups of mice are shown in Table 1, and the results of TG, TC, HDLC, and LDLC are shown in Table 2. Compared with the blank group, the model group showed significantly increased AST, ALT, TG, TC, and LDLC, indicating that the model caused liver damage and significantly affected liver lipid metabolism function. All groups in this invention improved lipid metabolism disorders to varying degrees, suggesting that this invention has a therapeutic effect on lipid metabolism disorders caused by chemical liver injury. The example groups improved liver damage, with Example 1 showing the most significant improvement. Although the comparative group showed some differences compared to the model group, it was not as effective as the examples. This suggests that Example 1 is the optimal formulation. Replacing any component and reformulating the formulation (Comparative Examples 1 and 2), or changing the ratio of raw materials (Comparative Example 5), did not improve liver damage as well as the formulation of this invention, confirming the rationality of the formulation and its significant liver-protective effect.

[0092] Furthermore, as can be seen from the technical effects of Comparative Examples 3 and 4, the effects of Polygonatum extract and Ganoderma lucidum extract are similar. However, in the overall composition of this invention, due to the interaction between Ganoderma lucidum and other components of the composition, it achieves a better effect in reducing AST, ALT, TG, and TC content than the composition of Comparative Example 1 which uses Polygonatum. The synergistic effect of the components in the composition of this application, resulting in a significant overall improvement in technical effect, was not expected.

[0093] Table 1

[0094]

[0095] In Table 1, ++++ P < 0.0001, indicating a significant difference compared to the control group; ## P < 0.01, ### P < 0.001, #### P < 0.0001, indicating a significant difference compared to the model group; & P < 0.05 && P < 0.01, &&& P < 0.001, &&&& P < 0.0001, indicating that each comparative group showed a significant difference compared to Example 1.

[0096] Table 2

[0097]

[0098] In Table 2, ++++ P < 0.0001, indicating a significant difference compared to the control group; ## P < 0.01, ### P < 0.001, #### P < 0.0001, indicating a significant difference compared to the model group; & P < 0.05 && P < 0.01, &&& P < 0.001, indicating that each comparative group showed a significant difference compared to Example 1; ns: no statistically significant difference compared to the model group.

[0099] 2.3.2 Effects on liver weight and liver index in mice

[0100] On the last day of the experiment, the mice were weighed, and their livers were quickly removed after the experiment. The liver weight and liver organ index were recorded, and the results are shown in Table 3. Compared with the blank group, the model group mice had heavier livers and higher liver indexes, indicating that the liver was severely damaged and enlarged after modeling. Compared with the model group, the example significantly reduced the liver weight and liver index of mice. Although the comparative example also reduced the liver weight and liver index of mice, there was no statistically significant difference. This suggests that the composition of the present invention can significantly improve the interference of drug-induced liver injury on the liver without causing liver damage.

[0101] Table 3

[0102]

[0103] In Table 3, + P < 0.05+++ P < 0.001, indicating a significant difference compared to the control group; # P < 0.05 ## P < 0.01, compared with the model group; ns: no statistically significant difference compared with the model group.

[0104] 2.3.3 Effects on liver histopathology of chemically induced liver injury

[0105] The fixed liver tissue was removed and stained with hematoxylin and eosin (HE) to observe the effects of Examples 1-3 on the chemical liver injury model. The results are shown in […]. Figure 1 Observation of the entire tissue section under a 40x objective lens revealed that in the control group, hepatocytes in the liver tissue were arranged in cords, radiating around the central vein. The hepatocyte cords were branched and anastomosed to form a network, with normal hepatocyte structure. Compared with the control group, the liver tissue sections of the model group showed increased hepatocyte volume and swelling, with prominent nuclei and indistinct cell boundaries; the hepatocyte nuclei were enlarged, located in the center of the cell, with loose cytoplasm, and some cytoplasm was empty and shiny, forming balloon-like lesions. Compared with the model group, Examples 1-3 alleviated the above conditions, with Example 1 showing the most significant reduction in balloon-like lesions in the liver tissue.

[0106] Experiment 2 Dosage Validation

[0107] The technical effects were verified according to the experimental methods under "Experiment 1 Animal Experiment".

[0108] The C57BL / 6J mice (weighing 18-22 grams), male, were purchased from Beijing Huafukang Biotechnology Co., Ltd., SPF grade, and all mice were approved by the Experimental Animal Management and Use Committee of the Affiliated Hospital of Inner Mongolia Medical University (Y.2025014). The mice were divided into 6 groups of 10 mice each: blank group, model group, low-dose group of Example 1, medium-dose group of Example 1, high-dose group of Example 1, and positive control group (silymarin). The positive control group (silymarin capsules) was purchased from Tianjin Tasly Shengte Pharmaceutical Co., Ltd.

[0109] In Example 1, the low-dose group of the composition was administered via gavage at a dose of 0.25 g / kg.

[0110] In Example 1, the dosage group of the composition administered via gavage was 0.5 g / kg;

[0111] In Example 1, the high-dose group of the composition was administered via gavage at a dose of 1.0 g / kg.

[0112] The oral dose for the positive control group was 54.6 mg / kg.

[0113] 1. Effects on serum AST, ALT, TG, TC, HDLC and LDLC levels in mice

[0114] The results of AST and ALT levels in different groups of mice are shown in Table 4, and the results of TG, TC, HDLC, and LDLC are shown in Table 5. Compared with the blank group, the model group showed significantly increased AST, ALT, TG, TC, and LDLC, indicating that the model caused liver damage and significantly affected liver lipid metabolism. Compared with the model group, the low, medium, and high doses of the composition in Example 1 and the positive control group improved the degree of liver damage to varying degrees. Among them, the medium dose and the positive control group in Example 1 showed the most significant improvement, and the effect of the medium dose group in Example 1 was comparable to that of the positive control group.

[0115] Table 4

[0116]

[0117] In Table 4, ++++ P < 0.0001, indicating a significant difference compared to the control group; ### P < 0.001, #### P < 0.0001, indicating a significant difference compared to the model group; ns: no statistically significant difference compared to the model group.

[0118] Table 5

[0119]

[0120] In Table 5, ++++ P < 0.0001, indicating a significant difference compared to the control group; ### P < 0.001, #### P < 0.0001, indicating a significant difference compared to the model group; ns: no statistically significant difference compared to the model group.

[0121] 2. Effects on liver weight and liver index in mice

[0122] On the last day of the experiment, the mice were weighed. After the experiment, the livers of the mice were quickly removed, and the liver weight and liver organ index were recorded. The results are shown in Table 6. Compared with the blank group, the livers of the model group mice were heavier and the liver index was higher, indicating that the liver was severely damaged and enlarged after modeling. Compared with the model group, each dose could significantly reduce the liver weight of mice. The medium and high doses of the composition could significantly reduce the liver index of mice, suggesting that the composition of the present invention can significantly improve the interference of drug-induced liver injury on the liver without causing damage to the liver.

[0123] Table 6

[0124]

[0125] In Table 6, ++ P < 0.01, +++ P < 0.001, indicating a significant difference compared to the control group; ## P < 0.01, ### P < 0.001, indicating a significant difference compared to the model group; ns: no statistically significant difference compared to the model group.

[0126] 3. Effects on liver histopathology in chemically induced liver injury

[0127] The fixed liver tissue was removed and stained with hematoxylin and eosin (HE) to observe the effects of different doses of the composition from Example 1 on a chemically induced liver injury model. The results are shown in [Figure Number]. Figure 2 Observation of the entire tissue section under a 40x objective lens revealed that in the control group, hepatocytes in the liver tissue of mice were arranged in cords, radiating around the central vein. The hepatocyte cords were branched and anastomosed to form a network, and the hepatocyte structure was normal. Compared with the control group, the liver tissue sections of the model group showed increased hepatocyte volume and swelling, with prominent nuclei and indistinct cell boundaries; the hepatocyte nuclei were enlarged, located in the center of the cell, and the cytoplasm was loose, with some cells exhibiting a clear cytoplasm, forming balloon-like lesions. Compared with the model group, each dose of Example 1 could alleviate the above conditions, with the dose in Example 1 showing the most significant reduction in balloon-like lesions in the liver tissue.

[0128] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A composition for adjuvant protection against chemically induced liver injury, characterized in that, By weight, the raw material components include: 20-30 parts kudzu root, 15-22 parts Ganoderma lucidum, 15-22 parts Hovenia dulcis, 6-10 parts Schisandra chinensis, 6-10 parts turmeric, 6-10 parts Dioscorea opposita, 3-5 parts licorice, 3-5 parts jujube, 3-5 parts dandelion, 1-3 parts chrysanthemum, 1-4 parts Polygonatum odoratum, and 1-3 parts dried tangerine peel.

2. The composition according to claim 1, characterized in that, By weight, the raw material components include: 25-28 parts kudzu root, 18-20 parts Ganoderma lucidum, 18-20 parts Hovenia dulcis, 6-8 parts Schisandra chinensis, 6-8 parts turmeric, 6-8 parts Dioscorea opposita, 3-4 parts licorice, 3-4 parts jujube, 3-4 parts dandelion, 2-3 parts chrysanthemum, 2-3 parts Polygonatum odoratum, and 2 parts dried tangerine peel.

3. The composition according to claim 2, characterized in that, By weight, the raw material components include: 28 parts kudzu root, 20 parts Ganoderma lucidum, 20 parts Hovenia dulcis, 8 parts Schisandra chinensis, 8 parts turmeric, 8 parts Dioscorea opposita, 4 parts licorice, 4 parts jujube, 4 parts dandelion, 3 parts chrysanthemum, 3 parts Polygonatum odoratum, and 2 parts dried tangerine peel.

4. A method for preparing the composition according to any one of claims 1-3, characterized in that, The steps include: decocting and extracting kudzu root, Ganoderma lucidum, Japanese raisin tree fruit, Schisandra chinensis, turmeric, Chinese yam, licorice, jujube, dandelion, chrysanthemum, Polygonatum odoratum, and dried tangerine peel; mixing and drying the extracts to obtain the composition.

5. The preparation method according to claim 4, characterized in that, The decoction extraction specifically involves extracting 1-3 times, with each extraction lasting 1-3 hours.

6. The preparation method according to claim 5, characterized in that, The decoction extraction process specifically involves two extractions, each lasting 2 hours.

7. The preparation method according to claim 4, characterized in that, The decoction extraction is performed using 8-15 times the weight of the raw material in water.

8. The use of the composition according to any one of claims 1-3 in the preparation of a medicament having the functions of assisting in the protection against chemically induced liver injury, improving damaged hepatocytes, reducing transaminase levels, reducing blood lipids, and improving liver morphology.

9. A health food product with auxiliary protective function against chemically induced liver damage, characterized in that, Composition comprising: the composition according to any one of claims 1-3.

10. A drug with auxiliary protective function against chemically induced liver injury, characterized in that, Composition comprising: the composition according to any one of claims 1-3.

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