Application of Radix Glehniae Herba in preparing medicine for preventing and / or treating liver injury

Abstract

The invention discloses the use of Radix Angelicae Pubescentis in preparing a drug for preventing and / or treating liver injury, and belongs to the technical field of traditional Chinese medicine. The application is the use of Radix Angelicae Pubescentis, its extract or its pharmaceutical composition in preparing a drug for preventing and / or treating liver injury. Experiments have shown that Radix Angelicae Pubescentis can reduce substantial damage such as hepatocyte inflammatory reaction and fatty degeneration, effectively reduce AST and AST levels in serum, and has a good preventive and / or therapeutic effect on liver injury.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicines, and relates to the use of Radix Glechomae Arnebiae in preparing a medicine for preventing and / or treating liver injury, and specifically relates to the use of Radix Glechomae Arnebiae, an extract thereof or a pharmaceutical composition thereof in preparing a medicine for preventing and / or treating liver injury. Background Art

[0002] The liver is the largest digestive gland and detoxification organ in the human body. It is the central link of the body's material and energy metabolism and an important organ for maintaining life activities. Liver disease is a common and extremely harmful disease in clinical practice, with a high mortality rate. Acute liver injury (ALI) refers to sudden liver cell damage and liver function abnormalities caused by various factors in a short period of time, characterized by metabolic disorders, imbalance of internal environment homeostasis and loss of liver function. Long-term or severe liver damage can form liver fibrosis, further develop into cirrhosis or liver cancer and endanger life. Therefore, it is of great significance to seek safe and effective drugs to protect the liver and intervene in the progression of acute injury.

[0003] Carbon tetrachloride (CCl4)-induced acute liver injury in rat models is widely used to study potential therapeutic strategies due to its similarities to acute chemical liver injury in humans.

[0004] Semiliquidambar cathayensis HTChang. is a plant of the Hamamelidaceae family. It is mainly distributed in southern and southeastern mountainous areas of China, such as Guangxi, Guangdong, Guizhou, etc. It has the effects of dispelling wind and dampness, relaxing muscles and activating blood circulation, and can be used to treat rheumatoid arthritis, traumatic injuries, postpartum paralysis, etc. Modern pharmacological studies have shown that Semiliquidambar cathayensis HTChang. has the effects of analgesia, anti-inflammatory, antioxidant, and blood circulation.

[0005] CN106822434A and CN106822426 disclose a traditional Chinese medicine formula, which includes 50-150 grams of flower snake, 10-50 grams of Chuanxiong, 10-50 grams of Sichuan papaya, 10-50 grams of Qiannianjian, 10-30 grams of processed Chuanwu, 10-50 grams of Gentiana macrophylla, 20-60 grams of Sichuan Cyathula, 30-80 grams of Banfenghe, and 50-150 grams of crystal sugar, and claims that it can treat hepatitis A and hepatitis B. CN103550513A discloses a Chinese medicine for treating cholestatic hepatitis, which is prepared from the following medicinal materials in weight ratio: 14 parts of Solanum cumini, 12 parts of Elsholtzia ciliata, 8 parts of Rhizoma Cibotii, 10 parts of Rhizoma Cibotii, 14 parts of Glehnia littoralis, 14 parts of Pinecones, 16 parts of Herba Catharinae, 8 parts of Herba Lycopodii, 14 parts of Herba Aurantii, 13 parts of Herba Danbulao, 16 parts of Herba Tamarindi, 15 parts of Herba Coriolus, 12 parts of Herba Bone-bone-dandruff, 14 parts of Herba Eupatorii, 8 parts of Herba Lycopodii, 10 parts of Herba Glechomae, and 12 parts of Herba Aurantii. It claims that the invention can safely and effectively treat cholestatic hepatitis.

[0006] CN103356886A discloses a Chinese medicine for treating primary biliary cirrhosis, which is prepared from the following medicinal materials in weight ratio: 12 parts of golden cherry, 14 parts of Scrophularia, 6 parts of wolfberry, 10 parts of mountain leech, 12 parts of kudzu, 14 parts of fragrant sedge, 10 parts of yellow cassia, 14 parts of bupleurum, 10 parts of cassia seed, 8 parts of cyperus, 14 parts of half maple, 10 parts of Baba flower, 14 parts of Baofuqi, 12 parts of Jinjinxiang, 14 parts of Ganhuangcao, 10 parts of Abacus, 8 parts of water fennel, 10 parts of radix amarantii, and 10 parts of white cabbage. It is claimed that the invention can safely and effectively treat primary biliary cirrhosis. CN105168353A discloses a Banfenghe wine, which is made from Banfenghe, cassia twigs and white wine. It claims that the Banfenghe wine provided by it has the effects of dispelling wind and dampness, promoting blood circulation and reducing swelling, nourishing yin and tonifying the kidney, nourishing the liver and spleen, and is mainly used to treat rheumatic joint pain, lumbar muscle strain, and liver and kidney yin deficiency.

[0007] At present, the research on the effect of Radix Glechomae Arnebiae on the liver is to use Radix Glechomae Arnebiae in combination with traditional Chinese medicine with liver protection effect, and there is a lack of corresponding mechanism and effect verification research, so it is difficult to measure its effectiveness. After searching, there are currently no relevant reports and applications of Radix Glechomae Arnebiae Arnebiae on liver damage. Summary of the invention

[0008] The purpose of the present invention is to provide the use of Radix Angelicae Pubescentis in the preparation of a drug for preventing and / or treating liver injury; in particular, the use of Radix Angelicae Pubescentis, its extract or its pharmaceutical composition in the preparation of a drug for preventing and / or treating liver injury. The Radix Angelicae Pubescentis of the present invention can reduce substantial liver damage such as hepatocyte inflammatory response and fatty degeneration, effectively reduce AST and AST levels in serum, and has a good preventive and therapeutic effect on liver injury.

[0009] In order to achieve the above object, the present invention adopts the following technical solutions:

[0010] The present invention provides application of Radix Angelicae Pubescentis, an extract thereof or a pharmaceutical composition thereof in preparing a drug for preventing and / or treating liver damage.

[0011] The herb of Hamamelidaceae comes from the herb of Hamamelidaceae (Semiliquidambarcathayensis HTChang.), and its roots, stems, branches or leaves are used as medicine, which are dried separately for later use.

[0012] The extract involved in the present invention is an extract of golden vine half maple, wherein the extraction comprises at least one of water decoction method, immersion method, percolation method, improved gelatin method, reflux method, solvent extraction method, steam distillation method, sublimation method, supercritical fluid extraction method, membrane separation technology, ultrafine grinding technology, traditional Chinese medicine flocculation separation technology, semi-bionic extraction method, ultrasonic extraction method, cyclone extraction method, pressurized countercurrent extraction method, enzyme method, macroporous resin adsorption method, ultrafiltration method and molecular distillation method.

[0013] In some specific embodiments, the extract is an alcohol extract of Herba Lycopodii.

[0014] Specifically, the preparation method of the alcohol extract is as follows: adding the Herba Lycopodii to alcohol for extraction, separating the solid and the liquid, and concentrating the filtrate to obtain the alcohol extract.

[0015] The volume fraction of the alcohol in this embodiment is 10-90%, preferably 30-80%, more preferably 45-75%, further preferably 50-75%, and most preferably 70%.

[0016] The alcohol in this embodiment is ethanol and / or methanol; preferably ethanol.

[0017] The solid-liquid separation described in this embodiment includes filtration and centrifugation, preferably filtration.

[0018] In some specific embodiments, the preparation method of the extract comprises the following steps: mixing the Herba Lycopodii with ethanol and refluxing for extraction, and concentrating the combined filtrate to obtain the extract.

[0019] Preferably, the amount of ethanol added is 5-25 times the mass of the Herba Lycopodii, preferably 15 times.

[0020] Preferably, the extraction is performed 1-3 times, each time for 0.5-2.5 hours; more preferably, the extraction is performed 2 times, each time for 1.5 hours.

[0021] The pharmaceutical composition of the present invention comprises Radix Glechomae var. sylvestris or its extract and pharmaceutically acceptable excipients.

[0022] The dosage form of the pharmaceutical composition of the present invention is granules, tablets, capsules, powders, pills, suspensions or liquid preparations.

[0023] In some specific embodiments, the pharmaceutical composition comprises other liver-protecting drugs, which serve as the second active ingredient and act together with the Radix Angelicae Pubescentis or its extract to exert a liver-protecting effect.

[0024] The other liver-protecting drugs described in the present invention are chemical drugs, traditional Chinese medicines or Chinese patent medicine preparations.

[0025] The pharmaceutical composition of the present invention is that the Herba Lysimachiae, its extract or its pharmaceutical composition and other liver-protecting drugs are directly mixed together or are combined after being independently packaged.

[0026] The liver injury described in the present invention is acute liver injury; preferably acute chemical liver injury, drug-induced liver injury and alcoholic liver injury.

[0027] In some specific embodiments, the prevention and / or treatment of liver injury includes at least one of the following features:

[0028] (a) Reduce liver coefficient;

[0029] (b) reduce serum ALT and / or AST levels;

[0030] (c) Reduce substantial liver damage, including hepatocellular inflammation and / or steatosis.

[0031] The beneficial effects of the present invention are:

[0032] Experiments have shown that the Herba Artemisiae Argyi of the present invention can reduce substantial damage such as inflammatory response and fatty degeneration of liver cells, effectively reduce AST and AST levels in serum, and has a good preventive and therapeutic effect on liver damage, especially acute chemical liver damage, drug-induced liver damage and alcoholic liver damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 The results are as follows: The effect of Radix Glehniae on serum ALT in rats induced by CCl4;

[0034] Figure 2 The results are as follows: the effect of Radix Glehniae on serum AST level in rats induced by CCl4;

[0035] Figure 3 The results of the pathological observation of liver tissue sections of rats induced by Radix Glehniae Banfenghe on CCl4;

[0036] Figure 4 This is the scoring result of Jinlu Banfenghe on the fatty degeneration grade of hepatocytes in rats induced by CCl4;

[0037] Figure 5 The results are as follows: the effect of Radix Glehniae on serum ALT in rats induced by alcohol;

[0038] Figure 6 The results are as follows: The effect of Radix Glehniae on serum AST level in rats induced by alcohol;

[0039] Figure 7 The results of the pathological observation of liver tissue sections of rats induced by alcohol by Jinlu Banfenghe.

[0040] Figure 8 This is the scoring result of Jinlu Banfenghe on the grade of fatty degeneration of hepatocytes in rats induced by alcohol;

[0041] Fig. 9 This is the effect of Radix Glehniae on serum ALT in drug-induced rats;

[0042] Fig.10 These are the results of the effects of Radix Glehniae on serum AST levels in drug-induced rats. DETAILED DESCRIPTION

[0043] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0044] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing specific embodiments rather than for limiting the scope of protection of the present invention.

[0045] When the embodiment gives a numerical range, it should be understood that, unless otherwise specified in the present invention, the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those of ordinary skill in the art to which the present invention belongs.

[0046] The present invention does not limit the sources of the raw materials used. Unless otherwise specified, the raw materials used in the present invention are common commercial products in the technical field.

[0047] The "pharmaceutical composition" described in the present invention comprises Herba Lycopodii or its extract and pharmaceutically acceptable excipients. In a specific embodiment, Herba Lycopodii or its extract described in the present invention is provided in the pharmaceutical composition in an effective amount (e.g., a therapeutically effective amount).

[0048] The "pharmaceutically acceptable" ingredients described in the present invention are substances that are suitable for human beings and / or animals without excessive adverse side effects (such as toxicity, irritation and allergic reactions), that is, substances with a reasonable benefit / risk ratio. The "pharmaceutically acceptable excipients" include inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or oils. Excipients (such as cocoa butter and suppository wax), colorants, coating agents, sweeteners and flavoring agents may also be present in the pharmaceutical composition.

[0049] The "pharmaceutical composition" described in the present invention can be prepared by any method known in pharmacy. Generally speaking, these preparation methods include associating the Herba Artemisiae Argyi or its extract (i.e., the only active ingredient) with a carrier or excipient and / or one or more other auxiliary ingredients, and then if necessary and / or desired, molding and / or packaging the product into a desired single dose or multiple dose unit.

[0050] The pharmaceutical composition of the present invention can be prepared according to known methods, such as the methods described in the general rules for preparation of the 16th edition of the Japanese Pharmacopoeia, the United States Pharmacopoeia, and the 9th edition of the European Pharmacopoeia. The pharmaceutical composition of the present invention can be appropriately administered to patients depending on the dosage form.

[0051] The sole active ingredient, pharmaceutically acceptable excipient in the "pharmaceutical composition" described herein will vary depending on the identity, size and / or condition of the subject being treated and further on the route of administration of the composition. The pharmaceutical composition may contain between 0.1% and 100% (w / w) of the first active ingredient.

[0052] The "pharmaceutical composition" described in the present invention may also optionally contain other therapeutic ingredients for use in combination, especially those additional therapeutic ingredients disclosed herein, such as "other liver protection drugs" (i.e., the second active ingredient), wherein the other liver protection drugs are chemical drugs, traditional Chinese medicines or Chinese patent medicine preparations.

[0053] "Treatment" as used herein, unless otherwise indicated, means reversing, alleviating, inhibiting the progression of, or preventing the condition or disorder or one or more symptoms of such condition or disorder to which the term applies, unless otherwise indicated. The term "treat" as used herein refers to the act of treating, as "treat" is defined immediately above.

[0054] The "effective amount" described in the present invention refers to an amount sufficient to induce a desired biological response. The effective amount of the active ingredient of the present invention may vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the mode of administration, and the age and health of the subject. In certain embodiments, the effective amount is a therapeutically effective amount. The effective amount is the amount of the only active ingredient described in the present invention in a single dose. In certain embodiments, the effective amount is the combined amount of the only active ingredient described in the present invention in multiple doses.

[0055] As used herein, the "therapeutically effective amount" is an amount sufficient to provide a therapeutic benefit in the treatment of a disorder or sufficient to delay or minimize one or more symptoms associated with the disorder. The therapeutically effective amount of Semiliquidambar cathayensis Hance and its extracts means the amount of the therapeutic agent that provides a therapeutic benefit alone or in combination with other therapies in the treatment of a disorder. The term "therapeutically effective amount" may encompass an amount that modifies the overall therapy, reduces or avoids the symptoms, signs or causes of a disorder, and / or enhances the therapeutic efficacy of another therapeutic agent. In certain embodiments, the therapeutically effective amount is an amount sufficient to treat any of the diseases or disorders described.

[0056] I. Therapeutic effect of Semiliquidambar cathayensis Hance on acute liver injury induced by carbon tetrachloride (CCl4)

[0057] In this invention, a model of acute liver injury induced by CCl4 in rats was used. Based on the comprehensive research results of liver tissue morphology observation, pathological evaluation, serum biochemical indexes, etc., the degree of liver injury in rats caused by intragastric administration of CCl4 and the protective effect of Semiliquidambar cathayensis Hance on it were explored.

[0058] 1.1 Experimental animals

[0059] 60 SPF-grade male SD rats, weighing 180 - 200 g, were provided by the Guangdong Provincial Laboratory Animal Center. The license number for the experimental unit's use is: SYXK(Yue)2022 - 0125, and the license number is: SCXK(Yue)2022 - 0002. All experimental animals were housed in a SPF-grade barrier environment animal house, with a temperature of 20 - 25 °C and a humidity of 50% - 70%.

[0060] 1.2 Experimental content

[0061] 1.2.1 Preparation of test samples

[0062] Preparation of Semiliquidambar cathayensis Hance medicinal liquid: Weigh 600 g of Semiliquidambar cathayensis Hance (purchased from Shanghai Hongyong Biotechnology Co., Ltd., production batch number 220029 - 202307), place it in an extraction container, add 15 times the amount of 70 v / v% ethanol aqueous solution, heat and reflux for extraction 2 times, each time for 1.5 hours, filter while it is hot, combine the filtrates, and concentrate to 600 mL to obtain a medicinal liquid concentration of approximately 1 g / mL as the stock medicinal liquid.

[0063] 1.2.2 Animal grouping and administration

[0064] 60 SD rats, 6-8 weeks old, weighing 180-200g, were fed ad libitum and adaptively fed for one week. The animals were randomly divided into 6 groups (n=10): blank control group (CON group), model control group (CCl4 group, MOD group), positive drug group (silybin group, SF group), low-dose group of Jinlu Banfenghe (JBFL group), medium-dose group of Jinlu Banfenghe (JBFM group) and high-dose group of Jinlu Banfenghe (JBFH group). The low, medium and high doses of Jinlu Banfenghe were 0.525g / kg, 1.575g / kg and 4.725g / kg, respectively. The recommended amount of silybin for rats is 100mg / (kg·d). The dose group and positive drug group were given the test samples according to the body weight of the rats every day, and the blank control group and model control group were given corresponding distilled water for 15 days. The animals were weighed twice a week and the results were recorded to adjust the dose of the test samples.

[0065] Starting from 18:00 on the evening of the 15th day of the experiment, the animals in each group were fasted overnight but not water-contained for 16 hours. At 10:00 the next morning, except for the blank control group, each group was given a corresponding dose of CCl4 based on the body weight of each rat to establish an acute liver injury model. Specifically, CCl4 was diluted with corn oil, and the concentration of CCl4 administered to rats was 2% (2ml CCl4 dissolved in 98ml corn oil), and the dosage was 5mL / kg BW. The blank control group was given the same volume of pure corn oil. Food can be resumed after the modeling is completed. All experimental groups were administered by gavage. Five hours after the modeling was completed (15:00), the dose group and the positive drug group continued to be given the test sample (with an interval of more than 4 hours from the gavage of CCl4). 12 hours after the modeling was completed (22:00), overnight fasting was continued for 12 hours without water-containment.

[0066] 1.2.3 Sample collection

[0067] The animals were anesthetized with isoflurane, and blood was collected from the abdominal aorta. The blood in the procoagulant blood collection tube was centrifuged at 3500 rpm for 10 minutes at 4°C, and the upper serum was separated and stored in a -80°C refrigerator. After the blood was collected, the rat heart, spleen, liver, kidney, colon and their contents were collected by dissection, quenched with liquid nitrogen and stored in a -80°C refrigerator.

[0068] 1.3 Experimental Results

[0069] 1.3.2 Effects of Radix Glehniae on rat liver coefficient

[0070] The results are shown in Table 1.

[0071] As shown in Table 1, compared with the CON group, the liver weight of rats in the MOD group increased significantly after modeling (P < 0.05), and the liver coefficient increased extremely significantly (P < 0.001). This shows that CCl4 induction can cause an increase in the liver weight and liver coefficient of rats.

[0072] Compared with the MOD group, the liver weight of the rats in the JBFH group was extremely significantly reduced (P < 0.01), and the liver coefficient was significantly reduced (P < 0.05), while there was no statistical difference in the liver weight and liver coefficient of the rats in other groups, indicating that oral administration of high-dose Radix Glehniae can reduce the effects of CCl4-induced liver disease in rats.

[0073] Table 1 Effects of Radix Glechomae Half-leaf Herb on the Liver Coefficient of Rats

[0074]

[0075] Note: * Compared with the CON group, P < 0.05, *** indicates P < 0.001 compared with the CON group; # Compared with the MOD group, P < 0.05, ## Compared with the MOD group, P<0.01.

[0076] 1.3.2 Effects of Radix Glehniae on serum ALT and AST levels in rats

[0077] The results are as follows Figure 1 and Figure 2 shown.

[0078] Depend on Figure 1 It can be seen that compared with the CON group, the ALT level in the serum of the rats in the MOD group was extremely significantly increased after modeling (P < 0.001), which shows that CCl4 induction can cause an increase in the ALT level in the serum of rats.

[0079] Compared with the MOD group, the ALT levels in the serum of rats in the SF group and JBFL group were significantly decreased (P < 0.01), and the ALT levels in the serum of rats in the JBFM group and JBFH group were extremely significantly decreased (P < 0.001), indicating that oral administration of silybin and different doses of Radix Angelicae Pubescentis can reduce the ALT level in the serum of rats.

[0080] Compared with the SF group, the ALT level in the serum of rats in the JBFH group was significantly decreased (P<0.05), indicating that the high-dose of Radix Angelicae Pubescentis was more effective than silybin in reducing the ALT level in serum.

[0081] Depend on Figure 2 It can be seen that compared with the CON group, the AST level in the serum of the rats in the MOD group was significantly increased after modeling (P < 0.01), which shows that CCl4 induction can cause an increase in the AST level in the serum of rats.

[0082] Compared with the MOD group, the serum AST level of rats in the SF group was significantly decreased (P < 0.05), the serum AST level of rats in the JBFL group and JBFM group was extremely significantly decreased (P < 0.01), and the serum AST level of rats in the JBFH group was extremely significantly decreased (P < 0.001), indicating that oral administration of silybin and different doses of Radix Codonopsis pilosulae can reduce the AST level in the serum of rats.

[0083] Compared with the SF group, the serum AST level of rats in the JBFH group was significantly decreased (P<0.05), indicating that high-dose Jinlu Banfenghe was more effective than silybin in reducing the serum AST level.

[0084] 1.3.3 Observation of liver tissue pathology

[0085] The results are as follows Figure 3 As shown, Figure 3 The length of the white scale is 500 μm. The pathological changes of the liver tissues of rats in each group were observed by HE staining. The results showed that the liver tissues of rats in the CON group were normal, the tissue cells were evenly distributed, the hepatic cords were clear, and there was no cell necrosis, fatty degeneration or inflammatory cell infiltration. Compared with the CON group, the nuclei of the hepatocytes in the MOD group induced by CCl4 were squeezed to one side, resulting in unclear nuclei, unclear cytoplasmic boundaries, fatty degeneration, diffuse distribution of fat vacuoles of different sizes, and large areas of focal necrosis accompanied by inflammatory cell infiltration and proliferation in the cytoplasm and hepatocytes. Compared with the MOD group, the necrotic cells in the liver of rats in each drug-treated group were significantly reduced, the inflammatory cell infiltration was significantly reduced, the hepatic cords were arranged more neatly, and there was a significant improvement, and the effect of the JBFH group was better than that of the SF group.

[0086] 1.3.4 Evaluation of hepatocyte fatty degeneration

[0087] According to the liver cell fatty degeneration grade score standard described in the "Health Food Function Testing and Evaluation Method (2023 Edition)", the area of ​​each type of fatty degeneration in each visual field is recorded separately, and the total score of the lesions in the observed visual field is accumulated. Figure 4 shown.

[0088] Depend on Figure 4 It can be seen that compared with the CON group, the grade of hepatic steatosis in rats in each group was significantly increased (P < 0.01). Compared with the MOD group, the grade of hepatic steatosis in the SF group was significantly reduced (P < 0.05), and the grade of hepatic steatosis in the JBHL group, JBFM group and JBFH group was significantly reduced (P < 0.01). This shows that silybin and golden vine half maple can alleviate CCl4-induced hepatic cell inflammation and steatosis and other substantial damage to a certain extent, so as to play a liver-protective role.

[0089] Meanwhile, compared with the SF group, the grade score of hepatocyte fatty degeneration in the JBFH group was significantly reduced (P<0.05), indicating that the high-dose Jinlu Banfenghe of the present invention has a better liver-protecting effect than silybin.

[0090] In summary, pretreatment with Radix Glehniae can alleviate CCl4-induced acute liver injury in rats and has a certain therapeutic effect on acute liver injury.

[0091] 2. Protective effect of Radix Glehniae on acute liver injury induced by alcohol

[0092] 2.1 Experimental Animals

[0093] 72 SPF male SD rats, weighing 180-200 g, were provided by Guangdong Experimental Animal Center.

[0094] 2.2 Experimental content

[0095] 2.2.1 Preparation of test samples

[0096] The preparation of the test samples is the same as that in 1.2.1 above.

[0097] 2.2.2 Animal grouping and drug administration

[0098] The animals were grouped and dosed as described in 1.2.2 above.

[0099] Starting from 18:00 on the evening of the 15th day of the experiment, all groups of animals were fasted overnight but not water for 16 hours. At 10:00 the next morning, except for the blank control group, each group was given a corresponding dose of alcohol solution based on the body weight of each rat to create an acute liver injury model. Anhydrous ethanol (analytical alcohol) was diluted with distilled water, and the rat alcohol modeling concentration was 50%, and the modeling volume was 12mL / kg BW. The blank control group was given a corresponding volume of distilled water. Food can be resumed after the modeling is completed. All experimental groups were administered by gavage. Five hours after the completion of the modeling (15:00), the dose group and the positive drug group continued to be given the test sample (with an interval of more than 4 hours from the alcohol gavage). 12 hours after the completion of the modeling (22:00), overnight fasting continued for 12 hours without water.

[0100] 2.2.3 Sample collection

[0101] Same as sample collection in 1.2.3 above.

[0102] 2.3 Experimental Results

[0103] 2.3.1 Effects of Radix Glehniae on rat liver coefficient

[0104] The results are shown in Table 2.

[0105] As shown in Table 2, compared with the CON group, the liver weight and body weight of the rats in the MOD group did not change significantly after modeling (P < 0.05), but the increase in the liver coefficient was extremely significant (P < 0.001), indicating that alcohol induction can cause an increase in the liver coefficient of rats.

[0106] Compared with the MOD group, the liver coefficient of rats in the JBFH group was significantly reduced (P < 0.05), while the liver coefficients of rats in other groups showed no significant changes (P > 0.05), indicating that oral administration of high doses of Radix Glehniae can reduce the effects of alcohol on the rat liver.

[0107] Table 2 Effects of Radix Glechomae Half Maple on rat liver coefficient

[0108]

[0109] Note: * Compared with the CON group, P < 0.05, ** indicates P < 0.01 compared with the CON group; *** indicates P < 0.001 compared with the CON group; # Compared with the MOD group, P < 0.05, ## Compared with the MOD group, P<0.01.

[0110] 2.3.2 Effects of Radix Glehniae on serum ALT and AST levels in rats

[0111] The results are as follows Figure 5 and Figure 6 shown.

[0112] Depend on Figure 5 It can be seen that compared with the CON group, the ALT level in the serum of the rats in the MOD group was extremely significantly increased after modeling (P < 0.001), which shows that alcohol induction can cause an increase in the ALT level in the serum of rats.

[0113] Compared with the MOD group, the ALT levels in the serum of rats in the SF group, JBFL group, JBFM group and JBFH group were significantly reduced (P < 0.01), indicating that oral administration of silybin and different doses of Radix Angelicae Pubescentis can reduce the ALT levels in the serum of rats.

[0114] Depend on Figure 6 It can be seen that compared with the CON group, the AST level in the serum of the rats in the MOD group was significantly increased after modeling (P < 0.01), which shows that alcohol induction can cause an increase in the AST level in the serum of rats.

[0115] Compared with the MOD group, the AST level in the serum of rats in the SF group, JBFL group and JBFH group was extremely significantly decreased (P < 0.01), and the AST level in the serum of rats in the JBFM group was significantly decreased (P < 0.05), indicating that oral administration of silybin and different doses of Radix Angelicae Pubescentis can reduce the AST level in the serum of rats.

[0116] 2.3.3 Observation of liver tissue pathology sections

[0117] The results are as follows Figure 7 As shown, Figure 7 The length of the white scale is 500 μm. The pathological changes of the liver tissues of rats in each group were observed by HE staining. The results showed that the liver tissues of rats in the CON group were normal, the tissue cells were evenly distributed, the hepatic cords were clear, and there was no cell necrosis, fatty degeneration or inflammatory cell infiltration. Compared with the CON group, the livers of rats in the alcohol-induced MOD group were scattered, the hepatic cords were disordered, the nuclei were squeezed to one side, the nuclei were blurred, the cytoplasm boundaries were unclear, fatty degeneration occurred to form vacuolar defects, and fat vacuoles of different sizes were diffusely distributed. Large areas of focal necrosis accompanied by inflammatory cell infiltration and proliferation were observed in the cytoplasm and hepatocytes. Compared with the MOD group, the cell damage in the livers of rats in each drug-treated group was alleviated, the necrotic cells were significantly reduced, the hepatic cords were arranged more neatly, the inflammatory cell infiltration was significantly reduced, and the vacuoles formed by degeneration were reduced, which was significantly improved, and the effect of the JBFH group was better than that of the SF group.

[0118] 2.3.4 Evaluation of hepatocyte fatty degeneration

[0119] According to the liver cell fatty degeneration grade score standard described in the "Health Food Function Testing and Evaluation Method (2023 Edition)", the area of ​​each type of fatty degeneration in each visual field is recorded separately, and the total score of the lesions in the observed visual field is accumulated. Figure 8 shown.

[0120] Depend on Figure 8 It can be seen that compared with the CON group, the hepatocyte steatosis grade score of the MOD group rats was significantly increased (P < 0.001), and there were significant differences in the other groups (P < 0.01). Compared with the MOD group, the hepatocyte steatosis grade score of the SF group, JBHL group, JBFM group and JBFH group was significantly reduced (P < 0.001). This shows that silybin and golden vine half maple can alleviate alcohol-induced hepatocyte inflammation and steatosis and other substantial damage to a certain extent, so as to play a liver-protective role.

[0121] Meanwhile, compared with the SF group, the grade of hepatic fatty degeneration in the JBFH group was significantly reduced (P<0.05), indicating that the high-dose Herba Artemisiae Argyi of the present invention has a better liver-protecting effect than silybin.

[0122] In conclusion, pretreatment with Radix Glehniae can alleviate alcohol-induced acute liver injury in rats.

[0123] 3. Protective effect of Radix Glehniae on drug-induced acute liver injury caused by acetaminophen (APAP)

[0124] 3.1 Experimental Animals

[0125] 50 SPF male SD rats weighing 180-200 g were provided by Guangdong Experimental Animal Center.

[0126] 3.2 Experimental content

[0127] 3.2.1 Preparation of test samples

[0128] The preparation of the test samples is the same as that in 1.2.1 above.

[0129] 3.2.2 Animal grouping and drug administration

[0130] 50 SD rats, 6-8 weeks old, weighing 180-200g, were fed ad libitum and adaptively fed for one week. The animals were randomly divided into 5 groups (n=10): blank control group (CON group), model control group (MOD group), positive drug group (silybin group, SF group), low-dose group of Jinlu Banfenghe (JBFL group) and high-dose group of Jinlu Banfenghe (JBFH group). The low and high doses of Jinlu Banfenghe were 1.575g / kg and 4.725g / kg, respectively. The recommended amount of silybin for rats is 100mg / (kg·d). The dose group and the positive drug group were given the test samples according to the body weight of the rats every day, and the blank control group and the model control group were given the corresponding distilled water for 15 days. The animals were weighed twice a week and the results were recorded to adjust the dose of the test samples.

[0131] Starting from 18:00 on the evening of the 15th day of the experiment, the animals in each group were fasted overnight but not water for 16 hours. At 10:00 the next morning, except for the blank control group, each group was given a corresponding dose of APAP solution according to the body weight of each rat, causing APAP-induced acute liver injury. APAP was prepared as a mixed solution with a concentration standard of 350mg / ml CMC-Na. Each rat was gavaged with 2g / Kg APAP once to create a liver injury model. The blank control group was given a corresponding volume of distilled water. Food can be resumed after the modeling is completed. All experimental groups were administered by gavage. 5 hours after the completion of the modeling (15:00), the dose group and the positive drug group continued to be given the test sample (with an interval of more than 4 hours from APAP gavage). 12 hours after the completion of the modeling (22:00), the rats continued to fast overnight but not water for 12 hours.

[0132] 3.2.3 Sample collection

[0133] Same as sample collection in 1.2.3 above.

[0134] 3.3 Experimental Results

[0135] The results are as follows Fig. 9 and Fig.10 shown.

[0136] Depend on Fig. 9 It can be seen that compared with the CON group, the ALT level in the serum of the rats in the MOD group was significantly increased after modeling (P < 0.001), which shows that drug induction can cause an increase in the ALT level in the serum of rats.

[0137] Compared with the MOD group, the ALT levels in the serum of rats in the SF group, JBFL group and JBFH group were significantly reduced (P < 0.001), indicating that oral administration of silybin and different doses of Radix Angelicae Pubescentis can reduce the ALT level in the serum of rats.

[0138] Depend on Fig.10 It can be seen that compared with the CON group, the AST level in the serum of the rats in the MOD group was significantly increased after modeling (P < 0.001), which shows that drug induction can cause an increase in the AST level in the serum of rats.

[0139] Compared with the MOD group, the decrease in serum AST levels in the SF, JBFL and JBFH groups was significantly different (P < 0.001), indicating that oral administration of silybin and different doses of Radix Angelicae Pubescentis could reduce the AST level in the serum of rats.

[0140] In summary, pretreatment with Radix Glehniae can alleviate drug-induced acute liver injury in rats.

[0141] The above is a further description of the present invention in conjunction with specific embodiments, but these embodiments are merely exemplary and do not constitute any limitation on the scope of the present invention. It should be understood by those skilled in the art that the details and forms of the technical solution of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the scope of protection of the present invention.

Claims

1. Use of a Herba Lycopodii Extract or a pharmaceutical composition thereof in the preparation of a drug for preventing and / or treating liver damage; The pharmaceutical composition comprises a sole active ingredient, a Herba Lysimachiae extract, and pharmaceutically acceptable excipients; The preparation method of the Herba Lycopodii Extract comprises: adding ethanol to Herba Lycopodii for extraction, separating the solid and the liquid, and concentrating the filtrate to obtain the extract; the amount of ethanol added is 5-25 times the mass of Herba Lycopodii; the extraction times are 1-3 times, each time for 0.5-2.5 hours; The volume fraction of the ethanol is 30-80%; The liver injury is chemical acute liver injury.

2. The use according to claim 1, characterized in that: The dosage form of the pharmaceutical composition is granules, tablets, capsules, powders, pills, suspensions or liquid preparations.

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