Use of semiliquidambar cathayensis h. t. chang. in preparing drug for preventing and / or treating liver injury

Abstract

Use of Semiliquidambar cathayensis H. T. Chang., an extract thereof, or a pharmaceutical composition thereof in preparing a drug for preventing and / or treating liver injury. Tests prove that Semiliquidambar cathayensis H. T. Chang. can relieve substantial damage such as liver cell inflammatory reaction and fat degeneration, effectively reduce AST and AST levels in serum, and have a good effect on preventing and / or treating liver injury.

Description

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

[0001] Priority claim: This application claims priority to Chinese patent application No. 2024101732566, filed on February 7, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present invention belongs to the technical field of traditional Chinese medicine, and relates to the use of Radix Glechomae and Radix Glechomae in preparing a medicine for preventing and / or treating liver injury, and specifically relates to the use of Radix Glechomae and Radix Glechomae, an extract thereof or a pharmaceutical composition thereof in preparing a medicine for preventing and / or treating liver injury. Background Art

[0003] The liver is the largest digestive gland and detoxification organ in the human body. It is the central link in the metabolism of substances and energy in the body and an important organ for maintaining life activities. Liver disease is a common and extremely harmful disease in clinical practice, characterized by 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 lead to liver fibrosis, further developing into cirrhosis or liver cancer, which is life-threatening. Therefore, it is of great significance to seek safe and effective drugs to protect the liver and intervene in the progression of acute damage.

[0004] Carbon tetrachloride (CCl4)-induced acute liver injury in rat models has been widely used to investigate potential therapeutic strategies due to its similarities to acute chemical liver injury in humans.

[0005] Semiliquidambar cathayensis HTChang., a member of the Hamamelidaceae family, is primarily distributed in the mountainous areas of southern and southeastern China, including Guangxi, Guangdong, and Guizhou. It dispels wind and dampness, relaxes muscles and activates blood circulation, and is used to treat rheumatoid arthritis, traumatic injuries, and postpartum paralysis. Modern pharmacological research has shown that Semiliquidambar cathayensis has analgesic, anti-inflammatory, antioxidant, and blood circulation-activating properties.

[0006] 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 Achyranthes, 30-80 grams of Banfenghe, and 50-150 grams of rock sugar, and claims that it can treat hepatitis A and hepatitis B. CN103550513A discloses a traditional Chinese medicine for treating cholestatic hepatitis. The medicine is prepared from the following medicinal materials in the following weight ratios: 14 parts of Solanum cubilense, 12 parts of Elsholtzia ciliata, 8 parts of Rhizoma Cibotii, 10 parts of Rhizoma Cibotii, 14 parts of Panax notoginseng, 14 parts of Pine cone, 16 parts of Herba Catharsis, 8 parts of Herba Lycopodii, 14 parts of Herba Auriculariae, 13 parts of Herba Danbulao, 16 parts of Herba Tamarindifoliae, 15 parts of Herba Coriariae, 12 parts of Herba Bogudan, 14 parts of Herba Eupatorii, 8 parts of Herba Lycopodii, 10 parts of Herba Glehniae, and 12 parts of Herba Gentianae. The invention claims to be a safe and effective treatment for cholestatic hepatitis.

[0007] CN103356886A discloses a traditional Chinese medicine for treating primary biliary cirrhosis. The medicine is prepared from the following medicinal materials in the following weight ratios: 12 parts of Rosa laevigata fruit, 14 parts of Scrophularia ningpoensis, 6 parts of Stellera chamaejasme, 10 parts of Dechiragina serrata, 12 parts of Longlong, 14 parts of Herba Cyperi, 10 parts of Cinnamomum villosum, 14 parts of Bupleurum chinense, 10 parts of Cassia seed, 8 parts of Cyperus rotundus, 14 parts of Hesperus rotundus, 10 parts of Baba flower, 14 parts of Herba Polygoni Multiflori, 12 parts of Herba Codonopsis pilosulae, 14 parts of Herba Atractylodes macrocephalae, 10 parts of Herba Atractylodes macrocephalae, 8 parts of Fennel, 10 parts of Herba Corydalis, and 10 parts of Herba Glycyrrhizae. The invention claims to be a safe and effective treatment for 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, and nourishing the liver and spleen. It is mainly used to treat rheumatic joint pain, lumbar muscle strain and liver and kidney yin deficiency.

[0008] Current research on the liver effects of Radix Glechomae Benth. (Glechomae Benth.) has focused on combining Radix Glechomae Benth. (Glechomae Benth.) with traditional Chinese medicines known for their liver-protective properties. However, lacking research examining the underlying mechanisms and efficacy, it is difficult to assess its effectiveness. A search revealed no reports or applications of Radix Glechomae Benth. (Glechomae Benth.) for the treatment of liver damage.

[0009] Summary of the Invention

[0010] The present invention aims to provide the use of Radix Glechomae var. jinlübanfenghe in preparing a medicament for preventing and / or treating liver injury; in particular, the use of Radix Glechomae var. jinlübanfenghe, an extract thereof, or a pharmaceutical composition thereof in preparing a medicament for preventing and / or treating liver injury. The Radix Glechomae var. jinlübanfenghe of the present invention can alleviate substantial liver damage such as hepatocyte inflammatory response and steatosis, effectively reduce serum AST and AST levels, and has a good preventive and therapeutic effect on liver injury.

[0011] In order to achieve the above object, the present invention adopts the following technical solutions:

[0012] The present invention provides use of Radix Glehniae chinensis, an extract thereof or a pharmaceutical composition thereof in preparing a drug for preventing and / or treating liver damage.

[0013] The herb Semiliquidambar cathayensis HTChang. of the Hamamelidaceae family is derived from the herb Semiliquidambar cathayensis HTChang., and its roots, stems, branches or leaves are used as medicine and are sun-dried for later use.

[0014] The extract of the present invention is an extract of golden vine half-leaf maple, wherein the extraction comprises at least one of water decoction method, maceration method, percolation method, modified 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.

[0015] In some specific embodiments, the extract is an alcohol extract of Radix Angelicae Dahuricae.

[0016] Specifically, the preparation method of the alcohol extract is as follows: adding the Herba Lycopodii chinensis to alcohol for extraction, separating the solid and liquid, and concentrating the filtrate to obtain the alcohol extract.

[0017] In this embodiment, the volume fraction of the alcohol is 10-90%, preferably 30-80%, more preferably 45-75%, further preferably 50-75%, and most preferably 70%.

[0018] The alcohol in this embodiment is ethanol and / or methanol; preferably ethanol.

[0019] The solid-liquid separation described in this embodiment includes filtration and centrifugation, preferably filtration.

[0020] In some specific embodiments, the preparation method of the extract comprises the following steps: mixing the Herba Lycopodii and ethanol and refluxing for extraction, and concentrating the combined filtrates to obtain the extract.

[0021] Preferably, the amount of ethanol added is 5-25 times the mass of the Herba Lycopodii, preferably 15 times.

[0022] 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.

[0023] The pharmaceutical composition of the present invention comprises Radix Glehniae or its extract and pharmaceutically acceptable excipients.

[0024] The dosage form of the pharmaceutical composition of the present invention is granules, tablets, capsules, powders, pills, suspensions or liquid preparations.

[0025] In some specific embodiments, the pharmaceutical composition comprises other liver-protecting drugs, which serve as the second active ingredient and act together with Radix Glehniae or its extract to exert a liver-protecting effect.

[0026] The other liver-protecting drugs described in the present invention are chemical drugs, traditional Chinese medicines or Chinese patent medicine preparations.

[0027] The pharmaceutical composition of the present invention is prepared by mixing Radix Glechomae var. truncatum, its extract or pharmaceutical composition and other liver-protecting drugs directly or combining them after being packaged separately.

[0028] 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.

[0029] In some specific embodiments, the prevention and / or treatment of liver injury includes at least one of the following features:

[0030] (a) Reduce liver coefficient;

[0031] (b) reducing serum ALT and / or AST levels;

[0032] (c) Reduce substantial liver damage, including hepatocellular inflammation and / or steatosis.

[0033] The beneficial effects of the present invention are:

[0034] Experiments have shown that the Herba Lycopodii 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 good preventive and therapeutic effects on liver damage, especially acute chemical liver damage, drug-induced liver damage and alcoholic liver damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] FIG1 shows the effect of Radix Glehniae Benth on serum ALT in rats induced by CCl4;

[0036] FIG2 shows the effect of Radix Glehniae Benth on serum AST levels in rats induced by CCl4;

[0037] FIG3 is the observation result of liver tissue pathology section of rats induced by Jinlu Banfenghe on CCl4;

[0038] FIG4 is the scoring result of Jinlu Banfenghe on the grade of hepatic steatosis in rats induced by CCl4;

[0039] FIG5 shows the effect of Radix Glehniae Banfenghe on serum ALT in alcohol-induced rats;

[0040] FIG6 shows the effect of Radix Glehniae Benth on serum AST levels in rats induced by alcohol;

[0041] FIG7 is the observation result of liver tissue pathological section of Jinlu Banfenghe on alcohol-induced rats;

[0042] FIG8 is the scoring result of Jinlu Banfenghe on the grade of hepatic steatosis in alcohol-induced rats;

[0043] FIG9 shows the effect of Radix Glehniae Banfenghe on serum ALT in drug-induced rats;

[0044] FIG10 shows the effect of Radix Glehniae Benth on serum AST levels in drug-induced rats. DETAILED DESCRIPTION

[0045] The following describes the embodiments of the present invention through specific examples. Those skilled in the art will readily understand the other advantages and benefits of the present invention from the disclosure herein. The present invention may also be implemented or applied through various other specific embodiments, and the details in this specification may be modified or altered based on different viewpoints and applications without departing from the spirit of the present invention.

[0046] 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.

[0047] When numerical ranges are given in the examples, it should be understood that, unless otherwise specified herein, 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 herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention belongs.

[0048] 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.

[0049] The "pharmaceutical composition" described in the present invention comprises Radix Angelicae Pubescentis or its extract and pharmaceutically acceptable excipients. In a specific embodiment, Radix Angelicae Pubescentis or its extract described in the present invention is provided in the pharmaceutical composition in an effective amount (e.g., a therapeutically effective amount).

[0050] As used herein, "pharmaceutically acceptable" ingredients are substances suitable for use in humans and / or animals without undue adverse side effects (e.g., toxicity, irritation, and allergic reactions), that is, substances with a reasonable benefit / risk ratio. "Pharmaceutically acceptable excipients" include inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants, and / or oils. Excipients (e.g., cocoa butter and suppository waxes), colorants, coating agents, sweeteners, and flavoring agents may also be present in the pharmaceutical composition.

[0051] 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 Lycopodii 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, shaping and / or packaging the product into a desired single-dose or multi-dose unit.

[0052] The pharmaceutical composition of the present invention can be prepared according to known methods, for example, the methods described in the general rules for preparation of the Japanese Pharmacopoeia, 16th edition, the United States Pharmacopoeia, and the European Pharmacopoeia, 9th edition. The pharmaceutical composition of the present invention can be appropriately administered to patients depending on the dosage form.

[0053] 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 depending 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.

[0054] 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-protecting drugs" (i.e., the second active ingredient), wherein the other liver-protecting drugs are chemical drugs, traditional Chinese medicines or Chinese patent medicine preparations.

[0055] As used herein, "treating," unless otherwise indicated, means reversing, alleviating, inhibiting the progression of, or preventing the condition or disorder to which the term applies, or one or more symptoms of such condition or disorder. The term "treating," as used herein, refers to the act of treating, as "treating" is defined immediately above.

[0056] As used herein, "effective amount" refers to an amount sufficient to elicit 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 status of the subject. In certain embodiments, the effective amount is a therapeutically effective amount. The effective amount is the amount of the sole active ingredient described in the present invention in a single dose. In certain embodiments, the effective amount is the combined amount of the sole active ingredient described in the present invention in multiple doses.

[0057] As used herein, "therapeutically effective amount" refers to an amount sufficient to provide a therapeutic benefit in the treatment of a condition or sufficient to delay or minimize one or more symptoms associated with the condition. The therapeutically effective amount of Semiliquidambar cathayensis Hance or its extract means an amount of the therapeutic agent that provides a therapeutic benefit in the treatment of a condition, either alone or in combination with other therapies. The term "therapeutically effective amount" may encompass an amount that modifies the overall therapy, reduces or avoids the symptoms, signs or causes of a condition, and / or enhances the therapeutic efficacy of another therapeutic agent. In certain embodiments, the therapeutically effective amount is an amount sufficient to treat any disease or condition described herein.

[0058] I. Therapeutic effect of Semiliquidambar cathayensis Hance on acute liver injury induced by carbon tetrachloride (CCl4)

[0059] 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 induced by intragastric administration of CCl4 and the protective effect of Semiliquidambar cathayensis Hance on it were explored.

[0060] 1.1 Experimental animals

[0061] 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 to use them is: SYXK (Guangdong) 2022 - 0125, and the license number is: SCXK (Guangdong) 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%.

[0062] 1.2. Experimental content

[0063] 1.2.1 Preparation of the test sample

[0064] Preparation of the 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 under reflux for extraction twice, each time for 1.5 hours, filter while it is hot, combine the filtrates, and concentrate to 600 mL to obtain a medicinal liquid with a concentration of approximately 1 g / mL as the stock medicinal liquid.

[0065] 1.2.2 Animal grouping and drug administration

[0066] Sixty SD rats, 6-8 weeks old, weighing 180-200 g, were fed ad libitum and adapted for one week. The animals were then randomly divided into six groups (n=10): a blank control group (CON group), a model control group (CCl4 group, MOD group), a positive drug group (silybin group, SF group), a low-dose Jinlu Banfenghe group (JBFL group), a medium-dose Jinlu Banfenghe group (JBFM group), and a high-dose Jinlu Banfenghe group (JBFH group). The low, medium, and high doses of Jinlu Banfenghe were 0.525 g / kg, 1.575 g / kg, and 4.725 g / kg, respectively. The recommended daily dose of silybin for rats is 100 mg / kg / day. The dose groups and the positive drug group were administered the test sample daily according to the rat's body weight. The blank and model control groups were given the corresponding distilled water for 15 days. The animals were weighed twice weekly and the results were recorded to adjust the test sample dosage.

[0067] Starting at 6:00 PM on the evening of the 15th day of the experiment, all animals in each group were fasted overnight but not watered for 16 hours. At 10:00 AM the following morning, except for the blank control group, each group was given a single 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 CCl4 concentration for rats was 2% (2 ml CCl4 dissolved in 98 ml corn oil), with a dosage of 5 mL / kg BW. The blank control group was given the same volume of pure corn oil. Food was resumed after modeling. All experimental groups were administered by gavage. Five hours after modeling (3:00 PM), the dose group and the positive drug group continued to receive the test sample (at least 4 hours after the CCl4 gavage). Twelve hours after modeling (10:00 PM), overnight fasting continued for another 12 hours, but not watered.

[0068] 1.2.3 Sample Collection

[0069] Animals were anesthetized with isoflurane, and blood was collected from the abdominal aorta. The blood in a procoagulant blood collection tube was centrifuged at 3500 rpm for 10 minutes at 4°C. The supernatant serum was separated and stored in a -80°C freezer. After blood collection, the rat heart, spleen, liver, kidney, colon, and their contents were collected by dissection, quenched with liquid nitrogen, and stored in a -80°C freezer.

[0070] 1.3 Experimental Results

[0071] 1.3.2 Effects of Jinlu Banfenghe on rat liver coefficient

[0072] The results are shown in Table 1.

[0073] 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.

[0074] Compared with the MOD group, the liver weight of 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 rats in other groups, indicating that oral administration of high-dose Radix Glehniae can reduce the effects of CCl4-induced liver disease in rats.

[0075] Table 1 Effects of Radix Glechomae Benth on the Liver Coefficient of Rats 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 Jinlu Banfenghe on serum ALT and AST levels in rats

[0077] The results are shown in Figures 1 and 2.

[0078] As shown in Figure 1, 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), indicating 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 Glehniae 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 high-dose Jinlu Banfenghe was more effective than silybin in lowering the ALT level in serum.

[0081] As shown in Figure 2, 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), indicating 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 Glehniae can reduce the AST level in rat serum.

[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 lowering the serum AST level.

[0084] 1.3.3 Observation of liver tissue pathology

[0085] The results are shown in Figure 3, where the white scale bar represents 500 μm. Hematoxylin and eosin staining revealed histopathological changes in the livers of rats in each group. The CON group showed no abnormalities in liver tissue, with evenly distributed cells and clear hepatic cords. There was no evidence of cellular necrosis, steatosis, or inflammatory cell infiltration. Compared with the CON group, the livers of the MOD group induced by CCl₄ showed hepatocyte nuclei squeezed to one side, resulting in blurred nuclei, poorly defined cytoplasm, and steatosis. Fatty vacuoles of varying sizes were diffusely distributed, and extensive focal necrosis, accompanied by inflammatory cell infiltration and proliferation, was observed in the cytoplasm and hepatocytes. Compared with the MOD group, all treatment groups showed significantly fewer necrotic cells and inflammatory cell infiltration, and more uniform hepatic cord arrangement, demonstrating significant improvement. The JBFH group showed superior efficacy to the SF group.

[0086] 1.3.4 Evaluation of hepatocyte steatosis

[0087] According to the hepatocyte steatosis grade scoring criteria described in the "Health Food Function Testing and Evaluation Methods (2023 Edition)", the area occupied by various types of steatosis in each visual field was recorded, and the total score of the lesions in the observed fields was accumulated. The results are shown in Figure 4.

[0088] As shown in Figure 4, compared with the CON group, the grade of hepatic steatosis in rats in all groups was significantly increased (P < 0.01). Compared with the MOD group, the grade of hepatic steatosis in the SF group was significantly decreased (P < 0.05), and the grade of hepatic steatosis in the JBHL, JBFM, and JBFH groups was significantly decreased (P < 0.01). This suggests that silybin and Jinlu Banfenghe can, to a certain extent, alleviate CCl4-induced hepatic cell inflammation and steatosis, thereby exerting a hepatoprotective effect.

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

[0090] In conclusion, 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 Effects of Jinlu Banfenghe on Alcohol-Induced Acute Liver Injury

[0092] 2.1 Experimental Animals

[0093] Seventy-two SPF male SD rats weighing 180-200 g were provided by the Guangdong Provincial Laboratory 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 in the same manner as in 1.2.2 above.

[0099] Starting at 6:00 PM on the evening of the 15th day of the experiment, all animals in each group were fasted overnight, but without water, for 16 hours. At 10:00 AM the following morning, except for the blank control group, each group was administered a single 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 to a 50% alcohol concentration for rats, with a modeling volume of 12 mL / kg BW. The blank control group was given a corresponding volume of distilled water. Food was resumed upon completion of modeling. All experimental groups were administered via gavage. Five hours after modeling (3:00 PM), the dose group and the positive drug group continued to receive the test sample (at least 4 hours after the alcohol gavage). Twelve hours after modeling (10:00 PM), overnight fasting continued for another 12 hours, but 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 Jinlu Banfenghe 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 liver coefficient increased with a very significant difference (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-dose Radix Glehniae can reduce the effects of alcohol-induced liver disease in rats.

[0107] Table 2 Effects of Radix Glechomae Benth on rat liver coefficient 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.

[0108] 2.3.2 Effects of Jinlu Banfenghe on serum ALT and AST levels in rats

[0109] The results are shown in Figures 5 and 6.

[0110] As shown in Figure 5, 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), indicating that alcohol induction can cause an increase in the ALT level in the serum of rats.

[0111] 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 decreased (P<0.01), indicating that oral administration of silybin and different doses of Radix Glehniae can reduce the ALT levels in the serum of rats.

[0112] As shown in Figure 6, compared with the CON group, the AST level in the serum of the rats in the MOD group was extremely significantly increased after modeling (P < 0.01), indicating that alcohol induction can cause an increase in the AST level in the serum of rats.

[0113] 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 Glehniae can reduce the AST level in rat serum.

[0114] 2.3.3 Observation of liver tissue pathology

[0115] The results are shown in Figure 7, where the white scale bar represents 500 μm. Hematoxylin and eosin staining revealed histopathological changes in the livers of rats in each group. The CON group showed no abnormalities in liver tissue, with evenly distributed cells and clear hepatic cords. There was no evidence of cellular necrosis, steatosis, or inflammatory cell infiltration. Compared with the CON group, the livers of the alcohol-induced MOD group showed scattered hepatocytes and disorganized hepatic cords. Nuclei were squeezed to one side, blurring the nuclei. The cytoplasm showed unclear boundaries, and steatosis with vacuolar defects was observed. Fatty vacuoles of varying sizes were diffusely distributed. Extensive focal necrosis, accompanied by inflammatory cell infiltration and proliferation, was observed in the cytoplasm and hepatocytes. Compared with the MOD group, all drug-treated groups showed reduced cellular damage, significantly fewer necrotic cells, more uniformly arranged hepatic cords, significantly reduced inflammatory cell infiltration, and fewer vacuoles due to degeneration, demonstrating significant improvement. The JBFH group showed superior results to the SF group.

[0116] 2.3.4 Evaluation of hepatocyte steatosis

[0117] According to the hepatocyte steatosis grade scoring criteria described in the "Health Food Function Testing and Evaluation Methods (2023 Edition)", the area occupied by various types of steatosis in each visual field was recorded, and the total score of the lesion in the observed field was accumulated. The results are shown in Figure 8.

[0118] As shown in Figure 8, compared with the CON group, the hepatocyte steatosis grade score in the MOD group was significantly increased (P < 0.001), and significant differences were also observed in the other groups (P < 0.01). Compared with the MOD group, the hepatocyte steatosis grade score in the SF, JBHL, JBFM, and JBFH groups was significantly decreased (P < 0.001). This suggests that silybin and ginseng syrup can, to a certain extent, alleviate alcohol-induced hepatocyte inflammation and steatosis, thereby exerting a hepatoprotective effect.

[0119] At the same time, compared with the SF group, the grade 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 hepatoprotective effect than silybin.

[0120] In conclusion, pretreatment with Radix Glehniae can alleviate alcohol-induced acute liver injury in rats.

[0121] 3. Protective effect of Jinlu Banfenghe on acetaminophen (APAP)-induced acute liver injury

[0122] 3.1 Experimental Animals

[0123] Fifty SPF male SD rats weighing 180-200 g were provided by Guangdong Provincial Laboratory Animal Center.

[0124] 3.2 Experimental Content

[0125] 3.2.1 Preparation of test samples

[0126] The preparation of the test samples is the same as that in 1.2.1 above.

[0127] 3.2.2 Animal grouping and drug administration

[0128] Fifty SD rats, 6-8 weeks old, weighing 180-200g, were fed ad libitum and fed for one week. The animals were then randomly divided into five groups (n=10): a blank control group (CON group), a model control group (MOD group), a positive drug group (silybin group, SF group), a low-dose Jinlu Banfenghe group (JBFL group), and a high-dose Jinlu Banfenghe group (JBFH group). The low and high doses of Jinlu Banfenghe were 1.575g / kg and 4.725g / kg, respectively. The recommended dose of silybin for rats is 100mg / (kg / d). The dose groups and the positive drug group were administered the test sample daily according to the rat's body weight, while the blank control and model control groups were administered distilled water for 15 days. The animals were weighed twice a week and the results were recorded to adjust the test sample dosage.

[0129] Starting at 6:00 PM on the 15th day of the experiment, all animals in each group were fasted overnight, but without water, for 16 hours. At 10:00 AM the following morning, all groups, except the blank control group, were administered a single dose of APAP solution based on their body weight, inducing APAP-induced acute liver injury. APAP was prepared at a standard concentration of 350 mg / ml CMC-Na. Each rat was gavaged once with 2 g / kg APAP to establish a liver injury model. The blank control group was given a corresponding volume of distilled water. Food was resumed upon completion of the modeling. All experimental groups were administered via gavage. Five hours after modeling (3:00 PM), the dosed and positive drug groups continued to receive the test sample (at least 4 hours after the APAP gavage). Twelve hours after modeling (10:00 PM), the rats were fasted overnight, but without water.

[0130] 3.2.3 Sample collection

[0131] Same as sample collection in 1.2.3 above.

[0132] 3.3 Experimental Results

[0133] The results are shown in Figures 9 and 10.

[0134] As shown in Figure 9, 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), indicating that drug induction can cause an increase in the ALT level in the serum of rats.

[0135] Compared with the MOD group, the ALT levels in the serum of rats in the SF, JBFL and JBFH groups were significantly decreased (P < 0.001), indicating that oral administration of silybin and different doses of Radix Glehniae can reduce the ALT levels in the serum of rats.

[0136] As shown in Figure 10, 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), indicating that drug induction can cause an increase in the AST level in the serum of rats.

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

[0138] In conclusion, pretreatment with Radix Glehniae can alleviate drug-induced acute liver injury in rats.

[0139] The above further describes the present invention in conjunction with specific embodiments. However, 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 solutions of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, and such modifications and replacements shall fall within the scope of protection of the present invention.

Claims

1. Use of Radix Glehniae, its extract or its pharmaceutical composition in the preparation of a drug for preventing and / or treating liver damage.

2. The use according to claim 1, characterized in that The extraction includes at least one of water decoction, maceration, percolation, modified gelatin method, reflux method, solvent extraction, steam distillation, sublimation, supercritical fluid extraction, membrane separation technology, ultrafine grinding technology, traditional Chinese medicine flocculation separation technology, semi-bionic extraction, ultrasonic extraction, cyclone extraction, pressurized countercurrent extraction, enzyme method, macroporous resin adsorption method, ultrafiltration and molecular distillation.

3. The use according to claim 1, characterized in that The extract is an alcohol extract of Radix Angelicae Dahuricae.

4. The use according to claim 3, characterized in that The preparation method of the alcohol extract comprises the following steps: adding the Radix Glehniae chinensis to alcohol for extraction, performing solid-liquid separation, and concentrating the filtrate to obtain the alcohol extract.

5. The use according to claim 4, characterized in that The volume fraction of the alcohol is 10-90%.

6. The use according to claim 4, characterized in that The alcohol is ethanol and / or methanol.

7. The use according to claim 1, characterized in that The pharmaceutical composition comprises Radix Glehniae or its extract and pharmaceutically acceptable excipients.

8. The use according to claim 7, characterized in that The dosage form of the pharmaceutical composition is granules, tablets, capsules, powders, pills, suspensions or liquid preparations.

9. The use according to claim 7, characterized in that The pharmaceutical composition comprises other liver-protecting drugs, wherein the liver-protecting drugs are chemical drugs, traditional Chinese medicines or Chinese patent medicine preparations; The pharmaceutical composition is prepared by mixing Radix Glechomae var. truncatum, its extract or pharmaceutical composition and other liver-protecting drugs directly or individually packaged and then combined together.

10. The use according to any one of claims 1 to 9, characterized in that: The liver injury is acute liver injury, preferably acute chemical liver injury, alcoholic liver injury and drug-induced liver injury.

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