CN118319972A - Indian rorifone herb or extract thereof, composition and application thereof - Google Patents

Abstract

The application discloses application of Indian rorifolia herb or an extract thereof in preparing a medicine or health care product for treating and/or preventing diseases caused by liver injury and application of the Indian rorifolia herb or an extract thereof in preparing a medicine or health care product for treating and/or preventing diseases and/or symptoms related to the diseases caused by liver injury. The liver injury-causing disease may be acute liver injury, chronic liver injury, liver fibrosis, liver cirrhosis, liver cancer, etc. The rorifone or the extract thereof has liver protection effect, and is proved to have the protection effect on diseases caused by liver injury through liver function index, oxidative stress level and liver pathological staining, thereby being hopeful to become a new therapeutic drug in the aspect of liver injury diseases.

Description

Indian rorifone herb or extract thereof, composition and application thereof

Technical Field

The application relates to the field of traditional Chinese medicine or pharmaceutical chemistry, in particular to Indian rorhiza herb or an extract thereof, a composition thereof and application thereof.

Background

The liver is the largest metabolic organ in the human body, and not only it is the largest gland in the digestive system. It plays a vital role in maintaining metabolism, eliminating toxins, and in the production and release of proteins. Liver injury is due to the combined action of various internal and external factors, which leads to abnormal structure and function of the liver. These factors may include hepatitis virus, drugs, alcohol, fat deposition, and some other disease or condition. If the liver injury is continuously worsened, fatty liver, cirrhosis, liver fibrosis and even liver cancer can be caused, so that the body health is seriously endangered.

With the rapid development of medicine technology, many medicines for treating liver injury have been researched and reported, and mainly comprise Chinese herbal medicines, biological medicines, chemical medicines, trace elements and the like. However, some studies have shown that some drugs against liver injury also inevitably produce biotoxicity. It can be seen that prevention and treatment of liver injury is still a serious problem at present.

The traditional Chinese medicine considers that acute liver injury is mostly related to improper diet, physical weakness, fatigue, emotional disorder and exogenous pathogenic toxin, and the pathogenesis of the acute liver injury is mainly caused by damp evils accumulated in the liver and spleen, and spleen and stomach disharmony is caused by liver and gallbladder disharmony, qi stagnation and blood stasis. Many traditional Chinese medicines and traditional Chinese medicine components with the effects of clearing heat and detoxicating, promoting diuresis and removing jaundice, activating blood and dissolving stasis, soothing liver and regulating qi and the like can directly treat liver injury, and have no obvious toxic or side effect, so that the research and development of the medicines have important practical significance for treating clinical liver injury.

Disclosure of Invention

The protection effect of the rorifolia on liver injury is not disclosed at present. A large number of experiments show that the Indian rorhiza herb or the extract thereof can be used for preventing and treating diseases caused by liver injury.

In particular, the application relates to the following aspects:

1. use of rorifone or its extract in the preparation of a medicament or health care product for treating and/or preventing diseases caused by liver injury.

2. Use of rorifone or its extract in the preparation of a medicament or health care product for the treatment and/or prevention of diseases and/or disorders related to diseases caused by liver damage.

3. The use according to item 1 or 2, wherein the disease caused by liver injury is selected from one of acute liver injury, chronic liver injury, liver fibrosis, liver cirrhosis, and liver cancer.

4. The use of item 1 or 2, wherein the cause of liver injury is selected from one of trauma, drug, alcohol, chemical toxic substance, virus or autoimmunity.

5. The use of item 1 or 2, wherein the medicament or health product is for use in a mammal.

6. The use according to item 1 or 2, wherein the single dose of the medicament or the health product corresponds to 10g to 200g of Indian rorhiza herb.

7. The use according to item 6, wherein the single dose of the medicament or the health product corresponds to 25g to 100g of Indian rorifolia herb.

8. The use of item 1 or 2, wherein the rorifone extract is rorifone water extract or rorifone alcohol extract.

9. The use of item 8, wherein the rorifolia water extract is extracted by:

Crushing: pulverizing herba rorippa into coarse powder;

water heating reflux extraction: adding water into coarse powder, reflux-extracting, filtering, adding water again, reflux-extracting again, filtering, and mixing the filtrates;

Concentrating the filtrate: concentrating the filtrate under reduced pressure to obtain the herba rorippa water extract.

10. The use of item 8, wherein the rorifone alcohol extract is extracted by:

Crushing: pulverizing herba rorippa into coarse powder;

heating and reflux extracting ethanol: adding ethanol into the coarse powder, reflux-extracting, filtering, adding ethanol again, reflux-extracting again, filtering, and mixing the two filtrates;

concentrating the filtrate: concentrating the filtrate under reduced pressure to obtain the rorifone ethanol extract.

11. The use of any one of items 1 to 10, wherein the medicament or health product further comprises one or more other active substances.

12. A composition for treating and/or preventing a disease caused by liver injury or a disease and/or disorder related to a disease caused by liver injury, wherein the composition comprises rorifolia or an extract thereof, the rorifolia or an extract thereof as the only active ingredient in the composition.

13. The composition of item 12, wherein the composition further comprises one or more pharmaceutically acceptable carriers or excipients, or further comprises one or more other active substances.

14. The composition of item 12 or 13, wherein the disease caused by liver injury is a disease related to item 3 or 4.

15. The composition of any one of claims 12-14, wherein the rorifolia extract is the rorifolia extract of any one of claims 8-10.

The rorifone or the extract thereof has the functions of improving liver function index, oxidative stress level and pathological liver injury condition, has the protection effect on diseases caused by liver injury, and is expected to become a new therapeutic drug in the aspect of liver injury diseases.

Drawings

The drawings are included to provide a better understanding of the application and are not to be construed as unduly limiting the application. Wherein:

FIG. 1 shows the variation of serum alanine Aminotransferase (ALT) in mice of different groups;

FIG. 2 shows the changes in serum aspartate Aminotransferase (AST) of different groups of mice;

FIG. 3 shows the change in reduced Glutathione (GSH) in liver tissue of different groups of mice;

FIG. 4 shows changes in the superoxide dismutase (SOD) of liver tissue of different groups of mice;

FIG. 5 is a graph of liver H & E staining of mice of different groups.

Detailed Description

The application will be further illustrated with reference to the following examples, which are to be understood as merely further illustrating and explaining the application and are not to be construed as limiting the application.

Unless defined otherwise, technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the materials and methods are described herein below. In case of conflict, the present specification, including definitions therein, will control and materials, methods, and examples, will control and be in no way limiting. The application is further illustrated below in connection with specific examples, which are not intended to limit the scope of the application.

The application provides an application of rorifone or an extract thereof in preparing a medicine or health care product for treating and/or preventing diseases caused by liver injury.

The application also provides an application of the rorifone or the extract thereof in preparing a medicine or health care product for treating and/or preventing diseases and/or symptoms related to diseases caused by liver injury.

Rorifolia (school name: rorippa indica (l.) Hiern), a plant of the genus rorifolia of the family cruciferae. The rorifolia is an annual herb, and has the effects of eliminating phlegm and relieving cough. Can be used for treating chronic bronchitis. Blood pressure lowering, urination promoting, blood cooling and hemostasis. Can be used for treating dizziness, hypertension, difficulty in urination, hematuria, and metrorrhagia. Not only has medicinal value, but also can be eaten as vegetables.

The liver injury mainly refers to the damage of liver cells, abnormal liver structure and function, and the liver injury can stimulate an organism to generate excessive high-activity oxygen free radicals, induce oxidative stress, trigger inflammatory reaction, cause the damage and apoptosis of the liver cells and release excessive transaminase into blood so as to influence normal liver function. If not prevented or treated in time, a series of complications may be induced or the development of other diseases may be promoted.

The innovative discovery of the application, the rorifone or the extract thereof has the effect of treating or preventing diseases caused by liver injury.

The liver injury is caused by a plurality of reasons, mainly including drug poisoning, virus infection, excessive alcohol intake, immune response, chemical toxic substances in contact environment, radiation injury and the like. The rorifone herb or the extract thereof of the present application has a therapeutic or prophylactic effect on diseases caused by liver injury without limiting the cause of liver injury, as long as liver injury caused by damage of liver cells, abnormal liver structure, function is within the scope of the present application, and in a preferred embodiment, the liver injury is liver injury caused by alcohol, virus, drug and chemical toxic substances.

The specific type of the disease caused by liver injury is not limited in the present application, and any disease caused by liver injury is within the scope of the present application, including but not limited to acute liver injury, chronic liver injury, liver fibrosis, liver cirrhosis, hepatitis, liver cancer, fatty liver, liver failure, etc., such as liver injury, fatty liver, hepatitis, liver cirrhosis, liver failure, etc., caused by factors such as alcohol. In a preferred embodiment, the disease is acute liver injury or chronic liver injury.

The diseases and/or symptoms related to the diseases caused by the liver injury refer to other diseases or symptoms caused by the liver injury, and the diseases or symptoms can be relieved or eliminated due to the liver injury or the liver injury after treatment.

As used herein, "treatment" refers to any improvement in any outcome of a disease, disorder, or condition, such as extending survival, lower incidence, and/or alleviating side effects caused by alternative modes of treatment. In some embodiments, treating comprises delaying or ameliorating the development of a disease, disorder, or condition (i.e., slowing or preventing or reducing the development of the disease or at least one clinical symptom thereof). In some embodiments, treating comprises delaying, alleviating, or ameliorating at least one physical parameter of a disease, disorder, or condition, including a physical parameter that may not be discernable by a patient. In some embodiments, the treatment comprises modulating the disease, disorder, or condition physically (e.g., stabilizing a discernible symptom), physiologically (e.g., stabilizing a physical parameter), or both.

As used herein, the term "preventing" refers to the prophylactic treatment of a disease or disorder; or delay the onset or progression of the disease, disorder, or condition.

The medicament or nutraceutical may be used in a patient or other animal receiving the medicament or nutraceutical of the application for treating, preventing, alleviating and/or alleviating the diseases or conditions described herein, and in a preferred embodiment the medicament or nutraceutical is used in a mammal. Including but not limited to humans, cows, horses, sheep, pigs, goats, rabbits, cats, dogs, mice, and any other mammal having a liver and which may be damaged.

In a preferred embodiment, the medicament or nutraceutical is for use in humans.

The amount of the drug or health product of the present application to be administered depends on many factors such as the nature and severity of the disease to be prevented or treated, the sex, age, weight and individual response of the patient or animal, the administration route and the number of administrations, etc. The above-mentioned doses may be administered in a single dosage form or divided into several, for example two, three or four dosage forms. The dosage level will be selected based on the particular route of administration, the severity of the condition being treated, the condition and past medical history of the patient to be treated, and the like.

It will be appreciated that the total daily dose of the medicament or health product of the application must be determined by the attending physician within the scope of sound medical judgment. For any particular patient, the particular therapeutically effective dose level will depend on a variety of factors including the disorder being treated and the severity of the disorder; the specific composition employed; age, weight, general health, sex and diet of the patient; time of administration, route of administration, and rate of excretion; duration of treatment; a drug used in combination or simultaneously; and similar factors well known in the medical arts. For example, it is common in the art to administer doses that begin at levels below that required to achieve the desired therapeutic effect and gradually increase until the desired effect is achieved.

In a further preferred embodiment, the medicament or nutraceutical is administered in a single dosage form, i.e. a single dosage form. In a specific embodiment, the single dose administered corresponds to 10g to 200g of Indian rorippa herb, for example 10g、15g、20g、25g、30g、35g、40g、45g、50g、55g、60g、65g、70g、75g、80g、85g、90g、95g、100g、110g、120g、130g、140g、150g、160g、170g、180g、190g、200g, and any value between these values. In a specific embodiment, the single dose administered corresponds to 25g to 100g of Indian rorippa herb. In a specific embodiment, the single dose administered corresponds to 25g to 50g of Indian rorippa herb. It will be appreciated by those skilled in the art that, typically, the weight of a person is 60kg, and if the weight of the person is not 60kg, the conversion may be made in accordance with this standard.

It will be appreciated by those skilled in the art that the number and frequency of administrations may be adjusted depending on the particular condition of the subject or the severity of liver injury.

In a specific embodiment, the single dose of the drug or nutraceutical is administered to a human 1-3 times per day. In a specific embodiment, the single dose of the drug or nutraceutical is administered to a human 1-3 times per day for 3 days to 1 month, e.g., 3 days, 5 days, 7 days, 10 days, 14 days, 20 days, 1 month, etc., in succession.

The medicament or health care product of the application can be administered in unit dosage form, and the administration route can be intestinal tract or parenteral tract, such as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritoneal or rectum, etc. For example, tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories, freeze-dried powder injection and the like. Can be common preparation, slow release preparation, controlled release preparation and various microparticle administration systems.

The herba rorippa extract can be herba rorippa water extract or herba rorippa alcohol extract, or mixture of water extract and alcohol extract.

The herba rorippae extract can be prepared from any part of whole strain of herba rorippae, such as root, stem, leaf, flower, etc.

The method for extracting the rorifone water extract can use any water extraction method in the field. In a preferred embodiment, the present application employs the following method for preparing aqueous extract of rorifone:

Crushing: pulverizing herba rorippa into coarse powder;

water heating reflux extraction: adding water into coarse powder, reflux-extracting, filtering, adding water again, reflux-extracting again, filtering, and mixing the filtrates;

Concentrating the filtrate: concentrating the filtrate under reduced pressure to obtain the herba rorippa water extract.

The water may be any form of water known in the art, such as purified water, deionized water, secondary water, and the like.

The number of reflux extractions is not limited in the present application, and may be one, two, three, four or more times.

The method for extracting the rorifone alcohol extract can use any alcohol extraction method in the field. In a preferred embodiment, the present application employs the following method for preparing the rorifone alcohol extract:

Crushing: pulverizing herba rorippa into coarse powder;

heating and reflux extracting ethanol: adding ethanol into the coarse powder, reflux-extracting, filtering, adding ethanol again, reflux-extracting again, filtering, and mixing the two filtrates;

concentrating the filtrate: concentrating the filtrate under reduced pressure to obtain the rorifone ethanol extract.

Wherein ethanol may be any other common alcohol in the art, such as methanol, propanol, butanol, etc.

The number of reflux extractions is not limited in the present application, and may be one, two, three, four or more times.

In a preferred embodiment, the medicament or health product optionally further comprises a pharmaceutically acceptable carrier or excipient. Including, but not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil and the like. The medicine or health care product of the present application may contain, in addition to the above-mentioned components, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.

In a preferred embodiment, the pharmaceutical or nutraceutical product of the present application may further comprise one or more additional active substances. The other active substances are substances which have activity and generate a certain function except for the rorhiza sativa or the extract thereof, and the active substances can be active substances for treating or preventing diseases caused by liver injury or diseases related to diseases caused by liver injury, can be active substances for assisting in treating or preventing diseases caused by liver injury and diseases related to diseases caused by liver injury, can also not treat diseases caused by liver injury and diseases related to diseases caused by liver injury, but can have positive effects or functions at other convenience, and can be one or more of tarragon, radix rehmanniae, cortex moutan, coptis chinensis, rehmannia root, bupleurum root, selfheal, achyranthes root, scutellaria baicalensis and the like. The other active substance may be a crude drug or an extract thereof (e.g., an aqueous extract, an alcoholic extract, or an extract of an aqueous extract and an alcoholic extract).

The present application further provides a composition for treating and/or preventing a disease caused by liver injury or a disease and/or disorder related to a disease caused by liver injury, wherein the composition comprises rorifolia or an extract thereof, and rorifolia or an extract thereof as the only active ingredient in the composition. I.e. rorifone or its extract is the only active ingredient for treating and/or preventing diseases caused by liver damage or diseases and/or disorders related to diseases caused by liver damage.

The term "composition" is intended to include products comprising the specified amounts of each specified ingredient as well as any product that results, directly or indirectly, from combinations of the specified amounts of each specified ingredient.

In a preferred embodiment, the composition further comprises one or more pharmaceutically acceptable carriers or excipients. The one or more pharmaceutically acceptable carriers or excipients are as described in any one of the above.

In a preferred embodiment, the composition further comprises one or more additional active substances. The one or more other active substances are other active substances as described in any one of the above.

The disease caused by liver injury is a disease caused by liver injury as described in any one of the above.

The rorifone extract is the rorifone extract as described in any one of the above.

The composition of any of the present application may be in the form of an injection, an oral liquid, a capsule, a tablet, a granule, or an extract remixed dosage form.

The composition according to any of the present application may be administered in a single dosage form by the enteral or parenteral route, such as oral, intramuscular, subcutaneous, nasal, oral mucosal, dermal, peritoneal or rectal, etc. For example, tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories, freeze-dried powder injection and the like. Can be common preparation, slow release preparation, controlled release preparation and various microparticle administration systems.

The dose of the composition according to any of the present application may be converted with reference to the dose of the rorifolia or extract thereof as described above.

In the present application, the terms "alanine aminotransferase (ALT, alanine aminotransferase)" and "aspartate aminotransferase (AST, ASPARTATE AMINOTRANSFERASE)" are enzymes whose blood values are increased, respectively, when the liver is damaged, that is, enzymes which use such characteristics as an index of liver function. The terms "reduced glutathione (GSH, reduced glutathione)" and "superoxide dismutase (SOD, superoxide dismutase)" are important constituent members of the antioxidant enzyme system in biological systems, i.e., enzymes that take advantage of this property for use as antioxidant stress levels.

The Indian rorifolia herb or the extract thereof has the efficacy of treating and/or preventing diseases caused by liver injury, and the effect of the Indian rorifolia herb or the extract thereof on AST, ALT, liver oxidative stress level and liver pathological structure in serum is verified through a mouse model experiment.

Specifically, biochemical detection shows that AST and ALT levels in serum are obviously increased after mice in a model group are injected into an abdominal cavity of CC1 ₄ for 16 hours; the AST and ALT indexes of the model group are obviously different from those of the control group (P < 0.001), the silybin administration group, the rorifolia water extraction high, medium and low dose groups and the alcohol extraction medium and low dose groups have improvement effects relative to the model group on ALT level, and the improvement effects of the rorifolia water extraction low dose group are most obvious (P < 0.001) and are superior to those of the positive control group; on AST level, the silybin administration group, the herba rorhizae water extraction high, medium and low dose group and the alcohol extraction high, medium and low dose group have improvement effects compared with the model group, and the herba rorhizae water and alcohol extraction low dose group has the most obvious improvement effect (P < 0.001) which is superior to that of the silybin administration positive control group.

The oxidation stress level detection shows that after the mice in the model group are injected into the abdominal cavity of CC1 ₄ for 16 hours, the GSH and SOD levels in the liver are obviously reduced, the GSH and SOD indexes of the model group are obviously different from those of the control group (P < 0.05), and the figures show that the high dose group, the medium dose group, the low dose group and the high dose group, the medium dose group and the low dose group of the rorifolia water extract can increase the content of GSH in liver tissues, especially the low dose group and the medium dose group and the low dose group of the rorifolia water extract can obviously improve the content of GSH in the mice, and the GSH and the SOD indexes of the model group are statistically obviously different from those of the model group (P < 0.05) and are far better than those of the positive control group to which silybin is administrated. The high dose group, the medium dose group and the low dose group of the rorifone water extract and the high dose group, the medium dose group and the low dose group of the rorifone alcohol extract can increase the SOD content of liver tissues, and particularly the high dose group, the medium dose group and the low dose group of the rorifone water extract can obviously increase the SOD content of liver tissues of mice, have obvious difference (P < 0.05) with a model group in statistics and are far better than a positive control group to which silybin is added.

The application further discovers through liver pathology and histological examination that a large number of inflammatory cell infiltration and hepatocyte necrosis or nucleus shrinkage and dissolution occur in the liver tissue of the model group; compared with the model group, the liver cell damage degree, liver necrosis or nucleus shrinkage and dissolution and inflammatory infiltration of the mice in the administration group are obviously reduced, and the cytoplasm is uniformly colored, which indicates that the rorifolia water/alcohol extract can improve the liver cell tissue structure and is superior to the positive control group to which the silybin is administered.

The medicines commonly used in the prior art for treatment or prevention, such as the positive medicine silybin used in the application, have low bioavailability, poor effect of being directly absorbed and utilized in intestinal tracts after taking, slow application curative effect and relatively long treatment course, while other medicines, such as bifendate, have the advantages of obviously and rapidly reducing transaminase, but have no obvious effect on the histological change of liver, and actually have poor effects of resisting inflammation and protecting liver cells, while the herba rorhizae or the extract thereof has obvious effect of obviously reducing transaminase, protecting the histological form of liver, can comprehensively regulate, and is mild and safe.

Examples

Experimental materials

Animals: SPF-class male C57BL/6 mice, body weight 22 (+ -2) g,8 weeks old. Purchased from zhuhai Bai Tong Biotechnology Co., ltd., animal license number: [ SCXK (Yue) 2022-0051].

Test article: rorippa, purchased in the market of guangxi Yulin.

Reagent: 95% ethanol (AR), shanghai microphone Biochemical technologies Co., ltd;

Cci ₄ (HPLC), shanghai microphone biochemistry technologies limited;

Olive oil (pharmaceutical grade), shanghai microphone Biochemical technology Co., ltd;

glutamic-oxaloacetic transaminase (AST), glutamic-pyruvic transaminase (ALT), reduced Glutathione (GSH), superoxide dismutase (SOD) detection kit, and Nanjing built biological engineering research all the companies.

Silybin (SY), shanghai Ala Biochemical technology Co., ltd;

example 1 preparation of Indian rorifone extract

In the embodiment, the Indian rorifolia herb is respectively subjected to alcohol extraction and water extraction, and the specific method is as follows,

1.1 Alcohol extraction

(1) Preliminary crushing: pulverizing herba rorippa into coarse powder.

(2) Heating and reflux extracting ethanol: taking dried coarse powder of traditional Chinese medicine, reflux-extracting with 95% ethanol at 80deg.C for 1 hr, filtering, adding 95% ethanol at 80deg.C for 1 hr, reflux-extracting at 80deg.C, filtering, and mixing the filtrates.

(3) Concentrating the filtrate: concentrating the filtrate at 60deg.C under reduced pressure until the relative content of the raw materials is 1g/mL to obtain herba rorippa extract.

1.2 Water extraction

(1) Preliminary crushing: pulverizing herba rorippa into coarse powder.

(2) Water heating reflux extraction: reflux-extracting dried coarse powder of Chinese medicinal materials with 8 times of water at 80deg.C for 1 hr, filtering, adding 8 times of water, reflux-extracting at 80deg.C for 1 hr, filtering, and mixing the filtrates.

(3) Concentrating the filtrate: concentrating the filtrate at 60deg.C under reduced pressure until the relative content of the raw materials is 1g/mL to obtain herba rorippa water extract.

Example 2 Experimental methods and results of the action of rorifone extract on carbon tetrachloride-induced acute liver injury in mice.

The aqueous and alcoholic rorifone extracts prepared in example 1 were used in the following experiments.

2.1 Experimental methods

2.1.1 Grouping and administration of animals

70 Mice were classified into a blank control group (NC), a model group, a herba rorhizae water extract low dose administration group (IRH-S-L), a herba rorhizae water extract medium dose administration group (IRH-S-M), a herba rorhizae water extract high dose administration group (IRH-S-H), a herba rorhizae alcohol extract low dose administration group (IRH-C-L), a herba rorhizae alcohol extract medium dose administration group (IRH-C-M), a herba rorhizae alcohol high dose administration group (IRH-C-H) and a positive control group (SY) according to a body weight random block design. Wherein, the positive control group is administrated with Silybin (SY) 150mg/kg body weight/day, the herba rorhizae low dose group is based on 3.25g/kg body weight crude drug concentration per day, the herba rorhizae medium dose group is based on 6.5g/kg body weight crude drug concentration per day, and the high dose group is based on 13g/kg body weight crude drug concentration per day, and the herba rorhizae low dose group is infused with stomach 1 time per day for 7 days.

2.1.2 Establishment of model of acute liver injury by CCl ₄

After the last 2 hours of administration, 10% (v: v) concentration of each group of disposable intraperitoneal injections cci ₄, except the blank group, was dissolved in olive oil (1 ml/kg) to induce an acute liver injury model; the blank group was intraperitoneally injected with an equal volume of olive oil.

2.1.3 Sample collection and handling

After anesthetizing mice with sodium pentobarbital 16 hours after cci ₄ injection, the mice were sacrificed by orbital sampling and the livers were isolated for subsequent detection. After the whole blood of the mice is stood for 30min at room temperature, the whole blood is centrifuged at 3000rpm for 15min at 4 ℃, the supernatant is taken for preservation, the same part of the liver of the mice is taken for histopathological examination, and the residual liver tissue is preserved at-80 ℃.

2.1.4 Determination of Biochemical indicators

Alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) were detected using the kit method and purchased from Nanjing's as a biological engineering research company.

2.1.5 Determination of oxidative stress index

Reduced Glutathione (GSH), superoxide dismutase (SOD) are detected by adopting a kit method and purchased from Nanjing built bioengineering research all companies.

2.1.6 Pathological section treatment

Taking liver tissues of the right side lobes (with the largest cross section, a portal area and a central vein area) of different mouse livers, and fixing the liver tissues in 4% paraformaldehyde; the paraffin section is prepared through the steps of washing, dehydrating, transparentizing, wax dipping, embedding, slicing and sticking, dewaxing, H & E dyeing, dehydrating, transparentizing, sealing and the like, and the histological lesion observation is carried out under an optical microscope.

2.1.7 Statistical data processing

All experimental data were statistically analyzed using GraphPad prism9.0 software, using one-wayANOVA with sample number n=4/5, #, P <0.05 when the metering data satisfied normal distribution; # and P <0.01; # #, P <0.001; p <0.05 compared to model group (CCl ₄); * P <0.01; * P <0.001.

2.2 Results

2.2.1 Serum ALT and AST levels in mice of each group

In fig. 1, the model group (CCl ₄) showed significantly higher ALT activity than the control group (NC) (P < 0.001), indicating that the modeling was successful and that administration of the aqueous or alcoholic rorifolia extract prepared in example 1 significantly reduced ALT activity. As can be seen from the graph, the high dose group (IRH-S-H), the medium dose group (IRH-S-M), the low dose group (IRH-S-H) and the medium dose group (IRH-C-M) of the rorifone alcohol extract have very good effects, ALT has a significant difference (P < 0.001) compared with the model group, and especially the effect of the low dose group (IRH-S-L) of the rorifone water extract is better than that of the positive control group of Silybin (SY).

In fig. 2, the model group (CCl ₄) has significantly higher AST activity than the control group (NC) (P < 0.001), indicating that the modeling was successful, and administration of the aqueous or alcoholic rorifolia extract prepared in example 1 can significantly reduce AST activity. As can be seen from the graph, the high dose group (IRH-S-H), the medium dose group (IRH-S-M), the low dose group (IRH-S-L) and the high dose group (IRH-C-H), the medium dose group (IRH-C-M) and the low dose group (IRH-C-L) of the rorifolia aqueous extract have very good effects, the AST values have significant differences (P < 0.05) compared with the model group, and especially, the effect of the rorifolia aqueous extract and the rorifolia alcoholic extract is better than that of the positive control group of Silybin (SY).

2.2.2 Liver GSH and SOD levels in mice of each group

In fig. 3, the liver tissue GSH content of the model group (CCl ₄) is significantly reduced (P < 0.01) compared with that of the control group (NC), the administration of the rorifolia extract prepared in example 1 can significantly increase the liver tissue GSH content, and as can be seen from the figure, the high dose group (IRH-S-H), the medium dose group (IRH-S-M), the low dose group (IRH-S-L) and the high dose group (IRH-C-H) of the rorifolia alcohol extract can significantly increase the liver tissue GSH content compared with the control group (NC), and particularly, the dose group (IRH-C-M) and the low dose group (IRH-C-L) of the rorifolia alcohol extract can significantly increase the in vivo H content of the mice, and the statistically significantly different (P < 0.05) from the model group, and far better than the positive control group (SY).

In fig. 4, the liver tissue SOD content of the model group (CCl ₄) is significantly reduced (P < 0.05) compared to the control group (NC), and administration of the rorifolia extract prepared in example 1 can increase the liver tissue SOD content. As can be seen from the graph, the high dose group (IRH-S-H), the medium dose group (IRH-S-M), the low dose group (IRH-S-L) and the high dose group (IRH-C-H) of the rorifone alcohol extract can increase the content of SOD in liver tissues, and particularly the high dose group (IRH-S-H) and the high dose group (IRH-C-H) of the rorifone alcohol extract can significantly increase the content of SOD in liver tissues of mice, and the low dose group (IRH-C-L) and the high dose group (IRH-S-H) of the rorifone alcohol extract can have significant differences (P < 0.05) with the model group in statistics and are far better than the positive control group for administration of Silybin (SY).

2.2.3 Liver pathological section conditions of mice in each group

Pathological sections are carried out on the liver tissues of mice in the normal group, the model group, the silybin group, the rorhiza water extract and the alcohol extract of the high, medium and low dose groups. As shown in fig. 5, the normal liver control group (NC) has a clear lobular structure, no obvious lesions on liver cells, abundant cytoplasm, and a clear nuclear structure. Model group (CCl ₄) liver, hepatocytes were denatured, and hepatocytes were necrotic or nuclei were dissolved by shrinkage and infiltration of inflammatory cells. As can be seen from the graph, the high dose group (IRH-S-H), the medium dose group (IRH-S-M), the low dose group (IRH-S-L) and the high dose group (IRH-C-H), the medium dose group (IRH-C-M) and the low dose group (IRH-C-L) of the rorifolia aqueous extract are administered, the liver lobule structure of the mice is clear, the damage degree of liver cells, the liver necrosis or nucleus shrinkage and dissolution, the inflammatory infiltration are obviously reduced, the cytoplasma coloring is even, and the mice are better than the positive control group (SY) to which the silybin is administered.

Claims (15)

1. Use of rorifone or its extract in the preparation of a medicament or health care product for treating and/or preventing diseases caused by liver injury.

2. Use of rorifone or its extract in the preparation of a medicament or health care product for the treatment and/or prevention of diseases and/or disorders related to diseases caused by liver damage.

3. The use according to claim 1 or 2, wherein the disease caused by liver injury is selected from one of acute liver injury, chronic liver injury, liver fibrosis, liver cirrhosis, liver cancer.

4. The use of claim 1 or 2, wherein the cause of liver injury is selected from one of trauma, drug, alcohol, chemical toxic substance, virus or autoimmunity.

5. The use of claim 1 or 2, wherein the medicament or nutraceutical is for use in a mammal.

6. The use according to claim 1 or 2, wherein the single dose of the medicament or health product corresponds to 10 g-200 g of rorifolia.

7. The use according to claim 6, wherein the single dose of the medicament or the health product corresponds to 25 g-100 g of Indian rorhiza herb.

8. The use of claim 1 or 2, wherein the rorifone extract is rorifone water extract or rorifone alcohol extract.

9. The use of claim 8, wherein the rorifone water extract is extracted by:

Crushing: pulverizing herba rorippa into coarse powder;

water heating reflux extraction: adding water into coarse powder, reflux-extracting, filtering, adding water again, reflux-extracting again, filtering, and mixing the filtrates;

Concentrating the filtrate: concentrating the filtrate under reduced pressure to obtain the herba rorippa water extract.

10. The use of claim 8, wherein the rorifone alcohol extract is extracted by:

Crushing: pulverizing herba rorippa into coarse powder;

heating and reflux extracting ethanol: adding ethanol into the coarse powder, reflux-extracting, filtering, adding ethanol again, reflux-extracting again, filtering, and mixing the two filtrates;

concentrating the filtrate: concentrating the filtrate under reduced pressure to obtain the rorifone ethanol extract.

11. The use according to any one of claims 1 to 10, wherein the medicament or health product further comprises one or more other active substances.

12. A composition for treating and/or preventing a disease caused by liver injury or a disease and/or disorder related to a disease caused by liver injury, wherein the composition comprises rorifolia or an extract thereof, the rorifolia or an extract thereof as the only active ingredient in the composition.

13. The composition of claim 12, wherein the composition further comprises one or more pharmaceutically acceptable carriers or excipients, or further comprises one or more other active substances.

14. The composition of claim 12 or 13, wherein the disease caused by liver injury is a disease according to claim 3 or 4.

15. The composition of any one of claims 12-14, wherein the rorifolia extract is the rorifolia extract of any one of claims 8-10.