CN118286310A - Application of nux Prinsepiae or extract and composition thereof in treating and/or preventing liver injury - Google Patents
Application of nux Prinsepiae or extract and composition thereof in treating and/or preventing liver injury Download PDFInfo
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- CN118286310A CN118286310A CN202410660206.0A CN202410660206A CN118286310A CN 118286310 A CN118286310 A CN 118286310A CN 202410660206 A CN202410660206 A CN 202410660206A CN 118286310 A CN118286310 A CN 118286310A
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- nux prinsepiae
- liver injury
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Abstract
The invention discloses application of nux Prinsepiae or extract thereof in preparing a product for treating and/or preventing diseases caused by liver injury. Through a large number of experiments, the invention creatively discovers that nux Prinsepiae or the extract thereof can be used for preventing or treating diseases caused by liver injury. Specifically, the invention verifies that nux Prinsepiae or the extract thereof has obvious improvement effect on AST and ALT in serum through a mouse model experiment, and further discovers that the serum biochemical level of nux Prinsepiae administration group is obviously reduced, the damage degree of mouse liver cells and inflammatory infiltration are obviously reduced, the lobular structure is clear, liver cables and liver sinuses have no obvious lesions, and the cytoplasm is uniformly colored through liver pathology and histological examination. The results show that the nux Prinsepiae or the extract thereof has obvious and rapid reduction of transaminase, protects the histological morphology of the liver, can comprehensively regulate, is mild and safe, and has application value for preventing and/or treating clinical liver injury.
Description
Technical Field
The invention belongs to the field of pharmaceutical chemistry, and particularly relates to application of nux Prinsepiae or extract and composition thereof in treating and/or preventing liver injury.
Background
The liver serves as a major site of metabolism and serves as an important defense against external toxins. Drug abuse, viral infection, and high volume drinking may induce liver damage. Meanwhile, chronic inflammation of the liver may become liver cancer and the like due to untimely intervention, and burden of patients is increased.
At present, a large number of liver protection medicines are researched and applied to clinical practice, mainly comprising Chinese herbal medicines, biological medicines, chemical medicines, trace elements and the like, but the medicines for treating or preventing liver injury commonly used in the prior art have some defects. For example, silymarin has low bioavailability, poor effect of being directly absorbed and utilized by intestinal tracts after being taken, slow application curative effect, relatively long treatment course, easy diarrhea and other problems. There are also studies showing that some drugs against liver injury inevitably produce biotoxicity. It can be seen that the current situation of preventing and treating liver injury is still severe.
The traditional Chinese medicine is the treasure of the traditional Chinese medicine in China, and a plurality of traditional Chinese medicines and traditional Chinese medicine components have the effects of clearing heat and detoxicating, promoting diuresis and removing jaundice, activating blood circulation and dissolving stasis, soothing liver and regulating qi and the like. Therefore, the traditional Chinese medicine capable of directly treating liver injury and free of obvious toxic and side effects has important practical significance for treating clinical liver injury.
Nux Prinsepiae, bairen, , semen Myristicae, fructus Pruni Prinsepiae, fructus Persicae, small Ma Ruzi, nux Prinsepiae. Is prepared from nux Prinsepiae PRINSEPIA of Rosaceae, or its variant teeth She Bian, PRINSEPIA of nux Trionycis, var. Serrata Rehd. Picking ripe fruits in summer and autumn, removing pulp, sun drying, crushing shell, and collecting kernel. Sweet and cold in nature and slightly bitter. Has the effects of dispelling wind and dissipating heat, nourishing liver and improving eyesight, and treating conjunctival congestion and swelling and pain, dim and photophobia, canthus rot and excessive tears and epistaxis. Prinsepia utilis Royle is mainly used for external application in modern clinical medicine, mainly used for treating ophthalmic diseases, and is also occasionally applied to headache, insomnia, dizziness and other diseases. At present, no report on the application of nux Prinsepiae or the extract thereof in preventing and treating liver injury is found.
Disclosure of Invention
The invention aims to overcome the defects and shortcomings in the prior art and provide application of nux Prinsepiae or extract and composition thereof in preparing products for treating and/or preventing liver injury.
The above object of the present invention is achieved by the following technical solutions:
the invention firstly provides application of nux Prinsepiae or extract thereof in preparing products for treating and/or preventing diseases caused by liver injury.
The invention also provides application of nux Prinsepiae or extract thereof in preparing products for treating and/or preventing diseases and/or symptoms related to diseases caused by liver injury.
Liver injury mainly refers to damage of liver cells, abnormal liver structure and function, and the liver injury stimulates the organism to generate excessive high-activity oxygen free radicals, induces oxidative stress, initiates inflammatory reaction, causes injury and apoptosis of the liver cells, and releases excessive transaminase into blood, thereby affecting 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. Through a large number of experiments, the invention innovatively discovers that nux Prinsepiae or the extract thereof can be used for preventing and/or treating liver injury, thereby having the effect of preventing and/or treating diseases caused by the liver injury.
In the present invention, 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.
Specifically, the influence of nux Prinsepiae or extract thereof on AST and ALT contents in serum is verified by a mouse model experiment. Biochemical examination shows that after the mice in the model group are subcutaneously injected with 16 h in CC1 4, the AST and ALT levels in serum are obviously increased; the 2 liver function indexes of the model group are obviously different from those of the control group (P < 0.01), and the silybin administration group, the nux Prinsepiae water extract high, medium and low dose groups have improvement effect (P < 0.01) compared with the model group. The invention further discovers through liver pathology and histological examination that the normal tissue structure of the model group liver disappears, the liver sinus disappears, and balloon-like degeneration is visible; liver cells are generally denatured, necrotic or nucleus is contracted and dissolved, and a large amount of inflammatory cells infiltrate; compared with the model group, the serum biochemical level of the nux Prinsepiae administration group is obviously reduced, the damage degree and inflammatory infiltration of liver cells of mice are obviously reduced, the lobular structure is clear, the hepatic chordae and the hepatic sinus have no obvious lesions, and the cytoplasm is uniformly colored. In conclusion, the nux Prinsepiae or the extract thereof has obvious and rapid reduction of transaminase, protects the histological morphology of the liver, can comprehensively regulate, and is mild and safe.
Further, liver injury is caused by a variety of factors, including drug poisoning, viral infection, excessive alcohol intake, immune response, chemical toxic substances in the contact environment, and radiation injury. The nux Prinsepiae or its extract of the present invention has therapeutic or prophylactic effect on diseases caused by liver injury without limiting the cause of liver injury, and liver injury caused by alcohol, virus, drug and chemical toxic substances is within the scope of the present invention as long as liver injury is caused by abnormal liver cell, liver structure and function.
Further, the specific type of the disease caused by liver injury is not limited in the present invention, and any disease caused by liver injury is within the scope of the present invention, 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.
The term "preventing and/or treating" means that the treatment, prevention, alleviation and/or alleviation of a disease or a disorder of the present invention is effected in a subject.
The term "disease and/or disorder" refers to a physical state of the subject that is associated with the disease and/or disorder of the present invention.
Further, the disease caused by the liver injury is selected from any one of acute liver injury, chronic liver injury, liver fibrosis, liver cirrhosis and liver cancer.
Further, the cause of the liver injury is selected from any one of trauma, medicine, alcohol, chemical toxic substance, virus or autoimmunity.
Further, the product is a medicine or a health product.
Further, the product is for use in mammals.
The medicament or health product may be for use in a patient or other animal receiving a medicament or health product of the invention for treating, preventing, alleviating and/or alleviating a disease or condition of the invention, in a preferred embodiment the medicament or health product is for use 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.
Preferably, the product is for use in humans.
The dosage of the pharmaceutical or nutraceutical product of the present invention 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 route and frequency of administration, 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 amount of the pharmaceutical or nutraceutical product of the present invention must be determined by the 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.
Further, the single dose of the product is equivalent to 15 g-200 g of the raw nux Prinsepiae.
In a further preferred embodiment, the pharmaceutical or nutraceutical product is administered in a single dosage form, i.e. in a single dosage form, and in a further preferred embodiment, the single dosage of administration corresponds to 15g to 200g of the green nux Prinsepiae, for example 16g, 17g, 18g, 19g, 20g, 30g, 40g, 50g, 60g, 70g, 80g, 90g, 100g, 110g, 120g, 130g, 140g, 150g, 160g, 170g, 180g, 190g, and more preferably 50g to 100g of the green nux Prinsepiae. 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.
The pharmaceutical or nutraceutical products of the present invention may be administered in unit dosage form by the enteral or parenteral route, such as oral, intramuscular, subcutaneous, nasal, oral mucosal, dermal, peritoneal or rectal. 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.
Further, the nux Prinsepiae extract is nux Prinsepiae water extract or nux Prinsepiae alcohol extract, or water-alcohol extract mixture.
Further, the nux Prinsepiae extract is nux Prinsepiae water extract.
Further, the nux Prinsepiae extract is prepared by using dried and mature nux Prinsepiae of plant nux Prinsepiae.
Further, any water extraction method in the art can be used for the extraction method of the nux Prinsepiae extract.
Further, the preparation method of the nux Prinsepiae extract comprises the steps of crushing nux Prinsepiae, adding water, heating and reflux-extracting, and concentrating filtrate to obtain the nux Prinsepiae water extract.
In a preferred embodiment, the present invention employs the following method for preparing nux Prinsepiae water extract:
S1, crushing: pulverizing nux Prinsepiae into coarse powder;
s2, water heating reflux extraction: adding pure water into the coarse powder, reflux-extracting, filtering, adding pure water again, reflux-extracting again, filtering, and combining the two filtrates;
s3, concentrating filtrate: concentrating the filtrate under reduced pressure to obtain the nux Prinsepiae water extract.
Preferably, the water may be any form of water known in the art, such as purified water, deionized water, secondary water, and the like.
Preferably, the number of the reflux extractions is not limited in the present invention, and may be one, two, three, four and more times.
Further, the product optionally contains 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 invention 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.
Further, one or more other active substances may be further included in the product for treating and/or preventing diseases caused by liver damage according to the present invention. The other active substances refer to substances which have activity and produce a certain function except the nux Prinsepiae or the extract thereof, the active substances can be active substances for treating or preventing diseases caused by liver injury, can be active substances for assisting in treating or preventing diseases caused by liver injury, can also not be active substances for treating diseases caused by liver injury, but can have positive effects or functions at other convenience, the active substances can be one or more of artemisia capillaris, radix rehmanniae, cortex moutan, coptis, rehmannia root, radix bupleuri, selfheal, achyranthes, radix scutellariae and the like or the extract thereof, and the other active substances can be crude drugs or the extract thereof (such as an aqueous extract, an alcohol extract or an alcohol-water mixture extract).
The present invention also provides a composition for treating and/or preventing a disease caused by liver injury, which comprises nux Prinsepiae or its extract as the only active ingredient.
Further, the composition further comprises one or more pharmaceutically acceptable carriers or excipients, or further comprises one or more other active substances.
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.
Preferably, the one or more pharmaceutically acceptable carriers or excipients are any of the carriers or excipients described above.
Preferably, the one or more further active substances are further active substances as described in any of the above.
Preferably, the nux Prinsepiae extract is any one of the above nux Prinsepiae extracts.
The composition according to any one of the present invention 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 invention 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 one of the present invention may be converted by referring to the dose of the nux Prinsepiae or its extract as described above.
Compared with the prior art, the invention has the following beneficial effects:
The invention provides application of nux Prinsepiae or extract thereof in preparing products for treating and/or preventing diseases caused by liver injury. Through a large number of experiments, the invention creatively discovers that nux Prinsepiae or the extract thereof can be used for preventing or treating diseases caused by liver injury. Specifically, the invention verifies that nux Prinsepiae or the extract thereof has obvious improvement effect on AST and ALT in serum through a mouse model experiment, and further discovers that the serum biochemical level of nux Prinsepiae administration group is obviously reduced, the damage degree of mouse liver cells and inflammatory infiltration are obviously reduced, the lobular structure is clear, liver cables and liver sinuses have no obvious lesions, and the cytoplasm is uniformly colored through liver pathology and histological examination. The results show that the nux Prinsepiae or the extract thereof has obvious and rapid reduction of transaminase, protects the histological morphology of the liver, can comprehensively regulate, is mild and safe, and has application value for preventing and/or treating clinical liver injury.
Drawings
FIG. 1 shows the changes in serum alanine Aminotransferase (ALT) in different groups of mice.
FIG. 2 shows the changes in serum aspartate Aminotransferase (AST) of different groups of mice.
FIG. 3 shows H & E sections of mice from different groups.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
Animals: SPF-class male C57BL/6 mice, body weight 22 (+ -2) g,7 weeks of age. Purchased from zhuhai Bai Tong Biotechnology Co., ltd., animal license number: [ SCXK (Yue) 2022-0051].
Test article: nux Prinsepiae is purchased from Guangzhou to the pharmaceutical industry Co., ltd.
Reagent:
95% ethanol (AR), shanghai microphone Biochemical technologies Co., ltd;
Carbon tetrachloride (HPLC), shanghai microphone Biochemical technologies Co., ltd;
olive oil (pharmaceutical grade), shanghai microphone Biochemical technology Co., ltd;
liver function and oxidative stress detection kit, nanjing builds biological engineering research all companies;
silybin (SY), shanghai Ala Biochemical technology Co., ltd.
Example 1 preparation of Prinsepia utilis Royle extract
In the embodiment, the nux Prinsepiae is extracted by water, and the specific method is as follows:
(1) Preliminary crushing: pulverizing nux Prinsepiae into coarse powder.
(2) And (3) heating and reflux extracting pure water: reflux-extracting dried coarse powder of Chinese medicinal materials with 8 times of pure water at 80deg.C for 1 hr, filtering, adding 8 times of pure 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.
EXAMPLE 2 Effect of Prinsepia extract on ALT and AST Activity of carbon tetrachloride-induced acute liver injury in mice
Alanine aminotransferase (ALT, alanine aminotransferase) and aspartate aminotransferase (AST, ASPARTATE AMINOTRANSFERASE) are enzymes whose blood values increase when the liver is damaged, respectively, that is, enzymes which use this property as an index of liver function. The invention evaluates the effect of nux Prinsepiae extract on the acute liver injury of mice induced by carbon tetrachloride through the indexes, and the specific method and the result are as follows:
1. Experimental method
1.1 Grouping and administration of animals
The 60 mice are divided into a blank control group, a model group, a nux Prinsepiae extract low-dose administration group, a nux Prinsepiae extract medium-dose administration group, a nux Prinsepiae extract high-dose administration group and a positive control group according to the design of a body weight random area. Wherein, the positive control group is administrated with silybin 100mg/kg body weight/day, the low dose group of nux Prinsepiae is administered according to the concentration of crude drug of 0.65g/kg body weight per day, the medium dose group is administered according to the concentration of crude drug of 1.3g/kg body weight per day, and the high dose group is administered by gastric lavage for 7 days according to the concentration of 2.6g/kg body weight per day.
1.2 Establishment of CCl 4 acute liver injury model
After 2h of last dose, 10% (v: v) concentration of single-shot subcutaneous injection CCl 4 in each group except the blank group was dissolved in olive oil (10 mL/kg) to induce an acute liver injury model; the blank group was subcutaneously injected with an equal volume of olive oil.
1.3 Sample collection and processing
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 ℃, and the supernatant is taken to measure liver function indexes (ALT, AST).
1.4 Determination of Biochemical index
Detecting alanine Aminotransferase (ALT) by a kit method, and purchasing all company Limited for biological engineering research in Nanjing, batch number 20230426;
Aspartic acid Aminotransferase (AST), purchased from south kyo construction bioengineering research, all company, lot 20230427.
1.5 Statistical data processing
All experimental data were statistically analyzed using GraphPad prism6.0 software, using one-way ANOVA when the metering data met normal distribution, using TAMHANES T (M) for testing if not, and n=5 for each group of numbers when data were analyzed, compared to control, #, P <0.05; # and P <0.01; # #, P <0.001; p <0.05 compared to model control; * P <0.01; * P <0.001.
2. Experimental results
In FIG. 1, the ALT activity of model group (CCl 4) is significantly higher than that of control group (NC), which indicates that the molding is successful, and the ALT activity of nux Prinsepiae extract prepared in example 1 can be significantly reduced, and it can be seen from the graph that the effects of nux Prinsepiae water extract high dose group (RR-H), medium dose group (RR-M) and low dose group (RR-L) are very good, and ALT value is significantly reduced compared with model group, especially ALT activity of nux Prinsepiae high dose group (RR-H) and nux Prinsepiae medium dose group (RR-M) is significantly reduced (P < 0.01) compared with model group, and the effect is much better than that of positive control group to which Silybin (SY) is administered.
In fig. 2, the AST activity of model group (CCl 4) is significantly higher than that of control group (NC), which indicates that the molding was successful, and it can be seen from the figure that administration of the nux Prinsepiae extract prepared in example 1 can reduce the AST activity, and that the nux Prinsepiae high dose group (RR-H) and the medium dose group (RR-M) have very good effects, and that AST values are significantly different (P < 0.01) compared with the model group.
Example 3 liver tissue Effect of Prinsepia extract on carbon tetrachloride-induced acute liver injury in mice
Liver tissue of the right lobe (with maximum cross section, portal area and central venous area) of the liver of the different mice in example 2 was taken and fixed 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.
Analysis of results:
The pathological section is carried out on the liver tissues of mice in the normal group, the model group, the silybin group, the nux Prinsepiae high, medium and low dose groups, and the result is shown in figure 3, and the lobule structure of the normal liver control group (NC) is clear, the liver cells have no obvious lesions, the cytoplasm is rich, and the nuclear structure is clear. Model group (CCl 4) liver, liver cells are commonly denatured, necrotic or nucleus is contracted and dissolved, and a large number of inflammatory cells infiltrate. As can be seen from the graph, the liver lobule structure of the mice after administration of the nux Prinsepiae extract is clear, and the liver chordae and the liver sinus have no obvious lesions and the cytoplasm is uniformly colored no matter the nux Prinsepiae extract is high-dose (RR-H) and medium-dose (RR-M) and low-dose (RR-L). And after the administration of the high dose group (RR-H) and the medium dose group (RR-M) and the low dose group (R-L), the liver lobule structure of the mice is clear, the liver cells are not obviously abnormal, the cytoplasm is rich, the nuclear structure is clear, and the positive control group is far better than the positive control group to which the Silybin (SY) is administered.
Although embodiments of the present invention have been described above in connection with the above, the present invention is not limited to the specific embodiments and fields of use described above, which are intended to be illustrative, instructive, and not limiting. Those skilled in the art, having the benefit of this disclosure, may effect numerous forms of the invention without departing from the scope of the invention as claimed.
Claims (8)
1. Application of nux Prinsepiae or its extract in preparing product for treating and/or preventing diseases caused by liver injury is provided.
2. The use according to claim 1, wherein the disease caused by liver injury is selected from any one or more of acute liver injury, chronic liver injury, liver fibrosis, cirrhosis or liver cancer.
3. The use according to claim 1, wherein the cause of the liver injury is selected from any one or more of trauma, drugs, alcohol, chemical toxins, viruses or autoimmunity.
4. The use according to claim 1, wherein the product is a pharmaceutical or health product.
5. The use according to claim 1, wherein the product is for use in mammals.
6. The use according to claim 1, wherein the single dose of said product corresponds to 15g to 200g of nux Prinsepiae.
7. The use according to claim 1, wherein said nux Prinsepiae extract is nux Prinsepiae water extract.
8. The use according to claim 7, wherein the nux Prinsepiae water extract is prepared by pulverizing nux Prinsepiae, adding water, heating and reflux extracting, collecting filtrate, and concentrating.
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