Application of rhizoma menispermi total alkaloids in preparation of medicine for preventing and treating non-alcoholic fatty liver disease
The treatment of non-alcoholic fatty liver with a total alkaloid of the larvae root has significantly improved the liver status of obese mice with high-fat and high-sugar diet, solved the problem of the lack of effective anti-NAFLD drugs in the prior art, and provided a potential therapeutic drug.
Patent Information
- Application Number
- CN202411896896.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-23
- Publication Date
- 2025-03-14
AI Technical Summary
The prior art has not yet effectively addressed the treatment of non-alcoholic fatty liver (NAFLD), especially in the absence of approved anti-NAFLD drugs in the context of rising prevalence worldwide.
Using the total alkaloid of the northern bean root as a traditional Chinese medicine extract, it was studied through experiments in obese mice with high-fat and high-sugar diets. It was found that it can significantly reduce weight, improve liver steatosis, reduce blood lipids, improve liver function, and inhibit the expression of inflammatory factors and liver fibrosis genes.
The total alkaloid of the larvae significantly improves the symptoms of non-alcoholic fatty liver, including reducing liver steatosis, lowering blood lipid levels, improving liver function, inhibiting inflammation and liver fibrosis, thus providing a potential therapeutic agent.
Smart Images
Figure CN119607053A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of biomedicine, and in particular to application of total alkaloids of Radix Sophorae Flavescentis in preparing medicines for preventing and treating non-alcoholic fatty liver disease. Background Art
[0002] In recent years, the incidence of non-communicable diseases has been increasing worldwide and has become a serious public health problem. Among them, the global prevalence of metabolic cardiovascular diseases such as diabetes, obesity, non-alcoholic fatty liver disease, hyperlipidemia and atherosclerosis has been rising year by year, causing a huge economic and social burden. Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease in the world, including a series of diseases ranging from simple steatosis or non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH), and then to NASH-related fibrosis, cirrhosis and hepatocellular carcinoma. If not effectively treated, NAFL may develop into NASH and increase the risk of liver failure. However, to date, no anti-NAFLD drugs have been approved by the US FDA or the European Medicines Agency (EMA). Therefore, there is an urgent need to find new NAFLD treatment drugs.
[0003] Natural products are the most common source of drugs. Since the development of natural medicines, 30%-40% of the drugs listed each year are directly or indirectly derived from natural products, which has great advantages for the research and development of future drugs. The root of northern bean is the dried rhizome of the Menispermum dauricum DC., a plant of the Menispermaceae family. The 2020 edition of the Chinese Pharmacopoeia records that the root of northern bean has a slight smell, bitter taste, and cold, and it belongs to the lung, stomach, and large intestine meridians. It has the effects of clearing heat and detoxifying, dispelling wind and relieving pain, and can be used to treat sore throat, enteritis, dysentery, rheumatism, and other symptoms. The alkaloid components in the root of northern bean are its characteristic components and bioactive components, with a total content of about 1.7% to 2.5%. In recent years, studies have found that the pharmacological effects of the total alkaloids of northern bean are very extensive. It not only has pharmacological activities such as antibacterial and anti-inflammatory based on traditional efficacy, but also has biological activities such as protecting the cardiovascular and cerebrovascular system, anti-tumor, anti-depression, and anti-Alzheimer's disease. However, whether it has the ability to prevent and treat liver-related diseases has not been reported at home and abroad. Summary of the invention
[0004] The purpose of the present invention is to provide an application of total alkaloids of Radix Sophorae Flavescentis in the preparation of a medicament for preventing and treating non-alcoholic fatty liver disease, so as to solve the problems existing in the above-mentioned prior art.
[0005] To achieve the above object, the present invention provides the following solutions:
[0006] One of the technical solutions of the present invention is the use of total alkaloids from Radix Sophorae Flavescentis in the preparation of medicines for preventing and treating non-alcoholic fatty liver disease.
[0007] The second technical solution of the present invention is a medicine for preventing and treating non-alcoholic fatty liver disease, comprising total alkaloids from Radix Sophorae Flavescentis.
[0008] Based on the above technical solution, the present invention has the following technical effects:
[0009] The present invention evaluates the effect of the Chinese medicine extract Menispermi Rhizoma Total Alkaloids (MRTA) in treating non-alcoholic fatty liver disease, and finds that the extract can significantly reduce the body weight of obese mice on a high-fat and high-sugar diet, effectively improve non-alcoholic fatty liver disease, effectively reduce liver fatty degeneration, reduce blood lipids, improve liver function, reduce the content of ALT, AST, and TG in the blood of obese mice on a high-fat and high-sugar diet, and inhibit the expression of inflammatory factors and liver fibrosis genes. This shows that the total alkaloids of Menispermi Rhizoma can be used as a potential drug for treating non-alcoholic fatty liver disease. BRIEF DESCRIPTION OF THE DRAWINGS
[0010] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0011] Figure 1 For use The high-resolution small animal ultrasound imaging system acquired the liver ultrasound images of mice in each group.
[0012] Figure 2 The effects of MRTA in the present invention on body weight and blood glucose in mice were shown in Figure 2. Each group of mice (n=5) was divided into a normal group (CON), a high-fat and high-sugar diet group (HFHSD), a low-dose group of total alkaloids from Radix Sophorae Flavescentis (MRTA 10 mg / kg), a high-dose group of total alkaloids from Radix Sophorae Flavescentis (MRTA 20 mg / kg), and a positive drug (Sulforaphane, SFN 15 mg / kg) control group; a is the change in body weight of mice, and b is the change in blood glucose of mice.
[0013] Figure 3The effects of MRTA in the present invention on serum-related indices of mice were studied. The mice in each group (n=5) were divided into a normal group (CON), a high-fat and high-sugar diet group (HFHSD), a low-dose of total alkaloids from Radix Sophorae Flavescentis (MRTA-L), a high-dose of total alkaloids from Radix Sophorae Flavescentis (MRTA-H), and a positive control group (SFN); a is the change in the content of alanine aminotransferase (ALT) in the serum of each group of mice, b is the change in the content of aspartate aminotransferase (AST) in the serum of each group of mice, and c is the change in the content of triglyceride (TG) in the serum of each group of mice.
[0014] Figure 4 The effects of MRTA in the present invention on relevant indicators in mouse liver, which are divided into a normal group (CON), a high-fat and high-sugar diet group (HFHSD), a low-dose of total alkaloids from northern bean root (MRTA-L), a high-dose of total alkaloids from northern bean root (MRTA-H), and a positive control group (SFN), with 5 mice in each group (n=5); a is the change in the content of reactive oxygen (ROS) in the liver of each group of mice, b is the change in the content of triglyceride (TG) in the liver, and c is the change in the content of malondialdehyde (MDA) in the liver.
[0015] Figure 5 The images are the liver appearance of mice in each group.
[0016] Figure 6 The liver tissues of mice in each group were frozen and sectioned and then stained with hematoxylin-eosin (HE).
[0017] Figure 7 The liver tissues of mice in each group were frozen and sectioned and then stained with Oil Red O for analysis.
[0018] Figure 8 qPCR analysis of the expression of liver fibrosis and hepatitis-related genes in the liver tissues of mice in each group (CON, HFHSD, MRTA-L, MRTA-H, SFN). Among them, a is the liver fibrosis gene Ctgf; b is the liver fibrosis gene Col3a1; c is the liver inflammation-related gene Tnf; d is the liver inflammation-related gene Il1b. DETAILED DESCRIPTION
[0019] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.
[0020] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.
[0021] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.
[0022] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.
[0023] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.
[0024] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.
[0025] The embodiment of the present invention provides the use of total alkaloids from Radix Sophorae Fructus in preparing a medicine for preventing and treating non-alcoholic fatty liver disease.
[0026] The present invention provides a Chinese medicine extract for preventing and treating non-alcoholic fatty liver disease, which is total alkaloids of Radix Sophorae Flavescentis. Total alkaloids of Radix Sophorae Flavescentis is a phenolic total alkaloid extracted from the dried rhizomes of the plant Puerariae ternata of the Menispermaceae family. The present invention provides a potential therapeutic drug for the prevention and treatment of fatty liver disease. The present invention is an extension of the clinical application of total alkaloids of Radix Sophorae Flavescentis. The present invention studies the effect of total alkaloids of Radix Sophorae Flavescentis in obese mice fed a high-fat diet, and confirms that total alkaloids of Radix Sophorae Flavescentis can significantly reduce the body weight, hepatitis, and liver fibrosis of obese mice fed a high-fat diet after treatment, and reduce the content of ALT and AST in the liver and blood of obese mice fed a high-fat diet. This shows that total alkaloids of Radix Sophorae Flavescentis have a therapeutic effect on fatty liver.
[0027] The embodiment of the present invention also provides a medicine for preventing and treating non-alcoholic fatty liver disease, comprising total alkaloids of Radix Sophorae Flavescentis.
[0028] In some specific embodiments, pharmaceutically acceptable excipients are also included.
[0029] In some specific embodiments, the pharmaceutically acceptable excipient is suitable for liquid dosage form, solid dosage form or paste dosage form.
[0030] In some specific embodiments, the non-alcoholic fatty liver disease is caused by a high-fat, high-sugar diet.
[0031] In some specific embodiments, the non-alcoholic fatty liver disease includes non-alcoholic simple fatty liver disease, non-alcoholic steatohepatitis and liver fibrosis.
[0032] In the application provided by the present invention, the pharmaceutical preparation for treating fatty liver is composed of an effective amount of total alkaloids of Radix Sophorae Fructus or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable adjuvant. Those skilled in the art can directly or indirectly add the total alkaloids of Radix Sophorae Fructus or a pharmaceutically acceptable salt thereof to various commonly used pharmaceutically acceptable adjuvants required for preparing different dosage forms, such as disintegrants, lubricants, emulsifiers, adhesives, etc., and prepare common dosage forms, including but not limited to capsules, microcapsules, tablets, granules, dispersed powders, injections, liposomes, oral solutions, and intravenous injections, by conventional pharmaceutical preparation methods.
[0033] Menispermi Rhizoma Total alkaloids (MRTA) was purchased from Chengdu Pusi Biotechnology Co., Ltd.
[0034] Example 1
[0035] This experiment used 25 SPF male 6-week-old C57BL / 6 mice, which were housed in the SPF Experimental Animal Center of Beijing University of Chinese Medicine and given standard diet and drinking water. The room temperature was controlled at 23±2°C. The experimental protocol involving mice in the experiment was approved by the Animal Experiment Ethics Committee of Beijing University of Chinese Medicine.
[0036] After 7 days of adaptation, the mice were randomly divided into groups, with 5 mice in each group, and divided into a normal group (CON) fed with normal diet, a high-fat and high-sugar diet model group (HFHSD), a low-dose MRTA group (10 mg / kg), a high-dose MRTA group (20 mg / kg) and a positive control group of sulforaphane (15 mg / kg, SFN). Except for the CON group fed with normal feed and ordinary drinking water, the mice in the other groups were given a high-fat diet (42% fat and 0.2% cholesterol) and high-sugar drinking water (high fructose-glucose solution, D-glucose: 18.9 g / L and D-fructose: 23.1 g / L).
[0037] The mice were gavaged with the drug every day. The normal and model groups were gavaged with the same dose of saline. The body weight of the mice was recorded every day. The gavage was continued for 8 weeks.
[0038] After the experiment, blood was collected from the tail tip of the mice using a blood glucose meter to measure blood glucose concentration. Then, the hair was removed from the abdomen of each group of mice, and the Vevo2100 ultra-high resolution small animal ultrasound imaging system was used to collect liver ultrasound images to detect and locate fatty liver. After the liver ultrasound image was acquired, the mice were anesthetized using a small animal inhalation gas anesthesia machine, and the eyeballs were removed to obtain blood and centrifuged to obtain plasma. The changes in blood-related indicators of the mice were measured using a fully automatic biochemical analyzer. After being killed by cervical dislocation, the liver was removed for subsequent experimental testing. SPSS11.5 statistical software was used for statistical analysis. Independent sample t-test was used for comparison between groups.
[0039] After the mouse abdomen was shaved, the operation table was placed horizontally. The probe was placed perpendicular to the mouse body and below the xiphoid process of the mouse sternum. Then the X-axis and Y-axis were fine-tuned to observe the low-echo liver image. The Y-axis was fine-tuned and the probe continued to move toward the tail to observe the liver and kidney sections. Compared with the normal group mice, the liver echo of the HFHSD model group mice was enhanced, showing fatty liver. The liver echo of the MRTA drug treatment group and the SFN positive control group was significantly weakened ( Figure 1 ), indicating that total alkaloids from Radix Sophorae Fructus have the effect of improving fatty liver.
[0040] Example 2
[0041] After 8 weeks of feeding and drug treatment, the mice in the normal group were responsive, with shiny hair and good mental state. The mice in the high-fat and high-sugar diet model group had rough and greasy hair and poor mental state. The hair of the mice in the group receiving oral administration of total alkaloids from Radix Sophorae Fructus and sulforaphane was significantly improved, their movements were free, and their mental state improved. The mice in the high-fat and high-sugar diet group without drug administration gained weight rapidly, while total alkaloids from Radix Sophorae Fructus and positive drugs could effectively inhibit the weight gain of mice, indicating that total alkaloids from Radix Sophorae Fructus could significantly reduce the weight of mice on a high-fat and high-sugar diet after treatment ( Figure 2 (a). Blood glucose test results showed that the blood glucose level of mice in the HFHSD model group was elevated, while the low-dose and high-dose groups of total alkaloids from Rhizoma Coptidis and the positive drug group could significantly reduce the blood glucose level ( Figure 2 (b)
[0042] Example 3
[0043] The results of liver function tests showed that the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum and malondialdehyde (MDA) in liver of the model group were significantly higher than those of the normal group, and decreased significantly after treatment with various doses of MRTA and positive drugs ( Figure 3 a, b and Figure 4c). Hepatic lipid accumulation is an important feature of non-alcoholic fatty liver disease. The results showed that after induction of a high-fat and high-sugar diet, serum and liver triglyceride levels in mice increased significantly ( Figure 3 Medium c and Figure 4 b), indicating that there is obvious lipid accumulation in the liver, and all doses of MRTA and positive drug treatment can significantly inhibit the triglyceride level in the liver ( Figure 3 Medium c and Figure 4 b), improve abnormal lipid accumulation in the liver and thus improve non-alcoholic fatty liver. Lipid metabolism disorders can lead to lipid accumulation in the liver, thereby affecting the generation of reactive oxygen species (ROS). Increased ROS levels and mitochondrial dysfunction promote the progression of non-alcoholic fatty liver. The results showed that after induction of a high-fat and high-sugar diet, the ROS level in the liver of mice increased significantly, and all doses of MRTA and positive drug treatment could significantly inhibit the ROS level in the liver ( Figure 4 Middle a), inhibiting the progression of non-alcoholic fatty liver disease.
[0044] Example 4
[0045] The liver photos showed that the livers of mice in the normal group were red, smooth, shiny, soft, with sharp edges and no swelling ( Figure 5 ), indicating that the blood supply is sufficient and the liver cells are healthy; while the liver of the HFHSD diet-induced model group mice was yellow, dull in color, and swollen with yellow spots, indicating excessive lipid accumulation in the liver of the mice, thus showing dull color and lipid accumulation. All drug treatment groups can significantly improve this phenomenon and reduce lipid accumulation in the liver ( Figure 5 ). The results of hematoxylin-eosin (H&E) staining and oil red O staining showed that the liver lobule structure of the normal group mice was intact, the outline was clearly visible, the liver sinusoids were normal, the hepatocyte cytoplasm was uniform and polygonal, and there was no degeneration; the liver parenchyma structure of the model group was severely damaged, with obvious vacuolar fatty degeneration, inflammatory cell infiltration and lipid accumulation in the portal area. The low-dose and high-dose MRTA groups and the positive drug group could significantly reduce the lipid degeneration of the liver and inhibit the size and number of lipid droplets in the liver ( Figure 6 , 7 ).
[0046] Example 5
[0047] Steatosis and steatohepatitis are considered to be prominent manifestations of NAFLD. In a high-fat, high-sugar diet-induced nonalcoholic fatty liver model, low- and high-dose MRTA treatment significantly inhibited the secretion of proinflammatory cytokines Tnf and Il1b in mice ( Figure 8 c, d); In the development of non-alcoholic fatty liver disease, fatty liver hepatitis is often accompanied by liver fibrosis. The results showed that drug treatment can significantly inhibit the expression of liver fibrosis genes Ctgf and Col3a1 in the liver ( Figure 8a, b).
[0048] In summary, the total alkaloids of northern bean root have a good preventive and therapeutic effect on non-alcoholic fatty liver disease, can effectively improve liver lipid accumulation, fatty hepatitis, and liver damage, can reduce liver and serum lipid levels, improve liver function, and reduce the secretion of inflammatory factors.
[0049] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation methods of the present invention. For ordinary technical users in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.
Claims
1. Application of total alkaloids from Radix Sophorae Flavescentis in the preparation of drugs for preventing and treating non-alcoholic fatty liver disease.
2. A drug for preventing and treating non-alcoholic fatty liver disease, characterized in that: Including total alkaloids from northern bean root.
3. The drug according to claim 2, characterized in that Also included are pharmaceutically acceptable excipients.
4. The drug according to claim 3, characterized in that The pharmaceutically acceptable excipients are suitable for liquid dosage forms, solid dosage forms or paste dosage forms.
5. The drug according to claim 3, characterized in that The non-alcoholic fatty liver disease is caused by a high-fat and high-sugar diet.
6. The drug according to claim 5, characterized in that The non-alcoholic fatty liver disease includes non-alcoholic simple fatty liver disease, non-alcoholic steatohepatitis and liver fibrosis.