CN110787196B - Application of tin leaf vine and its extracts in the preparation of drugs for the treatment and/or prevention of alcoholic liver injury - Google Patents
Application of tin leaf vine and its extracts in the preparation of drugs for the treatment and/or prevention of alcoholic liver injury Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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Abstract
本发明涉及药物应用领域，特别是锡叶藤及其提取物在制备治疗和/或预防酒精性肝损伤药物的应用。本发明所述锡叶藤提取物选自以下任一种或一种以上的混合：锡叶藤水提取物、锡叶藤醇提取物、锡叶藤乙酸乙酯提取物、锡叶藤正丁醇提取物。本发明提供的锡叶藤提取物能明显升高酒精性肝损伤小鼠血清的白蛋白含量，通过修复损伤细胞的内质网、高尔基体、核糖体等，改善肝合成白蛋白的能力，从而给肝脏提供保护。The invention relates to the field of pharmaceutical application, in particular to the application of tinea vine and its extract in preparing medicines for treating and/or preventing alcoholic liver damage. The extract of Cinnamon in the present invention is selected from any one or more of the following mixtures: water extract of Cinnamon, alcohol extract of Cinnamon, ethyl acetate extract of Cinnamon, n-butanol of Cinnamon Extract. The extract provided by the invention can significantly increase the serum albumin content of mice with alcoholic liver injury, and improve the ability of the liver to synthesize albumin by repairing the endoplasmic reticulum, Golgi body, ribosome, etc. Provides protection to the liver.
Description
Technical Field
The invention relates to the field of medicine application, in particular to application of cassis vine and an extract thereof in preparing a medicine for treating and/or preventing alcoholic liver injury.
Background
The Tinospora pareira is root of Tetracera asiatica (Lour.) Hoogland of Dilleniaceae. It is cool in nature and bitter in taste, and has the functions of astringing to arrest diarrhea, relieving swelling and alleviating pain. It is often used to treat diarrhea, dysentery, rectocele, etc. The medicinal materials are rich in various bioactive components such as tannin, flavone, terpenoids and the like, and have obvious activities such as bacteriostasis, anti-inflammation, anti-tumor and the like [ Chinese materia medica editorial Commission of Chinese traditional medicine administration, Chinese materia medica [ M ]. volume 3, Shanghai: shanghai science and technology press, 1999: 510-511.]. The Siberian cinquefoil herb has rich medicinal material resources, is produced in southern and southwest provinces of China, and is one of the common Chinese herbal medicines in folk.
For example, chinese patent application publication No. CN106236795A discloses a Chinese medicinal preparation with bacteriostatic action, in particular a Chinese medicinal preparation prepared from extract of cassis vine (root of cassis vine Tetracera asiatica (Lour.) Hoogland of the family of dillenaceae); research results show that the traditional Chinese medicine preparation has bacteriostatic activity on dysentery bacillus, Escherichia coli, staphylococcus aureus, bacillus subtilis and typhoid bacillus, and shows good bacteriostatic and anti-inflammatory effects.
At present, no report on the application of the cassis vine and the extract thereof in the preparation of the medicine for treating and/or preventing the alcoholic liver injury exists.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides the application of the cassiterite and the extract thereof in preparing the medicines for treating and/or preventing tumors.
The tinctoria vine is root of Tetracera asiatica (Lour.) Hoogland of Dilleniaceae.
The purpose of the invention can be realized by the following technical scheme:
application of herba Siphonostegiae in preparing medicine for treating and/or preventing liver injury is provided.
Preferably, the method comprises the step of extracting the cassiterite stems by adding a solvent to obtain the cassiterite stem extracts.
Preferably, the cassis vine extract is selected from any one or more of the following mixtures: water extract, alcohol extract, ethyl acetate extract and n-butyl alcohol extract of Siberian cinquefoil.
Preferably, the aqueous extract of cassis leaves is prepared by the following method: taking and crushing the cassiabarktree leaves, adding purified water according to the material-liquid ratio of 1: 20-30 every time, decocting for 1-2 h at 90-110 ℃, repeating for 2-4 times, and combining to obtain the cassiabarktree leaf tea.
Preferably, the ethanol extract of cassiteria japonica is prepared by the following method: taking and crushing the cassiabarktree leaves, adding 40-60% by volume of ethanol into the cassiabarktree leaves according to the material-liquid ratio of 1: 20-30 every time, performing reflux extraction on the cassiabarktree leaves in a water bath at the temperature of 70-90 ℃ for 1-2 hours, repeating the reflux extraction for 2-4 times, and combining the materials to obtain the cassiabarktree leaf tea.
Preferably, the cassis vine ethyl acetate extract is prepared by the following method: and (3) dispersing the Stachys sieboldii alcohol extract in 40-60% by volume of ethanol, continuously extracting with ethyl acetate for 10-20 times, wherein each time is 40-60 mL, and combining ethyl acetate extract to obtain the finished product.
Preferably, the n-butanol extract of cassis leaves is prepared by the following method: dispersing the Stachys sieboldii alcohol extract into 40-60% ethanol by volume percent, continuously extracting for 10-20 times by using ethyl acetate, wherein 40-60 mL of ethyl acetate extract is obtained each time, and discarding the ethyl acetate extract; and continuously extracting the residue with n-butyl alcohol for 10-20 times, wherein 40-60 mL of n-butyl alcohol is extracted each time, and mixing n-butyl alcohol extract liquor to obtain the compound.
Preferably, the aqueous extract of cassis leaves is prepared by the following method: pulverizing caulis et folium Stauntoniae, adding purified water at a material-liquid ratio of 1:25 each time, decocting at 100 deg.C for 1 hr, repeating for 3 times, and mixing.
Preferably, the ethanol extract of cassiteria japonica is prepared by the following method: pulverizing caulis et folium Stauntoniae, adding 50% ethanol at a ratio of 1:25, extracting under reflux in 80 deg.C water bath for 1 hr, repeating for 3 times, and mixing.
Preferably, the cassis vine ethyl acetate extract is prepared by the following method: and (3) dispersing the ethanol extract of the Stachys sieboldii in 50% ethanol by volume, continuously extracting with ethyl acetate for 15 times, wherein each time is 50mL, and combining ethyl acetate extract to obtain the finished product.
Preferably, the n-butanol extract of cassis leaves is prepared by the following method: dispersing the Stachys sieboldii alcohol extract in 50% ethanol by volume, continuously extracting with ethyl acetate for 15 times, each time 50mL, and discarding the ethyl acetate extract; extracting the residue with n-butanol for 15 times, each time 50mL, and mixing n-butanol extractive solutions.
Preferably, the liver injury comprises alcoholic liver injury or pharmaceutical liver injury.
Preferably, the liver injury comprises acute liver injury or chronic liver injury.
Preferably, the liver injury is a process before the liver tissue of the body develops into hepatitis, liver cirrhosis, liver fibrosis and liver cancer.
The invention also provides a pharmaceutical composition, which is prepared by mixing the cassytha leaves with pharmaceutically acceptable auxiliary components or therapeutic components.
The invention also provides a pharmaceutical composition, which is prepared by mixing any one of the cassiteria extract and pharmaceutically acceptable auxiliary components or therapeutic components.
Compared with the prior art, the method has the following advantages:
1. the Siberian cinquefoil herb extract provided by the invention has better effects on 6 indexes of ALT, ALB, MDA, NO, CAT and IL-6 than the positive medicament bifendate in ten indexes detected by alcohol.
2. The cassytha tenuifolia alcohol extract provided by the invention can obviously reduce the contents of ALT and AST in liver serum of mice with acute liver injury caused by alcohol, which indicates that cassytha tenuifolia has the functions of protecting liver and reducing enzyme; the ALB content is increased, which indicates that the Siberian cinquefoil herb can repair the ribosome, rough endoplasmic reticulum and Golgi apparatus in the damaged cells and restore the protein synthesis function of the liver; but also can reduce the content of MDA and NO, improve the activity of SOD and CAT, and prompt that the liver protection function of the cassis vine plays a role by resisting free radicals and inhibiting a lipid peroxidation mechanism; meanwhile, the content of IL-6, TNF-alpha and IFN-gamma in serum is reduced, and the Siberian cinquefoil herb can inhibit the generation of inflammatory factors, protect cell membranes and promote the regeneration and repair of cells.
3. The cassis vine extract provided by the invention can obviously reduce the transaminase content of the serum of a mouse with alcoholic liver injury, has good chemical resistance and alcoholic liver injury resistance, obviously reduces the degree of acute liver injury, provides a new and effective method for protecting the liver, and has research significance and application prospect.
4. The cassiterite extract provided by the invention can obviously increase the albumin content of the serum of an alcoholic liver injury mouse, and improve the capacity of synthesizing albumin by liver by repairing endoplasmic reticulum, Golgi apparatus, ribosome and the like of an injured cell, thereby protecting the liver.
5. The extract of each part of the cassiabarktree has certain protection effect on acute liver injury caused by alcohol, can enhance the activities of SOD and CAT, and reduce the contents of MDA and NO.
6. The extract of each part of the cassiabarktree has a certain protection effect on acute liver injury caused by alcohol, can reduce the content of IL-6, TNF-alpha and IFN-gamma in serum, reduce the generation of inflammatory factors, reduce the generation of inflammation, relieve the damage of toxic cells to liver cell membranes, maintain the structure and play a role in protecting the liver.
Drawings
FIG. 1 is a graph of HE staining of liver lobule tissue of a blank control group of mice.
FIG. 2 is a graph showing HE staining of liver lobular tissue of a mouse in a model group.
FIG. 3 is a positive drug group mouse liver lobular tissue HE staining diagram.
FIG. 4 is a graph of HE staining of liver lobular tissue of mice in the water extract high dose group.
FIG. 5 is a graph of HE staining of liver lobule tissue of mice in a high-dose group of n-butanol extract.
FIG. 6 is a graph of HE staining of liver lobular tissue of mice in the ethyl acetate extract high-dose group.
FIG. 7 is a graph of HE staining of liver lobule tissue of mice in a high-dose group of ethanol extracts.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Example 1 preparation of aqueous extract of Stachys sieboldii
The Ceylon leaf is collected in Guangxi Wuming peak forest lands, and is identified as the Tetracera asiatica (Lour.) Hoogland stem of Dilleniaceae plant Ceylon leaf by auxiliary professor of Teng Jianbei of Guangxi traditional Chinese medicine university. Taking 200g of pulverized Stachys sieboldii medicinal powder, adding 5000mL of pure water each time, decocting at 100 deg.C for 1h, repeating for 3 times, mixing the obtained extractive solutions for 3 times, and concentrating to obtain water extract containing 7.0771g crude drug per 1g of water extract.
Example 2 preparation of an alcohol extract of Stachys sieboldii
The Ceylon leaf is collected in Guangxi Wuming peak forest lands, and is identified as the Tetracera asiatica (Lour.) Hoogland stem of Dilleniaceae plant Ceylon leaf by auxiliary professor of Teng Jianbei of Guangxi traditional Chinese medicine university. Taking 100g of the crushed cassiterite powder, adding 2500mL of 50% ethanol each time, carrying out reflux extraction for 1h in a water bath at 80 ℃, repeating for 3 times, combining the obtained extracting solutions for 3 times, and concentrating to obtain cassiterite ethanol extract, wherein the crude drug content of 1g of the ethanol extract is 6.9638 g.
Example 3 preparation of Ethyl acetate extract, n-butanol extract of Stauntonia brachyanthera
Taking the alcohol extract obtained in the example 2, dispersing the alcohol extract in 50% ethanol, continuously extracting with ethyl acetate for 15 times, wherein each time is 50mL, and combining the extract liquor to obtain an ethyl acetate extract of the cassiterite stems; and continuously extracting the residue with n-butanol for 15 times, wherein each time is 50mL, and mixing the extract liquids to obtain the n-butanol extract of the cassiterite. Then concentrating at 80 deg.C to make crude drug content of 1g ethyl acetate extract 52.0833g and crude drug content of 1g n-butanol extract 41.6667 g.
Example 4 efficacy test
1. Laboratory animal
SPF-grade Kunming mice, 18-22g in body weight, half male and half female, were provided by the Experimental animals center of Guangxi medical university. Production license of experimental animal: SCXK Ossa 2014-. The license number of the experimental unit is SYXK Gui 2010-0001. The method is characterized by being bred in an SPF (specific pathogen free) animal house of Guangxi traditional Chinese medicine university, the room temperature is controlled to be 22-25 ℃, the humidity is controlled to be 60-70%, and the illumination is carried out for 12 hours every day.
2. Experimental reagent
Aspartate aminotransferase (AST/GOT) kit (96T, lys method): nanjing was established as bioengineering institute under the cargo number C010-1; alanine aminotransferase (ALT/GPT) test kit (96T, microplate method): nanjing was established as a bioengineering institute under the cargo number C009-2; albumin (ALB) assay kit (96T, colorimetric): nanjing was established as bioengineering institute under the product number A028-1; malondialdehyde (MDA) assay kit (96T, TBA method): nanjing was established as bioengineering institute under the cargo number A003-1; catalase (CAT) assay kit (96T, visible light method): nanjing was established as bioengineering institute under the product number A007-1; nitric Oxide (NO) assay kit (96T, one-step method): nanjing is built into a bioengineering institute, and has a product number of A013-2; total superoxide dismutase (SOD) assay kit (96T, WST-1 method): nanjing is built into a bioengineering institute, with a cargo number of A001-3; mouse interleukin-6 (IL-6), mouse tumor necrosis factor alpha (TNF-alpha), mouse gamma interferon (IFN-gamma): 96T, X-Y Biotechnology; paraffin, formaldehyde, xylene, absolute ethyl alcohol, ammonia water, ethanol, n-butanol and ethyl acetate (Shanghai national drug group chemical reagent Co., Ltd., analytical purity); hematoxylin, eosin (BASO, cat # 714094, BA 4099); neutral resin (beijing solibao, cat # G8590); bifendate dripping pill: beijing collaborates with the pharmaceutical factory, approved article number: the national standard of medicine H11020980; 52 percent of white spirit (red star Erguotou), peanut oil (luhua) and water are ultrapure water.
3. Laboratory apparatus
An enzyme-labeling instrument: U.S. BIO-TEK, model number: ELX 680;
ultraviolet spectrophotometer: agilent technologies, usa, model: agilent 8453;
an ultrasonic instrument: shanghai benezin ultrasound ltd, model: B2200S-T;
electronic analytical balance: beijing sydolis instruments systems ltd, model: BT 224S;
a desk-top high-speed centrifuge: beijing li centrifuge ltd, model: LG 16-WA;
a vacuum pump: zhengzhou great wall science and trade company, model: SHB-III;
electric heating constant temperature water bath: shanghai precision testing equipment Co., Ltd, type: DK-S26;
constant temperature incubator at 37 ℃: shanghai easily-expandable instruments, Inc., model number: 1-35 RPM;
a vortex mixer: shanghai globulus materialization instrument factory, model: WH-861;
an upright microscope: NIKON corporation, model number: ECLIPSE Ni;
a liquid transfer device: gilson P pipettor company, model: p2, P10, P20, P100, P200;
and (3) constant-temperature baking oven: shanghai hengyi scientific instruments ltd, model: DHG-9023A;
paraffin slicer: laike corporation, model: SQ 2125;
sheet spreading machine: laike corporation, model: PPTHK-21B;
microscopic image analysis system: NIKON corporation, model number: DS-Ri 2.
4. Experimental methods
(1) Grouping and administration:
taking 90 healthy mice, 20g +/-2 g, half of male and female, randomly dividing into 15 groups, 6 mice in each group, feeding the mice in a laboratory for 7 days, then, administering the drugs according to the table 1, intragastrically administering each group for 1 time every day, and administering equal volume of water to a blank control group and a model group for 14 days continuously.
TABLE 1 Alcoholic liver injury model grouping and dosing table
(2) Molding and material taking: 1h after the last dose: the liver injury models are manufactured by induced intragastric administration of 15mL/kg of body weight with 52% white spirit at one time; the blank control group was injected with an equal volume of saline intraperitoneally.
After fasting without water supply and 16 hours, removing eyeballs and taking blood, centrifuging at 3000r/min, taking supernatant, storing in a refrigerator at minus 20 ℃, separating serum, determining ALT, AST, ALB, IL-6, TNF-alpha and IFN-gamma indexes according to kit instructions, and determining SOD, MDA, NO and CAT biochemical indexes by 10% liver tissue homogenate solution.
(3) Sample preparation
1) Removing mouse eyeball to obtain blood, collecting blood with 1.5ml LEP tube, centrifuging at 3000rpm at-4 deg.C for 15min after blood coagulation, collecting serum with pipette gun in 0.5ml LEP tube, and storing in refrigerator at-80 deg.C for use.
2) And (3) quickly dissecting, taking out the thymus, the liver and the spleen of the mouse, washing accumulated blood with ice physiological saline, sucking water on filter paper, weighing and recording respectively.
3) Weighing mouse liver, taking about 0.3g, placing in 10mLEP tube, adding appropriate amount of ice physiological saline to prepare 10% liver homogenate, centrifuging at 3000rpm for 15min, sucking supernatant, placing in 1.5mLEP tube, and storing in refrigerator at-80 deg.C.
4) About 1 × 1cm liver left lobe 2 portions were cut and fixed in 4% paraformaldehyde for 48 hours.
(4) Index observation and detection
1) Before and after the experiment, the indexes of the hair, the spirit, the body type and the like of the mouse are observed once; after the test, the color and the shape of the liver tissue are observed.
2) And (3) biochemical immune index detection: after the medicine and the model are made, the fasting is not forbidden, after 16h, the eyeball is removed and blood is taken, centrifugation is carried out at 3000r/min, supernatant is taken, the supernatant is stored in a refrigerator at 80 ℃, serum is separated, ALT, AST, ALB, IL-6, TNF-alpha and IFN-gamma indexes are measured, SOD, MDA, NO and CAT biochemical indexes are measured by 10 percent liver tissue homogenate solution, and the protein concentration in the liver homogenate is measured by a BCA method. And (4) carrying out sample adding measurement according to the method of each index kit, thereby calculating the result.
3) Preparing pathological sections: quickly taking out the liver, fixing the liver tissue in 4% paraformaldehyde solution for 48h, performing HE staining, and storing at-80 ℃ for homogenizing and determining biochemical indexes. Preparing pathological sections of the liver, observing the pathological sections under an optical microscope, collecting images, and grading according to an Ishak grading system.
(5) Statistical treatment
Single factor analysis of variance was performed with SPSS 20.0 software, results in
Is represented by P<0.05 is a significant difference, P<A very significant difference was exhibited at 0.01. The variance is uniform, and every two LSDs are used for comparison; when the variance is irregular, Dunnett T3 method is used.
5. Results of the experiment
(1) Influence of Siberian cinquefoil on acute liver injury pathological changes
1) Observation with naked eyes
The skin and hair of the mouse are clean and tidy before the liver alcohol injury, the mouse is full of spirit, lively and well-moving, the body shape is normal, the liver color is bright red, and the mouse is soft and elastic. After being injured by alcohol, except the blank group, each group had phenomena of listlessness, slow response, significantly reduced activity, lusterless fur and the like in different degrees, and the model group was most obvious, the dissected liver was reddish, swollen and slightly rough in surface. The water extract, alcohol extract, ethyl acetate extract and n-butanol extract of the staphyliantha tinctoria have high dosage, and the bifendate group has drunkenness phenomenon, poor spirit, reduced activity, dull fur, no obvious change of body type, light red liver and slight swelling; the water extract, alcohol extract and ethyl acetate extract of the cassiteria indica, the middle and low dose groups of the n-butanol extract have the phenomenon of drunkenness, listlessness, reduced activity, dull fur, no obvious change of body types, and light red and swollen liver.
2) Microscopic observation
As shown in figure 1, the structure of liver lobules of blank mice is clear and complete, liver cells are arranged in the lobules regularly, the cell nucleus is large and round, the nucleolus is obvious, the liver cells are similar in size, the cytoplasm is rich, the radial arrangement is shown by taking a central vein as the center, no fibrous tissue hyperplasia is in a junction area, and no inflammatory cell appears under the observation of a microscope. The liver lobular structure of the model group mice is seriously damaged, the liver sinus structure and the liver funicle are unclear, disordered liver funicles are arranged, more fibroblasts appear, the proliferative fibrous connective tissue and the multiple junction area-central vein (P-C) are bridged, partial different degrees of focal necrosis and pseudolobules are visible, the liver junction area is enlarged, and obvious inflammatory cells infiltrate into the liver tissue. The bifendate group has structurally complete liver lobules and a sink area, false lobules are not formed, a small amount of inflammatory cells infiltrate into a part of the sink area, and the fibrous connective tissue hyperplasia is reduced and is obviously lighter than that of the model group in the overall lesion degree.
The liver lobule structures of mice with alcoholic liver damage are slightly disordered in a Siberian cinquefoil vine ethyl acetate high-dose group and a n-butyl alcohol high-dose group, are arranged in a normal liver cell cord manner, are less infiltrated by inflammatory cells, are not found in steatosis and obvious focal necrosis, reduce the fibrous connective tissue hyperplasia in a junction area, and are obviously lighter than the whole lesion degree of a model group. The mouse liver lobule structures are disordered in a Siberian cinquefoil water high dose group and a n-butanol medium dose group, the bridging necrosis of a part of region 3-central vein (P-C) appears, the formation part of steatosis and false lobule is visible, more obvious inflammatory cells are infiltrated in liver tissues, more fibrous connective tissues are proliferated, and the whole lesion degree is close to that of a model group. The disorder degree of the liver lobule structure of the mouse is more serious in the middle and low dose groups of the water extraction and the ethanol extraction of the staphylia stannifera, the multiple junction area-central vein (P-C) bridging appears, the steatosis and the formation part of false lobule are visible, obvious inflammatory cells are infiltrated in the liver tissue, more fibrous connective tissue is proliferated, and the whole lesion degree is more serious than that of the model group
3) Effect of HE staining on pathological section score results
As shown in table 2, the model group scores were significantly higher than the blank group compared to the blank group, with very significant differences (P <0.01) indicating successful model building. The bifendate group score has obvious difference (P <0.01) compared with the model group, and the results are different (P <0.05) when the water content is high, the ethanol content is high, the ethyl acetate content is high, and the ethanol medium score is compared with the model group.
TABLE 2 evaluation table of HE-stained pathological sections of liver of mice with alcoholic liver injury from different parts of Stachys sieboldii
Note: # P <0.05, # P <0.01, compared to the blank group;
p <0.05, P <0.01, compared to model group
(2) Biochemical index observation of cassis vine serum
1) Changes in serum ALT, AST, ALB
As can be seen from Table 3, the serum ALT activity exhibited a very significant increase (P <0.01) in the model group compared to the blank control group. Indicating that the model of the acute liver injury model is successfully made. Compared with the model group, the serum ALT levels of the cissampelos pareira water extract high-medium dose group, the n-butanol extract high-medium dose group, the ethyl acetate extract high-medium dose group and the ethanol extract high-medium dose group are reduced and show a dose-dependent trend, the statistical comparison difference shows a very significant (P is less than 0.01) and the ethanol extract low-dose group shows a significant difference (P is less than 0.05). The water extract dosage and ethanol dosage were not significant. As can be seen from Table 3, the serum AST activity exhibited a very significant increase (P <0.01) in the model group compared to the blank control group. Indicating that the model of the acute liver injury model is successfully made. Compared with the model group, the serum AST levels of the water extract high-dose group, the n-butanol extract high-medium dose group, the ethyl acetate extract high-medium dose group and the ethanol extract high-medium dose group of the staphylia sinensis are all reduced, the statistical comparison difference shows that the serum AST levels are extremely obvious (P is less than 0.01), and the dose group difference in the water extract is obvious (P is less than 0.05). The low dose group with water extract and the low dose group with ethanol extract are not significant (P is more than 0.05). The ALT data show that the Stachys sieboldii extract can relieve acute liver injury of mice caused by alcohol. As can be seen from Table 3, the ALB activity in the serum exhibited a very significant increase (P <0.01) in the model group compared to the blank control group. Indicating that the model of the acute liver injury model is successfully made. Compared with the model group, the serum ALB levels of the water extract high-dose group, the n-butanol extract high-medium-dose group, the ethyl acetate extract high-medium-dose group and the ethanol extract high-medium-dose group of the cissampelos pareira are obviously reduced, the statistical comparison difference shows that the serum ALB levels are extremely obvious (P is less than 0.01), and the difference of the water extract low-dose group and the ethanol extract low-dose group is not obvious (P is more than 0.05).
TABLE 3 Biochemical index table for serum of alcoholic liver modeling mouse
Note: # P <0.05, # P <0.01, compared to the blank group;
p <0.05, P <0.01, compared to model group
2) Changes in IL-6, TNF-alpha, IFN-gamma in serum
As can be seen from Table 4, the IL-6 content in the serum of the model group mice is higher than that of the blank group, and the results are significantly different (P <0.01), thereby showing that the model is successfully established. Compared with the model group, the content of IL-6 in serum of the water extract high and medium dosage group, the n-butanol extract high and medium dosage group, the ethyl acetate extract high and medium and low dosage group and the ethanol extract high and medium and low dosage group of the cassis leaf water extract is obviously reduced, and the results have significant difference or very significant difference (P < 0.05; P < 0.01). As can be seen from Table 4, compared with the blank group, the content of TNF-alpha in the serum of the mice in the model group is obviously higher than that in the blank group, and the result has very significant difference (P <0.01), thereby showing that the model is successfully established. Indicating that the model of the acute liver injury model is successfully made. Compared with the model group, the serum TNF-alpha level of the n-butyl alcohol extract of the staphylia sinensis, the ethyl acetate extract high and medium dose group and the ethanol extract high and medium dose group is obviously reduced, and the statistical comparison difference shows that the serum TNF-alpha level is very obvious (P is less than 0.01). As can be seen from Table 4, compared with the blank group, the content of IFN-gamma in the serum of the mice in the model group is obviously higher than that in the blank group, and the result has very significant difference (P <0.01), thereby showing that the model is successfully established. Compared with the model group, the serum IFN-gamma levels of the cissampelos pareira water extract high-dose group, the n-butanol extract high-medium dose group, the ethyl acetate extract high-medium dose group and the ethanol extract high-medium dose group are all obviously reduced, and the statistical comparison difference shows that the differences are extremely obvious (P is less than 0.01).
TABLE 4 ELISA biochemical index table for serum of alcoholic liver modeling mouse
Note: # P <0.05, # P <0.01, compared to the blank group;
p <0.05, P <0.01, compared to model group
(3) Biochemical index observation of liver tissue of Siberian cinquefoil
Change of SOD: as can be seen from Table 5, the SOD activity of liver tissue showed a very significant decrease (P <0.01) in the model group compared to the blank control group. Indicating that the model of the acute liver injury model is successfully made. Compared with the model group, the liver tissue SOD levels of the water extract high and medium dose group, the normal butanol, the ethyl acetate and the ethanol extract high and medium low dose group are all obviously increased (P is less than 0.01), which shows that the Siberian cinquefoil vine high dose extraction group can reduce the acute liver injury of mice caused by alcohol, and shows that the Siberian cinquefoil vine has certain antioxidation.
Change of MDA: as can be seen from Table 5, the MDA activity of the liver tissue showed a very significant increase (P <0.01) in the model group compared with the blank control group. Indicating that the model of the acute liver injury model is successfully made. Compared with the model group, the liver tissue MDA level of the water extract high-dose group, the n-butanol extract high-medium dose group, the ethyl acetate extract high-medium low-dose group and the ethanol extract high-medium dose group is obviously reduced (P is less than 0.01), which shows that the Siberian cinquefoil stem high-dose extract group can effectively reduce the acute liver injury of mice caused by alcohol, and the Siberian cinquefoil stem has a certain antioxidation effect.
Change in NO: as can be seen from Table 5, the NO activity of liver tissue showed a very significant increase (P <0.01) compared with that of the blank control group in the model group. Indicating that the model of the acute liver injury model is successfully made. Compared with the model group, the liver tissue NO levels of the cissampelos pareira water extract high-dose group, the n-butanol extract high-medium dose group, the ethyl acetate extract high-medium dose group and the ethanol extract high-medium dose group are obviously reduced (P is less than 0.01), and the n-butanol extract low-dose group is obviously different (P is less than 0.05).
Change in CAT: as can be seen from Table 5, the CAT activity in liver tissue showed a very significant decrease (P <0.01) in the model group compared with the blank control group. Indicating that the model of the acute liver injury model is successfully made. Compared with a model group, the CAT level of the hepatic tissue of each dose group of the staphylia stannifera is obviously increased (P is less than 0.01), and the difference is particularly obvious.
TABLE 5 Biochemical index table for liver tissue of alcoholic liver model mouse
Note: # P <0.05, # P <0.01, compared to the blank group;
p <0.05, P <0.01, compared to model group
Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (1)
1. The application of the cassiteria tinctoria extract in preparing the medicine for treating and/or preventing acute alcoholic liver injury is characterized in that the cassiteria tinctoria extract is cassiteria tinctoria ethyl acetate extract, cassiteria tinctoria n-butyl alcohol extract or a composition of the cassiteria tinctoria ethyl acetate extract and the cassiteria tinctoria n-butyl alcohol extract;
the cassis leaf vine extract is prepared by the following method: taking 100g of crushed stems of Tetracerasiatica (Lour.) Hoogland of Dilleniaceae, adding 2500mL of 50% ethanol each time, performing reflux extraction for 1h in a water bath at 80 ℃, repeating the reflux extraction for 3 times, combining the obtained extracting solutions for 3 times, concentrating to obtain an ethanol extract of Stephania sinensis, dispersing the obtained ethanol extract in 50% ethanol, continuously extracting for 15 times with ethyl acetate, each time extracting for 50mL, combining the extracting solutions, and concentrating to obtain an ethyl acetate extract of Stephania sinensis; extracting the residue with n-butanol for 15 times, each time 50mL, mixing the extractive solutions, and concentrating to obtain n-butanol extract of caulis et folium Stauntoniae.
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