CN113577179B - Pharmaceutical composition for liver injury protection and application thereof - Google Patents

Pharmaceutical composition for liver injury protection and application thereof Download PDF

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CN113577179B
CN113577179B CN202110806228.XA CN202110806228A CN113577179B CN 113577179 B CN113577179 B CN 113577179B CN 202110806228 A CN202110806228 A CN 202110806228A CN 113577179 B CN113577179 B CN 113577179B
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pharmaceutical composition
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liver injury
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唐志书
周瑞
宋忠兴
苏洁
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Shaanxi University of Chinese Medicine
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Abstract

The invention belongs to the field of medicines, and relates to a pharmaceutical composition for liver injury protection and application thereof. The invention provides a pharmaceutical composition for liver injury protection, which has good activity of promoting hepatocyte proliferation, activity of relieving oxidative damage of hepatocytes and activity of resisting hepatic fibrosis, and an application thereof.

Description

Pharmaceutical composition for liver injury protection and application thereof
Technical Field
The invention belongs to the field of medicines, relates to a pharmaceutical composition and application thereof, and particularly relates to a pharmaceutical composition for protecting liver injury and application thereof.
Background
Chronic liver disease is a disease with high worldwide morbidity and mortality. Hepatic fibrosis is a histopathological change caused by various acute or chronic liver diseases, such as viral hepatitis B or C, non-alcoholic fatty liver disease, alcoholic steatohepatitis, non-alcoholic steatohepatitis, biliary tract diseases, cholestatic liver diseases and the like. Liver damage leads to hyperactivity of Hepatic Stellate Cells (HSCs) and thus triggers increased extracellular matrix (ECM) synthesis, excessive collagen fibers are deposited in the extracellular space of hepatocytes, resulting in loss of blood supply to hepatocytes, which may further progress to cirrhosis and even liver cancer if proper attention and treatment is not given. It is a reversible scarring response, with nodule formation and organ retraction as important features, and almost all chronic liver diseases can cause liver fibrosis. Fortunately, the process of liver fibrosis is reversible and intervention at even advanced stages of cirrhosis can improve clinical outcome. At present, the treatment principle of hepatic fibrosis is to remove etiology and diminish inflammation, and the treatment strategy is to prevent HSCs (human hematopoietic stem cells) activation, collagen degradation and drug-targeted therapy, but no specific medicine is available for treating hepatic fibrosis at present. The traditional Chinese medicine has the characteristics of treatment based on syndrome differentiation and overall view, and has unique advantages in the aspect of anti-hepatic fibrosis.
According to the traditional Chinese medicine, the liver fibrosis belongs to the category of liver accumulation, and the liver fibrosis is formed by qi stagnation and blood stasis caused by deficiency of healthy qi and then invasion of pathogenic qi (Yi Zong Bi Shu), which is mostly caused by deficiency of healthy qi and six exogenous factors. The "the disease origin and disease symptoms" refers to the feature of the disease persisting for a long time. Therefore, liver injury can be caused by deficiency of healthy qi, damp-heat evil toxin, qi and blood stasis and the like, the traditional Chinese medicine is mainly used for treating diseases by strengthening healthy qi to eliminate pathogenic factors, and heavily activating blood and dissolving stasis when eliminating pathogenic factors, and dialectical administration is carried out by treatment methods of tonifying qi and yin, clearing damp-heat, activating blood and dissolving stasis and the like. Modern researches report that a plurality of Chinese herbal medicines have the effects of resisting hepatic fibrosis and viruses, relieving liver cell injury, repairing damaged liver cells, reducing transaminase and the like. Such as salvianolic acid, total glucosides of paeony, silymarin, radix bupleuri, astragaloside, astragalus polysaccharide, matrine, baicalin, rhein, angelica polysaccharide, deoxyschizandrin, panax notoginseng saponins, curcumin and the like, can relieve liver cell and tissue damage, repair damaged liver cells, promote regeneration of the damaged liver cells, improve inflammation, effectively reduce transaminase and restore liver function, and part of related preparations also have the effects of resisting hepatic fibrosis and viruses. The traditional Chinese medicine can be used by compatibility of medicines, has the effects of attenuation and synergism, plays the characteristics of multi-component, multi-target and multi-channel intervention on diseases, and has a certain effect of relieving chronic injuries and diseases needing long-term administration, so that the current research focuses on researching the action mechanism of protecting liver injuries, particularly reversing hepatic fibrosis, intervening and relieving liver cirrhosis, searching new treatment targets and developing novel medicines for treating liver injuries and having high safety.
Disclosure of Invention
In order to solve the technical problems in the background art, the invention provides a pharmaceutical composition for liver injury protection, which has good activity of promoting hepatocyte proliferation, activity of alleviating oxidative damage of hepatocytes and activity of resisting hepatic fibrosis, and an application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a pharmaceutical composition for liver injury protection, characterized by: the pharmaceutical composition for protecting liver injury is prepared from raw material medicines of American ginseng, hairyvein agrimonia herb and bud, Chinese angelica, paris rhizome, adenophora tetraphylla and platycodon grandiflorum.
The pharmaceutical composition for protecting liver injury is prepared from 20-30 parts of American ginseng, 20-30 parts of hairyvein agrimony, 10-20 parts of angelica sinensis, 10-20 parts of rhizoma paridis, 10-20 parts of adenophora tetraphylla and 10-20 parts of platycodon grandiflorum, and preferably, the pharmaceutical composition for protecting liver injury is prepared from 20 parts of American ginseng, 30 parts of hairyvein agrimony, 10 parts of angelica sinensis, 15 parts of rhizoma paridis, 15 parts of adenophora tetraphylla and 10 parts of platycodon grandiflorum.
An aqueous extract of a liver injury protecting pharmaceutical composition prepared based on the liver injury protecting pharmaceutical composition as described above.
A method of preparing an aqueous extract as hereinbefore described, the method comprising the steps of:
1) weighing radix Panacis Quinquefolii, herba et Gemma Agrimoniae, radix Angelicae sinensis, rhizoma paridis, radix Adenophorae and radix Platycodi according to the ratio of raw materials to obtain a raw material mixture;
2) decocting the mixture of the raw materials for 3 times, adding 10 times of water each time, decocting for 1.5 hours for the first two times and 1 hour for the third time, mixing the water decoctions, vacuum concentrating at 60 deg.C under reduced pressure to obtain soft extract with relative density of 1.05-1.15, spray drying, collecting fine powder and all medicinal powder adhered to the inner wall of spray dryer, and mixing to obtain water extract.
Use of an aqueous extract of a pharmaceutical composition for liver injury protection as hereinbefore described in the preparation of a medicament for liver injury protection.
Use of an aqueous extract of a pharmaceutical composition for liver injury protection as described above for the preparation of a medicament with hepatocyte proliferation promoting activity.
Use of an aqueous extract of a pharmaceutical composition for liver injury protection as hereinbefore described in the manufacture of a medicament active in reducing liver cell injury.
Use of an aqueous extract of a pharmaceutical composition for liver injury protection as hereinbefore described in the manufacture of a medicament active in reducing oxidative damage to hepatocytes.
The application of the aqueous extract of the pharmaceutical composition for protecting liver injury in preparing the anti-hepatic fibrosis medicine is disclosed.
The application of the aqueous extract of the pharmaceutical composition for protecting liver injury in preparing a medicament for relieving liver fibrosis.
The invention provides a pharmaceutical composition for protecting liver injury and application thereof, the pharmaceutical composition is composed of 6 medicines of American ginseng, hairyvein agrimony, adenophora tetraphylla, angelica, paris polyphylla and platycodon grandiflorum, and has the effects of strengthening body resistance and body resistance, strengthening body resistance and detoxifying, generating blood and stopping bleeding and the like, so the pharmaceutical composition is named as a 'body resistance prescription'. The traditional Chinese and western ginseng and the adenophora tetraphylla have the effects of tonifying qi and yin, tonifying vital qi, clearing away heat and toxic materials, enriching blood and activating blood, and eliminating phlegm and discharging pus. In addition, modern pharmacological studies show that compounds such as ginsenoside in American ginseng, angelica polysaccharide in angelica, platycodon polysaccharide, platycodin and adenophora polysaccharide in platycodon root have the effect of protecting liver, and hairyvein agrimonia herb and bud, platycodon root and rhizoma paridis have the effects of oxidation resistance and inflammation resistance. The invention discovers that the positive source formula has obvious proliferation promoting effect on normal human liver cells and uses hydrogen peroxide H2O2Inducing L02 liver cells to cause an oxidative damage model, and finding a positive source to H after administration2O2The damaged liver cells have a protective effect; by the action of the invention, the rat liver cell fatty degeneration phenomenon is obviously reduced, the number of fibroblasts and lymphocytes is reduced, the collagen deposition amount is reduced, and the liver fibrosis degree of the rat is greatly reduced, which shows that each dose group of the normal prescription has obvious repair effect on liver fibrosisGood anti-hepatic fibrosis activity, and is expected to be developed into new drugs for anti-hepatic fibrosis and liver injury protection.
Drawings
FIG. 1 is a histogram of the survival rate of L02 cells obtained in example 2;
FIG. 2 shows the positive source-side pair H of different concentrations in example 32O2Histogram of protective effects of induced oxidative damage to L02 hepatocytes;
FIG. 3 is a graph showing the results of Hoechst 33342/PI fluorescent staining in example 3;
FIG. 4 is a graph showing the change in body mass of rats with liver fibrosis in example 4;
FIG. 5 is a graph showing the liver/body mass ratio of liver fibrosis rats in example 4;
FIG. 6 is a graph of spleen/body weight ratio of liver fibrosis rats in example 4;
FIG. 7 is a schematic view of H & E, Masson staining of a pathological section in example 5;
FIG. 8 is a graph showing the results of the levels of glutamic-pyruvic transaminase, glutamic-oxalacetic transaminase and alkaline phosphatase in the liver fibrosis rats of example 6;
FIG. 9 is a graph showing the results of the levels of total serum protein and albumin in rats with liver fibrosis in example 7;
FIG. 10 is a graph showing the results of the total bile acid level in the serum of the liver fibrosis rat in example 8;
FIG. 11 is a graph showing the results of the total SOD level in liver tissues of liver fibrosis rats in example 9;
FIG. 12 is a graph showing the results of the malondialdehyde level in liver tissue of liver fibrosis rats in example 10;
FIG. 13 is a graph showing the results of the levels of hydroxyproline in serum from rats in liver fibrosis according to example 11;
FIG. 14 is a graph showing the results of the TGF-. beta.1 level in the serum of rats having liver fibrosis in example 12;
FIG. 15 is a graph showing the results of the serum TNF- α levels in rats with liver fibrosis according to example 13.
Detailed Description
The patent number 201310071107.0 issued by the applicant in 2013 and examined and authorized by the applicant is named as' a radiation-resistant productCompound medicine with protective effect, and provides a patent of compound medicine for resisting nuclear radiation protection60The influence of blood deficiency mice caused by Co ray radiation proves that the medicine for protecting the nuclear radiation damage can effectively protect the damage of the reproductive system of the hematopoietic system box caused by the nuclear radiation and improve the immunity. Through years of research, the original compound medicine formula is optimized, and the existing traditional Chinese medicine compound composition original source formula which is prepared from American ginseng, hairyvein agrimony, adenophora tetraphylla, angelica, rhizoma paridis and platycodon grandiflorum is determined. The research of the invention finds that the pharmaceutical composition has the function of promoting the proliferation of the normal human liver cells L02 and the function of resisting the oxidative damage of the liver cells, and preliminarily deduces that the pharmaceutical composition has the liver protection function. Chronic liver injury can cause hepatic fibrosis, and reports that the medicine composition is used for preventing and treating hepatic fibrosis diseases in a positive prescription are not found so far. After long-term experiments by the inventor, the finally obtained pharmaceutical composition for protecting liver injury is prepared from the bulk drugs of 20-30 parts of American ginseng, 20-30 parts of hairyvein agrimony, 10-20 parts of angelica, 10-20 parts of rhizoma paridis, 10-20 parts of adenophora tetraphylla and 10-20 parts of platycodon grandiflorum, and the pharmaceutical composition for protecting liver injury formed in the proportion range has very good or basically equivalent effects.
In order to illustrate the problem, the invention randomly adopts the raw materials of 20 parts of American ginseng, 30 parts of hairyvein agrimony, 10 parts of angelica, 15 parts of rhizoma paridis, 15 parts of adenophora tetraphylla and 10 parts of platycodon grandiflorum to prepare the pharmaceutical composition for protecting liver injury in the range, and carries out relevant tests and verifies the effect. The technical scheme provided by the invention is explained in detail by combining the specific embodiments as follows:
example 1 preparation of a Positive stock aqueous extract
The traditional Chinese medicine composition preparation is prepared from the following raw material medicines in parts by weight: 20 parts of American ginseng, 30 parts of hairyvein agrimony, 10 parts of angelica, 15 parts of rhizoma paridis, 15 parts of adenophora tetraphylla and 10 parts of platycodon grandiflorum, decocting for 3 times, adding 10 times of water each time, decocting for 1.5 hours each time in the first two times, decocting for 1 hour in the third time, combining water decoction liquids decocted for the three times, concentrating under vacuum and reduced pressure at 60 ℃ to obtain thick paste with the relative density of 1.05-1.15, spray drying, collecting fine powder and all medicinal powder adhered to the inner wall of a spray dryer, and mixing uniformly to obtain the water extract of the positive source formula.
The American ginseng adopted by the invention is cool in nature, sweet in taste and slightly bitter, enters heart, lung and kidney channels, and has the effects of tonifying qi and yin, clearing heat and promoting fluid production. The agrimony is neutral in nature and bitter and astringent in taste, and has the effects of astringing to stop bleeding, preventing malaria, stopping dysentery, detoxifying and tonifying deficiency. Chinese angelica root, radix Angelicae sinensis, warm in nature, sweet and pungent in flavor, enters liver, heart and spleen channels, and has the effects of enriching and activating blood, regulating menstruation, relieving pain, moistening intestines and relaxing bowels. Paris polyphylla, with slightly cold nature, bitter taste and little toxicity, enters liver meridian, clears away heat and toxic material, relieves swelling and pain, cools liver and calms convulsion. Adenophora tetraphylla is slightly cold in nature and sweet in taste, enters lung and stomach channels, nourishes yin to clear away the lung-heat, benefits the stomach to promote the production of body fluid, reduces phlegm and benefits qi. Platycodon grandiflorum, neutral in nature, bitter and pungent in taste, enters lung meridian, disperses lung, relieves sore throat, eliminates phlegm and expels pus.
Example 2 Effect of Positive Source formula aqueous extract on the viability of L02 human Normal hepatocytes
L02 cells were cultured at 1.5X 105The density of each/mL is inoculated in a 96-well plate and is kept overnight, the water extract of the positive source prescription is added, the working concentration of the water extract of the positive source prescription in each well is 20, 50, 100, 200, 500 and 1000 mu g/mL, MTT is added after 24h for cell viability measurement, the influence of the water extract of the positive source prescription with different concentrations on the viability of L02 human normal liver cells is measured, a bar chart of the viability of L02 cells is shown in detail in figure 1, the invention finds that the water extract of the positive source prescription is in the range of 100 mu g/mL to 500 mu g/mL, the proliferation promoting effect on human normal liver cells L02 is obvious, and the proliferation promoting effect is obvious at 500 mu g/mL.
Example 3 Positive Source formula aqueous extract vs. H2O2Protective effects of induced oxidative damage to L02 hepatocytes
The results of example 2 were used to screen the range of concentration of the appropriate positive source aqueous extract, further by H2O2Inducing and constructing a liver cell oxidative damage model, determining the survival rate of each group of cells by MTT, and finding that the positive source has obvious proliferation promoting effect on normal human liver cells and hydrogen peroxide H is used for promoting the proliferation of the normal human liver cells2O2Inducing L02 liver cells to cause oxidative damage model, and setting H after concentration exploration2O2Injury was performed at a concentration of 250 μ M, and a positive source was found to be peroxidized after administrationThe hydrogen-damaged liver cells have protective effect. Each group of living cells was stained by Hoechst 33342/PI double staining, the stained cells were observed under Hoechst 33342 fluorescence channel using fluorescence microscope, the total fluorescence number indicates the change of the number of L02 cells, the stained cells were necrotic cells when observed under PI fluorescence channel, and the staining results are shown in detail in FIG. 3. According to the invention, cells are dyed by the Hoechst 33342/PI kit, and the results show that in a bright field, compared with a blank group, the number of the cells in the model group is obviously reduced, the Hoechst 33342 fluorescence image shows that the nuclear matters of the model group are agglutinated and shrunk, which indicates that the liver cells are apoptotic after hydrogen peroxide damage, the PI fluorescence image shows that the cells in the model group have obvious necrotic cells, and the intervention administration of the positive source administration group can relieve the apoptosis and necrosis.
Example 4 Positive Source formula aqueous extract vs CCl4Liver protection effect of induced liver fibrosis rat
Male SD rats were randomly divided into 8 groups: control group, model group, positive source prescription low dose group, positive source prescription medium dose group, positive source prescription high dose group, positive source prescription negative control group, rhizoma paridis negative control group, each group contains 6. Except two negative control groups, the rats of the model group and the administration group are injected with 40 percent CCl in the abdominal cavity4(CCl4: olive oil 2: 3, 1mL/kg), control rats were injected intraperitoneally with an equal amount of olive oil solution 2 times a week for 8 weeks. Silymarin (200mg/kg) was administered to the positive group, and silymarin (1-2, 2-4, 4-6g/kg, in this and the following examples, 1.6 g/kg was administered to the positive low dose group, 3.2g/kg was administered to the medium low dose group, and 4.8g/kg was administered to the high dose group) for 8 weeks once a day. Continuously administering for 2 days after 8 weeks of last molding, fasting, performing abdominal aorta blood sampling after 3 days of anesthesia of all rats, and separating serum; taking a rat liver specimen, fixing the middle part of the left lobe of the liver in 10% paraformaldehyde fixing solution, and carrying out liver histopathological detection; taking specific part of liver, adding 9 times of physiological saline to carry out tissue homogenization, and centrifuging to extract supernatant for later use. The rest liver tissues are stored in a refrigerator at the temperature of 80 ℃ below zero and used for detecting the expression level of the target protein. Continuous monitoring of rat body massThe numerical values are shown in figure 4, and the liver and spleen organ indexes of each group are shown in figure 5 and figure 6. As can be seen from FIGS. 5 and 6, compared with the blank group, the liver and spleen organ indexes of the rats in the model group are both obviously increased, which indicates that the liver and spleen organ indexes of the rats in the model group are abnormal, and the liver and spleen organ indexes of the rats in the administration group are obviously reduced compared with those of the rats in the model group, which indicates that the water extract of the vital prescription has a certain improvement effect on the liver and spleen organ abnormality induced by carbon tetrachloride.
Example 5 Effect of Positive Source Water extract on improving degree of liver fibrosis in rat
As can be seen from the staining result of H & E, Masson in FIG. 7, the rat hepatocytes of hepatic fibrosis induced by carbon tetrachloride showed large-area steatosis, the pseudolobular structure was significantly increased, fibroblasts and lymphocytes were increased, and the number of collagen fibers was increased. The results show that: compared with a blank group, the model group has the phenomena of large-area hepatic cell fatty vacuole degeneration, fibroblast increase and lymphocyte increase; compared with the blank group, the collagen deposition quantity of the model group is obviously increased; and thirdly, the appearance of the normal liver of the rats in the blank group is red, the color of the liver of the rats in the model group is abnormal, and the surface of the rats is obviously hepatic fibrosis. The fat degeneration phenomenon of liver cells of rats in the administration group is obviously reduced, the number of fibroblasts and lymphocytes is reduced, the deposition amount of collagen is reduced, and the hepatic fibrosis degree of rats is greatly reduced, which shows that each dose group of the positive origin formula has obvious repair effect on hepatic fibrosis. The results show that the medicine composition is a positive source prescription, has good anti-hepatic fibrosis activity, and is expected to be developed into a new anti-hepatic fibrosis medicine.
Example 6 Effect of Positive-origin aqueous extracts on serum levels of glutamic-pyruvic transaminase, glutamic-oxaloacetic transaminase and alkaline phosphatase in liver fibrosis rats
Alanine Aminotransferase (ALT), aspartate Aminotransferase (AST) and alkaline phosphatase (ALP) are metabolic enzymes of the liver, and are indexes for clinical examination of liver injury. The expression levels of alanine Aminotransferase (ALT), aspartate Aminotransferase (AST) and alkaline phosphatase (ALP) in the serum of each rat group were measured using a full-automatic biochemical analyzer (Mindray, BS-330E). As can be seen from FIG. 8, consistent with the results of example 5, ALT, AST and ALP levels in serum of the administered group were all decreased compared with rats with hepatic fibrosis induced by carbon tetrachloride, which indicates that the hepatic fibrosis degree of the rats was significantly reduced after administration of the positive source aqueous extract.
Example 7 Effect of Positive Source Water extracts on serum Total protein, Albumin levels in rats with hepatic fibrosis
The expression levels of Total Protein (TP) and Albumin (ALB) in the serum of each rat group were measured using a fully automated biochemical analyzer (Mindray, BS-330E). As can be seen from fig. 9, both model TP and ALB levels were down-regulated compared to the blank control group. Compared with the model group, the TP level of the administration group is up-regulated, and the ALB level has no significant difference.
Example 8 Effect of Positive Source formula Water extracts on Total Cholesterol levels in serum from rats with hepatic fibrosis
Total Bile Acid (TBA) is a catabolic product of cholesterol in the liver, is secreted into bile by the liver and is discharged into an intestinal cavity along with the bile, acts on the digestion and absorption of fat, the generation and metabolism of the bile acid are closely related to the liver, and when liver cells are diseased, the serum total bile acid is easy to increase, and is a sensitive diagnostic index for detecting substantial liver injury. The expression level of Total Bile Acid (TBA) in the serum of each group of rats was measured using a fully automated biochemical analyzer (Mindray, BS-330E). The result shows that the TBA level of the model group is obviously increased compared with that of a blank control group, and compared with the model group, the positive source formula water extract administration group obviously reduces the TBA level, which indicates that the liver cells of rats in the model group are damaged and the metabolism of the liver function is abnormal, and the positive source formula administration group can reduce the damage degree of the liver.
Example 9 Effect of Positive stock Water extracts on Total superoxide dismutase levels in liver tissue of liver fibrosis rats
The total superoxide dismutase (SOD) level in rat liver tissue is measured by adopting a total superoxide dismutase detection kit (Nanjing Biotechnology Co., Ltd.), and the result shows that the SOD level of a model group is obviously reduced compared with that of a blank group, and compared with the model group, the SOD level of a positive source prescription administration group is obviously increased.
Example 10 Effect of Positive Source Water extracts on the level of malondialdehyde in liver tissue of liver fibrosis rats
The malondialdehyde detection kit (Nanjing Biotechnology Ltd.) is adopted to determine the level of Malondialdehyde (MDA) in rat liver tissues, and the results show that compared with a blank rat group, the MDA level in rat liver tissues of a hepatic fibrosis model group is obviously up-regulated, and an orthologous administration group is obviously down-regulated to abnormally increased MDA level of the model group. The results of example 9, combined with the analysis, show that the aqueous extract of the positive source has the function of reducing the oxidative stress level caused by hepatic fibrosis due to carbon tetrachloride. The results of the tests in example 9 and example 10 are mutually verified, which shows that the model group has oxidation damage phenomenon compared with the blank group, and the administration group can reverse the abnormal SOD and MDA levels in the model group, thus demonstrating that the positive source has certain effect on improving the liver oxidation damage.
Example 11 Effect of Positive Source Water extracts on serum hydroxyproline levels in rats with hepatic fibrosis
Hydroxyproline is an index for testing hepatic fibrosis degree, and Hydroxyproline (HYP) expression level in each group of rat serum is determined by hydroxyproline detection kit (Nanjing to Biotech limited). As can be seen from fig. 13, consistent with the results of example 5, fibrosis was relatively reduced and HYP levels in serum were significantly reduced after orthogenic administration compared to hepatic fibrosis rats.
Example 12 Effect of Positive Source Water extracts on serum TGF-beta 1 levels in rats with liver fibrosis
TGF-beta 1 enzyme-linked immunosorbent assay detection kit (Xinbo Sheng Biotechnology Co., Ltd.) is adopted to carry out in-vitro quantitative determination on the expression level of TGF-beta 1 in the serum of each group of rats. As can be seen from FIG. 14, consistent with the results of example 5, TGF- β 1 levels were significantly reduced in rats after administration of low and medium doses in a positive prescription, compared to hepatic fibrosis rats. TGF-beta 1 is transforming growth factor beta 1, and the key function is adjusting inflammation process, the experimental result shows that compared with the blank group, the TGF-beta 1 level in the blood serum of the hepatic fibrosis rat is obviously increased, and the TGF-beta 1 level in the model group can be obviously reduced by the positive group, the positive source prescription low and medium dosage administration group.
Example 13 Effect of Positive Source Water extracts on serum TNF- α levels in rats with liver fibrosis
The expression level of TNF-alpha in each group of rat serum was quantitatively determined in vitro by TNF-alpha enzyme-linked immunosorbent assay kit (Xinbo Sheng Biotech Co., Ltd.). The TNF-alpha is tumor necrosis factor alpha and is a cytokine related to systemic inflammation, and experimental results show that compared with a blank group, the TNF-alpha level in the serum of a hepatic fibrosis rat is obviously increased, and the TNF-alpha level in a model group can be obviously reduced by a positive group and a low-dose administration group of a positive source prescription, which shows that the positive source prescription has a certain effect on improving the inflammation level of the hepatic fibrosis. As can be seen from FIG. 15, consistent with the results of example 5, TNF-. alpha.levels were significantly down-regulated in rats after administration of low dose of the positive-source formulation compared to liver fibrosis rats.
The experimental results show that the positive source has a certain effect of anti-hepatic fibrosis on the level of animals from the aspects of body weight change, organ index, serum biochemical index, oxidation index and inflammatory index.

Claims (10)

1. A pharmaceutical composition for liver injury protection, characterized by: the pharmaceutical composition for protecting liver injury is prepared from the following raw materials of 20-30 parts of American ginseng, 20-30 parts of hairyvein agrimony, 10-20 parts of angelica, 10-20 parts of rhizoma paridis, 10-20 parts of adenophora tetraphylla and 10-20 parts of platycodon grandiflorum.
2. The pharmaceutical composition for liver injury protection according to claim 1, characterized in that: the pharmaceutical composition for protecting liver injury is prepared from the following raw materials of 20 parts of American ginseng, 30 parts of hairyvein agrimony, 10 parts of angelica, 15 parts of rhizoma paridis, 15 parts of adenophora tetraphylla and 10 parts of platycodon grandiflorum.
3. An aqueous extract of a pharmaceutical composition for liver injury protection prepared based on the pharmaceutical composition for liver injury protection of claim 1 or 2.
4. A process for the preparation of an aqueous extract according to claim 3, characterized in that: the preparation method comprises the following steps:
1) weighing radix Panacis Quinquefolii, herba et Gemma Agrimoniae, radix Angelicae sinensis, rhizoma paridis, radix Adenophorae and radix Platycodi according to the ratio of raw materials to obtain a raw material mixture;
2) decocting the mixture of the raw materials for 3 times, adding 10 times of water each time, decocting for 1.5 hours for the first two times and 1 hour for the third time, mixing the water decoctions, vacuum concentrating at 60 deg.C under reduced pressure to obtain soft extract with relative density of 1.05-1.15, spray drying, collecting fine powder and all medicinal powder adhered to the inner wall of spray dryer, and mixing to obtain water extract.
5. Use of an aqueous extract of the pharmaceutical composition for liver injury protection according to claim 3 for the preparation of a medicament for liver injury protection.
6. Use of an aqueous extract of the pharmaceutical composition for liver injury protection of claim 3 for the preparation of an active drug for promoting hepatocyte proliferation.
7. Use of an aqueous extract of the pharmaceutical composition for liver injury protection according to claim 3 for the preparation of an active agent for alleviating liver cell injury.
8. Use of an aqueous extract of the pharmaceutical composition for liver injury protection according to claim 3 for the preparation of an active agent for alleviating oxidative damage to hepatocytes.
9. Use of an aqueous extract of the pharmaceutical composition for liver injury protection according to claim 3 for the preparation of an anti-liver fibrosis medicament.
10. Use of an aqueous extract of the pharmaceutical composition for liver injury protection according to claim 3 for the preparation of a medicament for reducing liver fibrosis.
CN202110806228.XA 2021-07-16 2021-07-16 Pharmaceutical composition for liver injury protection and application thereof Active CN113577179B (en)

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