CN113713013A

CN113713013A - Composition for resisting liver injury and preparation method thereof

Info

Publication number: CN113713013A
Application number: CN202110958058.7A
Authority: CN
Inventors: 朱志铭; 赵大鹏; 张赞; 李丽丽; 康丽娟; 程晓东
Original assignee: Hebei Yuzhilin Pharmaceutical Co ltd; Hebei Yuzhilin Biotechnology Co ltd
Current assignee: Hebei Yuzhilin Pharmaceutical Co ltd
Priority date: 2021-08-20
Filing date: 2021-08-20
Publication date: 2021-11-30
Anticipated expiration: 2041-08-20
Also published as: CN113713013B

Abstract

The invention relates to an anti-liver injury composition, a clinical preparation and a preparation method thereof, wherein the composition comprises the following components in parts by weight: 100-300 parts of pseudo-ginseng powder, 80-120 parts of American ginseng powder, 75-90 parts of semen hoveniae extract and 60-75 parts of lucid ganoderma extract. The experiment proves that the composition used by the invention is safe to use, can effectively reduce the liver degeneration and has obvious effect of resisting the liver injury.

Description

Composition for resisting liver injury and preparation method thereof

Technical Field

The invention relates to a composition, in particular to a composition for resisting liver injury and a preparation method thereof.

Background

Liver damage is mainly caused by factors such as virus, medicine, alcohol, environment and the like, and is a common disease and a frequently encountered disease worldwide. The etiology of liver injury diseases is complex, the mechanism of the liver injury diseases is not clear up to now, but at present, the liver injury diseases mainly relate to organelle injury, change of cell structure, lipid peroxidation, lipodystrophy, change of enzyme activity, ion balance damage and the like, western medicine treatment is divided into medicine treatment and operation treatment, the medicine treatment mainly adopts expensive medicines such as interferon and the like, the operation treatment adopts artificial liver and liver transplantation, the operation cost is high, and the risk is large.

The traditional Chinese medicine has the advantages of treating multiple diseases and multiple target points, liver injury is clinically manifested by weakness, anorexia, liver pain, jaundice and the like and can be classified into the categories of hypochondriac pain, jaundice, accumulation and the like, pathogenesis is deficiency of vital qi, blood stasis accumulation, damp pathogen injury, damp-heat accumulation, liver depression and spleen deficiency, can be summarized into the categories of deficiency, heat, stasis and obstruction, and belongs to the categories of abdominal mass, collateral diseases and the like in the traditional Chinese medicine. Although some traditional Chinese medicine compositions have certain curative effect on liver injury, the formula is relatively complex, so that the invention needs to invent an anti-liver injury composition with simple formula and definite curative effect.

Disclosure of Invention

The invention aims to overcome the defects in the prior art, provide an anti-liver injury composition with simple formula and definite curative effect, and also provide a preparation method thereof.

In order to achieve the above object, the present invention provides the following technical solutions:

the composition for resisting liver injury comprises the following components in parts by weight:

100-300 parts of pseudo-ginseng powder, 80-120 parts of American ginseng powder, 75-90 parts of semen hoveniae extract and 60-75 parts of ganoderma lucidum extract.

Furthermore, the composition for resisting liver injury comprises the following components in parts by weight:

150-250 parts of pseudo-ginseng powder, 90-110 parts of American ginseng powder, 80-85 parts of semen hoveniae extract and 65-70 parts of lucid ganoderma extract.

190-210 parts of pseudo-ginseng powder, 95-105 parts of American ginseng powder, 82-84 parts of semen hoveniae extract and 66-68 parts of lucid ganoderma extract.

Preferably, in one embodiment of the present invention, a composition for resisting liver injury comprises the following components in parts by weight:

200 parts of pseudo-ginseng powder, 100 parts of American ginseng powder, 83 parts of hovenia dulcis thunb extract and 67 parts of lucid ganoderma extract.

The composition contains 100-200 meshes of pseudo-ginseng powder and 800 meshes of American ginseng powder.

In another aspect, the present invention provides a clinical preparation of the above composition for treating liver injury, wherein the dosage form of the preparation is tablet, powder, granule or hard capsule, preferably hard capsule.

The invention further provides a preparation method of the clinical preparation, which comprises the following steps: weighing Notoginseng radix powder and radix Panacis Quinquefolii powder, micronizing, weighing Ganoderma extract and semen Hoveniae extract, mixing, dry granulating, and filling with capsule filling machine with capsule weight of 0.1-0.5 g/granule.

In the anti-injury traditional Chinese medicine composition, the pharmacological actions of the components are as follows:

semen Hoveniae is fruit or seed of Hovenia dulcis Thunb of Rhamnaceae with fleshy fruit stem. Hovenia dulcis Thunb has sweet taste and mild nature and enters stomach meridian. Recorded in Bencao shiyi, the Chinese patent records "quenching thirst and relieving restlessness, removing diaphragm heat, moistening five internal organs, and facilitating urination and defecation. The Ming Dynasty's book of syndrome treatment records that the effect of alleviating hangover is just like the effect of Zhi Zhang. The Chinese materia medica records the effect of relieving alcoholism, and the compendium of materia medica records the effect of relieving vomiting. Is the key herb of traditional Chinese medicine for relieving alcoholism. Modern researches prove that the hovenia dulcis thunb has the effects of relieving alcoholism, protecting liver, resisting fibrosis, resisting tumor and aging, tranquilizing and easing pain, and pharmacological researches provide a good reading for the cognition of the hovenia dulcis thunb under the visual field of Chinese medicaments. Nowadays, hovenia dulcis thunb is randomly combined for treating alcoholic fatty liver and alcoholic liver disease caused by dampness, heat and stasis except for relieving alcoholism.

Notoginseng radix is dried root of Panax notoginseng (Burk.) F.H.Chen of Araliaceae. Has effects of removing blood stasis, stopping bleeding, promoting blood circulation, and relieving pain. The book Ben Cao Yu Zhen records that the herb enters liver and stomach. It also enters heart and large intestine. The Chinese medicine is named as Ginseng radix Notoginseng, which is the most precious one of the Chinese medicines, because it is recorded in Ben Cao gang mu Shi Yi (1765) that Ginseng radix tonifies qi first and Notoginseng radix tonifies blood first, with the same flavor and different potency. Long-term qi deficiency can lead to failure of blood circulation, while long-term failure of blood circulation can lead to blood stasis. The pseudo-ginseng has the characteristics of removing blood stasis and promoting tissue regeneration, reducing swelling and relieving pain, promoting blood circulation without damaging new tissue, stopping bleeding and retaining blood stasis. Modern pharmacology indicates that the panax notoginseng can achieve the effect of resisting hepatic fibrosis by inhibiting the secretion and deposition of collagen; further experiments show that the panax notoginseng saponins can reduce the content of hyaluronic acid, IV type collagen and propylene glycol; simultaneously, the content of superoxide dismutase is increased, and the hepatic fibrosis process of rats is slowed down; can inhibit the activation of hepatic stellate cells, reduce the generation of collagen fibers and improve the pathological grading of hepatic fibrosis.

American ginseng, which is the dried root of Panax quinquefolium L. American ginseng is cool in nature, sweet and slightly bitter in taste, enters heart, lung and spleen channels, has the effects of tonifying qi and yin, and is suitable for diseases such as deficiency of qi and yin, deficiency heat, vexation and tiredness and the like. The materia Medica of materia Medica: clear lung and kidney, cool heart and spleen to relieve summer heat and alleviate hangover. Modern pharmacological research proves that the American ginseng polysaccharide can weaken oxidative damage and enhance the immune function of an irradiated mouse; the liver ultrastructure of a rat is obviously improved, the oxidative stress of liver tissues is weakened, the expression quantity of ATM and gamma H2AX proteins is reduced, the American ginseng is prompted to be beneficial to repairing rat liver cell damage after electromagnetic radiation, and the mechanism of the American ginseng is possibly related to the improvement of the liver ultrastructure and the reduction of oxidative stress damage.

Ganoderma is fruiting body of Ganoderma lucidum of Polyporaceae. Sweet in nature and mild in taste. Has the effects of treating consumptive disease, cough, asthma, insomnia and dyspepsia. Scientific research shows that the efficacy and the function of the lucid ganoderma are obvious on protecting the liver, and the lucid ganoderma can be called 'immortal grass' for protecting the liver. Ganoderma has protective effect on liver injury caused by various physicochemical and biological factors. The administration of Ganoderma can protect liver and relieve liver injury before or after liver injury occurs. The ganoderma lucidum can also promote the metabolism of the liver to drugs and poisons, and has definite curative effect on toxic hepatitis. At present, the anti-inflammatory and liver-protecting action mechanism of ganoderan on acute alcoholic liver injury is not clear, and a large number of researches show that ganoderan can reduce the activity of Nitric Oxide Synthase (NOS) and inhibit the inflammatory reaction of organisms by inhibiting the formation of inflammatory reaction transmitters of liver tissues; can also protect the integrity of cell membranes by inhibiting free radical lipid peroxidation and play a role in protecting the liver. The ganoderma lucidum polysaccharide has lower contents of AST, ALT, triacylglycerol, total cholesterol, free fatty acid, IL-1 beta, IL-6 and TNF-alpha and oil red O staining score than a model group, and the ganoderma lucidum polysaccharide also has obvious effects of protecting liver, reducing fat, resisting inflammation, inhibiting fat deposition and the like on mice with acute alcoholic liver injury.

The composition can be added with one or more of dextrin, maltodextrin, beta-cyclodextrin, soluble starch, erythritol, mogroside, isomaltitol and the like as auxiliary materials and prepared into products in the forms of tablets, granules, hard capsules and the like according to a modern production method.

Compared with the prior art, according to the traditional Chinese medicine theory, the pseudo-ginseng, the American ginseng powder, the hovenia dulcis thunb extract and the lucid ganoderma extract are compatible, aiming at the pathogenesis of deficiency of vital qi, blood stasis internal accumulation, damp-heat internal accumulation and liver depression and spleen deficiency, the treatment method takes heat clearing, dampness removing, blood circulation promoting and blood stasis removing as main principles, wherein the pseudo-ginseng is the monarch drug and mainly plays the roles of promoting blood circulation and removing blood stasis, and dredging collaterals and promoting blood circulation; american ginseng powder and hovenia dulcis thunb extract are used as ministers to clear heat and promote fluid production, invigorate qi and nourish yin and moisten the five internal organs; the ganoderma lucidum extract is an adjuvant drug for tonifying qi and deficiency and benefiting liver and kidney. In the formula, the ganoderma lucidum extract, the pseudo-ginseng powder and the American ginseng powder are tonifying products, and are sweet in taste and slow in nature, so that dampness is often generated while tonifying. Therefore, the traditional Chinese medicine composition is matched with the hovenia dulcis thunb extract to induce diuresis and eliminate dampness, and makes up the defect that American ginseng, pseudo-ginseng and lucid ganoderma cannot treat dampness in reports. The formula has the effects of keeping multiple ducts down, combining treatment and treating both principal and secondary aspect of disease. The experiment proves that the composition used by the invention is safe to use, can effectively reduce the liver degeneration and has obvious effect of resisting the liver injury.

The technique adopts a superfine powder process to prepare the American ginseng powder, so that on one hand, the cell wall of the medicine is broken, the efficacy of the traditional Chinese medicine is improved, and the raw medicinal materials can be saved; on the other hand, the full components of the traditional Chinese medicine are still kept, the microbial content and dust of the product are controlled, the internal structure of the traditional Chinese medicine is not damaged by ultrafine grinding theoretically, the quality, the smell and the taste of the traditional Chinese medicine are kept, and four-smell and five-taste are especially kept. Can follow the principles of dialectical treatment and therapeutic method in traditional Chinese medicine.

Drawings

FIG. 1 auxiliary protection against chemical liver injury, negative control group liver (oil red O staining X200);

FIG. 2 is a graph showing the adjuvant protection against chemical liver injury, model control group liver (oil red O staining X200);

FIG. 3 auxiliary protection against chemical liver injury, example 1 liver (oil red O staining X200);

Detailed Description

The invention discloses a composition with a gastric mucosa injury protection effect and a preparation method thereof, and the composition can be realized by combining the relevant principles of traditional Chinese medicines and properly improving process parameters by taking the contents of the composition as reference by the technical personnel in the field. It is expressly intended that all such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the scope of the invention. While the invention has been described in terms of preferred embodiments, it will be apparent to those skilled in the art that variations may be applied, or changes and combinations may be made, in the methods and applications described herein to achieve and use the inventive techniques without departing from the spirit, scope, and content of the invention.

For a better understanding of the invention, and not as a limitation on the scope thereof, all numbers expressing quantities, percentages, and other numerical values used in this application are to be understood as being modified in all instances by the term "about". At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

The present invention is further illustrated by the following examples, which are not intended to limit the invention in any way.

The extracts of the part are all provided by Jianxi Jiahe Biotech company. The preparation method of the American ginseng powder comprises the following steps:

1. cleaning and selecting: placing radix Panacis Quinquefolii on a cleaning operation table, and removing impurities, foreign substances, moth-eaten parts, mildew and non-medicinal parts;

2. cleaning: placing the cleaned and qualified American ginseng in a bubble type medicine washing machine, and washing the raw materials with water until the surface is clean and has no soil;

3. and (3) sterilizing and drying: the cleaned pseudo-ginseng is flatly paved in a clean sterilization tray with the paving thickness of 1-3 cm, the front door is opened, the sterilization tray is placed in a sterilization cabinet, the sterilization temperature is 100-.

4. Crushing: crushing the sterilized and dried American ginseng into particles with the particle size of 80 meshes by using a crusher;

5. crushing: and carrying out airflow superfine grinding on the crushed American ginseng particles at a low temperature of between 5 ℃ below zero and 10 ℃ below zero by using a superfine grinder to obtain 800-mesh micropowder with the average particle size of 500 meshes.

Example 1: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

secondly, the preparation method comprises the following steps:

step two: weighing 200 parts of pseudo-ginseng powder and 100 parts of American ginseng powder, carrying out superfine grinding, wherein the pseudo-ginseng powder is 200 meshes, the American ginseng powder is 600 meshes, weighing 83 parts of lucid ganoderma extract and 67 parts of hovenia dulcis thunb extract, uniformly mixing, and carrying out dry granulation. And (3) filling the granules by using a capsule filling machine, controlling the weight of capsules to be 0.45 g/granule, and carrying out internal and external packaging after the capsule filling is finished to obtain a finished product.

Example 2: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

components	Parts by weight
		Notoginseng powder	200 portions of
American ginseng powder	100 portions of
		Semen Hoveniae extract	82 portions of
Ganoderma lucidum extract	68 portions of

Secondly, the preparation method comprises the following steps:

weighing 200 parts of pseudo-ginseng powder and 100 parts of American ginseng powder, carrying out superfine grinding, wherein the pseudo-ginseng powder is 200 meshes, the American ginseng powder is 600 meshes, weighing 68 parts of lucid ganoderma extract and 82 parts of hovenia dulcis thunb extract, uniformly mixing, and carrying out dry granulation. Filling the granules with a capsule filling agent, controlling the weight of capsules to be 0.5 g/granule, and carrying out internal and external packaging after the capsules are filled to obtain the finished product.

Example 3: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

components	Parts by weight
		Notoginseng powder	198 portions of
American ginseng powder	102 portions of
		Semen Hoveniae extract	83 portions of
Ganoderma lucidum extract	68 portions of

Secondly, the preparation method comprises the following steps:

198 parts of pseudo-ginseng powder and 102 parts of American ginseng powder are weighed and subjected to superfine grinding, the pseudo-ginseng powder is 200 meshes, the American ginseng powder is 600 meshes, 68 parts of lucid ganoderma extract and 83 parts of hovenia dulcis thunb extract are weighed and mixed uniformly, and dry granulation is carried out. Filling the granules with a capsule filling agent, controlling the weight of capsules to be 0.3 g/granule, and carrying out internal and external packaging after the capsules are filled to obtain the finished product.

Example 4: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

components	Parts by weight
		Notoginseng powder	202 portions of
American ginseng powder	98 portions of
		Semen Hoveniae extract	84 portions of
Ganoderma lucidum extract	66 portions of

Secondly, the preparation method comprises the following steps:

weighing 202 parts of pseudo-ginseng powder and 98 parts of American ginseng powder, carrying out superfine grinding, wherein the pseudo-ginseng powder is 200 meshes, the American ginseng powder is 600 meshes, weighing 66 parts of lucid ganoderma extract and 84 parts of hovenia dulcis thunb extract, uniformly mixing, and carrying out dry granulation. Filling the granules with a capsule filling agent, controlling the weight of capsules to be 0.2 g/granule, and carrying out internal and external packaging after the capsules are filled to obtain the finished product.

Example 5: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

components	Parts by weight
		Notoginseng powder	205 portions of
American ginseng powder	95 parts of
		Semen Hoveniae extract	82 portions of
Ganoderma lucidum extract	66 portions of

Secondly, the preparation method comprises the following steps:

weighing 205 parts of pseudo-ginseng powder and 95 parts of American ginseng powder, carrying out superfine grinding, wherein the pseudo-ginseng powder is 200 meshes, the American ginseng powder is 600 meshes, weighing 66 parts of lucid ganoderma extract and 82 parts of hovenia dulcis thunb extract, uniformly mixing, and carrying out dry granulation. Filling the granules with a capsule filling agent, controlling the weight of capsules to be 0.4 g/granule, and carrying out internal and external packaging after the capsule filling is finished to obtain a finished product.

Example 6: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

components	Parts by weight
		Notoginseng powder	190 portions of
American ginseng powder	105 portions of
		Semen Hoveniae extract	84 portions of
Ganoderma lucidum extract	68 portions of

Secondly, the preparation method comprises the following steps:

weighing 190 parts of pseudo-ginseng powder and 105 parts of American ginseng powder, carrying out superfine grinding, wherein the pseudo-ginseng powder is 200 meshes and the American ginseng powder is 600 meshes, weighing 68 parts of lucid ganoderma extract and 84 parts of hovenia dulcis thunb extract, uniformly mixing, and carrying out dry granulation. Filling the granules with a capsule filling agent, controlling the weight of capsules to be 0.3 g/granule, and carrying out internal and external packaging after the capsules are filled to obtain the finished product.

Comparative example 1: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

components	Parts by weight
		Notoginseng powder	200 portions of
American ginseng extract	100 portions of
		Semen Hoveniae extract	83 portions of
Ganoderma lucidum extract	67 portions of

Secondly, the preparation method comprises the following steps: the same as in example 1.

Wherein, the preparation of the American ginseng extract comprises the following steps:

pulverizing dried radix Panacis Quinquefolii, sieving with 60 mesh sieve, adding 8 times of deionized water, soaking for 1h, reflux extracting for 1h, centrifuging at 5000r/min for 10min to obtain supernatant, adding 5 times of deionized water into the residue, reflux extracting for 0.5h, centrifuging to obtain supernatant, mixing the supernatants, and freeze drying to obtain radix Panacis Quinquefolii extract.

Comparative example 2: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

components	Parts by weight
		Notoginseng radix extract	200 portions of
American ginseng powder	100 portions of
		Semen Hoveniae extract	83 portions of
Ganoderma lucidum extract	67 portions of

Wherein, the preparation of the pseudo-ginseng extract comprises the following steps:

adding 10 times of ethanol solution into Notoginseng radix powder, extracting at 25 deg.C for 4 hr, filtering, concentrating the filtrate, and freeze drying to obtain Notoginseng radix extract.

Comparative example 3: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

components	Parts by weight
		Notoginseng powder	200 portions of
Semen Hoveniae extract	83 portions of
		Ganoderma lucidum extract	67 portions of

Comparative example 4: composition comprising a metal oxide and a metal oxide

The method comprises the following steps:

components	Parts by weight
		Notoginseng powder	200 portions of
American ginseng powder	100 portions of
		Semen Hoveniae extract	83 portions of

Example 7: evaluation of the auxiliary Effect of the composition of the present invention on liver injury

1. Materials and methods

1.1 test substance: the compositions of examples 1 to 6 and the compositions of comparative examples 1 to 4 were prepared by weighing 6.75g of each composition, adding pure water to 100ml, and using 100ml of pure water as a blank control group and a model control group. Fully and evenly shaking before preparation and use.

1.2 Experimental animals: SPF grade SD rats, male, weighing 180-.

1.3 Experimental methods: adopting an alcoholic liver injury model: model for male mouse chemical liver injury caused by 50% ethanol

Dose grouping and test sample administration time: experiments assuming that 12 groups of examples 1-6, comparative examples 1-4, blank control group and model control group were used, 10 animals per group were administered with the corresponding test substances by oral gavage daily, the gavage capacity of mice was not 0.2ml/10g.BW, and the blank control group and the liver injury model control group were administered with the same amount of pure water. Animals were weighed twice a week and the gavage of the test subjects was adjusted according to body weight for a year.

1.3.2 Experimental procedures: after 30 days of gastric lavage, 14ml/kg of BW 50% ethanol is fed into the model group and each experimental group by one-time gastric lavage, pure water with equal volume is fed into the negative control group, animals are killed after weighing for 16 hours after fasting, the whole stomach is exposed, pylorus is tied, a proper amount of 10% formaldehyde solution is poured into the stomach, the stomach is fixed for 20min, then the stomach is cut along the greater curvature, the liver is weighed to calculate the ratio of viscera to body, 0.5g of liver is taken to prepare liver homogenate, and then the detection of various indexes (MDA, reduction type GSH and TG) and the pathological histology examination of the liver are carried out.

1.4 data processing: statistics are carried out by a pairwise comparison method of mean numbers between a plurality of experimental groups and a plurality of control groups.

2. Detecting the index

2.1 body weight

2.2 MDA, reduced GSH, TG in liver homogenate

2.3 histopathological examination of the liver

2.3.1 pathological examination material the livers of each group of animals were harvested from the middle of the left lobe of the liver as a cross section, frozen sectioned and stained with Sudan III. The degree of hepatocyte damage was observed under an optical microscope.

2.3.2 criteria for pathological diagnosis

2.3.1.1 evaluation method

Microscopic examination was performed to record pathological changes of cells starting from a segment of the liver field and the whole tissue section was observed continuously with a 40-fold objective lens. The distribution, extent and area of lipid droplets in the liver were mainly observed.

2.3.2.2 score

Recording the area of each lesion in each visual field occupied by the visual field in decibels, and accumulating the total lesion score of the observed visual field.

The lipid in the liver cells is scattered and is rarely divided into 0

The hepatic cells containing lipid droplets do not exceed 1/41 points

The hepatic cells containing lipid droplets do not exceed 1/22 points

The hepatic cells containing lipid droplets do not exceed 3/43 points

Liver tissue was almost replaced by lipid droplets for 4 points

2.3.2.3 determination of results

When any condition is satisfied, the tested sample can be judged to have the auxiliary protection effect on alcoholic liver injury.

2.3.2.3.1 liver MDA, reduction GSH and TG have positive detection indexes.

2.3.2.3.2 liver MDA, reduced GSH and TG three detection indexes including positive index and positive pathological histological examination result

3 results of the test

3.1 Effect on mouse body weight

The weight average of the mouse bodies of all groups is not obviously influenced by the test substance, and the differences are not significant (P is more than 0.05) compared with the model control group and the negative control group. (see Table 1)

TABLE 1 Effect of the compositions of the invention on mouse body weight (mean. + -. standard deviation)

Group of	Number of animals	0 week weight (g)	4 week heavy (g)
				Negative control group	10	19.5±1.0	43.7±2.1
Model control group	10	19.4±1.0	42.6±1.4
				Example 1	10	19.9±0.9	42.5±2.8
Example 2	10	19.8±1.1	43.0±2.4
				Example 3	10	20.0±1.1	43.5±2.4
Example 4	10	19.4±1.2	42.7±1.6
				Example 5	10	19.6±1.0	42.8±2.0
Example 6	10	19.7±0.9	43.0±2.5
				Comparative example 1 group	10	19.9±1.0	42.9±1.9
Comparative example 2 group	10	20.0±0.9	43.3±2.2
				Comparative example 3 group	10	19.7±1.0	42.5±1.7
Comparative example 4 group	10	20.1±1.2	42.9±2.1

3.2 Effect on MDA, reduced GSH and TG in liver homogenates

When the liver homogenate of a mouse in a liver injury model group is increased and the reduced GSH is reduced by comparing the liver homogenate with a negative control group after being given 50% alcohol for 16 hours according to the dose of 14ml/kg.BW (BW), the contents of MDA and TG are both obviously different (P is less than 0.01), which indicates that the experimental model is successful. Compared with a model control group, the contents of MDA and TG in liver homogenate of the mice of the examples 1 to 6 are obviously reduced; examples 1-6 mice showed a significant increase in reducing GSH in liver homogenates. Compared with experimental data of a comparative example, the indexes of MDA and TG values in liver homogenate of mice of the example are wholly lower than those of the comparative example, and the index of reduced GSH value is higher than that of the comparative example. The effect of the embodiment 1 is the best, and the embodiments 2 to 4 times are slightly worse than the embodiments 5 to 6. Through comparison of the experimental data of the embodiment and the comparative examples 1-4, the effect of the whole powder of the panax notoginseng and the American ginseng is further verified to be better than that of the panax notoginseng and the American ginseng extract, the compatible seeds lack the American ginseng or the lucid ganoderma, the effect is obviously poor, and the specific data result is shown in table 2.

Table 2: measurement of MDA, reduced GSH and TG values (mean. + -. standard deviation) in liver homogenates of mice with the compositions of the invention

Note: comparing each experimental group with model control group, P is less than 0.05; p < 0.01.

3.3 Effect on mouse liver histopathology

Compared with the negative control group, the animal test model is established when the hepatic steatosis score is statistically different (P is less than 0.01) by the significant difference between the model control group and the negative control group, and the immediate liver injury model of the test mouse is successfully reproduced. Examples 1-6 showed a significant reduction in the degree of hepatic steatosis as compared to the model group, with statistically significant differences (mean P < 0.05) and positive (see Table 3). As can be seen from the experimental data in Table 3, examples 1-6 exhibited greater reduction in hepatic steatosis than comparative examples 1-4. The experiments were ranked from large to small according to the degree of reduction in hepatocyte steatosis: example 1 > examples 2-4 > examples 5-6 > comparative examples 1-4. The negative control group, model control group and typical histopathological examination pictures of example 1 are shown in the attached tables 1-3, and the experimental results are shown in the table 3.

Table 3: effect of the compositions of the invention on histology of liver disease in mice (mean. + -. standard deviation)

Experimental group	Number of animals	X	P
				Blank control group	10	1.00±0.75	0.000
Model control group	10	3.05±0.48	--
				Example 1	10	2.55±0.64	0.042
Example 2	10	2.62±0.51	0.046
				Example 3	10	2.65±0.68	0.048
Example 4	10	2.69±0.74	0.045
				Example 5	10	2.67±0.73	0.047
Example 6	10	2.63±0.87	0.047
				Comparative example 1 group	10	2.78±0.62	0.161
Comparative example 2 group	10	2.75±0.72	0.157
				Comparative example 3 group	10	2.84±0.81	0.164
Comparative example 4 group	10	2.82±0.75	0.162

And (4) conclusion: (1) the weight gains of mice in all dose groups have no significant difference; (2) under the condition that the model is established, the MDA and TG contents in the liver homogenate of the mouse are both obviously reduced, and the reducing GSH content is obviously increased. (3) Pathology shows that examples 1-6 can remarkably reduce the liver steatosis, and the results of the animal experiments show that: the three indexes of liver MDA, GSH and TG and the pathological histological examination result are all positive.

Example 8: safety test

The hard capsules prepared in the embodiments 2 to 7 of the invention are respectively subjected to the mouse acute toxicity test, the bone marrow cell micronucleus test, the mouse teratospermia test, the rat 30-day feeding test and the Ames test, and the test results are as follows:

(1) in the acute oral toxicity test of SPF-grade Kunming mice of two sexes, the hard capsules obtained in examples 1-6 reach 12.0g/kg.bw (corresponding to 267 times of the human body recommended daily intake of 0.045 g/kg.bw) within 24h, no obvious toxic symptom and death of the animals are observed within an observation period of 14 days, the MTD value is measured to be more than 12.0g/kg.bw, and the evaluation standard of acute toxicity is specified and is of actual non-toxic grade.

(2) The results of the micronucleus test and the teratospermia test of the bone marrow cells of the mice are negative.

(3) The results of the 30-day feeding test show that: the samples were administered to SPF-class Wistar rats at doses of 1.125g/kg.bw, 2.250g/kg.bw, and 4.500g/kg.bw (corresponding to 25 times, 50 times, and 100 times, respectively, of the human recommended daily intake of 0.045 g/kg.bw) for 30 consecutive days without significant signs of toxicity and death. Compared with the negative control group, the indexes of the capsule prepared in each embodiment, such as rat weight, food intake, food utilization rate, hematology, visceral weight, visceral/body ratio, pathological histology and the like, in each dose group have no significant difference, no obvious abnormal change is seen in blood biochemistry, and no obvious toxicity is found under the test condition.

(4) Examples 1-6 for four test strains of salmonella typhimurium TA97a, TA98 and TA100\ TA104, when S9 is added or not added, 8, 40, 200, 1000 and 5000 mug/dish of 5 dosage groups, the tested substance has no obvious bacteriostasis to the test strain under the experimental condition, the number of the retrovariant colonies of each dosage group does not exceed 2 times of the number of the negative control bacteria, and no dosage-reaction relation exists, and the Ames test result is negative.

Claims

1. The composition for resisting liver injury is characterized by comprising the following components in parts by weight:

2. The composition for resisting liver injury according to claim 1, which is prepared from the following components in parts by weight:

3. The composition for resisting liver injury according to claim 1, which is prepared from the following components in parts by weight:

4. The composition for resisting liver injury according to claim 1, which is prepared from the following components in parts by weight:

5. The composition of claim 1, wherein the composition comprises 100-200 mesh panax notoginseng powder and 200-400 mesh panax quinquefolium powder.

6. A clinical preparation of the composition for treating liver injury according to any one of claims 1 to 5, wherein the preparation is in the form of a tablet, powder, granule or hard capsule.

7. The clinical formulation of claim 6, in the form of a hard gelatin capsule.

8. A method for preparing the clinical preparation according to claim 7, which comprises the following steps: weighing Notoginseng radix powder and radix Panacis Quinquefolii powder, micronizing, weighing Ganoderma extract and semen Hoveniae extract, mixing, dry granulating, and filling with capsule filling machine with capsule weight of 0.1-0.5 g/granule.