CN113616706A

CN113616706A - Liver-protecting immunity-enhancing Chinese herbal medicine preparation and preparation method thereof

Info

Publication number: CN113616706A
Application number: CN202111026451.9A
Authority: CN
Inventors: 赵剑
Original assignee: Jiangsu Shengshikanghe Biotechnology Co ltd
Current assignee: Jiangsu Shengshikanghe Biotechnology Co ltd
Priority date: 2021-09-02
Filing date: 2021-09-02
Publication date: 2021-11-09

Abstract

The invention relates to a liver-protecting and immunity-enhancing Chinese herbal medicine preparation and a preparation method thereof, belonging to the technical field of Chinese herbal medicine preparations. The liver-protecting and immunity-enhancing Chinese herbal medicine preparation is a capsule preparation which is prepared by mixing, granulating and filling kudzu root extract, silybum marianum extract, lucid ganoderma extract and schisandra chinensis extract serving as main raw materials. The raw materials in the preparation are medicinal and edible components and new resource food, so that the preparation has mild medicinal properties and high safety, and has good effects of protecting liver and enhancing immunity. The preparation can inhibit abnormal increase of Malondialdehyde (MDA) level in mouse liver, reduce Triglyceride (TG) level in liver, and has significant difference (P is less than 0.05) with model group in liver pathological evaluation; and simultaneously, the spleen lymphocyte transformation capacity, the antibody generation cell number and the serum hemolysin level of the mouse can be improved. The preparation method is simple in preparation process, can fully reserve the active ingredients in the raw materials of the preparation, and has strong practicability and higher social benefit.

Description

Liver-protecting immunity-enhancing Chinese herbal medicine preparation and preparation method thereof

Technical Field

The invention relates to a liver-protecting and immunity-enhancing Chinese herbal medicine preparation and a preparation method thereof, belonging to the technical field of Chinese herbal medicine preparations.

Background

Currently, chemical liver injury is caused by chemical substances including alcohol, toxic substances in the environment, and certain drugs. The liver, which is an important detoxification organ of the human body, has a dual blood supply of hepatic artery and hepatic vein, and chemicals can enter the liver through the portal vein of the gastrointestinal tract or systemic circulation for transformation, so that the liver is easily damaged by toxic substances among the chemicals. The natural and human industrial production processes have some substances which are toxic to the liver, the 'liver-philic toxicants' are generally susceptible in people, the incubation period is short, the pathological process is directly related to the infection dosage, and the liver cell necrosis, fat deformation, liver cirrhosis and liver cancer of different degrees of the liver can be caused. The traditional Chinese medicine considers that liver diseases are mainly caused by liver and spleen damage, qi stagnation, phlegm-damp stagnation and blood stasis internal contraction due to emotional depression, diet injury, external attack of cold evil and external invasion of toxin, so that the main pathogenesis of the liver and spleen diseases is liver depression and spleen deficiency and blood stasis internal stagnation, and the treatment is based on strengthening vital qi to eliminate pathogenic factors, soothing liver and strengthening spleen, removing blood stasis and promoting diuresis. Su Wen & Wuzang Generation cloud: the blood in the lying position of the deceased is due to the liver, the liver can be seen by blood, the feet can be moved by blood, the palm can be grasped by blood, and the fingers can be taken by blood. "the four saints heart source" also says: the liver governs storing blood, and blood in zang-fu organs and meridians is also perfused by the liver. The liver stores blood, so that one of the people can regulate qi and blood, and replenish essence, so that four people can use the liver. Therefore, regulating qi and blood can also improve immunity. From the perspective of traditional Chinese medicine, the liver protection and the enhancement of the immunity of the organism all need to tonify qi, activate blood, invigorate spleen and nourish yin.

Silybum marianum is native to Europe, Mediterranean regions, North Africa and central Asia, and its use for treating liver diseases has been recorded for thousands of years in Europe, and its extracts have been used for treating liver and gall diseases as early as the first century (23-79 years of Gong Yuan). Radix Puerariae has effects of expelling pathogenic factors from muscles and skin, relieving fever, promoting fluid production, quenching thirst, dredging meridian passage, and relieving alcoholism. Ganoderma has effects in invigorating qi, tranquilizing mind, relieving cough and asthma, and can be used for treating restlessness of heart-mind, insomnia, palpitation, cough and asthma due to lung deficiency, asthenia, short breath, and anorexia. Fructus Schisandrae has astringent, qi invigorating, salivation promoting, kidney invigorating, and heart calming effects. The market faces to the unilateral liver protection products which take silybum marianum, kudzuvine root, lucid ganoderma, Chinese magnoliavine fruit and the like as raw materials at home and abroad, but the research on the synergistic effect of the raw materials on the protection of chemical liver injury and the enhancement of immunity is less, and no single-side liver protection product is on the market.

In view of the above-mentioned drawbacks, the designer actively makes research and innovation to create a liver-protecting and immunity-enhancing Chinese herbal medicine preparation and a preparation method thereof, so that the Chinese herbal medicine preparation has industrial utilization value.

Disclosure of Invention

In order to solve the technical problems, the invention aims to provide a Chinese herbal medicine preparation for protecting liver and enhancing immunity and a preparation method thereof. Is prepared from pueraria root extract, silybum marianum extract, ganoderma extract and schisandra fruit extract through mixing, granulating and filling. The raw materials in the preparation are medicinal and edible components and new resource food, so that the preparation has mild medicinal properties and high safety, and has good effects of protecting liver and enhancing immunity. The preparation can inhibit abnormal increase of Malondialdehyde (MDA) level in mouse liver, reduce Triglyceride (TG) level in liver, and has significant difference (P is less than 0.05) with model group in liver pathological evaluation; meanwhile, the spleen lymphocyte transformation capacity, the antibody generation cell number and the serum hemolysin level of the mouse can be improved, and the spleen lymphocyte transformation capacity, the antibody generation cell number and the serum hemolysin level of the mouse have no obvious influence on the toe swelling degree, the abdominal cavity macrophage phagocytosis index, the abdominal cavity macrophage phagocytosis percentage, the carbon clearance phagocytosis index, the NK cell activity, the spleen/body weight ratio and the thymus/body weight ratio of the mouse. The preparation method is simple in preparation process, can fully reserve the active ingredients in the raw materials of the preparation, and has strong practicability and higher social benefit.

The liver-protecting immunity-enhancing Chinese herbal medicine preparation comprises the following raw and auxiliary materials in parts by weight:

35-39 parts of kudzu root extract;

5-9 parts of silybum marianum extract;

9-13 parts of ganoderma lucidum extract;

13-17 parts of schisandra extract;

24-28 parts of calcium carbonate;

2.3-2.7 parts of povidone K30;

0.8-1.2 parts of magnesium stearate;

the dosage of the liver-protecting immunity-enhancing Chinese herbal medicine preparation is 1.6g per day;

the silybum marianum extract is obtained by extracting silybum marianum with an ethanol solution;

the kudzu root extract is obtained by extracting kudzu root with an ethanol solution;

the schisandra extract is obtained by extracting schisandra with an ethanol solution;

the Ganoderma extract is obtained by extracting Ganoderma with ethanol solution.

Further, the feed additive comprises the following raw and auxiliary materials in parts by weight:

37 parts of kudzu root extract;

7 parts of silybum marianum extract;

11 parts of ganoderma lucidum extract;

15 parts of schisandra extract;

26 parts of calcium carbonate;

2.5 parts of povidone K30;

1.0 part of magnesium stearate.

Further, the extraction process conditions of the silybum marianum extract are as follows: the ethanol solution is 70% in volume, the time for extracting for 3 times is 1.5h, the extraction temperature is normal temperature, and the material-liquid ratio is 1: and 8, concentrating, extracting with ethyl acetate, and drying.

Further, the extraction process conditions of the kudzu root extract are as follows: the ethanol solution is 50% in volume, the time for 2 times of extraction is 2 hours, the extraction temperature is 80 ℃, and the material-liquid ratio is 1: 10, filtering, concentrating and spray drying after extraction.

Further, the extraction process conditions of the schisandra extract are as follows: the ethanol solution accounts for 60% by volume, the time for 2 times of extraction is 2 hours, the extraction temperature is normal temperature, and the material-liquid ratio is 1:8, filtering, concentrating and spray drying after extraction.

Further, the extraction process conditions of the ganoderma lucidum extract are as follows: the ethanol solution is 70% in volume, the extraction temperature is 76 ℃, the extraction time is 2 hours each time after 3 times of extraction, and the material-liquid ratio is 1: and 6, concentrating after extraction, and then spray-drying.

A preparation method of a Chinese herbal medicine preparation for protecting liver and enhancing immunity comprises the following specific preparation steps:

mixing the silybum marianum extract, the kudzuvine root extract, the schisandra extract, the lucid ganoderma extract and the calcium carbonate uniformly to obtain mixed powder, and mixing povidone K30 and 95% ethanol solution according to the mass ratio of 1:8, mixing to obtain a binder, uniformly bonding the mixed powder and the binder, and granulating and drying to obtain the traditional Chinese medicine granules;

and (3) uniformly mixing the traditional Chinese medicine granules with calcium carbonate and magnesium stearate to obtain total mixed granules, and making the total mixed granules into capsules by using a capsule filling machine to obtain the liver-protecting and immunity-enhancing Chinese herbal medicine preparation.

The mechanism of the invention is as follows:

silybum marianum: bitter and cool. It enters liver and gallbladder meridians. Silybum marianum is a multifunctional antioxidant, and has the effects of clearing away heat and toxic materials, soothing the liver and benefiting the gallbladder. Can be used for treating damp-heat in liver and gallbladder, hypochondriac pain, and jaundice. Silybum marianum is a multifunctional antioxidant, and has effects of resisting oxygen free radicals, resisting lipid peroxidation, protecting liver cell membrane, resisting hepatic fibrosis, and improving liver detoxication ability, so as to protect liver, increase the number of blood leukocyte, activate abdominal macrophage, enhance its swallowing ability, promote lymphocyte transformation, and improve lymphocyte transformation rate;

kudzu root: it has effects of invigorating spleen, stomach and lung channels, dredging meridian passage, and relieving alcoholism. Resisting lipid peroxidation, protecting liver cell, promoting proliferation of splenic T lymphocyte, and correcting T lymphocyte sub-group Th1/Th2 proportion disorder;

ganoderma lucidum: sweet and neutral. It enters heart, lung, liver and kidney meridians. The product has effects in invigorating qi, tranquilizing mind, improving liver function, purifying blood, activating cells, promoting metabolism, enhancing immunity, resisting oxidation, and scavenging free radicals, and can be used for treating restlessness, insomnia, palpitation, cough and asthma due to lung deficiency, asthenia, and short breath;

schisandra chinensis: wu Wei Zi | is sour, sweet and warm. It enters lung, heart and kidney meridians. Astringe, benefit qi, promote the production of body fluid, tonify kidney, calm heart; the schisandra fruit can reduce the degeneration and necrosis of liver cell, increase the mitochondria, RNA and glycogen of liver cell, and reversibly inhibit ALT activity in liver cell, thus reducing the level of serum transaminase, and the schisandra fruit has obvious improvement on immune function, mainly manifested by the increase of the content of the immunoglobulin secreted by the schisandra fruit and the enhancement of the capability of phagocyte and antibacterial substance.

By the scheme, the invention at least has the following advantages:

the liver-protecting and immunity-enhancing Chinese herbal medicine preparation is a capsule preparation which is prepared by mixing, granulating and filling kudzu root extract, silybum marianum extract, lucid ganoderma extract and schisandra chinensis extract serving as main raw materials. The raw materials in the preparation are medicinal and edible components and new resource food, so that the preparation has mild medicinal properties and high safety, and has good effects of protecting liver and enhancing immunity. The preparation can inhibit abnormal increase of Malondialdehyde (MDA) level in mouse liver, reduce Triglyceride (TG) level in liver, and has significant difference (P is less than 0.05) with model group in liver pathological evaluation; meanwhile, the spleen lymphocyte transformation capacity, the antibody-producing cell number and the serum hemolysin level of the mouse can be improved, and the toe swelling degree, the abdominal cavity macrophage phagocytosis index, the abdominal cavity macrophage phagocytosis percentage, the carbon clearance phagocytosis index, the NK cell activity, the spleen/body weight ratio and the thymus/body weight ratio of the mouse are not obviously influenced.

The foregoing is a summary of the present invention, and in order to provide a clear understanding of the technical means of the present invention and to be implemented in accordance with the present specification, the following is a detailed description of the preferred embodiments of the present invention.

Detailed Description

The following examples are given to further illustrate the embodiments of the present invention. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

Weighing 35-39 parts of kudzu root extract, 5-9 parts of silybum marianum extract, 9-13 parts of lucid ganoderma extract, 13-17 parts of schisandra chinensis extract, 24-28 parts of calcium carbonate, 2.3-2.7 parts of povidone K30 and 0.8-1.2 parts of magnesium stearate in parts by weight;

mixing herba Silybi Mariani extract, radix Puerariae extract, fructus Schisandrae extract, Ganoderma extract, and calcium carbonate uniformly to obtain mixed powder, adding polyvidone K30 into 95% ethanol until completely dissolved to obtain binder, uniformly binding the mixed powder and binder at a mass ratio of 1:8, granulating, and drying to obtain Chinese medicinal granule;

mixing the Chinese medicinal granules and magnesium stearate uniformly to obtain total mixed granules, and making the total mixed granules into capsules by using a capsule filling machine to obtain the liver-protecting and immunity-enhancing Chinese herbal medicine preparation;

the dosage of the liver-protecting and immunity-enhancing Chinese herbal medicine preparation is 1.6g per day.

The silybum marianum extract is obtained by extracting silybum marianum with an ethanol solution, wherein the ethanol solution accounts for 70% by volume, the extraction time is 1.5h every time for 3 times, the extraction temperature is normal temperature, and the material-liquid ratio is 1: and 8, concentrating, extracting with ethyl acetate, and drying.

The kudzu root extract is obtained by extracting kudzu roots with an ethanol solution, wherein the ethanol solution accounts for 50% by volume, the extraction time is 2 hours each time after 2 times, the extraction temperature is 80 ℃, and the material-liquid ratio is 1: 10, filtering, concentrating and spray drying after extraction;

the schisandra extract is obtained by extracting schisandra chinensis with an ethanol solution, wherein the volume part of the ethanol solution is 60%, the time for extracting 2 times is 2 hours, the extraction temperature is normal temperature, and the material-liquid ratio is 1:8, filtering, concentrating and spray drying after extraction;

the ganoderma lucidum extract is obtained by extracting ganoderma lucidum with an ethanol solution, wherein the ethanol solution accounts for 70% by volume, the extraction temperature is 76 ℃, the extraction time is 2 hours each time after 3 times, and the material-liquid ratio is 1: and 6, concentrating after extraction, and then spray-drying.

The selection and the dosage of the filling agent are as follows:

the raw materials of the product comprise kudzu root extract, schisandra extract, ganoderma extract and silybum marianum extract. In order to reduce hygroscopicity, the product should be granulated and then refilled. The product selects corn starch as a common auxiliary material as a filling agent, but the prepared sample has high moisture absorption rate and caking phenomenon. Therefore, substitution with calcium carbonate is considered. The calcium carbonate is white, odorless, tasteless, and superfine light powder or crystal, is safe in air, and is insoluble in water and alcohol. The moisture absorption is very low, and the anti-caking effect is achieved, wherein the moisture absorption is very low under 1% at the temperature of 25 ℃ and the relative humidity of 90%.

The types and amounts of fillers are now studied and the results are as follows:

item	Number 1	Number 2	No. 3	Number 4
					Raw materials	28.5g	28.5g	28.5g	28.5g
Corn starch	5g	10g	——	——
					Calcium carbonate	——	——	5g	10g
Rate of moisture absorption	18.56％	15.74％	13.29％	10.11％

Determination of moisture absorption rate: the mixed powder is filled into a weighing bottle with constant weight, the thickness is about 2mm, and the mixed powder is placed at the temperature of 25 ℃ and the relative humidity of 75% for 24h to measure the moisture absorption rate.

As is clear from the above table, when 10g of calcium carbonate was added to 28.5g of the raw material, the hygroscopicity of the mixed powder was small. In 1000 formulations, the total amount of raw materials is 285g, so the amount of calcium carbonate added is about 100g (the amount of calcium carbonate is finally adjusted to 101g according to the subsequent screening of the amounts of the binder and the lubricant). In addition, the recommended daily intake of calcium for adults in the dietary nutrient reference intake of Chinese residents is 800-1000mg, and the daily intake of calcium for adults in the nutrient supplement raw material catalog (I) for health food is 200-1000 mg. The calcium carbonate is used in the product in a daily intake of about 162mg, which is lower than the calcium carbonate. The calcium intake of other foods can just meet the standard, namely the optimal dosage of the calcium carbonate is 26 parts by weight.

The selection and the dosage of the adhesive are as follows:

the povidone K30 is widely applied in medicine and is one of three new medicinal auxiliary materials advocated internationally. The dose of povidone K30 was studied and the results are shown in the following table:

the test result shows that in 1000 preparations (raw material 285g), 10g of povidone K30 is added as a binder, and the prepared granules are easy to form. Therefore, the optimal usage amount of povidone K30 is 2.5 parts by weight.

Selection and dosage of the lubricant of the invention:

the auxiliary materials with one or more of the functions of flow aid, anti-adhesion and lubrication are generally called lubricants, and the common lubricants include magnesium stearate, aerosil, talcum powder and the like. Wherein, magnesium stearate is a lubricant with the widest application, belongs to hydrophobic lubricant, is easy to be uniformly mixed with particles, and is selected as the lubricant according to the actual situation.

Magnesium stearate is a lubricant which is widely applied, belongs to a hydrophobic lubricant, is easy to mix with granules, and the dosage of the magnesium stearate is generally 0.5-1%. Therefore, according to the past production experience, the formula amount of the calcium carbonate is slightly adjusted, the dosage of the magnesium stearate in the formula is 1%, and the angle of repose is detected and verified, and the results are as follows:

the angle of repose is generally less than or equal to 40 degrees, and the fluidity can meet the production requirement. As can be seen from the above table, the flowability of the material is good at a 1.0% magnesium stearate level. The amount of magnesium stearate used is therefore 1.0 part by weight.

Example 1

Weighing 35 parts of kudzu root extract, 5 parts of silybum marianum extract, 9 parts of lucid ganoderma extract, 13 parts of schisandra extract, 26 parts of calcium carbonate, 2.5 parts of povidone K30 and 1.0 part of magnesium stearate in parts by weight;

and (3) granulating the traditional Chinese medicines and uniformly mixing the traditional Chinese medicines with magnesium stearate to obtain total mixed granules, and preparing the total mixed granules into capsules by using a capsule filling machine to obtain the liver-protecting and immunity-enhancing Chinese herbal medicine preparation.

Example 2

Weighing 36 parts of kudzu root extract, 6 parts of silybum marianum extract, 10 parts of lucid ganoderma extract, 14 parts of schisandra extract, 26 parts of calcium carbonate, 2.5 parts of povidone K30 and 1.0 part of magnesium stearate in parts by weight;

Example 3

Weighing 37 parts of kudzu root extract, 7 parts of silybum marianum extract, 11 parts of lucid ganoderma extract, 15 parts of schisandra extract, 26 parts of calcium carbonate, 2.5 parts of povidone K30 and 1.0 part of magnesium stearate in parts by weight;

Example 4

Weighing 38 parts of kudzu root extract, 8 parts of silybum marianum extract, 12 parts of lucid ganoderma extract, 16 parts of schisandra extract, 26 parts of calcium carbonate, 2.5 parts of povidone K30 and 1.0 part of magnesium stearate in parts by weight;

Example 5

Weighing 39 parts of kudzu root extract, 9 parts of silybum marianum extract, 13 parts of lucid ganoderma extract, 17 parts of schisandra chinensis extract, 26 parts of calcium carbonate, 2.5 parts of povidone K30 and 1.0 part of magnesium stearate in parts by weight;

Example 6

The preparation method is the same as the example 1 of the invention, but the silybum marianum extract of the invention is not added, other raw materials and the preparation method are not changed, and the traditional Chinese medicine preparation is prepared in the same way;

example 7

The preparation method is the same as the example 1 of the invention, but the kudzuvine root extract of the invention is not added, other raw materials and the preparation method are not changed, and the traditional Chinese medicine preparation is prepared in the same way;

example 8

The preparation method is the same as the example 1 of the invention, but the schisandra extract of the invention is not added, other raw materials and the preparation method are not changed, and the traditional Chinese medicine preparation is prepared in the same way;

example 9

The preparation method is the same as the example 1 of the invention, but the ganoderma lucidum extract of the invention is not added, other raw materials and the preparation method are not changed, and the traditional Chinese medicine preparation is prepared in the same way;

comparative example 1

The preparation method is basically the same as the example 1 of the invention, except that the cassia seed with the same liver protection function is used for replacing the silybum marianum of the invention, other raw materials and the preparation method are not changed, and the traditional Chinese medicine preparation is also prepared;

firstly, safety test evaluation:

the Chinese herbal medicine preparation of the intermediate value example 3 in the examples 1 to 5 of the invention, which is calculated by weight parts of the main raw materials, is subjected to a safety test, and the results are as follows:

1. acute toxicity test of mice: the maximum tolerance test result shows that the MTD of the test object to mice with both sexes is more than 20.0g/kg. The sample was of the nontoxic grade according to the acute toxicity grading criteria.

2. And (3) genetic toxicity test:

inspection item	Test results
		Ames test	Negative of
Mouse marrow pleochromocyte micronucleus test	Negative of
		Mouse sperm aberration test	Negative of

3.30 days feeding trial: the animal growth and development of each experimental group in the experimental period are good, and indexes such as weight gain, food utilization rate, organ weight, organ coefficient and the like are all in the range of laboratory normal values when the animal growth and development of each experimental group are given to rats for 30 days at doses of 0.67, 1.33 and 2.67g/kg.bw (respectively equivalent to 25, 50 and 100 times of the recommended amount of a human body). The routine and biochemical indexes of the blood in the experimental group are within the normal value range of the laboratory. The examined organs of the histopathology examination experiment group have no significant pathological changes, and no harmful effect is observed.

II, evaluating a liver protection functional test:

the examples 1 to 9 of the present invention and the comparative example 1 were subjected to performance tests, and specific test results are shown in table 1:

1 materials and methods

1.1 sample: hard capsules/dark brown in color, recommended for human body at a dose of 1.6 g/person.day (adult body weight in 60 kg) are provided by Jiangsu Shengshi Kanghe Biotech Co.

1.2 Experimental animals: SPF-grade ICR male mice of the Jinanpunyue experimental animal breeding limited company are selected, the weight of the SPF-grade ICR male mice is 18.9-21.9 g, the SPF-grade ICR male mice are divided into 12 groups randomly, and the 12 groups are respectively 10 experimental groups, a model control group and a blank control group.

1.3 main instruments and reagents:

the main apparatus is as follows: SPECTRAMAX plus microplate reader, CPA2202S model electronic balance, PL203 model electronic balance, 2000GenoGrinder (TM) high throughput tissue grinder, centrifuge, vortex mixer, SW oscillating water bath, Leica CM1800 cryomicrotome, surgical instruments, glacial acetic acid (analytical purity).

The main reagents are as follows: malondialdehyde (MDA) kit, reduced Glutathione (GSH) kit, Triglyceride (TG) kit (GPO-PAD method).

1.4 dose selection and subject administration:

the recommended human intake of the sample is 1.6g/60kg.bw, and the experimental group setting measures 0.27g/kg.bw, which is equivalent to 10 times the recommended human intake. A blank control group was also set up. Taking purified water as a solvent to prepare samples to required concentrations respectively, namely taking 0.81g of the samples to prepare 60ml of the purified water respectively, performing intragastric gavage on a blank control group and a model control group, and after continuously performing oral gavage for 30 days according to 0.2ml/10g.bw per day, starting to measure various indexes.

1.5 test methods:

experiment 30d the experimental group was gavaged with 12ml/kg. bw of 50% ethanol (prepared from absolute ethanol with purified water) and the blank group was given purified water. Animals were sacrificed after a 16 hour overnight fast, livers were weighed, and liver coefficients were calculated. Then, MDA, GSH and TG contents in liver tissues are measured, and pathological and histological examination is carried out.

1.5.1 index detection: the liver is prepared into 10% homogenate, and the contents of MDA, GSH and TG are measured by a kit method.

1.5.2 histopathological examination: taking the left liver leaf of a mouse, taking a cross section from the middle part of the left liver leaf, freezing and slicing, and staining with oil red 0. During microscopic examination, pathological changes of cells are recorded from one end of the liver, and the whole tissue section is continuously observed by using a 40-fold objective lens. The distribution, extent and area of lipid droplets in the liver were mainly observed.

Grading standard:

1.6 statistics of test data: the database was built with excel 1 software and statistical analysis was performed with SPSS software. Performing anova, performing anova test, calculating F value, wherein F is less than F_0.05P > 0.05, conclusion: the difference between the average numbers of all groups has no significance; f value is more than or equal to F_0.05P is less than or equal to 0.05, and statistical analysis is carried out by using a pairwise comparison method (Dunnett test method) of the mean number between a plurality of experimental groups and a control group; carrying out appropriate variable conversion on the data with non-normal or uneven variance, and counting by using the converted data after meeting the homogeneity requirement of the normal or uneven variance; if the variable still does not reach the goal of being normal or uniform in variance after conversion, the statistics is carried out by using the rank sum test.

1.7 determination of results

Has auxiliary protection function on chemical liver injury (alcoholic liver injury model): the method comprises the following steps of firstly, obtaining positive results of three indexes of MDA, GSH and TG of the liver, judging that the sample to be tested has an auxiliary protection effect on the liver injury caused by ethanol, secondly, obtaining positive results of any two indexes of MDA, GSH and TG of the liver, obtaining positive pathological results of the liver, and judging that the sample to be tested has an auxiliary protection effect on the liver injury caused by ethanol.

1.8 detection results of various indexes in liver tissue

TABLE 1 results of MDA assay in liver tissue

Item	Animal number (only)	MDA (mu mol/g tissue)	P1	P2
					Blank control	10	0.27±0.03	/	/
Model control group	10	0.51±0.09	＜0.01	/
					Example 1	10	0.42±0.09	/	0.03
Example 2	10	0.40±0.06	/	0.03
					Example 3	10	0.39±0.08	/	0.02
Example 4	10	0.41±0.09	/	0.03
					Example 5	10	0.42±0.07	/	0.03
Example 6	10	0.52±0.03	/	0.08
					Example 7	10	0.55±0.04	/	0.06
Example 8	10	0.54±0.03	/	0.07
					Example 9	10	0.55±0.05	/	0.06
Comparative example 1	10	0.56±0.04	/	0.05

P1 value: p2: comparing with a model control group;

as can be seen from Table 1, the MDA content of the model control group is obviously higher than that of the blank control group, and the difference is very significant (P <0.01), which indicates that the liver injury model is established. The MDA contents of examples 1-5 have significant difference (P <0.05) compared with the model control group, the MDA contents of examples 6-9 have no significant difference (P > 0.05) compared with the model control group, and the significant difference of example 3 is larger, according to the judgment standard of health food inspection and evaluation technical specification, examples 1-5 have auxiliary protection effect on chemical liver injury, and the effect of example 3 is most obvious, but examples 6-9 have poor auxiliary protection effect on chemical liver injury, probably because the Chinese herbal medicine preparation has an optimal group distribution ratio, the best liver protection effect can be achieved only under the condition of the group distribution ratio of example 3 of the invention, and the drug effect is poor because of less Chinese herbal medicines in examples 6-9, thereby laterally verifying that each Chinese herbal medicine component of the invention has synergistic effect on the drug effect, in addition, in comparative example 1, the semen cassiae which is similar to the silybum marianum in property is used instead of the semen cassiae, and the Chinese herbal medicine preparation is prepared in the same way, but the final using effect is not good as that of the medicament which takes the silybum marianum as the effective component, probably because the synergistic effect can be generated only when the silybum marianum and the other three Chinese herbal medicine extracts are used together, and the best liver protection auxiliary effect is achieved.

TABLE 2 measurement results of GSH in liver tissue

Item	Animal number (only)	GSH (mu mol/g tissue)	P1	P2
					Blank control	10	7.19±0.83	/	/
Model control group	10	4.28±0.72	＜0.01	/
					Example 1	10	4.59±0.84	/	0.04
Example 2	10	4.60±0.69	/	0.04
					Example 3	10	4.65±0.72	/	0.03
Example 4	10	4.63±0.67	/	0.04
					Example 5	10	4.62±0.68	/	0.04
Example 6	10	4.31±0.56	/	0.06
					Example 7	10	4.35±0.65	/	0.07
Example 8	10	4.33±0.68	/	0.07
					Example 9	10	4.38±0.72	/	0.07

P1 value: p2: comparing with a model control group;

as can be seen from Table 2, the GSH content of the model control group is significantly lower than that of the blank control group, and there is a significant difference (P <0.01), indicating that the liver injury model is established. The GSH content in examples 1-5 is significantly different from that of a model control group (P is less than 0.05), and the GSH value is increased, wherein the GSH value of example 3 is the largest, which shows that the Chinese herbal medicine reagent of the invention has an auxiliary protection effect on liver injury, and corresponds to the MDA detection result, the optimal effect under the matching of example 3 verifies that the invention has an optimal matching, and examples 6-9 have poor drug effect because one less Chinese herbal medicine respectively, thereby laterally verifying that each Chinese herbal medicine component of the invention plays a beneficial role in the occurrence of drug effect. Similarly, in comparative example 1, the semen Cassiae with similar properties to that of Silybum marianum was used instead of the semen Cassiae, and the herb preparation was prepared, but the final effect was not as good as the herb preparation containing Silybum marianum as the effective ingredient, probably because only the extract of Silybum marianum and the extract of the other three herbs were used together to produce the synergistic effect, which served as the best liver protection aid.

TABLE 3 results of TG determination in liver tissue

P1 value: p2: comparing with a model control group;

as can be seen from Table 3, the TG content of the model control group is obviously higher than that of the blank control group, and the significant difference (P <0.01) is obtained, which indicates that the liver injury model is established. The TG content of examples 1-5 was significantly lower than the model control with significant differences (P <0.05), and the TG content of examples 6-9 was lower than the model control but no significant differences.

By comprehensively comparing the experimental results, the Malondialdehyde (MDA) and the Triglyceride (TG) of the model control group are obviously increased, the Glutathione (GSH) is obviously reduced, and the significant difference (P <0.01 or P <0.05) is obtained compared with the blank control group, which indicates that the liver injury model is established. The MDA content of the experimental group is lower than that of the model control group, and the obvious difference exists (P is less than 0.05); the GSH content is lower than that of a model control group, and the significant difference exists (P is less than 0.05); the TG content is obviously lower than that of a model control group, and the obvious difference exists (P < 0.05). According to the judgment standard of 'health food inspection and evaluation technical specification', the sample has auxiliary protection effect on chemical liver injury. Meanwhile, in the examples 1 to 5, each specific beneficial index is firstly improved and then reduced, and an optimal value is obviously provided, that is, the proportioning value of the example 3 of the invention is optimal, and by combining three groups of data, it can be seen that, in the comparative example 1, because semen cassiae with similar properties to silybum marianum are used for replacing, a Chinese herbal medicine preparation is prepared, but the final using effect is not good as a medicament with silybum marianum as an effective component, probably because the synergistic effect can be generated only when silybum marianum and three other Chinese herbal medicine extracting solutions are used together, the optimal liver protection auxiliary effect is achieved, and the proportioning ratio of the four Chinese herbal medicines of the invention is also laterally verified, the synergistic effect can be generated, and the optimal liver protection auxiliary effect is achieved.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, it should be noted that, for those skilled in the art, many modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.

Claims

1. A liver-protecting immunity-enhancing Chinese herbal medicine preparation is characterized in that: the material comprises the following raw and auxiliary materials in parts by weight:

35-39 parts of kudzu root extract;

5-9 parts of silybum marianum extract;

9-13 parts of ganoderma lucidum extract;

13-17 parts of schisandra extract;

24-28 parts of calcium carbonate;

2.3-2.7 parts of povidone K30;

0.8-1.2 parts of magnesium stearate;

2. The liver-protecting and immunity-enhancing Chinese herbal medicine preparation according to claim 1, characterized in that: the material comprises the following raw and auxiliary materials in parts by weight:

37 parts of kudzu root extract;

7 parts of silybum marianum extract;

11 parts of ganoderma lucidum extract;

15 parts of schisandra extract;

26 parts of calcium carbonate;

2.5 parts of povidone K30;

1.0 part of magnesium stearate.

3. The liver-protecting and immunity-enhancing Chinese herbal medicine preparation according to claim 1, characterized in that: the extraction process conditions of the silybum marianum extract are as follows: the ethanol solution is 70% in volume, the time for extracting for 3 times is 1.5h, the extraction temperature is normal temperature, and the material-liquid ratio is 1: and 8, concentrating, extracting with ethyl acetate, and drying.

4. The liver-protecting and immunity-enhancing Chinese herbal medicine preparation according to claim 1, characterized in that: the extraction process conditions of the kudzu root extract are as follows: the ethanol solution is 50% in volume, the time for 2 times of extraction is 2 hours, the extraction temperature is 80 ℃, and the material-liquid ratio is 1: 10, filtering, concentrating and spray drying after extraction.

5. The liver-protecting and immunity-enhancing Chinese herbal medicine preparation according to claim 1, characterized in that: the extraction process conditions of the schisandra extract are as follows: the ethanol solution accounts for 60% by volume, the time for 2 times of extraction is 2 hours, the extraction temperature is normal temperature, and the material-liquid ratio is 1:8, filtering, concentrating and spray drying after extraction.

6. The liver-protecting and immunity-enhancing Chinese herbal medicine preparation according to claim 1, characterized in that: the extraction process conditions of the ganoderma lucidum extract are as follows: the ethanol solution is 70% in volume, the extraction temperature is 76 ℃, the extraction time is 2 hours each time after 3 times of extraction, and the material-liquid ratio is 1: and 6, concentrating after extraction, and then spray-drying.

7. A preparation method of a Chinese herbal medicine preparation for protecting liver and enhancing immunity is characterized by comprising the following specific preparation steps: