CN112402511A

CN112402511A - Traditional Chinese medicine composition for liver health care and application thereof

Info

Publication number: CN112402511A
Application number: CN202011339147.5A
Authority: CN
Inventors: 吴长昱; 李世凯; 吴长新
Original assignee: Panjin Tianyuan Pharmaceutical Co ltd
Current assignee: Panjin Tianyuan Pharmaceutical Co ltd
Priority date: 2020-11-25
Filing date: 2020-11-25
Publication date: 2021-02-26

Abstract

The invention provides a traditional Chinese medicine composition for liver health care and application thereof, wherein the composition comprises the following components in parts by weight: 10-100 parts of silybum marianum extract, 10-50 parts of kudzu root extract, 10-50 parts of schisandra extract and 5-30 parts of ganoderma extract. The composition can be made into granule for liver health promotion, and its preparation method comprises: preparing herba Silybi Mariani extract, radix Puerariae extract, fructus Schisandrae extract and Ganoderma extract respectively; clathrating herba Silybi Mariani extract, radix Puerariae extract, and fructus Schisandrae extract with beta-cyclodextrin, adding water, stirring to obtain paste, and grinding for 20-30 min; drying the ground material, and crushing into powder of 80-100 meshes; mixing the powder with Ganoderma extract, maltodextrin and stevioside, adding ethanol to obtain soft material, granulating, and drying; and (4) carrying out size stabilization, quality inspection, inner packaging, sterilization, outer packaging and warehousing on the dried granules in sequence. The results of animal tests and clinical tests show that the granules have good repairing effect on chemical liver injury.

Description

Traditional Chinese medicine composition for liver health care and application thereof

Technical Field

The invention relates to the technical field of traditional Chinese medicine compositions, in particular to a traditional Chinese medicine composition for liver health care and application thereof.

Background

The development space of the Chinese health food market is large. In absolute terms, the consumption level of Chinese health food market is low. The national health food consumption accounts for only 1.47% of the total retail sales of the world total consumer goods; the consumption expenditure of the health-care food for urban and rural population is only 31 yuan per year, which is 1/17 in the United states and 1/12 in Japan. On average, Chinese consumers spend 0.07% of their total expenses for healthcare products, while consumers in European and American countries spend 2% of their total expenses, 29 times as much as China. Therefore, the development potential of the Chinese health food market is huge. According to the report of Chinese medicine, the growth situation of health-care food is as follows: in 2015, CAGR is 22.6%, in 2016, the yield reaches 2621.1 billion yuan, which is about 6 times of 2009, and the growth is very rapid. And at the end of 2016, more than 16000 health-care foods are approved in China, and more than 2500 health-care food production enterprises are available. The sales income of the Chinese nutrition and health care product industry is expected to keep increasing in the coming years.

The whole plant of silybum marianum can be used as a medicine, is a traditional Chinese medicine with obvious protective effect on liver, is bitter and cool in nature, and has the maximum effects of clearing heat and detoxicating, protecting liver and benefiting gallbladder. Meanwhile, the food has a certain protection effect on human brain, and can treat diseases and prevent external radiation source from damaging human body after being eaten. Silybum marianum has fewer adverse reactions. China is a big producing country of the silybum marianum extract, and the export is 1000 tons in 2019, but the silybum marianum extract is a crude product, and the silybum marianum extract is only a cheap raw material medicine trade conducted abroad, so the economic added value is different from that of a finished product medicine. The domestic scale of the finished medicine or health food products is not large, and only 4 health foods approved by the national drug administration and having auxiliary protection effect on chemical liver injury, namely the national food jian character type aqueous silybum marianum extract, are extremely different from the status of the producing country of the silybum marianum extract. In addition, China is a country with multiple liver diseases and has a large population base, the incidence rates of alcoholic fatty liver and non-alcoholic fatty liver disease reported in documents in China are respectively about 4.43% and 15%, the population numbers of the two patient groups can reach more than 2 hundred million, and in addition, the population numbers of liver injury caused by drug-induced chemical liver injury and other reasons are considered to be astonishing in the market, but the current liver health product market of China does not form a scale, and does not have a brand which occupies a certain scale, so that a health product which contains the silybum marianum extract and has a good liver health care effect and no toxic or side effect is developed, the economic added value of silybum marianum is improved, and the method is a way for benefiting the nation and seeking enterprise innovation under the new crown epidemic situation cage at present.

Disclosure of Invention

In view of the above, the present invention provides a traditional Chinese medicine composition for liver health care and an application thereof, aiming at the problems existing in the prior art. The technical scheme of the invention is as follows:

in a first aspect, the invention provides a traditional Chinese medicine composition for liver health care, which comprises the following components in parts by weight: 10-100 parts of silybum marianum extract, 10-50 parts of kudzu root extract, 10-50 parts of schisandra extract and 5-30 parts of ganoderma extract.

Preferably, the traditional Chinese medicine composition comprises the following components in parts by weight: 10-50 parts of silybum marianum extract, 10-15 parts of kudzu root extract, 10-15 parts of schisandra extract and 5-10 parts of ganoderma extract.

Optionally, the ganoderma lucidum extract is replaced by a salvia miltiorrhiza extract.

Optionally, the Chinese medicinal composition further comprises: 5-10 parts of wolfberry powder, 5-10 parts of codonopsis pilosula extract, 1-10 parts of pseudo-ginseng extract, 5-15 parts of coix seed powder, 1-5 parts of polygonum cuspidatum extract and 1-10 parts of liquorice extract.

In a second aspect, the invention provides a traditional Chinese medicine preparation for liver health care, which comprises a liquid preparation or a solid preparation prepared from the composition, wherein the composition accounts for 10-14% of the traditional Chinese medicine preparation by mass.

Further, the traditional Chinese medicine preparation also comprises pharmaceutically acceptable auxiliary materials.

In a third aspect, the present invention provides a granule for liver health, the preparation method of which comprises the following steps:

3) preparing the silybum marianum extract, the kudzu root extract, the schisandra chinensis extract and the ganoderma lucidum extract according to the weight parts;

4) clathrating herba Silybi Mariani extract, radix Puerariae extract, and fructus Schisandrae extract with beta-cyclodextrin, adding water, stirring to obtain paste, and grinding for 20-30 min;

3) drying the ground material, and crushing into powder of 80-100 meshes;

4) mixing the powder with Ganoderma extract, maltodextrin and stevioside, adding ethanol to obtain soft material, granulating, and drying;

5) and (4) granulating, inspecting quality, internally packaging, sterilizing, externally packaging and warehousing the dried granules in sequence to obtain the finished product.

Further, the preparation method of the silybum marianum extract comprises the following steps: cleaning and draining silybum marianum, crushing the silybum marianum into particles with the particle size of 2-3 mm, adding water with the dry weight of 6-8 times of the particle size of the silybum marianum, heating the particles to 100 ℃, decocting the particles for 2-3 hours, filtering the decoction, keeping the filtrate, and concentrating the filtrate in vacuum at the temperature of 40-50 ℃ and the vacuum degree of 0.095Mpa until the concentration of the filtrate is 0.6-0.8 g/ml to obtain the silybum mari. The preparation method of radix Puerariae extract, fructus Schisandrae extract and Ganoderma extract is the same as that of herba Silybi Mariani extract.

Furthermore, the addition amount of the beta cyclodextrin is 0.1-0.3% of the mass of the granules.

Further, the addition amount of the stevioside accounts for 0.01-0.06% of the mass of the granules.

Further, the drying temperature is 60-70 ℃.

Further, the preparation method also comprises the process of further adding medlar powder, codonopsis pilosula extract, panax notoginseng extract, coix seed powder, polygonum cuspidatum extract and liquorice extract into the mixture obtained in the step 4) and uniformly mixing. The preparation method of radix Codonopsis extract, Notoginseng radix extract, rhizoma Polygoni Cuspidati extract, Glycyrrhrizae radix extract and herba Silybi Mariani extract are the same.

Compared with the prior art, the invention can obtain the following technical effects:

the silybum marianum extract, the kudzuvine root extract, the schisandra chinensis extract and the lucid ganoderma extract are compounded to prepare the granules with the liver health care effect, and the granules have simple composition, reasonable proportion and convenient taking. And animal test and clinical test results show that the granules have good repairing effect on chemical liver injury.

Drawings

Fig. 1 is a flow chart of a preparation process of granules of the present invention.

Detailed Description

In the description of the present invention, it is to be noted that those whose specific conditions are not specified in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturers. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The present invention will now be described in further detail with reference to the following figures and specific examples, which are intended to be illustrative, but not limiting, of the invention.

Example 1

The embodiment provides a traditional Chinese medicine composition for liver health care, which comprises the following components in parts by weight: 40 parts of silybum marianum extract, 10 parts of kudzu root extract, 15 parts of schisandra extract and 10 parts of ganoderma extract.

Example 2

The embodiment provides a traditional Chinese medicine composition for liver health care, which comprises the following components in parts by weight: 100 parts of silybum marianum extract, 10 parts of kudzu root extract, 15 parts of schisandra extract and 5 parts of ganoderma extract.

Example 3

The embodiment provides a traditional Chinese medicine composition for liver health care, which comprises the following components in parts by weight: 10 parts of silybum marianum extract, 50 parts of kudzu root extract, 50 parts of schisandra extract and 30 parts of ganoderma extract.

Example 4

The embodiment provides a traditional Chinese medicine composition for liver health care, which comprises the following components in parts by weight: 50 parts of silybum marianum extract, 15 parts of kudzu root extract, 10 parts of schisandra extract, 5 parts of ganoderma extract, 10 parts of medlar powder, 10 parts of codonopsis pilosula extract, 5 parts of pseudo-ginseng extract, 10 parts of coix seed powder, 2 parts of polygonum cuspidatum extract and 10 parts of liquorice extract.

Example 5

The embodiment provides a traditional Chinese medicine composition for liver health care, which comprises the following components in parts by weight: 30 parts of silybum marianum extract, 12 parts of kudzu root extract, 12 parts of schisandra chinensis extract and 8 parts of salvia miltiorrhiza extract.

Example 6

The embodiment provides a traditional Chinese medicine composition for liver health care, which comprises the following components in parts by weight: 40 parts of silybum marianum extract, 35 parts of kudzu root extract, 25 parts of schisandra chinensis extract, 10 parts of salvia miltiorrhiza extract, 7 parts of medlar powder, 6 parts of codonopsis pilosula extract, 4 parts of pseudo-ginseng extract, 12 parts of coix seed powder, 3 parts of polygonum cuspidatum extract and 8 parts of liquorice extract.

Example 7

The present embodiment provides a granule for liver health care, which is prepared from the traditional Chinese medicine composition of embodiment 1, wherein the mass content of the traditional Chinese medicine composition in the granule is 12%, the preparation process is shown in fig. 1, and the preparation method comprises the following steps:

1) preparing the ingredients according to the weight parts of the traditional Chinese medicine composition in the embodiment 1, and then respectively preparing silybum marianum extract, kudzu root extract, schisandra chinensis extract and ganoderma lucidum extract; the preparation method of the silybum marianum extract comprises the following steps: cleaning and draining silybum marianum, crushing the silybum marianum into particles of 2-3 mm, adding water with the dry weight 6 times of that of the particles, heating the particles to 100 ℃, decocting the particles for 3 hours, filtering the decoction, keeping filtrate, and concentrating the filtrate in vacuum at the temperature of 45 ℃ and the vacuum degree of 0.095Mpa until the concentration of the filtrate is 0.7g/ml to obtain the silybum marianum extract. The preparation method of radix Puerariae extract, fructus Schisandrae extract and Ganoderma extract is the same as that of herba Silybi Mariani extract.

2) Clathrating herba Silybi Mariani extract, radix Puerariae extract, and fructus Schisandrae extract with beta-cyclodextrin, adding 1 time of water, stirring to obtain paste, and grinding for 30 min; the addition amount of cyclodextrin is 0.2% of the granule mass.

3) Drying the ground substance at 65 deg.C in oven, and pulverizing into 80-100 mesh powder.

4) Uniformly mixing the powder with the ganoderma lucidum extract, maltodextrin and stevioside, adding 80% ethanol to prepare a soft material, preparing the soft material into 18-mesh granules by using a swing granulator, and drying the 18-mesh granules by using an oven at 60-70 ℃; wherein, the addition amount of stevioside accounts for 0.02 percent of the mass of the granules, and the balance is maltodextrin.

5) Sorting the dried granules by using a 14-mesh sieve, sealing and packaging, sticking a barrel stick, sending the granules into an intermediate station for inspection, and preparing an intermediate for detection, wherein the water content is less than or equal to 5%;

6) taking the aluminum-plastic composite film qualified by inspection, and carrying out inner packaging on the semi-finished product of the granules qualified by inspection;

7) irradiation sterilization: 60CO radiation at a dose of 5 kGy.

8) Packaging and warehousing: and (5) externally packaging, and packaging and warehousing after the finished product is qualified. The storehouse is cool, ventilated and dry. The finished product is placed in an area marked with a qualified mark, and is required to be placed at a position which is more than 20cm away from the wall.

Example 8

The present example provides a granule for liver health care, which employs the traditional Chinese medicine composition of example 2, the mass content of the traditional Chinese medicine composition in the granule is 12%, and the preparation method is the same as that of example 7.

Example 9

The present example provides a granule for liver health care, which employs the traditional Chinese medicine composition of example 3, the mass content of the traditional Chinese medicine composition in the granule is 12%, and the preparation method is the same as that of example 7.

Example 10

The present embodiment provides a granule for liver health care, which employs the traditional Chinese medicine composition of embodiment 4, and the mass content of the traditional Chinese medicine composition in the granule is 12%, and the preparation method thereof is different from that of embodiment 7 in that: in the step 4), uniformly mixing the powder with the ganoderma lucidum extract, the medlar powder, the codonopsis pilosula extract, the panax notoginseng extract, the coix seed powder, the polygonum cuspidatum extract, the liquorice extract, the maltodextrin and the stevioside, adding ethanol to prepare soft materials, granulating and drying.

Example 11

The present embodiment provides a granule for liver health care, which employs the traditional Chinese medicine composition of embodiment 5, and the mass content of the traditional Chinese medicine composition in the granule is 12%, and the preparation method thereof is different from that of embodiment 7 in that: replacing Ganoderma extract with Saviae Miltiorrhizae radix extract.

Example 12

The present embodiment provides a granule for liver health care, which employs the traditional Chinese medicine composition of embodiment 6, and the mass content of the traditional Chinese medicine composition in the granule is 12%, and the preparation method thereof is different from that in embodiment 10 in that: replacing Ganoderma extract with Saviae Miltiorrhizae radix extract.

Example 13

The granules of examples 7 to 12 were subjected to property and quality tests

First, sensory performance index

As shown in table 1.

TABLE 1 sensory index of granules of examples 7 to 12

Second, physicochemical indexes

As shown in table 2.

TABLE 2 physicochemical indices of the granules of examples 7 to 12

Third, microorganism index

As shown in table 3.

TABLE 3 microbiological indicator of the granules of examples 7 to 12

Item	Index (I)	Detection method	Examples 7 to 12
				Total number of colonies, CFU/g	≤30000	GB 4789.2	Conform to
Coliform group, MPN/g	≤0.92	GB 4789.3 "MPN counting method"	Conform to
				Mold and Yeast, CFU/g	≤50	GB 4789.15	Conform to
Salmonella	≤0/25g	GB 4789.4	Conform to
				Staphylococcus aureus	≤0/25g	GB 4789.10	Conform to

Comparative example 1

The comparative example provides a granule for liver health care, the adopted traditional Chinese medicine composition only contains silybum marianum extract, and the preparation method is the same as that of example 7.

Comparative example 2

The comparative example provides a granule for liver health care, the adopted traditional Chinese medicine composition only comprises silybum marianum extract and ganoderma lucidum extract, and the preparation method is the same as that of example 7.

Comparative example 3

The comparative example provides a granule for liver health care, the adopted traditional Chinese medicine composition only comprises silybum marianum extract and salvia miltiorrhiza extract, and the preparation method is the same as that of example 11.

Animal experiments

First, toxicological test

(1) Acute oral toxicity test of rat and mouse, three genetic toxicity tests (Ames test, micronucleus test of mouse bone marrow cells and teratospermia test of mouse) and 30-day feeding test of rat.

Materials and methods

1) Sample preparation: the compositions of examples 1-6, the sample solvent was distilled water.

2) Experimental animals: SPF-level Kunming mice and SD rats bred by Liaoning Biotechnology Limited, and the license number of the experimental animal production is SCXK (Liao) 2010-0001. The sterile block mouse material is provided by Beijing Huafukang Biotechnology GmbH, and the production license number is SCXK (Jing) 2009-0008.

3) The test conditions are as follows: the barrier system animal laboratory has the temperature of 22 +/-1.5 ℃, the humidity of 50% +/-10%, the working illumination of 160-2801 x, the artificial illumination of 12h for 12h at night, the noise of less than 60dB, and the use license number SYXK (Liao) 2011-.

4) Acute toxicity test in mice (maximum tolerated dose method): 60 healthy mice (18-22 g) were selected, half of each male and female, and randomly divided into 6 groups, which respectively correspond to the compositions of examples 1-6, and the mice were fasted overnight for 16 hours before the test without restriction of water. The test was carried out at a dose of 24.00g/kgBW (corresponding to 272 times the recommended daily intake). Weighing 50.00g of sample, adding distilled water to 125mL, wherein the sample preparation concentration is 0.400g/mL, feeding the sample in a stomach irrigation mode, the stomach irrigation capacity is 20mL/kgBW, performing stomach irrigation for 3 times at intervals of 4 hours, continuously observing for 14 days, and determining the Maximum Tolerated Dose (MTD) if the mice do not die.

5) Acute toxicity test in rats (maximum tolerated dose method): 180-220 g of healthy rats 60 in each half of male and female were selected and randomly divided into 6 groups, which correspond to the compositions of examples 1-6, respectively, and the rats were fasted overnight for 16 hours before the test without restriction of water. The test was carried out at a dose of 24.00g/kgBW (corresponding to 272 times the recommended daily intake). Weighing 50.00g of sample, adding distilled water to 125mL, wherein the sample preparation concentration is 0.400g/mL, feeding the sample in a gastric lavage mode, carrying out gastric lavage for 3 times at intervals of 4 hours, continuously observing for 14 days if the rat does not die, and determining the Maximum Tolerated Dose (MTD).

6) Ames test: the four tested strains of Salmonella typhimurium with histidine deficiency TA97, TA98, TA100 and TA102 which are identified to meet the requirements are adopted for testing. According to the toxicity determination results, 5 dose groups of 5.000 mg/dish, 1.000 mg/dish, 0.200 mg/dish, 0.040 mg/dish and 0.008 mg/dish were set for the test. 2500mg, 500mg, 100mg, 20mg and 4mg of samples were taken, respectively, distilled water was added to 50mL, and the sample preparation concentrations were 50.0mg/mL, 10.0mg/mL, 2.0mg/mL, 0.4mg/mL and 0.08 mg/mL. The sample was sterilized at 0.103MPa (121 deg.C) for 20min and used as a positive control, a sample solvent control and an untreated control. 0.1mL of the enriched liquid of the test strain, 0.1mL of the sample solution for the test and 0.5mLS-9 of the mixed liquid (when metabolic activation is required) are added into the top layer culture medium, mixed uniformly and poured onto the bottom layer culture medium plate, and 3 plates are measured in each. The cells were incubated at 37. + -. 1 ℃ for 48h and the number of colonies transformed per dish was counted. The number of the retrogradation colonies of the dose group is more than doubled compared with that of the sample solvent control group, and the colony with the dose-response relationship is judged to be positive. The whole set of experiments was repeated under the same conditions. The compositions of examples 1 to 6 were all tested in the same manner as described above.

7) Micronucleus test of mouse bone marrow cells: the test was performed by oral gavage with two 24h intervals. 50 mice with weight of 25-30 g are randomly divided into 5 groups, each group comprises 10 mice, and the number of the mice is half of that of the male and female mice. The test was carried out with 3 dose groups of 8.00g/kgBW (one maximum gavage dose), 4.00g/kgBW, 2.00 g/kgBW. Samples of 40.00g, 20.00g and 10.00g are respectively taken, distilled water is respectively added to 100mL, the sample preparation concentration is 0.400g/mL, 0.200g/mL and 0.100g/mL, and fresh samples are prepared when in use. Cyclophosphamide (intraperitoneal injection) with the dose of 40mg/kgBW is used as a positive control, a sample solvent is used as a negative control, the intragastric perfusion volume is 20mL/kgBW, after the last sample is given for 6 hours, cervical vertebra dislocation is carried out to kill animals, sternum bone marrow is taken, a smear is diluted by calf serum, fixed by methanol, and stained by Giemsa. Under an oil microscope, 1000 pleochromophilic erythrocytes (PCE) are observed for each animal, the number of PCE containing microkernels is counted, and the microkernel cell rate (in a thousandth) is calculated. Each animal was observed for 200 PCEs, the number of mature red blood cells (NCE) was counted, and PCE/NCE was calculated. And statistical processing is performed. Due to the workload relationship, the test of the item was only performed on the Chinese medicinal compositions of example 4 and example 6, and the results were considered as representative. 8) Mouse teratospermia test: 25 sexually mature male mice weighing 25-35 g were used and randomly divided into 5 groups. The test was carried out with 3 dose groups of 8.00g/kgBW (one maximum gavage dose), 4.00g/kgBW, 2.00 g/kgBW. Samples of 40.00g, 20.00g and 10.00g are respectively taken, distilled water is respectively added to 100mL, the prepared concentration of the samples is 0.400g/mL, 0.200g/mL and 0.100g/mL, and the samples are prepared fresh when in use. Cyclophosphamide (intraperitoneal injection) with the dose of 40mg/kgBW is used as a positive control, a sample solvent is used as a negative control, the sample is given in a stomach irrigation mode, the stomach irrigation capacity is 20mL/kgBW, the stomach irrigation is carried out once a day for 5 days continuously, the animals are sacrificed 30 days after the last stomach irrigation, epididymis is taken for flaking, eosin staining is carried out, 1000 sperms with complete structures are observed and counted for each animal, the number of teratospermia is counted, the teratospermia rate (in percentage) is calculated, and statistical treatment is carried out. Due to the workload relationship, the test of the item was only performed on the Chinese medicinal compositions of example 4 and example 6, and the results were considered as representative.

9) Rat 30-day feeding trial: 80 rats with the weight of 70.85 +/-3.40 g are used, and the rats are male and female halves. According to the design of 100 times, 50 times and 25 times of the recommended daily intake, 3 dosage groups of 8.83g/kgBW, 4.42g/kgBW and 2.21gg/kgBW and a basal feed control group are set in the test, a feed step-by-step amplification and uniform mixing method is adopted to mix samples, 1767g, 883g and 442g are respectively sampled and uniformly mixed into the basal feed to 20kg, wherein 318g of casein is mixed into the high dosage group, the sample mixing amount is respectively 8.83%, 4.42% and 2.21%, the animals are raised in a single cage, and the general performance, the poisoning performance and the death condition of the animals are observed and recorded every day. Recording body weight and food intake twice a week, and calculating the weekly and total food utilization rate; removing eyeball and blood after fasting for 16h at the end of test, and measuring hemoglobin, erythrocyte count, leukocyte count and classification, serum glutamic-pyruvic transaminase, glutamic-oxalacetic transaminase, urea nitrogen, creatinine, blood sugar, total protein, albumin, total cholesterol, triglyceride, etc.; after head-broken bloodletting, general examination is carried out on all animals, absolute weights of organs such as liver, kidney, spleen and testis and the like are weighed, the organ body ratio is calculated, main organs are fixedly stored, when general examination on animals of each dosage group does not show obvious pathological changes, histopathological examination on liver, kidney, stomach, intestine, spleen, testis and ovary of a high-dosage group and a control group is carried out, and statistical treatment is carried out. Due to the workload relationship, the test of the item was only performed on the Chinese medicinal compositions of example 4 and example 6, and the results were considered as representative.

10) The statistical method comprises the following steps: and (4) adopting a toxicological inspection data entry statistical report integrated automatic analysis program to carry out statistical analysis, and directly importing the analysis result into an inspection report form. For the measurement data, a multi-sample anova test (Bartlett method) and a two-sample anova test of each experimental group and the solvent control group were performed.

11) The experimental results are as follows:

A. maximum tolerated dose test in mice (maximum use concentration and gavage volume): all samples were orally tolerated at maximum doses greater than 24.00g/kgBW by both sexes, and were of a nontoxic grade.

B. Maximum tolerated dose test in rats (maximum use concentration and gavage volume): all samples were orally tolerated at maximum doses greater than 24.00g/kgBW by both sexes, and were of a nontoxic grade.

C. And (3) genetic toxicity test: no genotoxic effect was observed in the samples.

D. Ames test: the result was negative.

E. Mouse marrow pleochromocyte micronucleus test: the result was negative.

F. Mouse teratospermia test: the result was negative.

G. Rat 30-day feeding trial: clinical examination, hematology examination, blood biochemistry examination and organ weighing result show that the difference is not significant when the test group and the control group of each test item are compared. The result of the pathological histological examination shows that no pathological histological change related to the sample is found in the liver, kidney, stomach, intestine, spleen, testis and ovary of the female rat and the male rat in the high-dose test group.

(2) Rat 90-day feeding test

1) Sample preparation: the compositions of examples 4 and 6.

2) Experimental animals: the license number of the experimental animal production of SPF SD rat bred by Liaoning Biotechnology Limited is SCXK (Liao) 2010-0001. The sterile block mouse material is provided by Beijing Huafukang Biotechnology GmbH, and the production license number is SCXK (Jing) 2009-0008.

3) The test conditions are as follows: the barrier system animal laboratory has the temperature of 22 +/-1.5 ℃, the humidity of 50% +/-10%, the working illumination of 200-2801 x, the artificial illumination of 12h for 12h at night, the noise of less than 60dB, and the use license number SYXK (Liao) 2011-.

4) Rat 90 days feeding trial: 80 rats with the weight of 72.76 +/-4.98 g are used, and the rats are male and female halves. 3 dosage groups of 8.00g/kgBW, 4.00g/kgBW and 2.00gg/kgBW and a basal feed control group are respectively mixed with the feed by a feed step-by-step amplification and mixing method, 8000g, 4000g and 2000g of samples are respectively mixed with the basal feed to 80kg, the mixing amount of the samples is respectively 10.00 percent, 5.00 percent and 2.50 percent, and 1740g of casein is mixed with the feed of the 8.00g/kgBW dosage group. Animals were housed in single cages and observed and recorded daily for general signs, signs of intoxication and mortality. Recording body weight and food intake twice a week, and calculating the weekly and total food utilization rate; after fasting for 16h in the middle test period (day 45) and the end test period (day 92), blood is taken from the intra-ocular angular venous plexus, hemoglobin, erythrocyte count, leukocyte count and classification are determined, after fasting for 16h in the end test period, blood is taken from the intra-ocular angular venous plexus, and serum glutamic pyruvic transaminase, glutamic-oxalacetic transaminase, urea nitrogen, creatinine, blood sugar, total protein, albumin, total cholesterol, triglyceride and the like are determined; taking each organ tissue after head breaking and bloodletting for general examination, weighing the absolute weight of organs such as liver, kidney, spleen, testis and the like, calculating the organ body ratio, fixedly storing main organs, carrying out histopathological examination on the liver, kidney, stomach, intestine, spleen, testis and ovary of a high-dose group and a control group when no obvious lesion is found in general examination of animals of each dose group, and carrying out statistical treatment.

5) The statistical method comprises the following steps: and (4) adopting a toxicological inspection data entry statistical report integrated automatic analysis program to carry out statistical analysis, and directly importing the analysis result into an inspection report form. For the measurement data, a multi-sample anova test (Bartlett method) and a two-sample anova test of each experimental group and the solvent control group were performed.

6) The experimental results are as follows:

clinical examination, hematology examination, blood biochemistry examination and organ weighing result show that the difference is not significant when the test group and the control group of each test item are compared. The result of the pathological histological examination shows that no pathological histological change related to the sample is found in the liver, kidney, stomach, intestine, spleen, testis and ovary of the female rat and the male rat in the high-dose test group.

Second, drug efficacy test

Effect on rat chemical liver injury model

1) Sample preparation: the compositions of examples 1-6 and the extracts of comparative examples 1-3, distilled water as a solvent.

2) Experimental animals: SPF 180-220 g male SD rats 110 bred by Liaoning Biotechnology Limited, and the production license number of the experimental animal is SCXK (Liao) 2010-0001. The sterile block mouse material is provided by Beijing Huafukang Biotechnology GmbH, and the production license number is SCXK (Jing) 2009-0008.

4) Grouping condition: test groups one to 9 correspond to the compositions of examples 1 to 6 and comparative examples 1 to 3 in sequence, the dosage is 2.65g/kgBW (30 times of the recommended dosage), distilled water is added to adjust to 100mL, the sample preparation concentration is 0.265g/mL, and the compositions are prepared fresh when in use. The samples are administrated in a gastric lavage mode, the sample solvent is administrated to the model control group and the blank control group, the gavage volume of the samples for the test is adjusted according to the weight (the gavage volume is calculated according to 10 mL/kgBW), and the gavage is performed for 1 time every day for 30 days continuously. After the experiment is finished, the experimental groups and the model control group are subjected to intragastric lavage once and are fed with 12mL/kgBW of 50% ethanol, the blank control group is fed with distilled water, the animals are killed after being fasted for 16h and the heads are cut off, and the contents of malondialdehyde, reduced glutathione and triglyceride in liver tissues and the pathological histological examination are measured.

5) The detection method comprises the following steps:

preparation of 10% liver homogenate: taking a certain amount of liver, washing with normal saline, wiping, weighing, shearing, placing in a homogenizer, adding cold normal saline, homogenizing at 15000r/min for 10s, intermittently suspending for 30s, repeating for 3 times to obtain 10% tissue homogenate (in ice water), centrifuging at 2000r/min for 15min, and taking the supernatant for testing.

② a method for measuring lipid peroxide degradation product Malondialdehyde (MDA) in liver homogenate:

A. the operating steps for measuring the MDA content of 10% liver tissue homogenate are as follows: the various reagents required for this step are given in table 1.

TABLE 1

Reagent	Standard tube	Standard blank tube	Measuring tube
				10nmol/L Standard (mL)	0.1	－	－
Anhydrous ethanol (mL)	－	0.1	－
				Test sample (mL)	－	－	0.1
Reagent one (mL)	0.1	0.1	0.1
				Reagent two (mL)	3.0	3.0	3.0
Reagent three (mL)	1.0	1.0	1.0

The reagents in the table 1 are mixed uniformly by a vortex mixer, the mouth of a test tube is tied by a preservative film, a small hole is punctured by a needle, boiling water bath is carried out for 40 minutes, the test tube is taken out and cooled by running water, and the test tube is centrifuged for 10 minutes at 4000 r/min. And (4) taking the supernatant, adjusting the optical path to 1cm at 532nm, adjusting the optical path to zero by using distilled water, and measuring the absorbance value of each tube.

B. Calculating the MDA content of the liver tissue:

MDA content (nmol/g tissue) ═ measurement tube absorbance-blank tube absorbance)/(standard tube absorbance-blank tube absorbance) × standard concentration (nmol/L)/liver homogenate concentration (g/mL)/1000.

C. And (4) judging a result: on the premise that the model is established, the MDA content of the test group is remarkably reduced compared with that of the model control group, the difference is remarkable, and the index is judged to be positive.

③ the method for measuring the liver homogenate reduced Glutathione (GSH):

A. preparing a supernatant fluid: taking 0.5mL of 10% homogenate supernatant, adding 2mL of reagent-application solution, mixing uniformly, centrifuging at 4000 rpm for 10 minutes, and taking 1mL of supernatant for color reaction.

B. The upper clear liquid GSH content determination operation steps: the reagents used are shown in table 2.

TABLE 2

Reagent	Standard tube	Standard blank tube	Measuring tube
				Reagent one (mL)	0.1	－	－
Anhydrous ethanol (mL)	－	0.1	－
				Test sample (mL)	－	－	0.1
Reagent one (mL)	0.1	0.1	0.1
				Reagent two (mL)	3.0	3.0	3.0
Reagent three (mL)	1.0	1.0	1.0

Mixing, standing for 5min at 420nm for 1cm, adjusting to zero with double distilled water, and measuring OD value of each tube.

C. Calculating the GSH content of the liver tissue:

GSH content (mgGSH/mg liver tissue) ═ concentration of standard (measured tube absorbance-blank tube absorbance)/(standard tube absorbance-blank tube absorbance) × (20 × 10)^-3mmol/L). times.GSH molecular weight (307). times.dilution factor before sample testing.times.liver homogenate concentration (mg/mL).

D. And (4) judging a result: under the premise that the model is established, the content of reduced GSH in the test group is obviously increased compared with that in the model control group, the difference is obvious, and the index is judged to be positive.

Fourthly, the method for measuring Triglyceride (TG) in the liver homogenate comprises the following steps: taking 10% liver homogenate, and determining with a full-automatic biochemical analyzer. On the premise that the model is established, the triglyceride content of the test group is remarkably reduced compared with that of the model control group, the difference is remarkable, and the index is judged to be positive.

6) Histological examination of liver cases

Firstly, material taking: the left lobe of the liver was harvested by cross-section, frozen and stained with Sudan IV.

Secondly, microscopic examination: pathological changes of cells were recorded starting from one end of the liver field and the whole tissue section was observed continuously with a 40-fold objective lens. Mainly observe the distribution, range and area of fat drops in the liver

③ grading standard:

judging the result: on the premise that the model is established, compared with a model control group, the steatosis of any test group is reduced, the difference is significant, and the index is judged to be positive.

7) Data processing: in a toxicological inspection data entry statistical report automatic analysis program on an Excel2003 platform, various statistical inspection methods simultaneously and automatically perform statistical analysis, and automatically transmit relevant data obtained by analysis to a designated position of an inspection report form. For the measured data, a multi-sample homogeneity test of variance (Bartlett method) and a two-sample homogeneity test of variance of each test group and a model control group are carried out. Uniform variance, adopting single-factor analysis of variance for multi-sample comparison, and adopting a minimum significant difference method, a Dunnett method and a new repolarization method for comparison of each test group and a model control group; variance, using approximate F test and double-sample variance t test or variable conversion (F test after conversion of percentage data by arcsine function) or rank-sum test (Wilcoxon two-sample comparison method)

8) Test results

The test results of the weight and the weight gain: as shown in table 3, the unit: g.

TABLE 3

As can be seen from Table 3, the body weight and the weight gain of each test group are not significantly different from those of the model control group (P > 0.05). The blank control group has no significant difference in body weight and body weight gain compared with the model control group (P > 0.05).

Measurement result of lipid peroxide degradation product Malondialdehyde (MDA) in liver homogenate: as shown in table 4.

TABLE 4

As shown in Table 4, the blank control group has significant difference (P <0.01) in MDA content compared with the model control group, which indicates that the alcoholic liver injury model is established. The MDA content of the test group I-six is obviously lower than that of the model control group (P <0.05), wherein the MDA content of the test group IV is most obviously reduced (P <0.01), and the MDA content of the test group six-seven is also reduced compared with that of the model control group, but the effect is obviously inferior to that of the test group I-six, which shows that the effect of the medicament in liver health care is reduced after the kudzu root extract and the schisandra extract are lacked. ③ determination result of liver homogenate reduced Glutathione (GSH): as shown in table 5.

TABLE 5

As can be seen from Table 5, the GSH content of the blank control group is significantly different from that of the model control group (P <0.01), which indicates that the alcoholic liver injury model is established. The GSH content of the test group I to six is obviously higher than that of the model control group (P <0.05), wherein the GSH content of the test group IV is increased most obviously (P <0.05), and the GSH content of the test group six to seven is also increased compared with that of the model control group, but the effect is obviously inferior to that of the test group I to six, which shows that the effect of the medicament without the kudzu root extract and the schisandra extract on liver health care is reduced.

Liver homogenate Triglyceride (TG) measurement: as shown in table 6.

TABLE 6

As shown in Table 6, the TG content in the blank control group was significantly different from that in the model control group (P <0.01), indicating that the model of alcoholic liver injury was established. The TG content of each test group was not significantly different from that of the model control group (P > 0.05).

The result of the liver histopathology examination: as shown in table 7, the unit: and (4) dividing.

TABLE 7

As can be seen from Table 7, the average of fat droplets in hepatocytes of the blank control group was significantly different from that of the model control group (P <0.01), indicating that the alcoholic liver injury model was established. The average distribution of fat droplets in the hepatocytes of each test group was significantly lower than that of the model control group (P < 0.01). And the average distribution of fat drops in the hepatocytes of the test groups I to six is obviously lower than that of the fat drops in the hepatocytes of the test groups seven to nine.

In conclusion, the test results show that: the traditional Chinese medicine composition has no obvious influence on the weight gain of male rats, can reduce the MDA content in liver homogenate of model rats, increase the GSH content in the liver homogenate of the model rats and reduce the average fat drop in liver cells of the model rats. Has no obvious influence on the TG content of rats.

Third, clinical trial

Test subjects: the male patients with alcoholic liver injury aged 41 years old and 63 years old respectively have symptoms of inappetence, fatigue, hypodynamia, nausea, abdominal distension and the like.

The test method comprises the following steps: the granule of example 5 is administered for 1 month 2 times daily, 5g each time. Liver function was examined in the same hospital before and after administration, and the results are shown in tables 4 and 5, respectively.

TABLE 4 results of liver function tests before taking two patients with alcohol liver injury

TABLE 5 results of liver function test 1 month after two patients with alcohol liver injury who took the granule of example 5

The data in table 4 show that two patients are typical alcoholic liver injury patients, and after 1 month of treatment, the ALT, AST, ALP, LDH and gamma-GT indexes beyond the normal range are all remarkably reduced, which indicates that the granule of the invention can play a better role in repairing alcoholic liver injury. In addition, after the medicine is taken for 1 month, the symptoms of fatigue, hypodynamia, nausea, abdominal distension and the like of the two patients are obviously improved, and the symptom of inappetence disappears.

In conclusion, the silybum marianum extract, the kudzuvine root extract, the schisandra chinensis extract and the lucid ganoderma extract are compounded to prepare the granules with the liver health care effect, and the granules have the advantages of simple composition, reasonable proportion and convenient taking. And animal test results show that the granules have good repairing effect on chemical liver injury. Clinical results show that the granules can obviously improve the liver function of a patient with liver injury after being taken for one month, and the symptoms of inappetence, fatigue, hypodynamia, nausea, abdominal distension and the like appearing during the illness period are greatly improved.

As used in the specification and claims, certain terms are used to refer to particular components or methods. As one skilled in the art will appreciate, different regions may refer to a component by different names. The present specification and claims do not intend to distinguish between components that differ in name but not in name. In the following description and in the claims, the terms "include" and "comprise" are used in an open-ended fashion, and thus should be interpreted to mean "include, but not limited to. "substantially" means within an acceptable error range, and a person skilled in the art can solve the technical problem within a certain error range to substantially achieve the technical effect. The description which follows is a preferred embodiment of the present application, but is made for the purpose of illustrating the general principles of the application and not for the purpose of limiting the scope of the application. The protection scope of the present application shall be subject to the definitions of the appended claims.

It is also noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a good or system that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such good or system. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other like elements in a commodity or system that includes the element.

While the foregoing description shows and describes several preferred embodiments of the invention, it is to be understood, as noted above, that the invention is not limited to the forms disclosed herein, but is not to be construed as excluding other embodiments and is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as expressed herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. A traditional Chinese medicine composition for liver health care is characterized by comprising the following components in parts by weight: 10-100 parts of silybum marianum extract, 10-50 parts of kudzu root extract, 10-50 parts of schisandra extract and 5-30 parts of ganoderma extract.

2. The traditional Chinese medicine composition for liver health care according to claim 1, which comprises the following components in parts by weight: 10-50 parts of silybum marianum extract, 10-15 parts of kudzu root extract, 10-15 parts of schisandra extract and 5-10 parts of ganoderma extract.

3. The traditional Chinese medicine composition for liver health care according to claim 1 or 2, wherein the ganoderma lucidum extract is replaced by a salvia miltiorrhiza extract.

4. The traditional Chinese medicine composition for liver health care according to claim 3, further comprising: 5-10 parts of wolfberry powder, 5-10 parts of codonopsis pilosula extract, 1-10 parts of pseudo-ginseng extract, 5-15 parts of coix seed powder, 1-5 parts of polygonum cuspidatum extract and 1-10 parts of liquorice extract.

5. A traditional Chinese medicine preparation for liver health care is characterized by comprising a liquid preparation or a solid preparation prepared from the composition of any one of claims 1 to 4, wherein the composition accounts for 10 to 14 percent of the traditional Chinese medicine preparation by mass.

6. A preparation method of granules for liver health care is characterized by comprising the following steps:

1) preparing the composition according to the weight parts of the claim 1, and then preparing the silybum marianum extract, the pueraria lobata extract, the schisandra chinensis extract and the ganoderma lucidum extract respectively;

2) clathrating herba Silybi Mariani extract, radix Puerariae extract, and fructus Schisandrae extract with beta-cyclodextrin, adding water, stirring to obtain paste, and grinding for 20-30 min;

3) drying the ground material, and crushing into powder of 80-100 meshes;

7. The method for preparing the granules for liver health care according to claim 6, wherein the method for preparing the silybum marianum extract comprises the following steps: cleaning and draining silybum marianum, crushing the silybum marianum into particles with the particle size of 2-3 mm, adding water with the dry weight of 6-8 times of the particle size of the silybum marianum, heating the particles to 100 ℃, decocting the particles for 2-3 hours, filtering the decoction, keeping the filtrate, and concentrating the filtrate in vacuum at the temperature of 40-50 ℃ and the vacuum degree of 0.095Mpa until the concentration of the filtrate is 0.6-0.8 g/ml to obtain a silybum mari; the preparation method of radix Puerariae extract, fructus Schisandrae extract and Ganoderma extract is the same as that of herba Silybi Mariani extract.

8. The preparation method of the granules for liver health care according to claim 6, wherein the addition amount of the beta-cyclodextrin is 0.1-0.3% of the mass of the granules.

9. The preparation method of the granules for liver health care according to claim 6, wherein the stevioside is added in an amount of 0.01 to 0.06% by mass of the granules.

10. The method for preparing granules for liver health according to any one of claims 6 to 9, wherein the method further comprises a process of further adding medlar powder, radix codonopsis pilosulae extract, panax notoginseng extract, coix seed powder, giant knotweed rhizome extract and liquorice extract into the mixture of step 4) and uniformly mixing.