A liver-protecting composition, preparation method and application thereof

Abstract

The present invention relates to a liver protection composition, a preparation method and an application thereof, and relates to the field of medical technology. The composition is prepared from the following raw materials by weight: 15 to 25 parts of hawthorn, 15 to 25 parts of kudzu root, 15 to 25 parts of Hovenia dulcis, 2 to 4 parts of ganoderma lucidum spores and 1 to 2 parts of water lotus, and the liver protection composition is obtained by extraction and freeze drying. The liver protection composition of the present invention can reduce the ALT, AST activity and MDA content in the serum of mice with alcoholic liver damage, increase the SOD activity in the serum, inhibit the expression of IL-1β and TNF-α in the liver, relieve liver enlargement, and show significant liver protection activity.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a liver-protecting composition, a preparation method and an application thereof. Background Art

[0002] The liver is the largest substantial organ in the human body. It is an important site for the metabolism of energy substances such as fat, protein and sugar. It is also the site for the metabolic transformation of most substances ingested into the body. It also has physiological functions such as detoxification, immunity, hematopoiesis and coagulation, and plays a very important role in maintaining the normal life activities of the organism. Factors such as viruses, alcohol, drugs and autoimmune abnormalities can cause liver damage such as liver fibrosis, cirrhosis and even cancer by producing liver oxidative stress and inducing inflammatory reactions. Clinical data show that the incidence of liver disease in China is gradually increasing, and the age of patients is showing a trend of younger development. Therefore, finding safe and efficient liver protection drugs from natural medicines has a very important application prospect.

[0003] Compound prescription is a major feature of Chinese medicine. It is a group of medicines that are properly matched according to the composition principle after the treatment method is determined by differentiation of symptoms and causes, and the appropriate dosage is selected. It is rich in chemical components, has the characteristics of multi-level and multi-target, and many chemical components have the ability to synergize and enhance synergy, which can reduce the dosage of medicines, save drug resources and treatment costs. For example, the authorization publication number is CN106074662B, which discloses a turmeric composition with alcohol sobering and liver protection effects. Safety and efficacy have been fully verified. However, it is still necessary to reasonably combine plants to find safer and more efficient liver protection combination drugs, which is of great significance. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a liver protection composition, a preparation method and its application. The purpose is to provide a composition of hawthorn, kudzu root, Hovenia dulcis, Ganoderma lucidum spores and water lily for preparing a liver protection drug.

[0005] In order to solve the above technical problems, the first purpose of the present invention is to provide a liver protection composition, which is prepared from the following medicinal raw materials in parts by weight: 15-25 parts of hawthorn, 15-25 parts of kudzu root, 15-25 parts of Hovenia dulcis, 2-4 parts of Ganoderma lucidum spores and 1-2 parts of Water Lily.

[0006] The beneficial effects of the present invention are:

[0007] (1) The liver protection composition of the present invention is composed of functional components of five plants, namely hawthorn, kudzu root, Hovenia dulcis, Ganoderma lucidum spores and Herba Polygoni Multiflori. Among them, hawthorn is sour and sweet, slightly warm, enters the spleen, stomach and liver meridians, and has the effects of strengthening the spleen and appetizing, digesting food and removing stagnation, promoting blood circulation and removing phlegm; kudzu root is cool in nature, flat in nature, and sweet in taste, and has the effects of clearing away heat, reducing fire, removing toxins, promoting body fluid and quenching thirst, and raising yang and stopping diarrhea; Hovenia dulcis is sweet in taste, flat in nature, and non-toxic, and has the effects of quenching thirst and relieving restlessness, removing heat from the diaphragm, and moistening the five internal organs; Ganoderma lucidum spores have the effects of protecting the liver and detoxifying, relieving cough and removing phlegm; Herba Polygoni Multiflori is sweet, slightly bitter, flat in nature, enters the liver, kidney and spleen meridians, and has the effects of nourishing, strengthening, removing dampness, and dispersing blood stasis.

[0008] (2) The present invention determines that the composition has the effects of protecting the liver, strengthening the spleen and nourishing the stomach based on the theory of traditional Chinese medicine. The pharmacodynamic studies in animals have confirmed that the hawthorn extract, kudzu root extract, Hovenia dulcis extract, Ganoderma lucidum spore extract, Herba Polygoni Multiflori extract and the combination of the five extracts of the present invention can reduce the ALT, AST activity and MDA content in the serum of mice with alcoholic liver injury, increase the SOD activity in the serum, inhibit the expression of IL-1β and TNF-α in the liver, relieve liver enlargement, and show significant liver protection activity.

[0009] Based on the above technical solution, the present invention can also be improved as follows.

[0010] Furthermore, it is prepared from the following medicinal raw materials in parts by weight: 18-22 parts of hawthorn, 18-22 parts of kudzu root, 18-22 parts of Hovenia dulcis, 2.5-3.5 parts of Ganoderma lucidum spores and 1-1.8 parts of Echinops lutea.

[0011] Further, it is prepared from the following medicinal raw materials in parts by weight: 20 parts of hawthorn, 20 parts of kudzu root, 20 parts of Hovenia dulcis, 3 parts of Ganoderma lucidum spores and 1 part of Polygonum aviculare.

[0012] Further, it is prepared from the following medicinal raw materials in parts by weight: 15 parts of hawthorn, 15 parts of kudzu root, 15 parts of hovenia dulcis, 2 parts of ganoderma lucidum spores and 1 part of water lotus.

[0013] Further, it is prepared from the following medicinal raw materials in parts by weight: 25 parts of hawthorn, 25 parts of kudzu root, 25 parts of hovenia dulcis, 4 parts of ganoderma lucidum spores and 1 part of water lotus.

[0014] The second object is to provide a method for preparing a liver-protecting composition, comprising the following steps:

[0015] Step 1, Extraction:

[0016] (1) Extraction of mixed coarse powder: Take corresponding mass fractions of hawthorn, kudzu root, and Hovenia dulcis, crush and mix them, and pass them through a 5-20 mesh sieve to obtain a mixed coarse powder. After adding 5-20 times the weight of water to the mixed coarse powder, heat and reflux at 95° C. for extraction for 0.5-1.5 hours, repeat the water extraction at least twice (the filter residue obtained after filtration is extracted with water again), filter after each extraction, collect and combine the filtrate I of the mixed coarse powder obtained by each filtration; add a volume fraction of 95% ethanol solution equivalent to 3 / 4 to 5 / 4 of the volume of the filtrate I of the mixed coarse powder, filter, and collect the filtrate II of the mixed coarse powder, which is the extract of the mixed coarse powder;

[0017] (2) Extraction of Ganoderma lucidum spores: crush the corresponding mass fraction of Ganoderma lucidum spores, pass through a 5-20 mesh sieve to obtain Ganoderma lucidum spore powder, add 65% to 85% ethanol in an amount of 5 to 20 times the weight by volume to the Ganoderma lucidum spore powder, perform ultrasonic extraction at 50° C. for 0.5 to 1.5 hours, repeat the water extraction at least twice, filter after each extraction, collect and combine the filtrate of the Ganoderma lucidum spore powder obtained by each filtration, and concentrate under reduced pressure to obtain the Ganoderma lucidum spore extract;

[0018] (3) Extraction of the yellow water lily: crush the roots of the yellow water lily in the above-mentioned mass fraction, pass through a 5-20 mesh sieve to obtain a yellow water lily crude powder, add 10-20 times the weight of water to the yellow water lily crude powder, heat and reflux at 95° C. for extraction for 0.5-1.5 hours, repeat the water extraction at least twice, filter after each extraction, collect and combine the filtrate of the yellow water lily crude powder obtained by each filtration, adsorb the filtrate of the yellow water lily crude powder with a macroporous resin, and then elute with 10-20% by volume ethanol and 65-75% by volume ethanol in turn, collect the eluate with 65-75% by volume ethanol, and concentrate under reduced pressure to obtain the yellow water lily extract;

[0019] Step 2, freeze drying:

[0020] The obtained extract of the mixed coarse powder, the extract of the Ganoderma lucidum spores and the extract of the Water Lily of the Valley are combined and freeze-dried to obtain a liver-protecting composition.

[0021] The beneficial effect of adopting the above scheme is that the method for preparing the liver-protecting composition provided by the present invention prepares a liver-protecting composition having a very significant liver-protecting effect.

[0022] Furthermore, when extracting Ganoderma lucidum spores in step 1, the power of the ultrasound is 200W to 300W; when extracting Water Lily in step 1, the macroporous resin is any one of DM-130, HP-10, HP-20, HPD-200, HPD-300, ADS-17 or a combination of at least two of them, the flow rate of the filtrate of the crude powder of Water Lily adsorbed by the macroporous resin is 0.05 column volumes / min, and the volume mass ratio of the filtrate of the crude powder of Water Lily to the macroporous resin is (10 to 20) mL / g; when extracting Ganoderma lucidum spores and Water Lily in step 1, the reduced pressure concentration is specifically to recover ethanol under reduced pressure at 50° C. until all the ethanol is recovered.

[0023] Among them, suitable types of macroporous resins such as DM-130, HP-10, HP-20, HPD-200, HPD-300, and ADS-17 can effectively enrich active ingredients such as saponins in the crude powder of the roots of the water lily.

[0024] Furthermore, in step 2, the specific conditions for freeze-drying are: at -55°C until the solvent in the combined extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the Water Lily of the Valley is completely removed.

[0025] The third object of the present invention is to provide an application of a liver protection composition, wherein any of the above liver protection compositions is used to prepare a liver protection drug.

[0026] The beneficial effect of adopting the above scheme is: the present invention has carried out a study on the protective effect of the composition on alcoholic liver damage in mice. The study shows that the composition can reduce the activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and malondialdehyde (MDA) content in the serum of mice with alcoholic liver damage, increase the activity of superoxide dismutase (SOD) in the serum, inhibit the expression of interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) in the liver, and relieve liver enlargement, confirming that the composition prepared by the present invention has the function of protecting the liver.

[0027] Furthermore, the dosage form of the liver-protecting drug is any one of tablets, pastes, powders, decoctions, granules, capsules or pills. DETAILED DESCRIPTION

[0028] The principles and features of the present invention are described below. The examples given are only used to explain the present invention and are not used to limit the scope of the present invention.

[0029] Example 1

[0030] The present embodiment relates to a liver-protecting composition, which is prepared from the following medicinal raw materials in parts by weight: 20 parts of hawthorn, 20 parts of kudzu root, 20 parts of Hovenia dulcis, 3 parts of Ganoderma lucidum spores and 1 part of Polygonum aviculare.

[0031] The present embodiment relates to a method for preparing a liver-protecting composition, comprising the following steps:

[0032] Step 1:

[0033] (1) Take 20 parts by weight of each of hawthorn coarse powder, kudzu root coarse powder and Hovenia dulcis coarse powder, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the filtrate I of the mixed coarse powder obtained by the two filtrations, add an equal volume of 95% ethanol, filter and collect the filtrate II of the mixed coarse powder, which is the extract of the mixed coarse powder.

[0034] (2) Take 3 parts by weight of Ganoderma lucidum spore powder, use 10 to 20 times the weight of 75% (v / v) ethanol, ultrasonically extract at 50°C for 1 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Add 5 to 15 times the weight of 75% (v / v) ethanol to the filter residue, ultrasonically extract at 50°C for 0.5 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Combine the filtrates of the Ganoderma lucidum spore powder obtained from the two filtrations, and recover the ethanol solvent under reduced pressure at 50°C to obtain the Ganoderma lucidum spore extract.

[0035] (3) Take 1 part by weight of crude powder of the root of the yellow flower water lily, add 10 to 20 times the weight of water, heat and reflux at 95°C for extraction for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for extraction for 0.5 hour, filter and collect the filtrate. Combine the two filtrates to obtain the filtrate of the crude powder of the yellow flower water lily, adsorb it on DM-130 macroporous resin, and elute it with 20% ethanol and 65% to 75% ethanol in turn, collect the eluate with 65% to 75% ethanol, and recover the ethanol solvent under reduced pressure at 50°C to obtain the extract of the yellow flower water lily.

[0036] Step 2: Combine the obtained extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus, and freeze-dry them at -55°C until the solvent in the combined extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus is completely removed to obtain a composition with a raw material weight ratio of 20:20:20:3:1.

[0037] Example 2

[0038] The present embodiment relates to a liver-protecting composition, which is prepared from the following medicinal raw materials in parts by weight: 15 parts of hawthorn, 15 parts of kudzu root, 15 parts of Hovenia dulcis, 4 parts of Ganoderma lucidum spores and 1 part of Polygonum aviculare.

[0039] The present embodiment relates to a method for preparing a liver-protecting composition, comprising the following steps:

[0040] Step 1:

[0041] (1) Take 15 parts by weight of each of hawthorn coarse powder, kudzu root coarse powder and Hovenia dulcis coarse powder, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the filtrate I of the mixed coarse powder obtained by the two filtrations, add an equal volume of 95% ethanol, filter and collect the filtrate II of the mixed coarse powder, which is the extract of the mixed coarse powder.

[0042] (2) Take 4 parts by weight of Ganoderma lucidum spore powder, use 10 to 20 times the weight of 65% (v / v) ethanol, ultrasonically extract at 50°C for 1 hour, the ultrasonic extraction power is 300W, filter, and collect the filtrate. Add 5 to 15 times the weight of 65% (v / v) ethanol to the filter residue, ultrasonically extract at 50°C for 0.5 hour, the ultrasonic extraction power is 300W, filter, and collect the filtrate. Combine the filtrates of the Ganoderma lucidum spore powder obtained from the two filtrations, and recover the ethanol solvent under reduced pressure at 50°C to obtain the Ganoderma lucidum spore extract.

[0043] (3) Take 1 part by weight of crude powder of the root of the yellow flower water lily, add 10 to 20 times the weight of water, heat and reflux at 95°C for extraction for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for extraction for 0.5 hour, filter and collect the filtrate. Combine the two filtrates to obtain the filtrate of the crude powder of the yellow flower water lily, adsorb it on HP-20 macroporous resin, and elute it with 15% ethanol and 65% to 75% ethanol in turn, collect the 65% to 75% ethanol eluate, and recover the ethanol solvent under reduced pressure at 50°C to obtain the extract of the yellow flower water lily.

[0044] Step 2: Combine the obtained extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus, and freeze-dry them at -55°C until the solvent in the combined extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus is completely removed to obtain a composition with a raw material weight ratio of 15:15:15:4:1.

[0045] Example 3

[0046] The present embodiment relates to a liver-protecting composition, which is prepared from the following medicinal raw materials in parts by weight: 15 parts of hawthorn, 15 parts of kudzu root, 15 parts of Hovenia dulcis, 2 parts of Ganoderma lucidum spores and 1 part of Polygonum aviculare.

[0047] The present embodiment relates to a method for preparing a liver-protecting composition, comprising the following steps:

[0048] Step 1:

[0049] (1) Take 15 parts by weight of each of hawthorn coarse powder, kudzu root coarse powder and Hovenia dulcis coarse powder, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the filtrate I of the mixed coarse powder obtained by the two filtrations, add an equal volume of 95% ethanol, filter and collect the filtrate II of the mixed coarse powder, which is the extract of the mixed coarse powder.

[0050] (2) Take 2 parts by weight of Ganoderma lucidum spore powder, use 10 to 20 times the weight of 85% (v / v) ethanol, ultrasonically extract at 50°C for 1 hour, the ultrasonic extraction power is 200W, filter, and collect the filtrate. Add 5 to 15 times the weight of 85% (v / v) ethanol to the filter residue, ultrasonically extract at 50°C for 0.5 hour, the ultrasonic extraction power is 200W, filter, and collect the filtrate. Combine the filtrates of the Ganoderma lucidum spore powder obtained from the two filtrations, and recover the ethanol solvent under reduced pressure at 50°C to obtain the Ganoderma lucidum spore extract.

[0051] (3) Take 1 part by weight of crude powder of the root of the yellow flower water lily, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. The filtrate of the crude powder of the yellow flower water lily obtained by combining the two filtrates is adsorbed on HPD-200 macroporous resin, and then eluted with 10% ethanol and 65% to 75% ethanol in turn, collect the 65% to 75% ethanol eluate, and recover the ethanol solvent under reduced pressure at 50°C to obtain the extract of the yellow flower water lily.

[0052] Step 2, combining the obtained extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus, and freeze-drying them at -55°C until the solvent in the combined extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus is completely removed to obtain a composition with a raw material weight ratio of 15:15:15:2:1.

[0053] Example 4

[0054] The present embodiment relates to a liver-protecting composition, which is prepared from the following medicinal raw materials in parts by weight: 25 parts of hawthorn, 25 parts of kudzu root, 25 parts of Hovenia dulcis, 2 parts of Ganoderma lucidum spores and 1 part of Polygonum aviculare.

[0055] The present embodiment relates to a method for preparing a liver-protecting composition, comprising the following steps:

[0056] Step 1:

[0057] (1) Take 25 parts by weight of each of hawthorn coarse powder, kudzu root coarse powder and Hovenia dulcis coarse powder, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the filtrate I of the mixed coarse powder obtained by the two filtrations, add an equal volume of 95% ethanol, filter and collect the filtrate II of the mixed coarse powder, which is the extract of the mixed coarse powder.

[0058] (2) Take 3 parts by weight of Ganoderma lucidum spore powder, use 10 to 20 times the weight of 75% (v / v) ethanol, ultrasonically extract at 50°C for 1 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Add 5 to 15 times the weight of 75% (v / v) ethanol to the filter residue, ultrasonically extract at 50°C for 0.5 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Combine the filtrates of the Ganoderma lucidum spore powder obtained from the two filtrations, and recover the ethanol solvent under reduced pressure at 50°C to obtain the Ganoderma lucidum spore extract.

[0059] (3) Take 1 part by weight of crude powder of the root of the water lily, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the two filtrates to obtain the filtrate of the crude powder of the water lily, adsorb it on DM-130 macroporous resin, elute it with 20% ethanol and 65% to 75% ethanol in turn, collect the 65% to 75% ethanol eluate, and recover the ethanol solvent under reduced pressure at 50°C to obtain the extract of the water lily.

[0060] Step 2: Combine the obtained extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus, and freeze-dry them at -55°C until the solvent in the combined extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus is completely removed to obtain a composition with a raw material weight ratio of 25:25:25:2:1.

[0061] Example 5

[0062] The present embodiment relates to a liver-protecting composition, which is prepared from the following medicinal raw materials in parts by weight: 25 parts of hawthorn, 25 parts of kudzu root, 25 parts of Hovenia dulcis, 4 parts of Ganoderma lucidum spores and 1 part of Polygonum aviculare.

[0063] The present embodiment relates to a method for preparing a liver-protecting composition, comprising the following steps:

[0064] Step 1:

[0065] (1) Take 25 parts by weight of each of hawthorn coarse powder, kudzu root coarse powder and Hovenia dulcis coarse powder, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the filtrate I of the mixed coarse powder obtained by the two filtrations, add an equal volume of 95% ethanol, filter and collect the filtrate II of the mixed coarse powder, which is the extract of the mixed coarse powder.

[0066] (2) Take 3 parts by weight of Ganoderma lucidum spore powder, use 10 to 20 times the weight of 75% (v / v) ethanol, ultrasonically extract at 50°C for 1 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Add 5 to 15 times the weight of 75% (v / v) ethanol to the filter residue, ultrasonically extract at 50°C for 0.5 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Combine the filtrates of the Ganoderma lucidum spore powder obtained from the two filtrations, and recover the ethanol solvent under reduced pressure at 50°C to obtain the Ganoderma lucidum spore extract.

[0067] (3) Take 1 part by weight of crude powder of the root of the water lily, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the two filtrates to obtain the filtrate of the crude powder of the water lily, adsorb it on DM-130 macroporous resin, elute it with 20% ethanol and 65% to 75% ethanol in turn, collect the 65% to 75% ethanol eluate, and recover the ethanol solvent under reduced pressure at 50°C to obtain the extract of the water lily.

[0068] Step 2: Combine the obtained extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus, and freeze-dry them at -55°C until the solvent in the combined extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus is completely removed to obtain a composition with a raw material weight ratio of 25:25:25:4:1.

[0069] Example 6

[0070] The present embodiment relates to a liver-protecting composition, which is prepared from the following medicinal raw materials in parts by weight: 20 parts of hawthorn, 20 parts of kudzu root, 20 parts of Hovenia dulcis, 3 parts of Ganoderma lucidum spores and 2 parts of Echinops xanthophylla.

[0071] The present embodiment relates to a method for preparing a liver-protecting composition, comprising the following steps:

[0072] Step 1:

[0073] (1) Take 20 parts by weight of each of hawthorn coarse powder, kudzu root coarse powder and Hovenia dulcis coarse powder, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the filtrate I of the mixed coarse powder obtained by the two filtrations, add an equal volume of 95% ethanol, filter and collect the filtrate II of the mixed coarse powder, which is the extract of the mixed coarse powder.

[0074] (2) Take 3 parts by weight of Ganoderma lucidum spore powder, use 10 to 20 times the weight of 75% (v / v) ethanol, ultrasonically extract at 50°C for 1 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Add 5 to 15 times the weight of 75% (v / v) ethanol to the filter residue, ultrasonically extract at 50°C for 0.5 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Combine the filtrates of the Ganoderma lucidum spore powder obtained from the two filtrations, and recover the ethanol solvent under reduced pressure at 50°C to obtain the Ganoderma lucidum spore extract.

[0075] (3) Take 1 part by weight of crude powder of the root of the water lily, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the two filtrates to obtain the filtrate of the crude powder of the water lily, adsorb it on DM-130 macroporous resin, elute it with 20% ethanol and 65% to 75% ethanol in turn, collect the 65% to 75% ethanol eluate, and recover the ethanol solvent under reduced pressure at 50°C to obtain the extract of the water lily.

[0076] Step 2: Combine the obtained extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus, and freeze-dry them at -55°C until the solvent in the combined extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the water lotus is completely removed to obtain a composition with a raw material weight ratio of 20:20:20:3:2.

[0077] Comparative Example 1

[0078] In this comparative example, the crude hawthorn powder was mixed with 10 to 20 times the weight of water, heated to 95°C for 1 hour, filtered, and the filtrate was collected. The filter residue was added with 5 to 15 times the weight of water, heated to 95°C for 0.5 hour, filtered, and the filtrate was collected. The two filtered solutions were combined, an equal volume of 95% ethanol was added, filtered, the filtrate was collected, and freeze-dried to obtain the hawthorn extract.

[0079] Comparative Example 2

[0080] In this comparative example, kudzu root coarse powder was added with 10 to 20 times the weight of water, heated to 95°C for 1 hour, filtered, and the filtrate was collected. The filter residue was added with 5 to 15 times the weight of water, heated to 95°C for 0.5 hour, filtered, and the filtrate was collected. The two filtered solutions were combined, an equal volume of 95% ethanol was added, filtered, the filtrate was collected, and freeze-dried to obtain kudzu root extract.

[0081] Comparative Example 3

[0082] In this comparative example, the crude powder of Hovenia dulcis was taken, mixed with 10 to 20 times the weight of water, heated under reflux at 95°C for 1 hour, filtered, and the filtrate was collected. The residue was added with 5 to 15 times the weight of water, heated under reflux at 95°C for 0.5 hour, filtered, and the filtrate was collected. The two filtered solutions were combined, an equal volume of 95% ethanol was added, filtered, the filtrate was collected, and freeze-dried to obtain the Hovenia dulcis extract.

[0083] Comparative Example 4

[0084] In this comparative example, spore powder of Ganoderma lucidum was extracted with 10 to 20 times the weight of 75% (v / v) ethanol at 50°C for 1 hour with an ultrasonic extraction power of 250W, filtered, and the filtrate was collected. 5 to 15 times the weight of 75% (v / v) ethanol was added to the filter residue, ultrasonically extracted for 0.5 hours at 50°C with an ultrasonic extraction power of 250W, filtered, and the filtrate was collected. The two filtered solutions were combined, the ethanol solvent was recovered under reduced pressure at 50°C, and then freeze-dried to obtain a Ganoderma lucidum spore extract.

[0085] Comparative Example 5

[0086] In this comparative example, the crude powder of the root of the yellow flower water lotus was added with 10 to 20 times the weight of water, heated to reflux at 95°C for 1 hour, filtered, and the filtrate was collected. The filter residue was added with 5 to 15 times the weight of water, heated to reflux at 95°C for 0.5 hour, filtered, and the filtrate was collected. The two filtrates were combined, adsorbed by DM-130 macroporous resin, and eluted with 20% ethanol and 65% to 75% ethanol in turn, and the 65% to 75% ethanol eluate was collected. After the ethanol solvent was recovered under reduced pressure at 50°C, it was freeze-dried to obtain the yellow flower water lotus extract.

[0087] Comparative Example 6

[0088] This comparative example relates to a liver-protecting composition, which is prepared from the following medicinal raw materials in parts by weight: 20 parts of hawthorn, 20 parts of kudzu root, 20 parts of Hovenia dulcis, and 3 parts of Ganoderma lucidum spores.

[0089] The present embodiment relates to a method for preparing a liver-protecting composition, comprising the following steps:

[0090] Step 1:

[0091] (1) Take 20 parts by weight of each of hawthorn coarse powder, kudzu root coarse powder and Hovenia dulcis coarse powder, add 10 to 20 times the weight of water, heat and reflux at 95°C for 1 hour, filter and collect the filtrate. Add 5 to 15 times the weight of water to the filter residue, heat and reflux at 95°C for 0.5 hour, filter and collect the filtrate. Combine the filtrate I of the mixed coarse powder obtained by the two filtrations, add an equal volume of 95% ethanol, filter and collect the filtrate II of the mixed coarse powder, which is the extract of the mixed coarse powder.

[0092] (2) Take 3 parts by weight of Ganoderma lucidum spore powder, use 10 to 20 times the weight of 75% (v / v) ethanol, ultrasonically extract at 50°C for 1 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Add 5 to 15 times the weight of 75% (v / v) ethanol to the filter residue, ultrasonically extract at 50°C for 0.5 hour, the ultrasonic extraction power is 250W, filter, and collect the filtrate. Combine the filtrates of the Ganoderma lucidum spore powder obtained from the two filtrations, and recover the ethanol solvent under reduced pressure at 50°C to obtain the Ganoderma lucidum spore extract.

[0093] Step 2: combining the obtained extract of the mixed coarse powder and the extract of the Ganoderma lucidum spores, freeze-drying, and obtaining a composition with a raw material weight ratio of 20:20:20:3.

[0094] Animal experiments

[0095] 1. Experimental Materials

[0096] Experimental animals: 90 SPF male Kunming mice, 6 to 8 weeks old, were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd. and adaptively raised for 7 days.

[0097] Animal model: Mice were randomly divided into 15 groups, 6 mice in each group, namely, blank control group, model group, bifendate group (Zhejiang Wanbangde Pharmaceutical Group Co., Ltd., 100 mg / kg / d), Example 1-6 groups (100 mg / kg / d) and Comparative Example 1-6 groups (100 mg / kg / d). Continuous gavage for 7 days, once a day, the blank control group and model group mice were gavaged with an equal amount of normal saline. From the 6th day of administration to the end of administration, except for the blank control group mice, 50% edible alcohol was immediately gavaged to the mice, 3 times, 12 hours apart, and the single alcohol dose was 5g / kg, to establish a mouse alcoholic liver injury model.

[0098] 2. Experimental methods

[0099] 2.1 Effect of the composition on serum SOD and MDA in mice with alcoholic liver injury

[0100] 4 hours after the last administration of the blank control group, model group, bifendate group, Example 1 to 6 groups and Comparative Example 1 to 6 groups of mice, a total of 15 groups, blood was collected from the fundus vein to prepare serum, and the SOD activity and MDA content in the serum of each group of mice were determined according to the instructions of the SOD detection kit (Cat. No.: A001-3) and the MDA detection kit (Cat. No.: A003-1) of Nanjing Jiancheng Bioengineering Institute.

[0101] 2.2 Effect of the composition on the expression of IL-1β and TNF-α in the liver of mice with alcoholic liver injury

[0102] 4 hours after the last administration of the blank control group, model group, bifendate group, Example 1 to 6 group and Comparative Example 1 to 6 group, a total of 15 groups of mice, the mice were killed by carbon dioxide asphyxiation, the liver was taken, weighed, and physiological saline was added at a ratio of 1:9 (g / mL), and fully homogenized on ice. Centrifuge for 10 minutes (13000g, 4°C), and the supernatant was taken. The interleukin IL-1β detection kit (Cat. No.: MM-0040M1) and TNF-α detection kit (Cat. No.: MM-0132M1) produced by Wuhan Enzyme Immunity Biotechnology Co., Ltd. were used to determine the IL-1β and TNF-α concentrations in the supernatant.

[0103] 2.3 Effects of the composition on liver index of mice with alcoholic liver injury

[0104] Four hours after the last administration of the drug, the mice in the blank control group, the model group, the bifendate group, the Example 1-6 groups and the Comparative Example 1-6 groups, a total of 15 groups, were killed by carbon dioxide asphyxiation and weighed. The livers were taken out and weighed, and the liver index was calculated according to the formula: liver index = liver weight / body weight × 100%.

[0105] 2.4 Effect of the composition on the activities of ALT and AST in the serum of mice with alcoholic liver injury

[0106] 4 hours after the last administration of the blank control group, model group, bifendate group, Example 1 to 6 groups and Comparative Example 1 to 6 groups of mice, a total of 15 groups, blood was collected from the fundus vein to prepare serum, and the ALT and AST activities in the serum of each group of mice were determined according to the instructions of the ALT detection kit (Cat. No.: C001-a) and the AST detection kit (Cat. No.: C002-a) of Changchun Huili Biotechnology Co., Ltd.

[0107] 3. Experimental results

[0108] 3.1 Effect of the composition on serum SOD and MDA in mice with alcoholic liver injury

[0109] Alcohol will produce a large amount of reactive oxygen species in the metabolic process, causing lipid peroxidation, thereby destroying the body's redox balance and mitochondrial function, inducing oxidative stress and lipid metabolism disorders, and finally leading to liver damage. As shown in Table 1, the SOD activity of the model group mice was significantly reduced, and the MDA content was significantly increased (p < 0.01), indicating that the reactive oxygen species produced by alcohol metabolism consumed a large amount of SOD, causing lipid peroxidation and producing a large amount of MDA. After being treated with 100mg / kg / d of Crataegus pinnatifida, Radix Puerariae, Fructus Hoveniae, Ganoderma Lucidum, and Herba Adenophorae extracts and compositions, the SOD activity in serum increased, and the MDA content decreased (p < 0.05), indicating that the extract and the composition can alleviate the oxidative emergency stress in the liver of alcoholic liver injury mice, and achieve the effect of protecting the liver. Wherein, the composition (embodiment 1) of the preferred embodiment has the most significant effect.

[0110] Table 1 Effect of the composition on SOD and MDA in serum of mice with alcoholic liver injury ( n=6)

[0111]

[0112]

[0113] Note: Compared with the blank control group, ## p<0.01; compared with the model group, * p<0.05, ** p<0.01

[0114] 3.2 Effect of the composition on the expression of IL-1β and TNF-α in the liver of mice with alcoholic liver injury

[0115] Studies have shown that alcohol-induced liver damage and endotoxins produced by alcohol metabolism can cause hepatitis, further aggravating liver damage. As shown in Table 2, the expression of two pro-inflammatory factors, IL-1β and TNF-α, in the liver of mice in the alcoholic liver damage model group increased significantly (p<0.01), indicating that the model mice had hepatitis. After treatment with 100mg / kg / d of hawthorn, kudzu root, Hovenia dulcis, Ganoderma lucidum spores, and yellow flower water lotus extract extracts and compositions, the expression of IL-1β and TNF-α in the mouse liver was significantly reduced (p<0.05), indicating that the extracts and compositions can inhibit the inflammatory response in the liver of mice with alcoholic liver damage and achieve the effect of protecting the liver. Among them, the composition of the preferred embodiment (Example 1) has the most significant effect.

[0116] Table 2 Composition inhibits the expression of IL-1β and TNF-α in the liver of mice with alcoholic liver injury ( n=6)

[0117]

[0118]

[0119] Note: Compared with the normal control group, ## p<0.01; compared with the model group, * p<0.05, ** p<0.01

[0120] 3.3 Effects of the composition on liver indexes of mice with alcoholic hepatitis

[0121] Liver damage is usually accompanied by symptoms such as liver congestion, edema, hypertrophy or atrophy, causing the liver index to deviate from the normal value. Therefore, the liver index can reflect the degree of liver damage. As shown in Table 3, the liver index of mice in the alcoholic liver damage model group increased significantly (p < 0.01), indicating that the liver of the model mice was damaged. After being treated with 100mg / kg / d of hawthorn, kudzu root, Hovenia dulcis, Ganoderma lucidum spores, and water lotus extracts and compositions, the liver index was significantly reduced (p < 0.05), indicating that the extracts and compositions can alleviate alcohol-induced liver damage in mice. Among them, the composition of the preferred embodiment (Example 1) has the most significant effect.

[0122] Table 3 Effects of the composition on liver index of mice with alcoholic hepatitis ( n=6)

[0123] Group Liver index (%) Blank control group 4.08±0.14 Model Group <![CDATA[4.88±0.09 ## ]]> Bifendate group <![CDATA[4.19±0.05 ** ]]> Example 1 Group <![CDATA[4.25±0.09 ** ]]> Example 2 Group <![CDATA[4.51±0.11 ** ]]> Example 3 Group <![CDATA[4.33±0.12 ** ]]> Example 4 Group <![CDATA[4.55±0.10 ** ]]> Example 5 Group <![CDATA[4.29±0.11 ** ]]> Example 6 Group <![CDATA[4.40±0.08 ** ]]> Comparative Example 1 <![CDATA[4.67±0.07 * ]]> Comparative Example 2 <![CDATA[4.64±0.10 * ]]> Comparative Example 3 <![CDATA[4.60±0.13 ** ]]> Comparative Example 4 <![CDATA[4.69±0.08 * ]]> Comparative Example 5 <![CDATA[4.72±0.06 * ]]> Comparative Example 6 <![CDATA[4.59±0.12 ** ]]>

[0124] Note: Compared with the normal control group, ## p<0.01; compared with the model group, * p<0.05, ** p<0.01

[0125] 3.4 Effect of the composition on ALT and AST activities in the serum of mice with alcoholic hepatitis

[0126] ALT and AST in the blood are important indicators of liver damage. As shown in Table 4, the ALT and AST activities in the serum of mice in the alcoholic liver damage model group increased significantly (p<0.01), indicating that the liver of the model mice was damaged. After treatment with 100% of the extracts of Crataegus pinnatifida, Puerariae lobata, Hovenia dulcis, Ganoderma lucidum spores, and Water Lily of the Valley of the Dead, and the composition, the ALT and AST activities in the serum of mice decreased significantly (p<0.01), indicating that the extracts and the composition can alleviate alcohol-induced liver damage in mice. Among them, the composition of the preferred embodiment (Example 1) has the most significant effect.

[0127] Table 4 Compositions reduce the activity of ALT and AST in the serum of mice with alcoholic hepatitis ( n=6)

[0128] Group ALT(U / L) AST(U / L) Blank control group 34.2±7.4 38.8±5.3 Model Group <![CDATA[186.0±9.9 ## ]]> <![CDATA[197.0±14.3 ## ]]> Bifendate group <![CDATA[79.8±5.2 ** ]]> <![CDATA[87.4±12.9 ** ]]> Example 1 Group <![CDATA[87.9±6.4 ** ]]> <![CDATA[98.3±8.8 ** ]]> Example 2 Group <![CDATA[129.1±5.8 ** ]]> <![CDATA[136.5±10.6 ** <!-- 10 -->]]> Example 3 Group <![CDATA[97.3±7.1 ** ]]> <![CDATA[105.4±9.7 ** ]]> Example 4 Group <![CDATA[132.5±8.5 ** ]]> <![CDATA[140.9±10.4 ** ]]> Example 5 Group <![CDATA[91.4±6.9 ** ]]> <![CDATA[107.9±9.8 ** ]]> Example 6 Group <![CDATA[95.5±7.2 ** ]]> <![CDATA[104.5±11.2 ** ]]> Comparative Example 1 <![CDATA[145.9±8.3 ** ]]> <![CDATA[152.7±12.5 ** ]]> Comparative Example 2 <![CDATA[142.8±7.5 ** ]]> <![CDATA[149.6±9.4 ** ]]> Comparative Example 3 <![CDATA[140.6±8.1 ** ]]> <![CDATA[146.8±8.9 ** ]]> Comparative Example 4 <![CDATA[157.7±7.6 ** ]]> <![CDATA[168.4±9.2 ** ]]> Comparative Example 5 <![CDATA[141.4±8.6 ** ]]> <![CDATA[148.3±7.9 ** ]]> Comparative Example 6 <![CDATA[134.7±7.8 ** ]]> <![CDATA[142.9±12.1 ** ]]>

[0129] Note: Compared with the normal control group, ## p<0.01; compared with the model group, * p<0.05, ** p<0.01

[0130] In summary, the present invention has carried out a study on the protective effect of the composition on alcoholic liver damage in mice. The study shows that the composition of the present invention can reduce the activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and malondialdehyde (MDA) content in the serum of mice with alcoholic liver damage, increase the activity of superoxide dismutase (SOD) in the serum, inhibit the expression of interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) in the liver, and relieve liver enlargement, confirming that the composition prepared by the present invention has the function of protecting the liver.

[0131] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0132] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A liver-protecting composition, characterized in that: The product is prepared from the following raw materials by weight: 15-25 parts of hawthorn, 15-25 parts of kudzu root, 15-25 parts of Hovenia dulcis, 2-4 parts of ganoderma lucidum spores and 1-2 parts of water lotus; The yellow flower water lily is the root of the yellow flower water lily.

2. A liver-protecting composition according to claim 1, characterized in that: The medicine is prepared from the following medicinal raw materials in parts by weight: 18-22 parts of hawthorn, 18-22 parts of kudzu root, 18-22 parts of Hovenia dulcis, 2.5-3.5 parts of ganoderma lucidum spores and 1-1.8 parts of water lotus.

3. A liver-protecting composition according to claim 1, characterized in that: The medicine is prepared from the following medicinal raw materials in parts by weight: 20 parts of hawthorn, 20 parts of kudzu root, 20 parts of Hovenia dulcis, 3 parts of Ganoderma lucidum spores and 1 part of Polygonum aviculare.

4. A liver-protecting composition according to claim 1, characterized in that: The medicine is prepared from the following medicinal raw materials in parts by weight: 15 parts of hawthorn, 15 parts of kudzu root, 15 parts of Hovenia dulcis, 2 parts of Ganoderma lucidum spores and 1 part of Polygonum aviculare.

5. A liver-protecting composition according to claim 1, characterized in that: The medicine is prepared from the following medicinal raw materials in parts by weight: 25 parts of hawthorn, 25 parts of kudzu root, 25 parts of Hovenia dulcis, 4 parts of Ganoderma lucidum spores and 1 part of Polygonum aviculare.

6. A method for preparing a liver-protecting composition according to any one of claims 1 to 5, characterized in that: The steps include: Step 1, Extraction: (1) Extraction of mixed coarse powder: Take corresponding weight portions of hawthorn, kudzu root and Hovenia dulcis, crush and mix them, and pass them through a 5-20 mesh sieve to obtain a mixed coarse powder. After adding 5-20 times the weight of water to the mixed coarse powder, heat and reflux at 95° C. to extract for 0.5-1.5 hours, repeat the water extraction at least twice, filter after each extraction, and collect and combine the filtrate I of the mixed coarse powder obtained by each filtration; Adding a 95% ethanol solution in an amount equivalent to 3 / 4 to 5 / 4 of the volume of the filtrate I of the mixed coarse powder, filtering, and collecting the filtrate II of the mixed coarse powder, wherein the filtrate II of the mixed coarse powder is the extract of the mixed coarse powder; (2) Extraction of Ganoderma lucidum spores: crush the corresponding mass fraction of Ganoderma lucidum spores, pass through a 5-20 mesh sieve to obtain Ganoderma lucidum spore powder, add 65% to 85% ethanol in an amount of 5 to 20 times the weight by volume to the Ganoderma lucidum spore powder, perform ultrasonic extraction at 50° C. for 0.5 to 1.5 hours, repeat the water extraction at least twice, filter after each extraction, collect and combine the filtrate of the Ganoderma lucidum spore powder obtained by each filtration, and concentrate under reduced pressure to obtain the Ganoderma lucidum spore extract; (3) Extraction of the yellow water lily: crush the roots of the yellow water lily in the above-mentioned mass fraction, pass through a 5-20 mesh sieve to obtain a yellow water lily crude powder, add 10-20 times the weight of water to the yellow water lily crude powder, heat and reflux at 95° C. for extraction for 0.5-1.5 hours, repeat the water extraction at least twice, filter after each extraction, collect and combine the filtrate of the yellow water lily crude powder obtained by each filtration, adsorb the filtrate of the yellow water lily crude powder with a macroporous resin, and then elute with 10-20% by volume ethanol and 65-75% by volume ethanol in turn, collect the eluate with 65-75% by volume ethanol, and concentrate under reduced pressure to obtain the yellow water lily extract; Step 2, freeze drying: The obtained extract of the mixed coarse powder, the extract of the Ganoderma lucidum spores and the extract of the Water Lily of the Valley are combined and freeze-dried to obtain a liver-protecting composition.

7. The method for preparing a liver-protecting composition according to claim 6, characterized in that: When extracting Ganoderma lucidum spores in step 1, the power of the ultrasonic wave is 200W-300W; when extracting Water Lily in step 1, the macroporous resin is any one of DM-130, HP-10, HP-20, HPD-200, HPD-300, ADS-17 or a combination of at least two of them, the flow rate of the filtrate of the crude powder of Water Lily adsorbed by the macroporous resin is 0.05 column volumes / min, and the volume mass ratio of the filtrate of the crude powder of Water Lily to the macroporous resin is (10-20) mL / g; when extracting Ganoderma lucidum spores and Water Lily in step 1, the reduced pressure concentration is specifically to recover ethanol under reduced pressure at 50° C. until all the ethanol is recovered.

8. The method for preparing a liver-protecting composition according to claim 6, characterized in that: In step 2, the specific conditions for freeze drying are: at -55°C until the solvent in the combined extract of the mixed crude powder, the extract of the Ganoderma lucidum spores and the extract of the Water Lily of the Valley is completely removed.

9. Use of a liver-protecting composition, characterized in that: The liver protection composition according to any one of claims 1 to 5 is used for preparing a liver protection drug.

10. The use of a liver protection composition according to claim 9, characterized in that: The dosage form of the liver-protecting drug is any one of tablets, pastes, powders, decoctions, granules, capsules or pills.

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