Composition for preventing and treating liver cancer and preparation method thereof

Abstract

The present invention proposes a composition for preventing and treating liver cancer and a preparation method thereof, and belongs to the field of medical technology. Salvia miltiorrhiza, Psoralea corylifolia, and Cordyceps militaris are extracted with water, filtered, the filter residue is retained, ethanol precipitation is used to obtain active polysaccharides, and the filtrate is dried to obtain a water extract; selenium-enriched yeast exopolysaccharides are obtained and mixed with active polysaccharides to obtain an organic selenium-active polysaccharide complex; the filter residue is added to water, inoculated with saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis, fermented, cysteine ​​is added, fermented again, freeze-dried, and a fermentation product is obtained, and a water extract, an organic selenium-active polysaccharide complex, a fermentation product, curcumin, and oligofructose are mixed and embedded to obtain a composition for preventing and treating liver cancer. The composition for preventing and treating liver cancer obtained by the present invention plays a good role in preventing liver cancer and has broad application prospects.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a composition for preventing and treating liver cancer and a preparation method thereof. Background Art

[0002] Hepatocellular carcinoma has a poor prognosis and a low cure rate. Traditional Chinese medicine treatment can maximize the survival of patients, improve the quality of life, and increase the survival rate. There is currently no specific medicine or specific treatment method. Western medicine surgery can quickly remove early and mid-stage cancer, but it is often prone to recurrence; at the same time, for a patient with liver cancer, surgery will risk death because he also suffers from quite serious liver cirrhosis. In this case, using good anti-cancer preparations is a better treatment method. Although radiotherapy can shrink and degenerate cancer cells, it can only be used for local treatment, and only 20% of patients are suitable; although chemotherapy can be used for systemic treatment, patients often cannot persist due to serious toxic side effects and adverse reactions.

[0003] Traditional Chinese medicine believes that liver cancer is a disease with deficiency of qi and blood as the root cause, dampness, heat, blood stasis and toxicity as the symptoms, and the treatment should be to strengthen the body and eliminate evil, and treat both the symptoms and the root cause. Salvia miltiorrhiza is a commonly used anti-tumor drug, which was first seen in Shennong's Herbal Classic. It has the effects of promoting blood circulation and removing blood stasis, nourishing blood and calming the mind, cooling blood and eliminating carbuncle. The Classic says: "It can cure evil qi in the heart and abdomen, rumbling in the intestines like running water, cold and heat accumulation; it can eliminate lesions and masses, stop irritability and fullness, and replenish qi."

[0004] Nanchang University's 2011 master's thesis "Tanshinone IIA induces apoptosis of liver cancer cell SMMG-7721 and its mechanism study" disclosed that Tanshinone IIA can inhibit the growth of liver cancer cells by inhibiting the cell cycle and inducing apoptosis. Its inhibitory effect is time- and dose-dependent. Tanshinone IIA can promote the activation of apoptosis protein caspase-3, the cleavage of PARP protein, and downregulate apoptosis inhibitor protein XIAP, but has no effect on the expression of Survivin mRNA.

[0005] Patent document CN101596202A, published on December 9, 2009, discloses the use of Tanshinone IIA emulsion in the treatment of liver diseases. Specifically, after intravenous administration of Tanshinone IIA emulsion, the drug can be rapidly targeted and distributed to the liver, and is used to treat liver diseases such as fatty hepatitis, viral hepatitis, liver fibrosis and liver cancer, with good results.

[0006] The paper "Study on the Mechanism of Icariin in Inhibiting HepG2 Cell Migration" published in the journal "Lishizhen Traditional Chinese Medicine" No. 9, 2011, used different concentrations of icariin to act on HepG2 cells for 24 hours, and then detected the cell adhesion rate through adhesion experiment and the migration speed through scratch injury experiment. It was concluded that icariin inhibited the adhesion and movement of HepG2 cells and played an inhibitory role in tumor metastasis.

[0007] Patent document CN106995829A, published on August 1, 2017, discloses that icariin prepared by enzymatic conversion of total flavonoids from epimedium has a significant inhibitory effect on the proliferation of liver cancer, lung cancer, colon cancer and breast cancer.

[0008] In recent years, as Chinese medicine treatment of tumor diseases has gained wide attention, the research on the anti-tumor effect of Chinese medicine compound prescriptions has provided a new opportunity for the treatment of malignant tumors. The effect of effective Chinese medicine prescriptions is better than that of decoction pieces, because effective prescriptions eliminate the interference of non-drug substances. Summary of the invention

[0009] The purpose of the present invention is to provide a composition for preventing and treating liver cancer and a preparation method thereof, which has the functions of protecting the mucosal barrier function, reducing the toxicity of carcinogens, reducing endotoxin levels, restoring the body's immune response to normal, alleviating liver cell damage caused by inflammatory reactions, improving symptoms of viral hepatitis, inhibiting further development of cirrhosis, promoting cancer cell apoptosis, playing a good role in preventing liver cancer, and having broad application prospects.

[0010] The technical solution of the present invention is achieved in this way:

[0011] The invention provides a preparation method of a composition for preventing and treating liver cancer. The method comprises the following steps: extracting salvia miltiorrhiza, trillium and cordyceps militaris with water, filtering, retaining the filter residue, precipitating with ethanol to obtain active polysaccharides, and drying the filtrate to obtain a water extract; obtaining selenium-enriched yeast extracellular polysaccharides and mixing them with the active polysaccharides to obtain an organic selenium-active polysaccharide complex; adding the filter residue into water, inoculating saccharomyces cerevisiae, lactobacillus salivarius and bacillus subtilis, fermenting, adding cysteine, fermenting again, freeze-drying to obtain a fermentation product, and mixing and embedding the water extract, the organic selenium-active polysaccharide complex, the fermentation product, curcumin and oligofructose to obtain a composition for preventing and treating liver cancer.

[0012] As a further improvement of the present invention, the following steps are included:

[0013] S1. Water extraction: Wash, dry and crush Salvia miltiorrhiza, Psoralea corylifolia and Cordyceps militaris respectively to obtain a mixed powder, add it into water, heat and boil it for extraction, repeat the operation 2-3 times, filter, retain the filter residue, combine the filtrate, concentrate, add ethanol to precipitate, collect the solid by centrifugation, and dry it to obtain the active polysaccharide; recover ethanol from the filtrate, dry it, and obtain a water extract;

[0014] S2. Preparation of selenium-enriched yeast exopolysaccharide: inoculating selenium-enriched yeast into a culture medium, fermenting and culturing, centrifuging, adding a mixed solvent to the supernatant, precipitating, centrifuging, adding ethanol to the supernatant, and drying to obtain selenium-enriched yeast exopolysaccharide;

[0015] S3. Preparation of organic selenium-active polysaccharide complex: adding the selenium-enriched yeast extracellular polysaccharide obtained in step S2 to water, adding the active polysaccharide obtained in step S1, stirring and mixing evenly, and drying to obtain an organic selenium-active polysaccharide complex;

[0016] S4. Activation of strains: Saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis were inoculated into Gould's medium, respectively, and activated and cultured to obtain strain seed liquid;

[0017] S5 fermentation: the filter residue in step S1 is added to water, sterilized, inoculated with the saccharomyces cerevisiae, Lactobacillus salivarius, Bacillus subtilis seed solution obtained in step S4, fermented for a first period of time, adding additives, fermented for a second period of time, to obtain a fermentation mixture;

[0018] S6. Promoting glutathione synthesis fermentation: adding cysteine ​​to the fermentation mixture in step S5, continuing the fermentation culture for a third period of time, freeze-drying, and obtaining a fermentation product;

[0019] S7. Encapsulation: Add the water extract obtained in step S1, the organic selenium-active polysaccharide complex obtained in step S3, the fermentation product obtained in step S6, curcumin, oligofructose, sodium alginate and sodium carboxymethyl cellulose into water, mix well to obtain an aqueous phase; add the aqueous phase and lecithin into fish oil, emulsify, add calcium chloride solution dropwise, solidify at room temperature, centrifuge, wash, and freeze-dry to obtain a composition for preventing and treating liver cancer.

[0020] As a further improvement of the present invention, the mass ratio of Salvia miltiorrhiza, Psoralea corylifolia and Cordyceps militaris in step S1 is 3-5:1-3:5-7, the solid-liquid ratio of the mixed powder and water is 1:5-7 g / mL, the time of the heating and boiling extraction is 1-2h, the ethanol is added until the ethanol content of the system is 60-70wt%, and the precipitation time is 3-5h.

[0021] As a further improvement of the present invention, the formula of the culture medium in step S2 is: 20-40 parts by weight of glucose, 3-5 parts by weight of peptone, KH 2 PO 4 2-3 parts by weight, MgSO 4 7H 2O 1-2 weight parts, vitamin B1 0.01-0.02 weight parts, water 1000 weight parts, the inoculation amount of the selenium-enriched yeast is 2-3w / v%, the fermentation culture conditions are 50-55℃, 50-70r / min, the time is 24-36h, the mixed solvent is a mixture of chloroform and n-butanol, the volume ratio is 4:1, the volume ratio of the supernatant and the mixed solvent is 3-5:1, the precipitation time is 1-2h, ethanol is added to the supernatant until the ethanol content of the system is 60-70wt%, and the precipitation time is 3-5h; the mass ratio of the selenium-enriched yeast extracellular polysaccharide and the active polysaccharide in step S3 is 5-7:3-5.

[0022] As a further improvement of the present invention, the activation culture conditions in step S4 are 40-42°C, 50-70r / min, activation culture 18-24h, and the bacterial content of the bacterial seed liquid is 10 8 -10 9 cfu / mL; the mass ratio of the filter residue and water in step S5 is 2-5:12-15, the inoculation amounts of the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solutions are 2-3v / v%, 1-2v / v% and 2-3v / v%, respectively; the fermentation conditions are 37-40°C, 50-70r / min, the first time period is 24-36h, the second time period is 12-18h, the additive is a mixture of calcium chloride and vitamin B1 in a mass ratio of 3-5:1-2, and the additive is added in an amount of 3-5wt% of the total mass of the system.

[0023] As a further improvement of the present invention, in step S6, the mass ratio of the fermentation mixture to cysteine ​​is 100-120:5-7, the fermentation culture conditions are 37-40°C, 50-70r / min, and the third time period is 18-24h.

[0024] As a further improvement of the present invention, the mass ratio of the water extract, the organic selenium-active polysaccharide complex, the fermentation product, curcumin, oligofructose, sodium alginate, sodium carboxymethyl cellulose and water in step S7 is 3-5:7-10:12-15:1-2:0.5-1:20-25:7-10:200-250, the mass ratio of the aqueous phase, lecithin and fish oil is 100-120:2-3:200-250, the concentration of the calcium chloride solution is 3-5wt%, and the time for room temperature curing is 30-40min.

[0025] As a further improvement of the present invention, the present invention specifically comprises the following steps:

[0026] S1. Water extraction: 3-5 parts by weight of Salvia miltiorrhiza, 1-3 parts by weight of Trillium gracile, and 5-7 parts by weight of Cordyceps militaris were washed, dried, and crushed to obtain a mixed powder, which was added to water. The solid-liquid ratio of the mixed powder to water was 1:5-7 g / mL. The mixture was heated to boiling and extracted for 1-2 h. The operation was repeated 2-3 times. The residue was retained. The filtrate was combined and concentrated. Ethanol was added to the ethanol content of the system to 60-70 wt %. The mixture was precipitated for 3-5 h. The solid was collected by centrifugation and dried to obtain an active polysaccharide. The filtrate was recovered with ethanol and dried to obtain a water extract.

[0027] S2. Preparation of selenium-enriched yeast extracellular polysaccharides: inoculate selenium-enriched yeast into the culture medium at an inoculation amount of 2-3 w / v%, 50-55°C, 50-70 r / min, ferment and culture for 24-36 hours, centrifuge, add a mixed solvent to the supernatant, the volume ratio of the supernatant to the mixed solvent is 3-5:1, precipitate for 1-2 hours, centrifuge, add ethanol to the supernatant until the ethanol content of the system is 60-70wt%, precipitate for 3-5 hours, dry, and obtain selenium-enriched yeast extracellular polysaccharides;

[0028] The formula of the culture medium is: 20-40 parts by weight of glucose, 3-5 parts by weight of peptone, KH 2 PO 4 2-3 parts by weight, MgSO 4 7H 2 O 1-2 parts by weight, vitamin B1 0.01-0.02 parts by weight, water 1000 parts by weight;

[0029] The mixed solvent is a mixture of chloroform and n-butanol in a volume ratio of 4:1;

[0030] S3. Preparation of organic selenium-active polysaccharide complex: add 5-7 parts by weight of selenium-enriched yeast extracellular polysaccharide obtained in step S2 to 100 parts by weight of water, add 3-5 parts by weight of active polysaccharide obtained in step S1, stir and mix evenly, and dry to obtain an organic selenium-active polysaccharide complex;

[0031] S4. Activation of strains: Saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis were inoculated into Gould's medium respectively, and activated and cultured for 18-24 hours at 40-42°C and 50-70r / min to obtain strain seed liquid with a bacterial content of 10 8 -10 9 cfu / mL;

[0032] S5. Fermentation: adding 20-50 parts by weight of the filter residue in step S1 to 120-150 parts by weight of water, sterilizing, inoculating the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution prepared in step S4, wherein the inoculation amount of the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution is 2-3v / v%, 1-2v / v% and 2-3v / v%, respectively, at 37-40°C, 50-70r / min, fermenting for 24-36h, adding additives, wherein the additive is added in an amount of 3-5wt% of the total mass of the system, and fermenting for 12-18h to obtain a fermentation mixture;

[0033] The additive is a mixture of calcium chloride and vitamin B1, with a mass ratio of 3-5:1-2;

[0034] S6. Promoting glutathione synthesis fermentation: adding 5-7 parts by weight of cysteine ​​to 100-120 parts by weight of the fermentation mixture in step S5, 37-40°C, 50-70r / min, fermenting for 18-24h, freeze-drying, and obtaining a fermentation product;

[0035] S7. Encapsulation: add 3-5 parts by weight of the water extract obtained in step S1, 7-10 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 12-15 parts by weight of the fermentation product obtained in step S6, 1-2 parts by weight of curcumin, 0.5-1 parts by weight of oligofructose, 20-25 parts by weight of sodium alginate, and 7-10 parts by weight of sodium carboxymethyl cellulose to 200-250 parts by weight of water, mix well, and obtain an aqueous phase; add 100-120 parts by weight of the aqueous phase and 2-3 parts by weight of lecithin to 200-250 parts by weight of fish oil, emulsify, add 50 parts by weight of 3-5wt% calcium chloride solution dropwise, solidify at room temperature for 30-40 minutes, centrifuge, wash, and freeze-dry to obtain a composition for preventing and treating liver cancer.

[0036] The present invention further protects a composition for preventing and treating liver cancer obtained by the above preparation method.

[0037] The present invention further protects the use of the above-mentioned composition for preventing and treating liver cancer in the preparation of products for preventing and treating liver cancer, liver failure and hepatitis.

[0038] The present invention has the following beneficial effects:

[0039] Danshen is the dried root and rhizome of the plant Salvia miltiorrhiza of the Labiatae family. It is slightly cold in nature and bitter in taste. It enters the heart and liver meridians and has the effects of promoting blood circulation and removing scars, relieving pain, clearing the heart and eliminating vexation, cooling blood and eliminating carbuncle. It has pharmacological activities such as antioxidant, antibacterial, anti-inflammatory, improving microcirculation, analgesic, and anti-cardiovascular and cerebrovascular diseases. Trillium has multiple pharmacological activities such as anti-inflammatory, antibacterial, anti-tumor, and anti-angiogenesis, and can be used for anti-cancer. Cordyceps sinensis, the main active ingredient, is a unique active ingredient in Cordyceps sinensis. It has obvious sedative, anti-fatigue, anti-tumor, cancer cell inhibition, and androgen secretion effects, which can enhance the body's immune function.

[0040] The present invention extracts salvia miltiorrhiza, trillium and Cordyceps militaris through water, and the obtained active components include active substances such as salvianolic acid B, cordycepin, alkaloids, saponins, flavonoids, cordyceps polysaccharides, salvia miltiorrhiza polysaccharides and trillium polysaccharides, wherein salvianolic acid B is polymerized by 3 molecules of salvia miltiorrhiza and 1 molecule of caffeic acid, and is a component with the highest content and the strongest activity among the water-soluble effective components of salvia miltiorrhiza, and salvianolic acid B has an in vitro growth inhibition effect on cancer cells. Salvianolic acid B can alleviate the effect of acute liver injury by inhibiting the TLR4 / MyD88 / NF-κB signaling pathway, and the TLR4 / MyD88 / NF-κB signaling pathway is a key pathway widely present in various cells and involved in inflammatory reactions, and is an important regulatory mechanism for mediating tumorigenesis. Salvianolic acid B can affect the composition of intestinal flora, increase the relative abundance of beneficial intestinal bacteria, reduce the relative abundance of harmful bacteria, and improve the intestinal environment. Trillium polysaccharide is the main bioactive component of Trillium, which can improve the body's immunity, inhibit PI3K / AKT / mTOR activation, induce autophagy, and then inhibit angiogenesis and promote tumor cell apoptosis. Cordycepin is a natural nucleoside antibiotic and a natural anticancer antibiotic. Cordycepin is 3-deazaadenosine, a nucleic acid derivative of nitrogen-containing glycosides, a purine alkaloid, and has antiviral and anti-tumor effects, especially a strong inhibitory effect on a variety of solid malignant tumors.

[0041] Selenium can participate in the occurrence and development of liver diseases in humans or animals, and can effectively prevent liver damage induced by various reasons. It is the main component of glutathione peroxidase, which is an important antioxidant enzyme in the liver of humans or animals. At the same time, selenium also removes various free radicals in the body together with superoxide dismutase, catalase, etc., thereby reducing the occurrence of body diseases. Selenium can also increase the level of glutathione in the liver to resist liver cell damage caused by free radicals. Selenium supplementation can significantly reduce the activation of hepatic stellate cells and the expression of genes related to liver fibrosis.

[0042] In the present invention, after fermentation with selenium-enriched yeast, the bacteria are removed by centrifugation, and the protein is removed by the Sevag method to obtain a selenium-enriched yeast extracellular polysaccharide liquid, which is mixed with active polysaccharides obtained by extracting salvia miltiorrhiza, trillium and Cordyceps militaris, and an organic selenium-active polysaccharide complex is formed through chelation. Compared with inorganic selenium, the organic selenium produced by the fermentation of selenium-enriched yeast is more easily absorbed by the human body and is safe and non-toxic, thereby playing a good role in anti-oxidation and anti-inflammatory, increasing the level of glutathione in the liver, reducing the activation of hepatic stellate cells and the expression of genes related to liver fibrosis, thereby playing a good role in protecting the liver, preventing the progression of liver disease, and preventing and treating liver disease, cirrhosis, liver failure and the like.

[0043] Under normal circumstances, the intestinal flora of the human body will form a protective barrier against pathogens. When the intestinal protective barrier is damaged, it may lead to increased intestinal mucosal permeability and cause intestinal bacterial translocation. Intestinal flora imbalance leads to reduced colonization resistance of the flora and weakened barrier protection, causing a large number of pathogens in the intestine to invade, colonize, and reproduce. Intestinal bacterial translocation will accelerate the hyperdynamic circulation state, increase portal vein pressure, and increase intestinal mucosal damage. At the same time, intestinal flora imbalance leads to reduced colonization resistance of the small intestine, causing overgrowth of small intestinal bacteria, and then a large number of pro-inflammatory cytokines are secreted, causing severe inflammatory reactions, destroying the microbial barrier, and ultimately causing bile acid metabolism disorders, leading to liver failure, and then developing into liver cancer.

[0044] Probiotics can effectively inhibit the apoptosis of intestinal mucosal epithelial cells, thereby protecting the mucosal barrier function, reducing the toxicity of carcinogens, reducing endotoxin levels, and improving intestinal flora disorders, thereby maintaining the balance of the microecology, restoring the body's immune response to normal, reducing liver cell damage caused by inflammatory reactions, improving symptoms of viral hepatitis, inhibiting the further development of cirrhosis, and playing a good role in preventing liver cancer.

[0045] The probiotics added by the present invention include saccharomyces cerevisiae, lactobacillus salivarius, and bacillus subtilis, wherein lactobacillus salivarius has the effects of improving liver function, protecting intestinal barrier, and alleviating liver fibrosis, can prevent or treat liver cirrhosis and liver failure, and its metabolites can protect liver function, play a positive role in preventing liver cancer, such as indolepropionic acid, which is a product of tryptophan decomposition by bacteria, can inhibit NF-κB signaling pathway, reduce the level of proinflammatory cytokines, antagonize liver inflammation and liver damage, and can also improve the inherent immunity of liver and intestine, and then enhance the ability of liver to clear hepatitis virus, alleviate liver inflammation, and strengthen intestinal barrier function at the same time, so as to play an anti-cancer effect. Saccharomyces cerevisiae can promote the growth of bifidobacteria (probiotics) in intestinal flora, and inhibit the number of Candida albicans (pathogenic bacteria); it can also protect mucosal barrier function by reducing apoptosis of intestinal mucosal epithelial cells, and prevent further development of liver cirrhosis by inhibiting the release of inflammatory mediators such as TNF-α and IL-6. Bacillus subtilis can degrade aflatoxin and prevent liver cancer by reducing the toxicity of carcinogens. It can also effectively improve intestinal flora imbalance and restore the body's normal immune response to HBV, while reducing the release of inflammatory mediators and liver cell damage, thereby improving the symptoms of viral hepatitis. At the same time, it can stimulate Prevotella to secrete anti-inflammatory substances, effectively inhibiting the level of Th17 cells in the tumor. Th17 cells are a subset of helper T cells that can characteristically secrete IL-17, which can inhibit tumor immunity and stimulate tumor angiogenesis, participate in the occurrence and development of tumors, and exert anti-cancer effects by inhibiting the level of Th17 cells.

[0046] Probiotics can not only produce beneficial substances through their own fermentation, but also inhibit the colonization of harmful bacteria, improve viral hepatitis and cirrhosis, strengthen liver and intestinal inherent immunity, reduce the carcinogenicity of toxic substances and prevent the occurrence of liver cancer. They can also stimulate the secretion of anti-inflammatory substances, inhibit the level of Th17 cells in tumors and enhance anti-tumor immune responses to play an anti-cancer effect. They have the advantages of being safe, harmless and inexpensive, and are expected to become an emerging prevention and treatment method for liver cancer.

[0047] The present invention supplements calcium chloride and vitamin B2 during the probiotic fermentation process, increases the resistance of the probiotics through the regulation of trace elements, and increases the time period of the probiotics in the stable period, thereby promoting the probiotics to produce more beneficial substances.

[0048] Glutathione is an important antioxidant in the body. It can remove excessive free radicals in the body, combine with free radicals to reduce the level of oxidative stress in the body, and participate in the glycolysis pathway and tricarboxylic acid cycle in the body, so that the body can obtain higher energy and activate various enzymes in the body, thereby promoting the synthesis and decomposition of carbohydrates, fats and proteins, and thus improving the nutritional metabolism of cells. Glutathione also participates in a variety of biochemical reactions in human or animal cells; it protects the integrity of liver cell membranes and increases the activity of related enzymes in the liver. In addition, glutathione also has a strong detoxification effect, and has a good detoxification effect on substances that are harmful to the body, such as organic solvents, carbon monoxide, alcohol, heavy metals, carcinogens, etc.

[0049] The brewer's yeast in the fermentation bacteria of the present invention can accumulate more glutathione in the cell, thereby greatly increasing the amount of glutathione in the composition. Cysteine ​​is an important raw material for the synthesis of glutathione. The present invention promotes the production of glutathione produced by the fermentation of brewer's yeast by adding cysteine, thereby playing a good role in protecting the liver and playing a good role in preventing and treating liver cancer.

[0050] In addition, the curcumin added in the present invention can reduce liver damage by increasing the glutathione content in the liver. At the same time, curcumin and oligofructose are also a good prebiotic, which can promote the proliferation of intestinal probiotics in a targeted manner, thereby inhibiting the growth of harmful bacteria, regulating liver function through the liver-intestine axis, promoting the recovery of liver function, inhibiting the formation of fatty liver, and playing a role in protecting the liver.

[0051] The composition for preventing and treating liver cancer prepared by the present invention protects the mucosal barrier function, reduces the toxicity of carcinogens, reduces the level of endotoxin, restores the body's immune response to normal, alleviates liver cell damage caused by inflammatory reactions, improves symptoms of viral hepatitis, inhibits further development of cirrhosis, promotes apoptosis of cancer cells, plays a good role in preventing liver cancer, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0053] Figure 1 This is a comparison chart of the survival rates of each group in Test Example 1 of the present invention;

[0054] Figure 2 This is a comparison chart of the release rates of each group in Test Example 1 of the present invention. DETAILED DESCRIPTION

[0055] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0056] Selenium-enriched yeast, 10 billion cfu / g, was purchased from Angel Yeast Co., Ltd.; Saccharomyces cerevisiae, a highly active dry yeast for brewing, was purchased from Angel Yeast Co., Ltd.; Lactobacillus salivarius, 350 billion cfu / g, was purchased from Guangzhou Huijian Biotechnology Co., Ltd.; Bacillus subtilis, 20 billion cfu / g, was purchased from Beihai Yeshengwang Biotechnology Co., Ltd.

[0057] Example 1

[0058] This embodiment provides a method for preparing a composition for preventing and treating liver cancer, which specifically comprises the following steps:

[0059] S1. Water extraction: 3 parts by weight of Salvia miltiorrhiza, 1 part by weight of Trillium gracile, and 5 parts by weight of Cordyceps militaris were washed, dried, and crushed to obtain a mixed powder, which was added to water. The solid-liquid ratio of the mixed powder to water was 1:5 g / mL. The mixture was heated and boiled for extraction for 1 h. The operation was repeated twice. The residue was retained. The filtrate was combined and concentrated to a relative density of 1.05. Ethanol was added to the system until the ethanol content was 60 wt %. The mixture was precipitated for 3 h. The solid was collected by centrifugation and dried to obtain an active polysaccharide. The filtrate was used to recover ethanol and dried to obtain a water extract.

[0060] S2. Preparation of selenium-enriched yeast extracellular polysaccharides: inoculate selenium-enriched yeast into the culture medium at an inoculation amount of 2w / v%, 50°C, 50r / min, ferment and culture for 24h, centrifuge, add a mixed solvent to the supernatant, the volume ratio of the supernatant to the mixed solvent is 3:1, precipitate for 1h, centrifuge, add ethanol to the supernatant until the ethanol content of the system is 60wt%, precipitate for 3h, dry, and obtain selenium-enriched yeast extracellular polysaccharides;

[0061] The formula of the culture medium is: 20 parts by weight of glucose, 3 parts by weight of peptone, KH 2 PO 4 2 parts by weight, MgSO 4 7H 2 O 1 weight part, vitamin B1 0.01 weight part, water 1000 weight parts;

[0062] The mixed solvent is a mixture of chloroform and n-butanol in a volume ratio of 4:1;

[0063] S3. Preparation of organic selenium-active polysaccharide complex: 5 parts by weight of selenium-enriched yeast extracellular polysaccharide prepared in step S2 was added to 100 parts by weight of water, and 3 parts by weight of active polysaccharide prepared in step S1 was added, stirred and mixed for 20 minutes, and dried to obtain an organic selenium-active polysaccharide complex;

[0064] S4. Activation of strains: Saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis were inoculated into Gao's medium respectively, and activated and cultured for 18 hours at 40°C and 50 r / min to obtain strain seed liquid with a bacterial content of 10 8 cfu / mL;

[0065] S5. Fermentation: 20 parts by weight of the filter residue in step S1 was added to 120 parts by weight of water, sterilized, inoculated with the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution prepared in step S4, wherein the inoculation amount of the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution was 2v / v%, 1v / v% and 2v / v%, respectively, at 37°C, 50r / min, and fermented for 24h, and an additive was added, wherein the additive was added in an amount of 3wt% of the total mass of the system, and fermented for 12h to obtain a fermentation mixture;

[0066] The additive is a mixture of calcium chloride and vitamin B1, with a mass ratio of 3:1;

[0067] S6. Promoting glutathione synthesis fermentation: adding 5 parts by weight of cysteine ​​to 100 parts by weight of the fermentation mixture in step S5, culturing at 37°C, 50 r / min, fermenting for 1 to 8 hours, and freeze-drying to obtain a fermentation product;

[0068] S7. Encapsulation: Add 3 parts by weight of the water extract obtained in step S1, 7 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 12 parts by weight of the fermentation product obtained in step S6, 1 part by weight of curcumin, 0.5 parts by weight of oligofructose, 20 parts by weight of sodium alginate, and 7 parts by weight of sodium carboxymethyl cellulose to 200 parts by weight of water, and mix for 20 minutes to obtain an aqueous phase; add 100 parts by weight of the aqueous phase and 2 parts by weight of lecithin to 200 parts by weight of fish oil, emulsify at 10,000 r / min for 10 minutes, add 50 parts by weight of 3wt% calcium chloride solution dropwise, solidify at room temperature for 30 minutes, centrifuge, wash, and freeze-dry to obtain a composition for preventing and treating liver cancer.

[0069] Example 2

[0070] This embodiment provides a method for preparing a composition for preventing and treating liver cancer, which specifically comprises the following steps:

[0071] S1. Water extraction: 5 parts by weight of Salvia miltiorrhiza, 3 parts by weight of Trillium gracile, and 7 parts by weight of Cordyceps militaris were washed, dried, and crushed to obtain a mixed powder, which was added to water. The solid-liquid ratio of the mixed powder to water was 1:7 g / mL. The mixture was heated and boiled for extraction for 2 h. The operation was repeated 3 times. The residue was retained. The filtrate was combined and concentrated to a relative density of 1.12. Ethanol was added to the system until the ethanol content was 70 wt %. The mixture was precipitated for 5 h. The solid was collected by centrifugation and dried to obtain an active polysaccharide. The filtrate was recovered with ethanol, dried, and a water extract was obtained.

[0072] S2. Preparation of selenium-enriched yeast extracellular polysaccharides: inoculate selenium-enriched yeast into the culture medium at an inoculation amount of 3w / v%, 55°C, 70r / min, ferment and culture for 36h, centrifuge, add a mixed solvent to the supernatant, the volume ratio of the supernatant to the mixed solvent is 5:1, precipitate for 2h, centrifuge, add ethanol to the supernatant until the ethanol content of the system is 70wt%, precipitate for 5h, dry, and obtain selenium-enriched yeast extracellular polysaccharides;

[0073] The formula of the culture medium is: 40 parts by weight of glucose, 5 parts by weight of peptone, KH 2 PO 4 3 parts by weight, MgSO 4 7H 2 O 2 parts by weight, vitamin B1 0.02 parts by weight, water 1000 parts by weight;

[0074] The mixed solvent is a mixture of chloroform and n-butanol in a volume ratio of 4:1;

[0075] S3. Preparation of organic selenium-active polysaccharide complex: 7 parts by weight of selenium-enriched yeast extracellular polysaccharide prepared in step S2 was added to 100 parts by weight of water, and 5 parts by weight of active polysaccharide prepared in step S1 was added, stirred and mixed for 20 minutes, and dried to obtain an organic selenium-active polysaccharide complex;

[0076] S4. Activation of strains: Saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis were inoculated into Gould's medium respectively, and activated and cultured for 24 h at 42°C and 70 r / min to obtain strain seed liquid with a bacterial content of 10 9 cfu / mL;

[0077] S5. Fermentation: 50 parts by weight of the filter residue in step S1 was added to 150 parts by weight of water, sterilized, inoculated with the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution prepared in step S4, wherein the inoculation amount of the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution was 3v / v%, 2v / v% and 3v / v%, respectively, at 40°C, 70r / min, and fermented for 36h, adding additives, wherein the additive was added in an amount of 5wt% of the total mass of the system, and fermented for 18h to obtain a fermentation mixture;

[0078] The additive is a mixture of calcium chloride and vitamin B1, with a mass ratio of 5:2;

[0079] S6. Promoting glutathione synthesis fermentation: adding 7 parts by weight of cysteine ​​to 120 parts by weight of the fermentation mixture in step S5, culturing at 40°C, 70r / min, fermenting for 24h, freeze-drying, and obtaining a fermentation product;

[0080] S7. Encapsulation: Add 5 parts by weight of the water extract obtained in step S1, 10 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 15 parts by weight of the fermentation product obtained in step S6, 2 parts by weight of curcumin, 1 part by weight of oligofructose, 25 parts by weight of sodium alginate, and 10 parts by weight of sodium carboxymethyl cellulose to 250 parts by weight of water, and mix for 20 minutes to obtain an aqueous phase; add 120 parts by weight of the aqueous phase and 3 parts by weight of lecithin to 250 parts by weight of fish oil, emulsify at 10,000 r / min for 10 minutes, add 50 parts by weight of a 5wt% calcium chloride solution dropwise, solidify at room temperature for 40 minutes, centrifuge, wash, and freeze-dry to obtain a composition for preventing and treating liver cancer.

[0081] Example 3

[0082] This embodiment provides a method for preparing a composition for preventing and treating liver cancer, which specifically comprises the following steps:

[0083] S1. Water extraction: 4 parts by weight of Salvia miltiorrhiza, 2 parts by weight of Trillium gracile, and 6 parts by weight of Cordyceps militaris were washed, dried, and crushed to obtain a mixed powder, which was added to water. The solid-liquid ratio of the mixed powder to water was 1:6 g / mL. The mixture was heated and boiled for extraction for 1.5 h. The operation was repeated 3 times. The residue was retained. The filtrate was combined and concentrated to a relative density of 1.07. Ethanol was added to the system to obtain an ethanol content of 65 wt %. The mixture was precipitated for 4 h. The solid was collected by centrifugation and dried to obtain an active polysaccharide. The filtrate was used to recover ethanol and dried to obtain a water extract.

[0084] S2. Preparation of selenium-enriched yeast extracellular polysaccharides: inoculate selenium-enriched yeast into the culture medium at an inoculation amount of 2.5 w / v%, 52°C, 60 r / min, ferment and culture for 30 h, centrifuge, add a mixed solvent to the supernatant, the volume ratio of the supernatant to the mixed solvent is 4:1, precipitate for 1.5 h, centrifuge, add ethanol to the supernatant until the ethanol content of the system is 65 wt%, precipitate for 4 h, dry, and obtain selenium-enriched yeast extracellular polysaccharides;

[0085] The formula of the culture medium is: 30 parts by weight of glucose, 4 parts by weight of peptone, KH 2 PO 4 2.5 parts by weight, MgSO 4 7H 2O 1.5 parts by weight, vitamin B1 0.015 parts by weight, water 1000 parts by weight;

[0086] The mixed solvent is a mixture of chloroform and n-butanol in a volume ratio of 4:1;

[0087] S3. Preparation of organic selenium-active polysaccharide complex: 6 parts by weight of selenium-enriched yeast extracellular polysaccharide prepared in step S2 are added to 100 parts by weight of water, and 4 parts by weight of active polysaccharide prepared in step S1 are added, stirred and mixed for 20 minutes, and dried to obtain an organic selenium-active polysaccharide complex;

[0088] S4. Activation of strains: Saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis were inoculated into Gould's medium respectively, and activated and cultured for 21 hours at 41°C and 60 r / min to obtain strain seed liquid with a bacterial content of 10 9 cfu / mL;

[0089] S5. Fermentation: 35 parts by weight of the filter residue in step S1 was added to 135 parts by weight of water, sterilized, inoculated with the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution prepared in step S4, the inoculation amounts of the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution were 2.5 v / v%, 1.5 v / v%, 2.5 v / v%, 38 ° C, 60 r / min, fermented for 30 h, added additives, the additive was added in an amount of 4wt% of the total mass of the system, fermented for 15 h, and a fermentation mixture was obtained;

[0090] The additive is a mixture of calcium chloride and vitamin B1, with a mass ratio of 4:1.5;

[0091] S6. Promoting glutathione synthesis fermentation: adding 6 parts by weight of cysteine ​​to 110 parts by weight of the fermentation mixture in step S5, culturing at 38°C, 60r / min, fermenting for 21h, freeze-drying, and obtaining a fermentation product;

[0092] S7. Encapsulation: Add 4 parts by weight of the water extract obtained in step S1, 8.5 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 13.5 parts by weight of the fermentation product obtained in step S6, 1.5 parts by weight of curcumin, 0.7 parts by weight of oligofructose, 22 parts by weight of sodium alginate, and 8.5 parts by weight of sodium carboxymethyl cellulose to 225 parts by weight of water, and mix for 20 minutes to obtain an aqueous phase; add 110 parts by weight of the aqueous phase and 2.5 parts by weight of lecithin to 225 parts by weight of fish oil, emulsify at 10,000 r / min for 10 minutes, add 50 parts by weight of 4wt% calcium chloride solution dropwise, solidify at room temperature for 35 minutes, centrifuge, wash, and freeze-dry to obtain a composition for preventing and treating liver cancer.

[0093] Example 4

[0094] Compared with Example 3, the difference is that the additive is single calcium chloride.

[0095] Example 5

[0096] Compared with Example 3, the difference is that the additive is a single vitamin B1.

[0097] Comparative Example 1

[0098] Compared with Example 3, the difference is that Salvia miltiorrhiza is not added in step S1.

[0099] The details are as follows:

[0100] S1. Water extraction: 2 parts by weight of Trillium gracile and 6 parts by weight of Cordyceps militaris were washed, dried and crushed to obtain a mixed powder, which was added into water with a solid-liquid ratio of the mixed powder to water of 1:6 g / mL. The mixture was heated and boiled for extraction for 1.5 h. The operation was repeated 3 times. The residue was retained and the filtrate was combined and concentrated to a relative density of 1.07. Ethanol was added to the system to reach an ethanol content of 65 wt %. The mixture was precipitated for 4 h, and the solid was collected by centrifugation and dried to obtain an active polysaccharide. Ethanol was recovered from the filtrate and dried to obtain a water extract.

[0101] Comparative Example 2

[0102] Compared with Example 3, the difference is that no Tripterygium wilfordii is added in step S1.

[0103] The details are as follows:

[0104] S1. Water extraction: 4 parts by weight of Salvia miltiorrhiza and 6 parts by weight of Cordyceps militaris were washed, dried, and crushed to obtain a mixed powder, which was added to water with a solid-liquid ratio of 1:6 g / mL. The mixture was heated to boiling and extracted for 1.5 h. The operation was repeated 3 times. The residue was retained and the filtrate was combined and concentrated to a relative density of 1.07. Ethanol was added to the system to reach an ethanol content of 65 wt %. The mixture was precipitated for 4 h, and the solid was collected by centrifugation and dried to obtain an active polysaccharide. Ethanol was recovered from the filtrate and dried to obtain a water extract.

[0105] Comparative Example 3

[0106] Compared with Example 3, the difference is that Cordyceps militaris is not added in step S1.

[0107] The details are as follows:

[0108] S1. Water extraction: 4 parts by weight of Salvia miltiorrhiza and 2 parts by weight of Trillium gracile were washed, dried and crushed to obtain a mixed powder, which was added into water with a solid-liquid ratio of the mixed powder to water of 1:6 g / mL. The mixture was heated to boiling and extracted for 1.5 h. The operation was repeated 3 times. The residue was retained and the filtrate was combined and concentrated to a relative density of 1.07. Ethanol was added to the system to reach an ethanol content of 65 wt %. The mixture was precipitated for 4 h, and the solid was collected by centrifugation and dried to obtain an active polysaccharide. Ethanol was recovered from the filtrate and dried to obtain a water extract.

[0109] Comparative Example 4

[0110] Compared with Example 3, the difference is that steps S2 and S3 are not performed, and the organic selenium-active polysaccharide complex in step S7 is replaced by the active polysaccharide prepared in step S1.

[0111] The details are as follows:

[0112] S7. Encapsulation: 4 parts by weight of the water extract prepared in step S1, 8.5 parts by weight of the active polysaccharide prepared in step S1, 13.5 parts by weight of the fermentation product, 1.5 parts by weight of curcumin, 0.7 parts by weight of oligofructose, 22 parts by weight of sodium alginate, and 8.5 parts by weight of sodium carboxymethyl cellulose are added to 225 parts by weight of water, and mixed for 20 minutes to prepare an aqueous phase; 110 parts by weight of the aqueous phase and 2.5 parts by weight of lecithin are added to 225 parts by weight of fish oil, emulsified at 10000 r / min for 10 minutes, 50 parts by weight of 4wt% calcium chloride solution is added dropwise, solidified at room temperature for 35 minutes, centrifuged, washed, and freeze-dried to prepare a composition for preventing and treating liver cancer.

[0113] Comparative Example 5

[0114] Compared with Example 3, the difference is that in step S5, brewer's yeast is not inoculated and step S6 is not performed.

[0115] The details are as follows:

[0116] S5. Fermentation: Add 35 parts by weight of the filter residue in step S1 to 135 parts by weight of water, sterilize, and inoculate with the brewer's yeast, Lactobacillus salivarius, and Bacillus subtilis seed solutions prepared in step S4, wherein the inoculation amounts of the brewer's yeast, Lactobacillus salivarius, and Bacillus subtilis seed solutions are 2.5 v / v%, 1.5 v / v%, and 2.5 v / v%, respectively. Ferment for 30 hours at 38°C and 60 r / min, add additives, wherein the additive amount is 4wt% of the total mass of the system, and ferment for 15 hours to obtain a fermentation mixture.

[0117] Comparative Example 6

[0118] Compared with Example 3, the difference is that Lactobacillus salivarius is not inoculated in step S5.

[0119] The details are as follows:

[0120] S5. Fermentation: Add 35 parts by weight of the filter residue in step S1 to 135 parts by weight of water, sterilize, and inoculate with the brewer's yeast and Bacillus subtilis seed solutions prepared in step S4, wherein the inoculation amounts of the brewer's yeast and Bacillus subtilis seed solutions are 2.5 v / v% and 4 v / v%, respectively. Ferment for 30 hours at 38°C and 60 r / min, add additives, wherein the additive amount is 4wt% of the total mass of the system, and ferment for 15 hours to obtain a fermentation mixture.

[0121] Comparative Example 7

[0122] Compared with Example 3, the difference is that Bacillus subtilis is not inoculated in step S5.

[0123] The details are as follows:

[0124] S5. Fermentation: 35 parts by weight of the filter residue in step S1 are added to 135 parts by weight of water, sterilized, and inoculated with the brewer's yeast and Lactobacillus salivarius seed solutions prepared in step S4, wherein the inoculation amounts of the brewer's yeast and Lactobacillus salivarius seed solutions are 2.5 v / v% and 4 v / v%, respectively. The mixture is fermented at 38°C, 60 r / min, for 30 hours. An additive is added, wherein the additive is added in an amount of 4wt% of the total mass of the system, and the fermentation is carried out for 15 hours to obtain a fermentation mixture.

[0125] Comparative Example 8

[0126] Compared with Example 3, the difference is that the seed liquid of Lactobacillus salivarius and Bacillus subtilis is not inoculated in step S5.

[0127] The details are as follows:

[0128] S5. Fermentation: Add 35 parts by weight of the filter residue in step S1 to 135 parts by weight of water, sterilize, and inoculate with the brewer's yeast seed solution prepared in step S4, wherein the inoculation amount of the brewer's yeast seed solution is 2.5 v / v%, 38°C, 60 r / min, ferment for 30 hours, add additives, the additive amount of which is 4wt% of the total mass of the system, and ferment for 15 hours to obtain a fermentation mixture.

[0129] Comparative Example 9

[0130] Compared with Example 3, the difference is that no additive is added in step S5.

[0131] The details are as follows:

[0132] S5. Fermentation: 35 parts by weight of the filter residue in step S1 are added to 135 parts by weight of water, sterilized, and inoculated with the saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis seed solutions prepared in step S4, wherein the inoculation amounts of the saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis seed solutions are 2.5 v / v%, 1.5 v / v%, and 2.5 v / v%, respectively, at 38°C, 60 r / min, and fermented for 45 hours to obtain a fermentation mixture.

[0133] Comparative Example 10

[0134] Compared with Example 3, the difference is that steps S4, S5 and S6 are not performed.

[0135] The details are as follows:

[0136] S1. Water extraction: 4 parts by weight of Salvia miltiorrhiza, 2 parts by weight of Trillium gracile, and 6 parts by weight of Cordyceps militaris were washed, dried, and crushed to obtain a mixed powder, which was added to water. The solid-liquid ratio of the mixed powder to water was 1:6 g / mL. The mixture was heated and boiled for extraction for 1.5 h. The operation was repeated 3 times. The residue was retained. The filtrate was combined and concentrated to a relative density of 1.07. Ethanol was added to the system to obtain an ethanol content of 65 wt %. The mixture was precipitated for 4 h. The solid was collected by centrifugation and dried to obtain an active polysaccharide. The filtrate was used to recover ethanol and dried to obtain a water extract.

[0137] S2. Preparation of selenium-enriched yeast extracellular polysaccharides: inoculate selenium-enriched yeast into the culture medium at an inoculation amount of 2.5 w / v%, 52°C, 60 r / min, ferment and culture for 30 h, centrifuge, add a mixed solvent to the supernatant, the volume ratio of the supernatant to the mixed solvent is 4:1, precipitate for 1.5 h, centrifuge, add ethanol to the supernatant until the ethanol content of the system is 65 wt%, precipitate for 4 h, dry, and obtain selenium-enriched yeast extracellular polysaccharides;

[0138] The formula of the culture medium is: 30 parts by weight of glucose, 4 parts by weight of peptone, KH 2 PO 4 2.5 parts by weight, MgSO 4 7H 2 O 1.5 parts by weight, vitamin B1 0.015 parts by weight, water 1000 parts by weight;

[0139] The mixed solvent is a mixture of chloroform and n-butanol in a volume ratio of 4:1;

[0140] S3. Preparation of organic selenium-active polysaccharide complex: 6 parts by weight of selenium-enriched yeast extracellular polysaccharide prepared in step S2 are added to 100 parts by weight of water, and 4 parts by weight of active polysaccharide prepared in step S1 are added, stirred and mixed for 20 minutes, and dried to obtain an organic selenium-active polysaccharide complex;

[0141] S4. Encapsulation: 4 parts by weight of the water extract obtained in step S1, 8.5 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 1.5 parts by weight of curcumin, 0.7 parts by weight of oligofructose, 22 parts by weight of sodium alginate, and 8.5 parts by weight of sodium carboxymethyl cellulose are added to 225 parts by weight of water, and mixed for 20 minutes to obtain an aqueous phase; 110 parts by weight of the aqueous phase and 2.5 parts by weight of lecithin are added to 225 parts by weight of fish oil, emulsified at 10,000 r / min for 10 minutes, 50 parts by weight of 4wt% calcium chloride solution is added dropwise, and the mixture is cured at room temperature for 35 minutes. The mixture is centrifuged, washed, and freeze-dried to obtain a composition for preventing and treating liver cancer.

[0142] Comparative Example 11

[0143] Compared with Example 3, the difference is that cysteine ​​is not added in step S6.

[0144] The details are as follows:

[0145] S6. Re-fermentation: ferment the fermentation mixture in step S5 at 38°C, 60r / min, for 21h, and freeze-dry to obtain a fermentation product.

[0146] Comparative Example 12

[0147] Compared with Example 3, the difference is that no embedding is performed in step S7, and the components are simply mixed.

[0148] The details are as follows:

[0149] S7. Encapsulation: 4 parts by weight of the water extract obtained in step S1, 8.5 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 13.5 parts by weight of the fermentation product obtained in step S6, 1.5 parts by weight of curcumin, and 0.7 parts by weight of oligofructose are mixed for 20 minutes to obtain a composition for preventing and treating liver cancer.

[0150] Comparative Example 13

[0151] Compared with Example 3, the difference is that curcumin is not added in step S7.

[0152] The details are as follows:

[0153] S7. Encapsulation: add 4 parts by weight of the water extract obtained in step S1, 8.5 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 13.5 parts by weight of the fermentation product obtained in step S6, 2.2 parts by weight of oligofructose, 22 parts by weight of sodium alginate and 8.5 parts by weight of sodium carboxymethyl cellulose to 225 parts by weight of water, mix for 20 minutes to obtain an aqueous phase; add 110 parts by weight of the aqueous phase and 2.5 parts by weight of lecithin to 225 parts by weight of fish oil, emulsify at 10,000 r / min for 10 minutes, add 50 parts by weight of 4wt% calcium chloride solution dropwise, solidify at room temperature for 35 minutes, centrifuge, wash and freeze-dry to obtain a composition for preventing and treating liver cancer.

[0154] Comparative Example 14

[0155] Compared with Example 3, the difference is that oligofructose is not added in step S7.

[0156] The details are as follows:

[0157] S7. Encapsulation: Add 4 parts by weight of the water extract obtained in step S1, 8.5 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 13.5 parts by weight of the fermentation product obtained in step S6, 2.2 parts by weight of curcumin, 22 parts by weight of sodium alginate, and 8.5 parts by weight of sodium carboxymethyl cellulose to 225 parts by weight of water, and mix for 20 minutes to obtain an aqueous phase; add 110 parts by weight of the aqueous phase and 2.5 parts by weight of lecithin to 225 parts by weight of fish oil, emulsify at 10,000 r / min for 10 minutes, add 50 parts by weight of 4wt% calcium chloride solution dropwise, solidify at room temperature for 35 minutes, centrifuge, wash, and freeze-dry to obtain a composition for preventing and treating liver cancer.

[0158] Comparative Example 15

[0159] Compared with Example 3, the difference is that curcumin and oligofructose are not added in step S7.

[0160] The details are as follows:

[0161] S7. Encapsulation: add 4 parts by weight of the water extract obtained in step S1, 8.5 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 13.5 parts by weight of the fermentation product obtained in step S6, 22 parts by weight of sodium alginate and 8.5 parts by weight of sodium carboxymethyl cellulose to 225 parts by weight of water, mix for 20 minutes to obtain an aqueous phase; add 110 parts by weight of the aqueous phase and 2.5 parts by weight of lecithin to 225 parts by weight of fish oil, emulsify at 10,000 r / min for 10 minutes, add 50 parts by weight of 4wt% calcium chloride solution dropwise, solidify at room temperature for 35 minutes, centrifuge, wash, and freeze-dry to obtain a composition for preventing and treating liver cancer.

[0162] Test Example 1: Sustained-release test

[0163] 1g of the composition for preventing and treating liver cancer prepared in Examples 1-3 of the present invention and Comparative Example 12 was added to 9mL of artificial simulated gastric juice and 9mL of artificial simulated intestinal juice, respectively, and reacted at 37°C and 50r / min for 2h and 3h, respectively. In addition, an equal amount of the composition for preventing and treating liver cancer was added to 9mL of artificial simulated gastric juice, reacted at 37°C and 50r / min for 2h, centrifuged, and then added to 9mL of artificial simulated intestinal juice, and continued to react at 37°C and 50r / min for 3h. After the reaction, the probiotic cell count was performed. The survival rate was calculated according to the following formula:

[0164] Survival rate (%) = N t / N 0 ×100%

[0165] Where N t is the concentration of probiotics that survive after a certain period of incubation (cfu / g), N 0 is the initial probiotic concentration (cfu / g).

[0166] The release rate was calculated according to the following formula:

[0167] Release rate (%) = (W t -W 0 ) / W 0 ×100%

[0168] Where W t is the initial weight of the sample; W 0 The weight of the sample was measured after a certain period of incubation.

[0169] Results Figure 1 and Figure 2 .

[0170] As can be seen from the figure, the composition for preventing and treating liver cancer prepared in Examples 1-3 of the present invention has good integrity in artificial simulated gastric juice, and releases active substances after being transferred to artificial simulated intestinal juice, indicating that the composition for preventing and treating liver cancer has the characteristics of pH responsiveness and resistance to gastric acid, and has the characteristics of targeted delivery and controlled release of active components in the intestine.

[0171] Test Example 2 Inhibition rate of cell proliferation

[0172] The compositions for preventing and treating liver cancer prepared in Examples 1-5 and Comparative Examples 1-15 were added into water and stirred at 37° C. for 10 h to prepare 50 μg / mL sample solutions.

[0173] Mouse liver cancer H22 cells in logarithmic growth phase were prepared into cell suspension using RPMI1640 culture medium and the cell density was adjusted to 10 4100 μL per well was inoculated into a 96-well plate. The plate was divided into 20 groups, with 5 replicate wells in each group. 50 μL of sample solution was added to each well and placed at 37°C and 5 v / v% CO 2 Culture in a saturated humidity cell culture incubator for 24 hours, add 5 μg / mL MTT solution, incubate in the incubator for 4 hours, centrifuge, remove the liquid in the wells and add 150 μL DMSO, shake for 10 minutes, and measure the absorbance OD value at a wavelength of 490 nm using an enzyme marker. Repeat 3 times for each group.

[0174] Inhibition rate = (1-OD value of test well / OD value of control well) × 100%

[0175] The results are shown in Table 1.

[0176] Table 1

[0177]

[0178]

[0179] It can be seen from the above table that the compositions for preventing and treating liver cancer prepared in Examples 1-3 of the present invention have a good effect of inhibiting the proliferation of liver cancer cells in vitro.

[0180] Test Example 3

[0181] The liver cancer H22 cell line in Test Example 1 was collected and centrifuged three times at 10000r / rain for 5 min to adjust the cell density to 1×10 6 / mL, the cell (survival rate ≥ 95%) suspension 0.2mL was injected into the right axillary subcutaneous of BALB / c mice, and the whole process was strictly aseptic. After 24h, it was randomly divided into 22 groups, namely the model group, the cyclophosphamide group, the embodiment 1-5 group, and the comparative example 1-15 group. At the same time, the 1st group was set as the normal group without tumor inoculation, the cyclophosphamide group was given 20mg / kg cyclophosphamide, the normal group and the model group were given the same volume of physiological saline by gavage, and each group was given 1g / kg of the composition for preventing and treating liver cancer prepared by the corresponding group by gavage, and the administration was continued for 14d, and the mice were weighed before and after the test. On the 15th day, anesthesia was performed, blood was taken from the orbit to separate the serum for standby use, and the mice were killed by myelotomy, and the tumor was peeled off and weighed.

[0182] Weight growth rate (%) = (weight gain of drug administration group - weight gain of model group) / weight gain of model group × 100%;

[0183] Tumor inhibition rate (%) = (average tumor weight of model group - average tumor weight of drug-treated group) / average tumor weight of model group × 100%).

[0184] Serum IL-2 and TNF-α detection: Blood was collected from mice in each group without adding anticoagulant, centrifuged, and determined by ELISA method. The operating steps were carried out according to the instructions of the kit.

[0185] The results are shown in Table 2.

[0186] Table 2

[0187]

[0188]

[0189] Note: * compared with the blank group, P < 0.05; # compared with the model group, P < 0.05.

[0190] It can be seen from the above table that the compositions for preventing and treating liver cancer prepared in Examples 1-3 of the present invention have a significant effect of inhibiting the growth of liver cancer cells, do not damage the physique of mice, and have a protective and improving effect on the damage of immune indicators.

[0191] Compared with Example 3, the additives in Examples 4 and 5 are single calcium chloride or vitamin B1. Compared with Example 3, no additive is added in step S5 in Comparative Example 9. The inhibition rate of cell proliferation is slightly decreased, the tumor inhibition rate is slightly decreased, and the contents of IL-2 and TNF-α are decreased. The present invention supplements calcium chloride and vitamin B2 during the probiotic fermentation process, increases the resistance of probiotics through the regulation of trace elements, and increases the time period of the probiotics in the stable period, thereby promoting the production of more beneficial substances.

[0192] Comparative Examples 1, 2, and 3 are compared with Example 3, and step S1 does not add salvia miltiorrhiza, Psoralea corylifolia or Cordyceps militaris. The inhibition rate of cell proliferation decreases, the tumor inhibition rate decreases, the weight growth rate decreases, and the IL-2 and TNF-α contents decrease. The present invention extracts salvia miltiorrhiza, Psoralea corylifolia, and Cordyceps militaris through water, and the active components obtained include active substances such as salvianolic acid B, cordycepin, alkaloids, saponins, flavonoids, cordyceps polysaccharides, salvia miltiorrhiza polysaccharides, and Psoralea corylifolia polysaccharides, wherein salvianolic acid B is polymerized by 3 molecules of salvia miltiorrhiza and 1 molecule of caffeic acid, and is a component with the highest content and the strongest activity in the water-soluble effective ingredient salvianolic acid of salvia miltiorrhiza, and salvianolic acid B has an in vitro growth inhibitory effect on cancer cells. Salvianolic acid B can reduce the effect of acute liver injury by inhibiting the TLR4 / MyD88 / NF-κB signaling pathway, and the TLR4 / MyD88 / NF-κB signaling pathway is a key pathway widely present in various cells to participate in inflammatory response, and is an important regulatory mechanism for mediating tumorigenesis. Tannic acid B can affect the composition of intestinal flora, increase the relative abundance of beneficial intestinal bacteria, reduce the relative abundance of harmful bacteria, and improve the intestinal environment. Trillium polysaccharide is the main bioactive component of Trillium, which can improve the body's immunity, inhibit P13K / AKT / mTOR activation, induce autophagy, and then inhibit angiogenesis and promote tumor cell apoptosis. Cordycepin is a natural nucleoside antibiotic and a natural anticancer antibiotic. Cordycepin is 3-deazaadenosine, a nucleic acid derivative of nitrogen-containing glycosides, a purine alkaloid, with antiviral and anti-tumor effects, especially a strong inhibitory effect on a variety of solid malignant tumors.

[0193] Comparative Example 4 Compared with Example 3, steps S2 and S3 are not carried out, and in step S7, organic selenium-active polysaccharide complex is replaced by the active polysaccharide obtained by step S1. The inhibition rate of cell proliferation decreases, the tumor inhibition rate decreases, the weight growth rate decreases, and the IL-2 and TNF-α contents decrease. In the present invention, after selenium-enriched yeast fermentation, the cell is removed by centrifugation, and the Sevag method removes protein to obtain selenium-enriched yeast extracellular polysaccharide liquid, and after mixing with the active polysaccharide obtained by extracting through salvia miltiorrhiza, Psoralea corylifolia, and Cordyceps militaris, by chelation, organic selenium-active polysaccharide complex can be formed, compared to inorganic selenium, the organic selenium produced by selenium-enriched yeast fermentation is more easily absorbed by the human body, and safe and non-toxic, thereby playing a good anti-oxidation, anti-inflammatory, improving the level of glutathione in the liver, reducing the activation of hepatic stellate cells and the expression of liver fibrosis-related genes, thereby playing a good protection of the liver, preventing liver lesion progression, and preventing and treating the effects of liver, cirrhosis, liver failure, etc.

[0194] Comparative Example 5 Compared with Example 3, Saccharomyces cerevisiae was not inoculated in step S5, and step S6 was not performed. The inhibition rate of cell proliferation decreased, the tumor inhibition rate decreased, the weight growth rate decreased, and the IL-2 and TNF-α contents decreased. Saccharomyces cerevisiae in the fermentation bacteria of the present invention can promote the growth of bifidobacteria (probiotics) in the intestinal flora and inhibit the number of Candida albicans (pathogenic bacteria); it can also protect the mucosal barrier function by reducing the apoptosis of intestinal mucosal epithelial cells, and prevent the further development of cirrhosis by inhibiting the release of inflammatory mediators such as TNF-α and IL-6. Saccharomyces cerevisiae can also accumulate more glutathione in the cell, thereby greatly increasing the amount of glutathione in the composition.

[0195] Compared with Example 3, in Comparative Examples 6 and 7, Lactobacillus salivarius or Bacillus subtilis was not inoculated in step S5. Compared with Example 3, in Comparative Example 8, Lactobacillus salivarius and Bacillus subtilis seed liquid were not inoculated in step S5. Compared with Example 3, in Comparative Example 10, steps S4, S5 and S6 were not performed. The inhibition rate of cell proliferation decreased, the tumor inhibition rate decreased, the weight growth rate decreased, and the IL-2 and TNF-α contents decreased. The Lactobacillus salivarius added by the present invention has the effects of improving liver function, protecting the intestinal barrier, and alleviating liver fibrosis, and can prevent or treat cirrhosis and liver failure, but its metabolites can protect liver function and play a positive role in preventing liver cancer, such as indolepropionic acid, which is a product of tryptophan decomposition by bacteria, which can inhibit NF-κB signaling pathway, reduce the level of proinflammatory cytokines, antagonize liver inflammation and liver damage, and can also improve the inherent immunity of the liver and intestines, thereby enhancing the ability of the liver to clear hepatitis viruses, reduce liver inflammation, and strengthen the intestinal barrier function, thereby playing an anti-cancer effect. Bacillus subtilis can degrade aflatoxin and prevent liver cancer by reducing the toxicity of carcinogens. It can also effectively improve intestinal flora imbalance and restore the body's normal immune response to HBV, while reducing the release of inflammatory mediators and liver cell damage, thereby improving the symptoms of viral hepatitis. At the same time, it can stimulate Prevotella to secrete anti-inflammatory substances, effectively inhibiting the level of Th17 cells in the tumor. Th17 cells are a subset of helper T cells that can characteristically secrete IL-17, which can inhibit tumor immunity and stimulate tumor angiogenesis, participate in the occurrence and development of tumors, and exert anti-cancer effects by inhibiting the level of Th17 cells, with a synergistic effect.

[0196] Comparative Example 11 Compared with Example 3, cysteine ​​was not added in step S6. The tumor inhibition rate decreased, the weight growth rate decreased, and the IL-2 and TNF-α contents decreased. Cysteine ​​is an important raw material for the synthesis of glutathione. The present invention promotes the production of glutathione, a product of brewer's yeast fermentation, by adding cysteine, thereby playing a good role in protecting the liver and preventing and treating liver cancer.

[0197] Compared with Example 3, in Comparative Example 12, no embedding was performed in step S7, and the components were simply mixed. The inhibition rate of cell proliferation decreased, the tumor inhibition rate decreased, the weight growth rate decreased, and the IL-2 and TNF-α contents decreased. By embedding, the resistance of probiotics to gastric acid can be improved, and the polysaccharide active components can be prevented from being oxidized and decomposed. It has the characteristics of targeted delivery and controlled release of active components in the intestine, thereby greatly improving the activity of the active components.

[0198] Compared with Example 3, in Comparative Examples 13 and 14, curcumin or oligofructose was not added in step S7. Compared with Example 3, in Comparative Example 15, curcumin and oligofructose were not added in step S7. The inhibition rate of cell proliferation decreased, and the tumor inhibition rate decreased. The curcumin also added in the present invention can reduce liver damage by increasing the glutathione content in the liver. At the same time, curcumin and oligofructose are also a kind of good prebiotics, which can promote the proliferation of intestinal probiotics in a targeted manner, thereby inhibiting the growth of harmful bacteria, regulating liver function through the liver-intestine axis, promoting the recovery of liver function, inhibiting the formation of fatty liver, and protecting the liver.

[0199] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a composition for preventing and treating liver cancer, characterized in that: Extracting salvia miltiorrhiza, trillium and cordyceps militaris with water, filtering, retaining the filter residue, ethanol precipitation to obtain active polysaccharides, and drying the filtrate to obtain a water extract; obtaining selenium-enriched yeast extracellular polysaccharides and mixing them with active polysaccharides to obtain an organic selenium-active polysaccharide complex; adding the filter residue to water, inoculating saccharomyces cerevisiae, lactobacillus salivarius and bacillus subtilis, fermenting, adding cysteine, fermenting again, freeze-drying to obtain a fermentation product, and mixing and embedding the water extract, the organic selenium-active polysaccharide complex, the fermentation product, curcumin and oligofructose to obtain a composition for preventing and treating liver cancer; The following steps are involved: S1. Water extraction: Wash, dry and crush Salvia miltiorrhiza, Psoralea corylifolia and Cordyceps militaris respectively to obtain a mixed powder, add it into water, heat and boil it for extraction, repeat the operation 2-3 times, filter, retain the filter residue, combine the filtrate, concentrate, add ethanol to precipitate, collect the solid by centrifugation, and dry it to obtain the active polysaccharide; recover ethanol from the filtrate, dry it, and obtain a water extract; S2. Preparation of selenium-enriched yeast exopolysaccharide: inoculating selenium-enriched yeast into a culture medium, fermenting and culturing, centrifuging, adding a mixed solvent to the supernatant, precipitating, centrifuging, adding ethanol to the supernatant, and drying to obtain selenium-enriched yeast exopolysaccharide; S3. Preparation of organic selenium-active polysaccharide complex: adding the selenium-enriched yeast extracellular polysaccharide obtained in step S2 to water, adding the active polysaccharide obtained in step S1, stirring and mixing evenly, and drying to obtain an organic selenium-active polysaccharide complex; S4. Activation of strains: Saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis were inoculated into Gould's medium, respectively, and activated and cultured to obtain strain seed liquid; S5. Fermentation: adding the filter residue in step S1 to water, sterilizing, inoculating the saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis seed solution obtained in step S4, fermenting for a first period of time, adding an additive, and fermenting for a second period of time to obtain a fermentation mixture; the additive is a mixture of calcium chloride and vitamin B1; S6. Promoting glutathione synthesis fermentation: adding cysteine ​​to the fermentation mixture in step S5, continuing the fermentation culture for a third period of time, freeze-drying, and obtaining a fermentation product; S7. Encapsulation: Add the water extract obtained in step S1, the organic selenium-active polysaccharide complex obtained in step S3, the fermentation product obtained in step S6, curcumin, oligofructose, sodium alginate and sodium carboxymethyl cellulose into water, mix well to obtain an aqueous phase; add the aqueous phase and lecithin into fish oil, emulsify, add calcium chloride solution dropwise, solidify at room temperature, centrifuge, wash, and freeze-dry to obtain a composition for preventing and treating liver cancer.

2. The preparation method according to claim 1, characterized in that: In step S1, the mass ratio of Salvia miltiorrhiza, Psoralea corylifolia and Cordyceps militaris is 3-5:1-3:5-7, the solid-liquid ratio of the mixed powder and water is 1:5-7 g / mL, the heating and boiling extraction time is 1-2 h, ethanol is added until the ethanol content of the system is 60-70 wt%, and the precipitation time is 3-5 h.

3. The preparation method according to claim 1, characterized in that: The formula of the culture medium in step S2 is: 20-40 weight parts of glucose, 3-5 weight parts of peptone, 2-3 weight parts of KH2PO4, 1-2 weight parts of MgSO4·7H2O, 0.01-0.02 weight parts of vitamin B1, and 1000 weight parts of water. The inoculation amount of the selenium-enriched yeast is 2-3w / v%, the fermentation culture conditions are 50-55°C, 50-70r / min, and the time is 24-36h. The mixed solvent is a mixed solvent of chloroform and n-butanol, and the volume ratio is 4:

1. The volume ratio of the supernatant and the mixed solvent is 3-5:

1. The precipitation time is 1-2h, ethanol is added to the supernatant until the ethanol content of the system is 60-70wt%, and the precipitation time is 3-5h. The mass ratio of the selenium-enriched yeast extracellular polysaccharide and the active polysaccharide in step S3 is 5-7:3-5.

4. The preparation method according to claim 1, characterized in that: The activation culture conditions in step S4 are 40-42°C, 50-70r / min, activation culture for 18-24h, and the bacterial content of the bacterial seed liquid is 10 8 -10 9 cfu / mL; the mass ratio of the filter residue and water in step S5 is 2-5:12-15, the inoculation amounts of the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solutions are 2-3v / v%, 1-2v / v% and 2-3v / v%, respectively; the fermentation conditions are 37-40°C, 50-70r / min, the first time period is 24-36h, the second time period is 12-18h, the mass ratio of calcium chloride and vitamin B1 in the additive is 3-5:1-2, and the amount of the additive added is 3-5wt% of the total mass of the system.

5. The preparation method according to claim 1, characterized in that: In step S6, the mass ratio of the fermentation mixture to cysteine ​​is 100-120:5-7, the fermentation culture conditions are 37-40° C., 50-70 r / min, and the third time period is 18-24 h.

6. The preparation method according to claim 1, characterized in that: The mass ratio of the water extract, the organic selenium-active polysaccharide complex, the fermentation product, curcumin, oligofructose, sodium alginate, sodium carboxymethyl cellulose and water in step S7 is 3-5:7-10:12-15:1-2:0.5-1:20-25:7-10:200-250, the mass ratio of the aqueous phase, lecithin and fish oil is 100-120:2-3:200-250, the concentration of the calcium chloride solution is 3-5wt%, and the time for room temperature curing is 30-40min.

7. The preparation method according to claim 1, characterized in that: The specific steps include: S1. Water extraction: 3-5 parts by weight of Salvia miltiorrhiza, 1-3 parts by weight of Trillium gracile, and 5-7 parts by weight of Cordyceps militaris were washed, dried, and crushed to obtain a mixed powder, which was added to water. The solid-liquid ratio of the mixed powder to water was 1:5-7 g / mL. The mixture was heated and boiled for extraction for 1-2 h. The operation was repeated 2-3 times. The residue was retained. The filtrate was combined and concentrated. Ethanol was added until the ethanol content of the system was 60-70 wt %. The mixture was precipitated for 3-5 h. The solid was collected by centrifugation and dried to obtain an active polysaccharide. The filtrate was recovered by ethanol and dried to obtain a water extract. S2. Preparation of selenium-enriched yeast extracellular polysaccharides: inoculate selenium-enriched yeast into the culture medium at an inoculation amount of 2-3 w / v%, 50-55°C, 50-70 r / min, ferment and culture for 24-36 hours, centrifuge, add a mixed solvent to the supernatant, the volume ratio of the supernatant to the mixed solvent is 3-5:1, precipitate for 1-2 hours, centrifuge, add ethanol to the supernatant until the ethanol content of the system is 60-70wt%, precipitate for 3-5 hours, dry, and obtain selenium-enriched yeast extracellular polysaccharides; The formula of the culture medium is: 20-40 parts by weight of glucose, 3-5 parts by weight of peptone, 2-3 parts by weight of KH2PO4, 1-2 parts by weight of MgSO4·7H2O, 0.01-0.02 parts by weight of vitamin B1, and 1000 parts by weight of water; The mixed solvent is a mixture of chloroform and n-butanol in a volume ratio of 4:1; S3. Preparation of organic selenium-active polysaccharide complex: add 5-7 parts by weight of selenium-enriched yeast extracellular polysaccharide obtained in step S2 to 100 parts by weight of water, add 3-5 parts by weight of active polysaccharide obtained in step S1, stir and mix evenly, and dry to obtain an organic selenium-active polysaccharide complex; S4. Activation of strains: Saccharomyces cerevisiae, Lactobacillus salivarius, and Bacillus subtilis were inoculated into Gould's medium respectively, and activated and cultured for 18-24 hours at 40-42°C and 50-70r / min to obtain strain seed liquid with a bacterial content of 10 8 -10 9 cfu / mL; S5. Fermentation: adding 20-50 parts by weight of the filter residue in step S1 to 120-150 parts by weight of water, sterilizing, inoculating the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution prepared in step S4, wherein the inoculation amount of the saccharomyces cerevisiae, Lactobacillus salivarius and Bacillus subtilis seed solution is 2-3v / v%, 1-2v / v% and 2-3v / v%, respectively, at 37-40°C, 50-70r / min, fermenting for 24-36h, adding additives, wherein the additive is added in an amount of 3-5wt% of the total mass of the system, and fermenting for 12-18h to obtain a fermentation mixture; The additive is a mixture of calcium chloride and vitamin B1, with a mass ratio of 3-5:1-2; S6. Promoting glutathione synthesis fermentation: adding 5-7 parts by weight of cysteine ​​to 100-120 parts by weight of the fermentation mixture in step S5, 37-40°C, 50-70r / min, fermenting for 18-24h, freeze-drying, and obtaining a fermentation product; S7. Encapsulation: add 3-5 parts by weight of the water extract obtained in step S1, 7-10 parts by weight of the organic selenium-active polysaccharide complex obtained in step S3, 12-15 parts by weight of the fermentation product obtained in step S6, 1-2 parts by weight of curcumin, 0.5-1 parts by weight of oligofructose, 20-25 parts by weight of sodium alginate, and 7-10 parts by weight of sodium carboxymethyl cellulose to 200-250 parts by weight of water, mix well, and obtain an aqueous phase; add 100-120 parts by weight of the aqueous phase and 2-3 parts by weight of lecithin to 200-250 parts by weight of fish oil, emulsify, add 50 parts by weight of 3-5wt% calcium chloride solution dropwise, solidify at room temperature for 30-40 minutes, centrifuge, wash, and freeze-dry to obtain a composition for preventing and treating liver cancer.

8. A composition for preventing and treating liver cancer obtained by the preparation method according to any one of claims 1 to 7.

9. Use of the composition for preventing and treating liver cancer as claimed in claim 8 in preparing a drug for preventing and treating liver cancer.

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