A selenomethionine-containing composition and its preparation method and application

Abstract

The present invention relates to a selenomethionine-containing composition and a preparation method and application thereof, belonging to the field of biomedical technology. The composition is compounded with L-selenomethionine, L-selenomethylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymolyte, and each component has a good synergistic effect, and can achieve a very excellent liver fibrosis treatment effect. Among them, the combination of L-selenomethionine and L-selenomethylselenocysteine ​​can promote the absorption rate of selenium, can control and appropriately increase the liver enrichment of selenium, achieve a higher immune ability, improve the liver's antiviral property, is safe and reliable, and will not cause selenium poisoning. L-theanine can promote the effective absorption of L-selenomethionine and L-selenomethylselenocysteine. Monkfish liver peptide and Tribulus terrestris enzymolyte are prepared by a special process, both have a good liver fibrosis treatment effect, and effectively alleviate the development process of liver fibrosis.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to a selenomethionine-containing composition, a preparation method and an application thereof. Background Art

[0002] Liver fibrosis is a common manifestation in the progression of various chronic liver diseases and is a key link in the transition to cirrhosis and liver cancer. The main pathogenesis of liver fibrosis is the excessive increase and abnormal deposition of liver extracellular matrix. Among them, hepatitis virus infection is one of the main factors leading to liver fibrosis. Among viral hepatitis, chronic hepatitis caused by viruses such as type A and type B mainly causes early liver damage and liver fibrosis. Although more and more attention is paid to hepatitis virus vaccination for infants and young children, and the infection rate of viral hepatitis in non-adults has been significantly reduced, the overall infection rate of viral hepatitis is still high.

[0003] Among them, hepatitis B virus (HBV) infection is a major public health problem, and the infection rate of hepatitis B virus is very high. Once chronic liver disease develops from liver fibrosis to cirrhosis or even ascites, various treatments are difficult to work, so research on how to prevent and treat liver fibrosis has become a hot topic in the medical community today.

[0004] At present, there are many effective ingredients for intervening in the treatment of liver fibrosis, including Patent 2021111959888, a liver fibrosis prevention and treatment drug; Patent 2020102564748, a Chinese medicine compound for anti-liver fibrosis and its use; Patent 2021103558753, the use of phellodendrine as a drug for treating liver fibrosis; and Patent 202211082124X, the use of isoliensinine and methyl neferine in the preparation of anti-liver fibrosis drugs; their treatment mechanisms and effects vary, and they mainly play an auxiliary role in promoting treatment; the main treatment method still relies on antiviral treatment to remove pathogenic factors and stop the development of liver fibrosis.

[0005] In addition to the above-mentioned drugs, selenium can also improve the treatment of liver fibrosis. Selenium is an essential trace element in the body. Selenium and its metabolites have antioxidant, immune-enhancing, and oxygen free radical scavenging functions in the body. They can also regulate related gene expression and basal metabolism. They play an important role in the prevention and treatment of digestive system diseases, cardiovascular system diseases, and respiratory system diseases. There is a special correlation between the severity of liver disease and the serum selenium content. Selenium, as an antioxidant, can also improve the body's immunity, and is of great significance in the treatment of liver disease. Selenium can significantly reduce the degree of liver cell degeneration and fibrosis. Transforming growth factor β1 (TGF-β1) is the cytokine in the body that is currently known to be most closely related to the formation of tissue fibrosis. Abnormal activation of the TGF-β1 / Smad-3 signaling pathway is closely related to the occurrence and development of diseases such as benign biliary stenosis and liver fibrosis. Selenium can inhibit the expression of TGF-β1 and the activation of the TGF-β1 / Smad-3 signaling pathway. Therefore, the existing technology usually uses selenium-rich drugs or foods to prevent and delay the progression of liver fibrosis.

[0006] Although selenium can improve liver fibrosis, the dietary supplementation of selenium alone is not very ideal, and can only be used as a nutritional supplement. Therefore, in the prior art, selenium is often combined with other material components to improve the improvement and treatment effect of liver fibrosis, such as patent No. 2015800724114 liver fibrosis improver, which discloses the use of selenium with zinc, chromium, magnesium, potassium, royal jelly and vitamin C. However, the effect of the current selenium-containing composition drug treatment of liver fibrosis is also limited, and it also plays an auxiliary role in promoting treatment; in addition, the existing selenium components used are generally selenium-enriched yeast, selenium-enriched edible fungi, selenite, selenoprotein, etc., which are cytotoxic (such as cell wall detachment, increased intracellular vacuoles, cell membrane rupture, cell necrosis, etc.) when exceeded, so the dosage cannot be too high, and the dosage for patients with different physiques is not easy to grasp, and the safety is poor. Therefore, the technology in the treatment of liver fibrosis needs to be improved. Summary of the invention

[0007] The curative effects of various existing drug components for the treatment of liver fibrosis are still unsatisfactory, and in the pharmaceutical composition containing selenium, improper dosage of organic or inorganic selenium can cause cytotoxicity, and low-content selenium alone can improve liver fibrosis and has the problem of low effect. The present invention provides a selenomethionine-containing composition and its preparation method and application, which are used in combination with L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymolyte, and have a good synergistic effect, and can achieve a very excellent liver fibrosis treatment effect. Among them, the combination of L-selenomethionine and L-seleno-methylselenocysteine ​​can promote the absorption rate of selenium, control and appropriately increase the liver enrichment of selenium, achieve a higher immune ability, improve the liver's antiviral property, and be safe and reliable without selenium poisoning. L-theanine can promote the effective absorption of L-selenomethionine and L-seleno-methylselenocysteine. Anglerfish liver peptide and Tribulus terrestris enzymatic hydrolysate are prepared by special process, both of which have good therapeutic effect on liver fibrosis and effectively alleviate the development of liver fibrosis. The specific technical scheme is as follows:

[0008] A selenomethionine-containing composition is prepared from the following raw materials in parts by mass: 0.02 to 0.05 parts of L-selenomethionine, 0.03 to 0.08 parts of L-seleno-methylselenocysteine, 0.02 to 0.05 parts of L-theanine, 1 to 3 parts of monkfish liver peptide, and 4 to 8 parts of tribulus terrestris enzymatic hydrolysate.

[0009] In the above technical scheme, the preparation method of the anglerfish liver peptide is:

[0010] S1: add anglerfish liver protein to a phosphate buffer solution of pH 7.5-8.0, 40 mM-50 mM, add trypsin, perform enzymatic hydrolysis at 36°C-38°C for 2h-3h, inactivate the enzyme at high temperature, cool to room temperature, and use a centrifugal ultrafiltration tube with an ultrafiltration membrane of 5 kDa and 3 kDa molecular weight cutoff to collect the filtrate between 3 kDa and 5 kDa by centrifugal filtration;

[0011] S2: Add phosphate buffer solution of pH 7.5-8.0 and 40mM-50mM to the filtrate, add subtilisin, perform enzymolysis at 36°C-38°C for 1.5h-2h, inactivate the enzyme at high temperature, cool to room temperature, add papain, perform enzymolysis at 50°C-55°C for 1.5h-2h, inactivate the enzyme at high temperature, cool to room temperature, use a centrifugal ultrafiltration tube with a 2kDa ultrafiltration membrane to perform centrifugal filtration to collect the filtrate less than 2kDa, vacuum freeze-dry, and break up to obtain anglerfish liver peptide.

[0012] In S1 of the above-mentioned method for preparing anglerfish liver peptide, the added mass of the phosphate buffer is 8 to 10 times that of the anglerfish liver protein; the added mass of the trypsin is 0.5% to 1.5% of the anglerfish liver protein; the temperature of the high-temperature enzyme inactivation is 90°C to 100°C, and the time is 10 min to 15 min.

[0013] In S2 of the above-mentioned method for preparing monkfish liver peptide, the added mass of the phosphate buffer is 8 to 10 times that of the monkfish liver protein; the added mass of the subtilisin is 1% to 1.5% of the monkfish liver protein; the added mass of the papain is 1% to 1.5% of the monkfish liver protein; the temperature of the high-temperature enzyme inactivation is 90°C to 100°C, and the time is 10 min to 15 min.

[0014] In the above-mentioned method for preparing the monkfish liver peptide, the method for extracting the monkfish liver protein is as follows: taking a freshly washed monkfish liver, crushing it, adding it to 8 to 10 times the mass of the monkfish liver in 30°C to 35°C warm water, soaking it for 6h to 10h, removing the water layer above the monkfish liver precipitation, draining the monkfish liver precipitation, adding 0.12mol / L to 0.15mol / L saline with a mass of 10 to 15 times the mass of the monkfish liver, and 20kHz to 24kHz Ultrasonic immersion for 1h to 2h, centrifugal separation at 2000r / min to 3000r / min for 10min to 20min, and collecting the supernatant; adding 3.5mol / L to 4.0mol / L ammonium sulfate aqueous solution to the supernatant, and continuously stirring until the protein is completely precipitated; centrifugal separation at 4000r / min to 5000r / min for 10min to 20min, collecting the precipitate, freeze-drying, and breaking up to obtain anglerfish liver protein.

[0015] In the above technical scheme, the preparation method of the Tribulus terrestris enzymatic hydrolysate is:

[0016] Fresh Tribulus terrestris is crushed, a phosphate buffer solution with a pH value of 5.0 to 5.5 and a value of 40 mM to 50 mM is added, hemicellulase and pectinase are added, enzymolysis is carried out at 45 to 50° C. for 2 to 3 hours, the enzyme is inactivated at high temperature, the temperature is lowered to room temperature, the pH value is adjusted to 6.0 to 6.5, β-glucosidase and β-galactosidase are added, enzymolysis is carried out at 35 to 40° C. for 3 hours to 4 hours, the enzyme is inactivated at high temperature, the temperature is lowered to room temperature, bromelain and papain are added, enzymolysis is carried out at 50 to 55° C. for 2 hours to 3 hours, the enzyme is inactivated at high temperature, the temperature is lowered to room temperature, the pH value is adjusted to 7.5 to 8.0, subtilisin is added, enzymolysis is carried out at 36 to 38° C. for 1.5 hours to 2 hours, the enzyme is inactivated at high temperature, the temperature is lowered to room temperature, a centrifugal ultrafiltration tube with a 3 kDa ultrafiltration membrane is used to perform centrifugal filtration to collect a filtrate with a content less than 3 kDa, and the filtrate is freeze-dried in vacuum and broken up to obtain a Tribulus terrestris hydrolysate.

[0017] In the preparation method of the above-mentioned enzymatic hydrolysate of Tribulus terrestris, the added mass of the phosphate buffer is 10 to 12 times that of Tribulus terrestris; the added mass of the hemicellulase is 1.0% to 2.0% of that of Tribulus terrestris, and the added mass of the pectinase is 0.5% to 1.0% of that of Tribulus terrestris; the added mass of the β-glucosidase is 0.5% to 1.0% of that of Tribulus terrestris, and the added mass of the β-galactosidase is 0.5% to 1.0% of that of Tribulus terrestris; the added mass of the bromelain is 1.0% to 2.0% of that of Tribulus terrestris, and the added mass of the papain is 1.0% to 2.0% of that of Tribulus terrestris; the added mass of the subtilisin is 0.5% to 1.5% of that of Tribulus terrestris; the temperature of the high-temperature enzyme inactivation is 90° C. to 100° C., and the time is 10 min to 15 min.

[0018] In the preparation method of the above-mentioned enzymatic hydrolysate of Tribulus terrestris, the enzymatic activities of the hemicellulase, β-glucosidase, bromelain and papain are all 100,000 U / g to 200,000 U / g; the enzymatic activity of the pectinase is 60,000 U / g to 100,000 U / g; the enzymatic activities of the β-galactosidase and subtilisin are all 10,000 U / g to 20,000 U / g.

[0019] The method for preparing the above-mentioned selenomethionine-containing composition comprises the following steps:

[0020] L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymatic hydrolysate are uniformly mixed according to mass proportions to obtain a composition.

[0021] The selenomethionine-containing composition is used for preparing a medicine for treating liver fibrosis.

[0022] The selenomethionine-containing composition provided by the present invention and its preparation method and application have the following beneficial effects compared with the prior art:

[0023] 1. The metabolism of selenium in the human body depends on its specific chemical form. Inorganic and organic forms are incorporated into proteins through different pathways and rates, thereby affecting cell function. The present invention uses L-selenomethionine and L-seleno-methylselenocysteine ​​as selenium sources, which are the safest organic selenium components, are more easily absorbed by cells, improve the absorption amount and absorption rate, increase the enrichment in the liver, and control the intake to ensure that the selenium content of the human body is not exceeded, and selenium poisoning will not occur. Among them, L-selenomethionine is absorbed quickly, but L-selenomethionine cannot be non-specifically incorporated into proteins, and methionine-tRNA cannot distinguish methionine from L-selenomethionine. When methionine intake is restricted, a large proportion of L-selenomethionine replaces methionine and is non-specifically incorporated into proteins in the animal body. Therefore, the amount of L-selenomethionine is difficult to control, and there is a risk of selenium poisoning. The amount of L-selenomethionine used in the present invention is relatively small, which can effectively prevent L-selenomethionine poisoning; but the small amount of L-selenomethionine will lead to a small amount of liver enrichment, and a good improvement effect of liver fibrosis cannot be achieved. Therefore, L-seleno-methylselenocysteine ​​is added as a good supplement to achieve orderly absorption. L-seleno-methylselenocysteine ​​is directly converted into methyl selenide by degradation by β-lyase, which is easy to control absorption. Compared with L-selenomethionine, L-seleno-methylselenocysteine ​​has higher safety, can well control absorption, and prevent selenium poisoning.

[0024] 2. L-theanine is added to the composition of the present invention. L-theanine is a unique amino acid found in tea trees, which can relieve stress, promote relaxation, improve organ vitality and increase mental energy. L-theanine is used in combination with L-selenomethionine and L-seleno-methylselenocysteine ​​to promote the absorption of L-selenomethionine and L-seleno-methylselenocysteine, thereby promoting an increase in selenium content in serum and liver, and improving the therapeutic effect of liver fibrosis.

[0025] 3. Anglerfish liver is a non-toxic fish liver that can promote metabolism, accelerate the liver's excretion of toxins in the body, and play a role in clearing the liver, improving eyesight, and replenishing qi and blood. The addition of Anglerfish liver peptides in the composition of the present invention is to extract part of the Anglerfish liver protein, use trypsin to enzymatically hydrolyze, then extract peptides between 3kDa and 5kDa, and then use subtilisin and papain for graded enzymatic hydrolysis to extract small molecule peptides less than 2kDa. The obtained Anglerfish liver peptides have a good therapeutic effect on liver fibrosis, which is incomparable to general liver extracts.

[0026] Four, Tribulus terrestris is a medicine for calming the liver and relieving depression, promoting blood circulation and dispelling wind, and its ingredients can be used to treat symptoms such as liver qi stagnation, chest and flank discomfort, liver carbuncle, and hepatic atrophy. But the treatment methods for Tribulus terrestris are different, and the treatment direction and effect are also different. The present invention uses hemicellulase and pectinase to hydrolyze Tribulus terrestris, then uses β-glucosidase and β-galactosidase to hydrolyze, then uses bromelain and papain to hydrolyze, and finally uses subtilisin to hydrolyze, and obtains a small molecule extract less than 2kDa, wherein it contains various small molecule active substances, and can effectively eliminate the undesirable components of liver in Tribulus terrestris. Through experimental detection, the Tribulus terrestris hydrolysate prepared by the present invention has an outstanding liver fibrosis treatment effect, and the Tribulus terrestris hydrolysate prepared by different enzymolysis methods has different liver fibrosis treatment effects.

[0027] 5. The present invention adopts L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymatic hydrolysate for use, which has a good synergistic effect. Low-dose treatment can also achieve a very excellent liver fibrosis treatment effect, can significantly reduce the change degree of ALT, AST, ALP, HA, PC-III, LN, LPS content, effectively alleviate the development of liver fibrosis, and Se in serum and liver 4+ Properly increasing the content can achieve higher immunity and improve the liver's antiviral ability. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a photo of the liver status of a mouse in the test group of Example 1 after 12 weeks;

[0029] Figure 2 This is a photo of the liver status of a mouse in the model control group after 12 weeks. DETAILED DESCRIPTION

[0030] The present invention is further described below in conjunction with specific implementation cases and drawings, but the present invention is not limited to these embodiments.

[0031] Example 1

[0032] A selenomethionine-containing composition is prepared from the following raw materials in parts by mass: 0.03 parts of L-selenomethionine, 0.05 parts of L-seleno-methylselenocysteine, 0.03 parts of L-theanine, 2 parts of monkfish liver peptide, and 6 parts of tribulus terrestris enzymatic hydrolysate.

[0033] Wherein, the preparation method of anglerfish liver peptide is:

[0034] S1: add the monkfish liver protein to a pH 7.8, 45 mM phosphate buffer solution with a mass of 9 times the monkfish liver protein, add 1.0% trypsin with the mass of the monkfish liver protein, perform enzymatic hydrolysis at 37°C for 2.5 h, inactivate the enzyme at 95°C for 12 min, cool to room temperature, and use a centrifugal ultrafiltration tube with a molecular weight cutoff of 5 kDa and 3 kDa ultrafiltration membrane to collect the filtrate between 3 kDa and 5 kDa by centrifugal filtration;

[0035] S2: Add 9 times the amount of monkfish liver protein in pH7.8, 50mM phosphate buffer to the filtrate, add 1.2% of the amount of monkfish liver protein in subtilisin, perform enzymolysis at 37°C for 1.5h, inactivate the enzyme at 95°C for 10min, cool to room temperature, add 1.2% of the amount of monkfish liver protein in papain, perform enzymolysis at 50°C for 2h, inactivate the enzyme at 95°C for 10min, cool to room temperature, use a centrifugal ultrafiltration tube with a 2kDa ultrafiltration membrane to centrifuge and collect the filtrate less than 2kDa, freeze-dry in vacuum, and break up to obtain monkfish liver peptide.

[0036] Among them, the method for extracting monkfish liver protein is as follows: take freshly washed monkfish liver, crush it, add it into 32°C warm water with a mass of 9 times that of monkfish liver, soak it for 8 hours, remove the water layer above the monkfish liver precipitate, drain the monkfish liver precipitate, add 0.15mol / L saline with a mass of 12 times that of monkfish liver, soak it for 1.5 hours under 20kHz ultrasound, centrifuge it at 2500r / min for 15 minutes, collect the supernatant; add 4.0mol / L ammonium sulfate aqueous solution to the supernatant, continue stirring until the protein is completely precipitated; centrifuge it at 4500r / min for 15 minutes, collect the precipitate, freeze-dry it, and break it up to obtain monkfish liver protein.

[0037] Wherein, the preparation method of Tribulus terrestris enzymolysate is:

[0038] Fresh Tribulus terrestris was crushed, 11 times the mass of Tribulus terrestris was added with pH 5.2, 45 mM phosphate buffer, 1.5% hemicellulase by mass of Tribulus terrestris and 0.8% pectinase by mass of Tribulus terrestris were added, enzymolysis was carried out at 48°C for 2.5 h, enzyme was inactivated at 95°C for 10 min, the temperature was lowered to room temperature, pH was adjusted to 6.2, 0.8% β-glucosidase by mass of Tribulus terrestris and 0.8% β-galactosidase by mass of Tribulus terrestris were added, enzymolysis was carried out at 37°C for 3.5 h, enzyme was inactivated at 95°C for 10 min, the temperature was lowered to room temperature, and the temperature was adjusted to 5.2. The mixture was heated to 40 ℃, 10 ℃ of temperature was added, 1.5% bromelain by weight of Tribulus terrestris and 1.5% papain by weight of Tribulus terrestris were added, and the mixture was enzymolyzed at 52 ℃ for 2.5 h, and the enzyme was inactivated at 95 ℃ for 10 min. The mixture was cooled to room temperature, and the pH was adjusted to 7.8. 1.0% subtilisin by weight of Tribulus terrestris was added, and the mixture was enzymolyzed at 37 ℃ for 1.5 h, and the enzyme was inactivated at 95 ℃ for 10 min. The mixture was cooled to room temperature, and a centrifugal ultrafiltration tube with a 3 kDa ultrafiltration membrane was used to collect the filtrate with a content less than 3 kDa by centrifugal filtration. The filtrate was freeze-dried in vacuum and broken up to obtain a Tribulus terrestris hydrolyzate.

[0039] The enzymatic activities of the hemicellulase, β-glucosidase, bromelain and papain used in this embodiment are all 100,000 U / g; the enzymatic activity of the pectinase is 60,000 U / g; the enzymatic activities of the β-galactosidase and the subtilisin are all 10,000 U / g.

[0040] The method for preparing the above-mentioned selenomethionine-containing composition comprises the following steps:

[0041] L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymatic hydrolysate are uniformly mixed according to mass proportions to obtain a composition.

[0042] The selenomethionine-containing composition is used for preparing a medicine for treating liver fibrosis.

[0043] Example 2

[0044] A selenomethionine-containing composition is prepared from the following raw materials in parts by mass: 0.02 parts of L-selenomethionine, 0.03 parts of L-seleno-methylselenocysteine, 0.02 parts of L-theanine, 1 part of monkfish liver peptide, and 4 parts of tribulus terrestris enzymatic hydrolysate.

[0045] Wherein, the preparation method of anglerfish liver peptide is:

[0046] S1: add the monkfish liver protein to a pH 7.5, 40 mM phosphate buffer solution with a mass of 8 times the monkfish liver protein, add trypsin with a mass of 0.5% of the monkfish liver protein, perform enzymatic hydrolysis at 36°C for 2 hours, inactivate the enzyme at 90°C for 10 minutes, cool to room temperature, and use a centrifugal ultrafiltration tube with a molecular weight cutoff of 5 kDa and 3 kDa ultrafiltration membrane to collect the filtrate between 3 kDa and 5 kDa by centrifugal filtration;

[0047] S2: Add 8 times the amount of monkfish liver protein in pH7.5, 40mM phosphate buffer to the filtrate, add 1% of the amount of monkfish liver protein in subtilisin, perform enzymolysis at 36°C for 1.5h, inactivate the enzyme at 90°C for 10min, cool to room temperature, add 1% of the amount of monkfish liver protein in papain, perform enzymolysis at 50°C-55°C for 1.5h, inactivate the enzyme at 90°C for 10min, cool to room temperature, use a centrifugal ultrafiltration tube with a 2kDa ultrafiltration membrane to centrifuge and collect the filtrate less than 2kDa, freeze-dry in vacuum, and break up to obtain monkfish liver peptide.

[0048] Among them, the method for extracting monkfish liver protein is as follows: take freshly washed monkfish liver, crush it, add it into 30°C warm water with a mass of 8 times that of monkfish liver, soak it for 6 hours, remove the water layer above the monkfish liver precipitate, drain the monkfish liver precipitate, add 0.12mol / L saline with a mass of 10 times that of monkfish liver, soak it for 1 hour under 20kHz ultrasound, centrifuge it at 2000r / min for 10 minutes, collect the supernatant; add 3.5mol / L ammonium sulfate aqueous solution to the supernatant, continue stirring until the protein is completely precipitated; centrifuge it at 4000r / min for 10 minutes, collect the precipitate, freeze-dry it, and break it up to obtain monkfish liver protein.

[0049] Wherein, the preparation method of Tribulus terrestris enzymolysate is:

[0050] Fresh Tribulus terrestris was crushed, 10 times the mass of Tribulus terrestris with pH 5.0 and 40 mM phosphate buffer was added, 1.0% hemicellulase by mass of Tribulus terrestris and 0.5% pectinase by mass of Tribulus terrestris were added, enzymolysis was carried out at 45°C for 2 h, enzyme was inactivated at 90°C for 10 min, the temperature was lowered to room temperature, pH was adjusted to 6.0, 0.5% β-glucosidase by mass of Tribulus terrestris and 0.5% β-galactosidase by mass of Tribulus terrestris were added, enzymolysis was carried out at 35°C for 3 h, enzyme was inactivated at 90°C for 10 min, the temperature was lowered to room temperature , add 1.0% bromelain and 1.0% papain by weight of Tribulus terrestris, enzymolysis at 50°C for 2h, inactivate the enzyme at 90°C for 10min, cool to room temperature, adjust pH to 7.5, add 0.5% subtilisin by weight of Tribulus terrestris, enzymolysis at 36°C for 1.5h, inactivate the enzyme at 90°C for 10min, cool to room temperature, use a centrifugal ultrafiltration tube with a 3kDa ultrafiltration membrane to centrifuge and collect the filtrate less than 3kDa, vacuum freeze-dry, and break up to obtain Tribulus terrestris hydrolysate.

[0051] The enzymatic activities of the hemicellulase, β-glucosidase, bromelain and papain used in this embodiment are all 100,000 U / g; the enzymatic activity of the pectinase is 60,000 U / g; the enzymatic activities of the β-galactosidase and the subtilisin are all 10,000 U / g.

[0052] The method for preparing the above-mentioned selenomethionine-containing composition comprises the following steps:

[0053] L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymatic hydrolysate are uniformly mixed according to mass proportions to obtain a composition.

[0054] The selenomethionine-containing composition is used for preparing a medicine for treating liver fibrosis.

[0055] Example 3

[0056] A selenomethionine-containing composition is prepared from the following raw materials in parts by mass: 0.03 parts of L-selenomethionine, 0.04 parts of L-seleno-methylselenocysteine, 0.03 parts of L-theanine, 1.5 parts of monkfish liver peptide, and 5 parts of tribulus terrestris enzymatic hydrolysate.

[0057] Wherein, the preparation method of anglerfish liver peptide is:

[0058] S1: add the monkfish liver protein to a pH 7.5, 40 mM phosphate buffer solution with a mass of 8 times the monkfish liver protein, add trypsin with a mass of 0.8% of the monkfish liver protein, perform enzymatic hydrolysis at 37°C for 2 h, inactivate the enzyme at 90°C for 10 min, cool to room temperature, and use a centrifugal ultrafiltration tube with a molecular weight cutoff of 5 kDa and 3 kDa ultrafiltration membrane to collect the filtrate between 3 kDa and 5 kDa by centrifugal filtration;

[0059] S2: Add 8 times the amount of monkfish liver protein in pH7.6, 50mM phosphate buffer to the filtrate, add 1% of the amount of monkfish liver protein in subtilisin, perform enzymolysis at 37°C for 1.5h, inactivate the enzyme at 90°C for 10min, cool to room temperature, add 1% of the amount of monkfish liver protein in papain, perform enzymolysis at 55°C for 2h, inactivate the enzyme at 90°C for 10min, cool to room temperature, use a centrifugal ultrafiltration tube with a 2kDa ultrafiltration membrane to centrifuge and collect the filtrate less than 2kDa, freeze-dry in vacuum, and break up to obtain monkfish liver peptide.

[0060] Among them, the method for extracting monkfish liver protein is as follows: take freshly washed monkfish liver, crush it, add it into 30°C warm water with a mass of 8 times that of monkfish liver, soak it for 7 hours, remove the water layer above the monkfish liver precipitate, drain the monkfish liver precipitate, add 0.15mol / L saline with a mass of 12 times that of monkfish liver, soak it for 1 hour under 20kHz ultrasound, centrifuge it at 2000r / min for 10 minutes, collect the supernatant; add 3.5mol / L ammonium sulfate aqueous solution to the supernatant, continue stirring until the protein is completely precipitated; centrifuge it at 4000r / min for 10 minutes, collect the precipitate, freeze-dry it, and break it up to obtain monkfish liver protein.

[0061] Wherein, the preparation method of Tribulus terrestris enzymolysate is:

[0062] Fresh Tribulus terrestris was crushed, 10 times the mass of Tribulus terrestris was added with pH 5.0, 40 mM phosphate buffer, 1.2% hemicellulase by mass of Tribulus terrestris and 0.6% pectinase by mass of Tribulus terrestris were added, enzymolysis was carried out at 45°C for 2 hours, enzyme was inactivated at 90°C for 10 minutes, the temperature was lowered to room temperature, pH was adjusted to 6.0, 0.6% β-glucosidase by mass of Tribulus terrestris and 0.6% β-galactosidase by mass of Tribulus terrestris were added, enzymolysis was carried out at 35°C for 3 hours, enzyme was inactivated at 90°C for 10 minutes, the temperature was lowered to room temperature, Add 1.2% bromelain and 1.2% papain by weight of the tribulus terrestris, perform enzymolysis at 52°C for 2.5 hours, inactivate the enzyme at 90°C for 10 minutes, cool to room temperature, adjust the pH to 7.5, add 0.8% subtilisin by weight of the tribulus terrestris, perform enzymolysis at 37°C for 1.5 hours, inactivate the enzyme at 90°C for 10 minutes, cool to room temperature, use a centrifugal ultrafiltration tube with a 3kDa ultrafiltration membrane to perform centrifugal filtration to collect a filtrate with a content less than 3kDa, perform vacuum freeze drying, and break up to obtain a tribulus terrestris hydrolysate.

[0063] The enzymatic activities of the hemicellulase, β-glucosidase, bromelain and papain used in this embodiment are all 100,000 U / g; the enzymatic activity of the pectinase is 60,000 U / g; the enzymatic activities of the β-galactosidase and the subtilisin are all 10,000 U / g.

[0064] The method for preparing the above-mentioned selenomethionine-containing composition comprises the following steps:

[0065] L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymatic hydrolysate are uniformly mixed according to mass proportions to obtain a composition.

[0066] The selenomethionine-containing composition is used for preparing a medicine for treating liver fibrosis.

[0067] Example 4

[0068] A selenomethionine-containing composition is prepared from the following raw materials in parts by mass: 0.05 parts of L-selenomethionine, 0.08 parts of L-seleno-methylselenocysteine, 0.05 parts of L-theanine, 3 parts of monkfish liver peptide, and 8 parts of tribulus terrestris enzymatic hydrolysate.

[0069] Wherein, the preparation method of anglerfish liver peptide is:

[0070] S1: add the monkfish liver protein to a pH 8.0, 50 mM phosphate buffer solution with a mass 10 times that of the monkfish liver protein, add trypsin with a mass 1.5% of the monkfish liver protein, perform enzymatic hydrolysis at 38°C for 3 h, inactivate the enzyme at 100°C for 15 min, cool to room temperature, and use a centrifugal ultrafiltration tube with a molecular weight cutoff of 5 kDa and 3 kDa ultrafiltration membrane to collect the filtrate between 3 kDa and 5 kDa by centrifugal filtration;

[0071] S2: Add 10 times the amount of monkfish liver protein in pH 8.0, 50 mM phosphate buffer to the filtrate, add 1.5% of the amount of monkfish liver protein in subtilisin, perform enzymolysis at 38°C for 2 hours, inactivate the enzyme at 100°C for 15 minutes, cool to room temperature, add 1.5% of the amount of monkfish liver protein in papain, perform enzymolysis at 55°C for 2 hours, inactivate the enzyme at 100°C for 15 minutes, cool to room temperature, use a centrifugal ultrafiltration tube with a 2 kDa ultrafiltration membrane to perform centrifugal filtration to collect the filtrate less than 2 kDa, perform vacuum freeze drying, and break up to obtain monkfish liver peptide.

[0072] The method for extracting the monkfish liver protein is as follows: take the freshly washed monkfish liver, crush it, add it into 35°C warm water with a mass 10 times that of the monkfish liver, soak it for 10 hours, remove the water layer above the monkfish liver precipitate, drain the monkfish liver precipitate, add 0.15mol / L saline with a mass 15 times that of the monkfish liver, soak it for 2 hours under 24kHz ultrasound, centrifuge it at 3000r / min for 20 minutes, collect the supernatant; add 4.0mol / L ammonium sulfate aqueous solution to the supernatant, continue stirring until the protein is completely precipitated; centrifuge it at 5000r / min for 20 minutes, collect the precipitate, freeze-dry it, and break it up to obtain the monkfish liver protein.

[0073] Wherein, the preparation method of Tribulus terrestris enzymolysate is:

[0074] Fresh Tribulus terrestris was crushed, 12 times the mass of Tribulus terrestris was added with pH 5.5, 50 mM phosphate buffer, 2.0% hemicellulase by mass of Tribulus terrestris and 1.0% pectinase by mass of Tribulus terrestris were added, enzymolysis was carried out at 50°C for 3 h, enzyme was inactivated at 100°C for 15 min, the temperature was lowered to room temperature, pH was adjusted to 6.5, 1.0% β-glucosidase by mass of Tribulus terrestris and 1.0% β-galactosidase by mass of Tribulus terrestris were added, enzymolysis was carried out at 40°C for 4 h, enzyme was inactivated at 100°C for 15 min, the temperature was lowered to room temperature, the temperature was adjusted to 5.5, the pH value was adjusted to 6.5, 1.0% β-glucosidase by mass of Tribulus terrestris and 1.0% β-galactosidase by mass of Tribulus terrestris were added, the temperature was adjusted to 5.5, the temperature was adjusted to 5.5, the temperature was adjusted to 6.5, the pH value was adjusted to 6.5, 1.0% β-glucosidase by mass of Tribulus terrestris and 1.0% β-galactosidase by mass of Tribulus terrestris were added, the temperature was adjusted to 5.5 ... The mixture was heated to 40 ℃ and heated to 80 ℃, 2.0% bromelain and 2.0% papain by weight of Tribulus terrestris were added, and the mixture was enzymolyzed at 55 ℃ for 3 h, and the enzyme was inactivated at 100 ℃ for 15 min. The mixture was cooled to room temperature, and the pH value was adjusted to 8.0. Subtilisin with a weight of 1.5% of the Tribulus terrestris was added, and the mixture was enzymolyzed at 38 ℃ for 2 h, and the enzyme was inactivated at 100 ℃ for 15 min. The mixture was cooled to room temperature, and a centrifugal ultrafiltration tube with a 3 kDa ultrafiltration membrane was used to collect the filtrate with a weight less than 3 kDa by centrifugal filtration, and the filtrate was freeze-dried in vacuum and broken up to obtain a Tribulus terrestris hydrolyzate.

[0075] The enzymatic activities of the hemicellulase, β-glucosidase, bromelain and papain used in this embodiment are all 200,000 U / g; the enzymatic activity of the pectinase is 100,000 U / g; the enzymatic activities of the β-galactosidase and the subtilisin are all 20,000 U / g.

[0076] The method for preparing the above-mentioned selenomethionine-containing composition comprises the following steps:

[0077] L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymatic hydrolysate are uniformly mixed according to mass proportions to obtain a composition.

[0078] The selenomethionine-containing composition is used for preparing a medicine for treating liver fibrosis.

[0079] Example 5

[0080] A selenomethionine-containing composition is prepared from the following raw materials in parts by mass: 0.04 parts of L-selenomethionine, 0.07 parts of L-seleno-methylselenocysteine, 0.04 parts of L-theanine, 2.5 parts of monkfish liver peptide, and 7 parts of tribulus terrestris enzymatic hydrolysate.

[0081] Wherein, the preparation method of anglerfish liver peptide is:

[0082] S1: add the monkfish liver protein to a pH 8.0, 50 mM phosphate buffer solution with a mass 10 times that of the monkfish liver protein, add trypsin with a mass 1.4% of the monkfish liver protein, perform enzymatic hydrolysis at 37°C for 3 h, inactivate the enzyme at 100°C for 15 min, cool to room temperature, and use a centrifugal ultrafiltration tube with a molecular weight cutoff of 5 kDa and 3 kDa ultrafiltration membrane to collect the filtrate between 3 kDa and 5 kDa by centrifugal filtration;

[0083] S2: Add 10 times the amount of monkfish liver protein in pH7.5, 40mM phosphate buffer to the filtrate, add 1.4% of the amount of monkfish liver protein in subtilisin, perform enzymolysis at 37°C for 2h, inactivate the enzyme at 100°C for 15min, cool to room temperature, add 1.4% of the amount of monkfish liver protein in papain, perform enzymolysis at 50°C for 1.5h, inactivate the enzyme at 100°C for 15min, cool to room temperature, use a centrifugal ultrafiltration tube with a 2kDa ultrafiltration membrane to perform centrifugal filtration to collect the filtrate less than 2kDa, freeze-dry in vacuum, and break up to obtain monkfish liver peptide.

[0084] Among them, the method for extracting monkfish liver protein is as follows: take freshly washed monkfish liver, crush it, add it into 35°C warm water with a mass 10 times that of the monkfish liver, soak it for 10 hours, remove the water layer above the monkfish liver precipitate, drain the monkfish liver precipitate, add 0.15mol / L saline with a mass 15 times that of the monkfish liver, soak it for 2 hours under 24kHz ultrasound, centrifuge it at 3000r / min for 10 minutes, collect the supernatant; add 4.0mol / L ammonium sulfate aqueous solution to the supernatant, continue stirring until the protein is completely precipitated; centrifuge it at 4000r / min for 10 minutes, collect the precipitate, freeze-dry it, and break it up to obtain monkfish liver protein.

[0085] Wherein, the preparation method of Tribulus terrestris enzymolysate is:

[0086] Fresh Tribulus terrestris was crushed, 10 times the mass of Tribulus terrestris was added with pH 5.3, 50 mM phosphate buffer, 1.8% hemicellulase by mass of Tribulus terrestris and 0.8% pectinase by mass of Tribulus terrestris were added, enzymolysis was carried out at 50°C for 3 h, enzyme was inactivated at 100°C for 10 min, the temperature was lowered to room temperature, pH was adjusted to 6.4, 0.8% β-glucosidase by mass of Tribulus terrestris and 0.8% β-galactosidase by mass of Tribulus terrestris were added, enzymolysis was carried out at 40°C for 3 h, enzyme was inactivated at 100°C for 10 min, the temperature was lowered to room temperature , add 1.8% bromelain and 1.6% papain by weight of Tribulus terrestris, enzymolysis at 55°C for 2h, inactivate the enzyme at 100°C for 10min, cool to room temperature, adjust pH to 7.6, add 1.2% subtilisin by weight of Tribulus terrestris, enzymolysis at 37°C for 1.5h, inactivate the enzyme at 100°C for 10min, cool to room temperature, use a centrifugal ultrafiltration tube with a 3kDa ultrafiltration membrane to centrifuge and collect the filtrate less than 3kDa, vacuum freeze-dry, and break up to obtain Tribulus terrestris hydrolysate.

[0087] The enzymatic activities of hemicellulase, β-glucosidase, bromelain and papain used in this embodiment are all 200,000 U / g; the enzymatic activity of pectinase is 60,000 U / g; the enzymatic activities of β-galactosidase and subtilisin are both 20,000 U / g.

[0088] The method for preparing the above-mentioned selenomethionine-containing composition comprises the following steps:

[0089] L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymatic hydrolysate are uniformly mixed according to mass proportions to obtain a composition.

[0090] The selenomethionine-containing composition is used for preparing a medicine for treating liver fibrosis.

[0091] Comparative Example 1

[0092] L-selenomethionine and L-seleno-methylselenocysteine ​​were replaced by selenium-enriched yeast (effective substance content 2000 ppm); other parameters and methods were the same as in Example 1.

[0093] Comparative Example 2

[0094] No L-theanine was added, and the L-theanine component was replaced by starch; other parameters and methods were the same as in Example 1.

[0095] Comparative Example 3

[0096] No monkfish liver peptide was added, and the monkfish liver peptide acid component was replaced by starch; other parameters and methods were the same as in Example 1.

[0097] Comparative Example 4

[0098] No Tribulus terrestris hydrolysate was added, and the Tribulus terrestris hydrolysate component was replaced by starch; other parameters and methods were the same as in Example 1.

[0099] Comparative Example 5

[0100] The preparation method of anglerfish liver peptide is as follows:

[0101] S1: Add the monkfish liver protein to a pH 7.8, 45 mM phosphate buffer solution with a mass of 9 times that of the monkfish liver protein, add trypsin with a mass of 1.0% of the monkfish liver protein, perform enzymatic hydrolysis at 37° C. for 2.5 h, inactivate the enzyme at 95° C. for 12 min, cool to room temperature, use a centrifugal ultrafiltration tube with a molecular weight cutoff of 5 kDa and 3 kDa ultrafiltration membrane, perform centrifugal filtration to collect the filtrate between 3 kDa and 5 kDa, vacuum freeze-dry, and break up to obtain monkfish liver peptide; other parameters and methods are the same as those in Example 1.

[0102] Comparative Example 6

[0103] The preparation method of Tribulus terrestris enzymatic hydrolysate is as follows:

[0104] Fresh Tribulus terrestris was crushed, 11 times the mass of Tribulus terrestris with a pH of 5.2 and 45 mM phosphate buffer was added, 1.5% of the mass of Tribulus terrestris hemicellulase and 0.8% of the mass of Tribulus terrestris pectin were added, and the mixture was hydrolyzed at 48° C. for 2.5 h, the enzyme was inactivated at 95° C. for 10 min, the mixture was cooled to room temperature, the pH was adjusted to 7.8, 1.0% of the mass of Tribulus terrestris subtilisin was added, the mixture was hydrolyzed at 37° C. for 1.5 h, the enzyme was inactivated at 95° C. for 10 min, the mixture was cooled to room temperature, a centrifugal ultrafiltration tube with a 3 kDa ultrafiltration membrane was used to centrifuge and collect the filtrate with a mass less than 3 kDa, the mixture was freeze-dried in vacuum, and the mixture was broken up to obtain a Tribulus terrestris hydrolyzate; other parameters and methods were the same as those in Example 1.

[0105] The following experimental tests were performed on the compositions of the above embodiments and the compositions of the comparative examples.

[0106] 1. Experimental Animals:

[0107] 104 8-week-old male C57 mice (purchased from Wuhan Shulaibao Biotechnology Co., Ltd.), weighing 20±2.0g.

[0108] 2. Experimental instruments and equipment:

[0109] Electronic balance, freezing table, dehydration, embedding machine, pathology slicer, tissue spreader, oven, cover slip, slide, refrigerated centrifuge, vortex mixer, pipette gun, decolorization shaker, histochemical pen, microscope, polarizing microscope, imaging system.

[0110] 3. Modeling method:

[0111] C57 mice were randomly divided into 13 groups (8 mice in each group): a blank control group, a model control group and 12 test groups (each group corresponds to the composition of the gavage embodiment and the comparative example), all mice were free to drink water and take standard mouse food at room temperature, and the experiment was conducted after 7 days of adaptive feeding. Thioacetamide (TAA) was prepared into a solution with a mass concentration of 40 mg / ml with normal saline. The mice in the blank control group were raised normally for 8 weeks. The model control group and the test group were intraperitoneally injected with 200 mg / kg TAA every other day for 8 weeks.

[0112] 4. Treatment methods:

[0113] Intragastric administration and modeling were carried out simultaneously. For 8 weeks, the test group was intragastrically administered with 0.02 mg / kg of the composition sample twice a day; the blank control group and the model control group were not intragastrically administered with the composition sample.

[0114] After 8 weeks, 2 mice were randomly selected from each group for HE staining to evaluate whether the mice in the model control group had liver fibrosis. The model was established and the respective intervention measures were continued. After the 12th week, all mice in each group were killed to obtain serum and liver samples for the following index detection.

[0115] V. Testing indicators and methods:

[0116] Microplate assay: changes in serum ALT, AST, ALP, HA, PC-III, and LN;

[0117] Elisa method detection: Inflammatory factors in serum: LPS;

[0118] IPC-MS detection: serum and liver tissue selenium ion Se 4+ Content situation.

[0119] 6. Statistics of test results:

[0120] The test results of various indicators are shown in Table 1 below.

[0121] Table 1 Test results (average value)

[0122]

[0123]

[0124] From the above results, it can be seen that the compositions of Examples 1 to 5 can effectively improve the development of liver fibrosis. Compared with the model control group, the changes in the contents of ALT, AST, ALP, HA, PC-III, LN, and LPS were significantly reduced, and the levels of Se in serum and liver were significantly reduced. 4+The content is appropriately increased, and within a reasonable range, there is no selenium poisoning phenomenon, proving that organic Se is easily absorbed, has a high absorption rate and is safe and reliable. As shown in the results of Comparative Example 1, selenium-enriched yeast shows a low absorption rate of selenium compared to L-selenomethionine and L-seleno-methylselenocysteine, and the selenium content in serum and liver is low, which affects the reduction of the therapeutic effect of liver fibrosis to a certain extent. As shown in the results of Comparative Example 2, L-theanine can promote liver vitality, promote the absorption of L-selenomethionine and L-seleno-methylselenocysteine, and then promote the increase of selenium content in serum and liver, and promote the improvement of the therapeutic effect of liver fibrosis. As shown in the results of Comparative Examples 3 and 5, the anglerfish liver peptide prepared by graded enzymolysis of a specific enzyme in a specific order has a good effect on improving the therapeutic effect of liver fibrosis; and the anglerfish liver peptides prepared by different enzymolysis methods have a large difference in the effect of treating liver fibrosis, that is, the anglerfish liver peptide after omitting the enzymolysis step has little effect on the treatment of liver fibrosis. From the results of Comparative Examples 4 and 6, it can be seen that the Tribulus terrestris hydrolysate prepared by graded enzymolysis with various enzymes in a specific order has an outstanding therapeutic effect on liver fibrosis, and the therapeutic effects of the Tribulus terrestris hydrolysate prepared by different enzymolysis methods are different, that is, the Tribulus terrestris hydrolysate after omitting the enzymolysis step is significantly reduced in the treatment of liver fibrosis, and does not reach the therapeutic effect of the composition of Example 1. From the comparison of the results of Example 1 and each comparative example, it can be seen that

[0125] The combination of L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris hydrolysate has a good synergistic effect, and low-dose treatment (0.02 mg / kg composition sample) can also achieve very excellent liver fibrosis treatment effects.

[0126] in addition, Figure 1 The liver status of a mouse in the test group of Example 1 after 12 weeks is shown in FIG. Figure 2 The liver status of a mouse in the model control group after 12 weeks is shown in the figure; by comparing the two, it can be seen that the composition has a very obvious therapeutic effect on liver fibrosis and can significantly delay the progression of liver fibrosis.

Claims

1. A selenomethionine-containing composition, characterized in that: The composition is prepared from the following raw materials in parts by weight: 0.02 to 0.05 parts of L-selenomethionine, 0.03 to 0.08 parts of L-seleno-methylselenocysteine, 0.02 to 0.05 parts of L-theanine, 1 to 3 parts of monkfish liver peptide, and 4 to 8 parts of tribulus terrestris enzymatic hydrolysate; The preparation method of the anglerfish liver peptide is as follows: S1: add anglerfish liver protein to a phosphate buffer solution of pH 7.5-8.0, 40 mM-50 mM, add trypsin, perform enzymatic hydrolysis at 36°C-38°C for 2h-3h, inactivate the enzyme at high temperature, cool to room temperature, and use a centrifugal ultrafiltration tube with an ultrafiltration membrane of 5 kDa and 3 kDa molecular weight cutoff to collect the filtrate between 3 kDa and 5 kDa by centrifugal filtration; The method for extracting the monkfish liver protein comprises the following steps: taking a freshly washed monkfish liver, crushing it, adding it into 30-35°C warm water with a mass of 8-10 times the mass of the monkfish liver, soaking it for 6-10 hours, removing the water layer above the monkfish liver precipitation, draining the monkfish liver precipitation, adding 0.12mol / L-0.15mol / L saline with a mass of 10-15 times the mass of the monkfish liver, and soaking it for 1 hour under 20kHz-24kHz ultrasound. 2h, centrifuge at 2000r / min-3000r / min for 10min-20min, collect the supernatant; add 3.5mol / L-4.0mol / L ammonium sulfate aqueous solution to the supernatant, continue stirring until the protein is completely precipitated; centrifuge at 4000r / min-5000r / min for 10min-20min, collect the precipitate, freeze-dry, and break it up to obtain anglerfish liver protein; S2: Add phosphate buffer solution of pH 7.5-8.0 and 40mM-50mM to the filtrate, add subtilisin, perform enzymolysis at 36°C-38°C for 1.5h-2h, inactivate the enzyme at high temperature, cool to room temperature, add papain, perform enzymolysis at 50°C-55°C for 1.5h-2h, inactivate the enzyme at high temperature, cool to room temperature, use a centrifugal ultrafiltration tube with a 2kDa ultrafiltration membrane to centrifuge and collect the filtrate less than 2kDa, freeze-dry in vacuum, and break up to obtain anglerfish liver peptide; The preparation method of the Tribulus terrestris enzymolysate is: Fresh Tribulus terrestris is crushed, a phosphate buffer solution with a pH value of 5.0 to 5.5 and a value of 40 mM to 50 mM is added, hemicellulase and pectinase are added, enzymolysis is carried out at 45 to 50° C. for 2 to 3 hours, the enzyme is inactivated at high temperature, the temperature is lowered to room temperature, the pH value is adjusted to 6.0 to 6.5, β-glucosidase and β-galactosidase are added, enzymolysis is carried out at 35 to 40° C. for 3 hours to 4 hours, the enzyme is inactivated at high temperature, the temperature is lowered to room temperature, bromelain and papain are added, enzymolysis is carried out at 50 to 55° C. for 2 hours to 3 hours, the enzyme is inactivated at high temperature, the temperature is lowered to room temperature, the pH value is adjusted to 7.5 to 8.0, subtilisin is added, enzymolysis is carried out at 36 to 38° C. for 1.5 hours to 2 hours, the enzyme is inactivated at high temperature, the temperature is lowered to room temperature, a centrifugal ultrafiltration tube with a 3 kDa ultrafiltration membrane is used to perform centrifugal filtration to collect a filtrate with a content less than 3 kDa, and the filtrate is freeze-dried in vacuum and broken up to obtain a Tribulus terrestris hydrolysate.

2. A selenomethionine-containing composition according to claim 1, characterized in that: In S1 of the method for preparing anglerfish liver peptide, the added mass of the phosphate buffer is 8 to 10 times that of the anglerfish liver protein; the added mass of the trypsin is 0.5% to 1.5% of the anglerfish liver protein; the temperature of the high-temperature enzyme inactivation is 90° C. to 100° C., and the time is 10 min to 15 min.

3. A selenomethionine-containing composition according to claim 1, characterized in that: In S2 of the method for preparing monkfish liver peptide, the added mass of the phosphate buffer is 8 to 10 times that of the monkfish liver protein; the added mass of the subtilisin is 1% to 1.5% of the monkfish liver protein; the added mass of the papain is 1% to 1.5% of the monkfish liver protein; the temperature of the high-temperature enzyme inactivation is 90°C to 100°C, and the time is 10 min to 15 min.

4. A selenomethionine-containing composition according to claim 1, characterized in that: In the preparation method of the enzymatic hydrolysate of Tribulus terrestris, the added mass of the phosphate buffer is 10 to 12 times that of Tribulus terrestris; the added mass of the hemicellulase is 1.0% to 2.0% of that of Tribulus terrestris, and the added mass of the pectinase is 0.5% to 1.0% of that of Tribulus terrestris; the added mass of the β-glucosidase is 0.5% to 1.0% of that of Tribulus terrestris, and the added mass of the β-galactosidase is 0.5% to 1.0% of that of Tribulus terrestris; the added mass of the bromelain is 1.0% to 2.0% of that of Tribulus terrestris, and the added mass of the papain is 1.0% to 2.0% of that of Tribulus terrestris; the added mass of the subtilisin is 0.5% to 1.5% of that of Tribulus terrestris; the temperature of the high-temperature enzyme inactivation is 90° C. to 100° C., and the time is 10 min to 15 min.

5. A selenomethionine-containing composition according to claim 1, characterized in that: In the preparation method of the enzymatic hydrolysate of Tribulus terrestris, the enzymatic activities of the hemicellulase, β-glucosidase, bromelain and papain are all 100,000 U / g to 200,000 U / g; the enzymatic activity of the pectinase is 60,000 U / g to 100,000 U / g; and the enzymatic activities of the β-galactosidase and subtilisin are all 10,000 U / g to 20,000 U / g.

6. The method for preparing a selenomethionine-containing composition according to claim 1, characterized in that: The steps include: L-selenomethionine, L-seleno-methylselenocysteine, L-theanine, monkfish liver peptide and Tribulus terrestris enzymatic hydrolysate are uniformly mixed according to mass proportions to obtain a composition.

7. Use of the selenomethionine-containing composition according to claim 1 in preparing a medicament for treating liver fibrosis.

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