Preparation method of ganoderma lucidum ergothioneine extract and application of ganoderma lucidum ergothioneine extract in preparation of health care products with liver protection effect

Abstract

The invention relates to the technical field of functional foods and health-care products, in particular to a preparation method of a ganoderma lucidum ergothioneine extract and application of the ganoderma lucidum ergothioneine extract in preparation of health-care products with liver protection effects. Comprising the following steps: mixing ganoderma lucidum powder with a buffer solution, and adding enzymatic hydrolysate for enzymolysis; centrifuging the enzymolysis reaction liquid, and collecting supernate containing ergothioneine; concentrating the supernate containing the ergothioneine, performing alcohol precipitation treatment, and collecting alcohol precipitation supernate; enabling the alcohol precipitation supernate to pass through a macroporous adsorption resin column, eluting and then collecting eluent; and concentrating and drying the eluent to obtain the ganoderma lucidum ergothioneine extract. The ganoderma lucidum ergothioneine extract prepared by the invention can be used for preparing a composition with a liver protection effect, and a brand new composition which is high in bioavailability and can be distributed in a liver in a targeted manner is provided, so that multi-channel synergistic liver protection is realized, and the stability, the absorption efficiency and the actual liver protection effect of a product are remarkably improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of functional food and health products, and particularly relates to a preparation method of ganoderma ergothioneine extract and application of the ganoderma ergothioneine extract in preparation of health products with liver protection efficacy. BACKGROUND

[0002] As the most important metabolic and detoxification organ of the human body, the liver is continuously challenged by endogenous and exogenous factors. Alcohol abuse, irrational drug use, high-fat diet and environmental pollutants can all induce the liver to produce excessive reactive oxygen species, triggering oxidative stress, which is a common core pathological link of alcoholic liver disease, drug-induced liver injury and non-alcoholic fatty liver disease. Continuous oxidative stress attacks the cell membrane, proteins and DNA of liver cells, leading to inflammation, fatty degeneration and apoptosis of liver cells, and if not intervened, may gradually develop into liver fibrosis and cirrhosis, seriously endangering human health.

[0003] At present, the efficacy components of oral liver protection products on the market are mostly dependent on silybinin, glutathione, bicyclol or multiple plant extracts. These components have certain liver protection effects, but generally have some inherent defects. For example, the star component glutathione is easily degraded in the gastrointestinal tract after oral administration, and the liver cell uptake efficiency is limited, resulting in unsatisfactory bioavailability; and some active ingredients from plants have complex chemical structures, large fluctuations in effective content, and unclear mechanisms of action. These limitations make it difficult for some existing products to achieve the desired stable and potent effects.

[0004] In the process of seeking a solution, ergothioneine, a naturally occurring chiral amino acid base in some mushrooms and grains, shows great application potential. It is an extraordinary cell protector that not only has the ability to directly scavenge hydroxyl radicals, the strongest reactive oxygen species, but also effectively quenches singlet oxygen, with an antioxidant capacity far superior to many common antioxidants. More uniquely, ergothioneine can be efficiently taken up by cells through the specific transporter OCTN-1 and enriched in organs with high energy consumption and high oxidative stress risk, such as the liver, precisely exerting protective effects at the cellular and mitochondrial levels.

[0005] However, despite the stable nature and excellent efficacy of ergothioneine, its successful development as an efficient oral liver protection product still faces key challenges. First, ergothioneine is limited in its natural sources and is costly. Second, the action pathways of a single component are relatively limited, making it difficult to fully cover the multiple needs of liver protection. Finally, how to organically combine ergothioneine with other auxiliary components through scientific formula design and sophisticated preparation process to build a liver protection system that synergistically enhances absorption, targeting and multiple pharmacological activities is a technical problem that must be solved to improve the core competitiveness of the product and achieve industrialization. SUMMARY

[0006] The present application aims to solve the technical problems of low bioavailability, single target point, limited antioxidant effect and unstable core efficacy component of existing oral liver protection products, and provides a new composition with high bioavailability and ergothioneine as the core, which can be targeted to the liver, to realize multi-pathway synergistic liver protection and significantly improve the stability, absorption efficiency and actual liver protection effect of the product.

[0007] In order to achieve the above-mentioned application purposes, the present application provides the following technical solutions: A preparation method of ganoderma lucidum ergothioneine extract, comprising the following steps: (a) mixing ganoderma lucidum powder with buffer solution, adding enzyme solution, performing enzyme hydrolysis reaction, heating to inactivate the enzyme, and obtaining enzyme hydrolysis reaction liquid; (b) centrifuging the enzyme hydrolysis reaction liquid obtained in step (a) to collect supernatant containing ergothioneine; (c) after concentrating the supernatant containing ergothioneine, adding ethanol for alcohol precipitation treatment, and collecting alcohol precipitation supernatant; (d) passing the alcohol precipitation supernatant through a macroporous adsorption resin column, first eluting with water, then eluting with an ethanol solution with a concentration of 20-40%, and collecting the ethanol eluate; (e) concentrating and drying the collected ethanol eluate to obtain ganoderma lucidum ergothioneine extract.

[0008] Preferably, the enzyme solution in step (a) is: endo-β-1, 3-glucanase, endo-chitinase, β-glucosidase and subtilisin, and the mass ratio is 2.5-3.5:1.5-2.5:1-2:1. Preferably, the enzyme hydrolysis time in step (a) is 2-3h.

[0009] Preferably, the buffer solution in step (a) is acetic acid-sodium acetate buffer solution or citric acid-disodium hydrogen phosphate buffer solution.

[0010] Preferably, the mass ratio of ganoderma lucidum powder to buffer solution in step (a) is 1:6-12, and the mass ratio of buffer solution to enzyme hydrolysis solution is 100:2-3.

[0011] Preferably, the type of macroporous adsorption resin column in step (d) is one of HP20 type, HPD100 type, HPD300 type, D101 type or 816 type.

[0012] The application provides an oral liver-protecting composition containing ergothioneine, which comprises the following components in mass fractions: Ganoderma lucidum ergothioneine extract 35-45 parts, silymarin 4-7 parts, ginsenoside Rb1 2-5 parts, pueraria extract 4-7 parts, salvia miltiorrhiza extract (alcohol extract) 4-7 parts, turmeric extract (extracted by steam distillation) 2-5 parts, schisandrin B 2-5 parts and apioin 2-5 parts.

[0013] The application provides a preparation method of the oral liver-protecting composition containing ergothioneine.

[0014] The application provides an application of the oral liver-protecting composition containing ergothioneine in the preparation of health products with liver-protecting efficacy.

[0015] Preferably, the health products with liver-protecting efficacy are one or more of alcoholic liver injury maintenance tablets, chemical liver injury protection capsules, hyperlipidemia fatty liver conditioning pills, non-alcoholic fatty liver compound tablets, liver fibrosis early intervention products, liver protection essence capsules for staying up late, pre-drinks for liver protection, liver repair granules after drinking, drug liver auxiliary sustained-release capsules, liver and gallbladder comprehensive conditioning soft capsules, liver qi stagnation relieving pills, liver fire clearing tea, liver and eye homologous maintenance tablets, liver cirrhosis auxiliary nutritional powder and liver detoxification health-care pills.

[0016] Compared with the prior art, the application has the following beneficial effects: The preparation method of the Ganoderma lucidum ergothioneine extract provided by the application adopts a process route combining multi-enzyme synergistic enzymolysis and multi-step purification, and realizes efficient and directional extraction of ergothioneine in Ganoderma lucidum. The method adopts a composite enzymolysis solution composed of endo-β-1, 3-glucanase, endo-chitinase, β-glucosidase and subtilisin in a specific mass ratio, and performs enzymolysis on Ganoderma lucidum powder under suitable buffer system and time conditions, effectively destroys the cell wall structure of Ganoderma lucidum, and promotes the release of ergothioneine. The enzymolysis strategy not only improves the extraction efficiency, but also avoids the damage to active ingredients caused by traditional high-temperature or strong solvent treatment, and has the characteristics of mild conditions and strong specificity.

[0017] Further, the application realizes high-selectivity enrichment and purification of ergothioneine through a combined process of alcohol precipitation treatment and macroporous adsorption resin column purification. The alcohol precipitation step effectively removes impurities such as polysaccharides, and then a specific type of macroporous resin is used in combination with gradient ethanol elution to accurately capture and separate ergothioneine, solving the technical problems of low content and difficult extraction of ergothioneine in natural raw materials.

[0018] The oral liver-protecting composition of the present application is not a simple superposition of each component, but a synergistic defense and repair network with ergothioneine as the core. The core mechanism is that ganoderma ergothioneine can enter liver cells efficiently by virtue of its unique OCTN-1 transporter channel, directly acting on the core of oxidative stress such as mitochondria, and providing the most basic antioxidant protection for cells. On this basis, silymarin and ginsenoside Rb1 and other components play a strong role in promoting liver cell regeneration and stabilizing cell membranes. This means that after ergothioneine resists most free radical attacks, silymarin and other ginsenosides can better perform the "repair" task, and the two form an efficient link in "prevention" and "treatment", together maintaining the health and integrity of liver cells.

[0019] In addition, other active ingredients in the composition provide key synergistic support in terms of anti-inflammatory and metabolic regulation. For example, turmeric extract and schisandrin B have significant anti-inflammatory activity and can effectively inhibit the inflammatory signaling pathways such as NF-κB triggered by oxidative stress, thereby synergizing with ergothioneine to "inhibit the release of inflammatory factors". At the same time, pueraria extract and apigenin and other components help improve liver lipid metabolism and reduce liver burden, creating a more favorable in vivo environment for antioxidant and anti-inflammatory effects. This multi-level, multi-target synergistic cooperation ensures that the composition can intervene in all aspects of the pathological process from free radical clearance, damage reduction, inflammation inhibition to repair promotion, thereby achieving a comprehensive and strong liver-protecting effect that single components cannot achieve.

[0020] The ergothioneine-containing oral liver-protecting composition of the present application realizes a synergistic liver-protecting mechanism of multiple targets and multiple pathways through scientific formula design. The composition takes ganoderma ergothioneine extract as the core, synergizes with silymarin, ginsenoside Rb1, pueraria extract and other active ingredients, and acts on multiple key links of liver protection. This design breaks through the bottleneck of traditional liver-protecting products with single component and limited effect, and constructs a comprehensive and efficient liver-protecting system.

[0021] In the preparation process, the present application uses a composite enzymatic solution composed of specific types and proportions to enzymatically hydrolyze ganoderma, and combines subsequent alcohol precipitation and macroporous adsorption resin purification steps to significantly improve the extraction efficiency and purity of ergothioneine in ganoderma. Animal experiments have confirmed that the complete extraction process is crucial to ensuring the liver-protecting effect of the final product, and no mice died during the mouse experiment, proving the safety of the composition of the present application.

[0022] Further, the animal experiment data powerfully prove the actual liver protection efficacy of the composition. In the carbon tetrachloride-induced acute liver injury model, the serum ALT and AST levels of the experimental group mice taking the composition are significantly lower than those of the model group and all the control groups. This not only verifies the effectiveness of the composition as a whole, but also highlights the irreplaceable synergistic value of the core components and the complete process, and shows the liver protection ability significantly better than the products missing or replacing any key component.

[0023] In summary, the oral liver protection composition provided by the present application successfully integrates the targeted antioxidant components with high bioavailability and various auxiliary liver protection active substances, and guarantees the effectiveness and stability of the components through an optimized preparation process. The final product has the outstanding beneficial effects of clear mechanism of action, strong targeting, high bioavailability, and multi-pathway synergistic effect, and provides a reliable technical solution for developing a new generation of highly effective liver protection health products. BRIEF DESCRIPTION OF DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, a brief introduction will be given below to the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings in the following description are only embodiments of the present application, and those skilled in the art can also obtain other drawings according to the provided drawings without creative labor.

[0025] Figure 1 Figure 1 is a graph of experimental data statistics results for Example 1. DETAILED DESCRIPTION

[0026] The technical solutions provided by the present application will be described in detail below in conjunction with the embodiments, but they should not be understood as limiting the scope of protection of the present application.

[0027] Example 1

[0028] A preparation method of an oral liver protection composition containing ergothioneine, 35 parts of ganoderma ergothioneine extract, 4 parts of silymarin, 2 parts of ginsenoside Rb1, 4 parts of pueraria extract, 4 parts of salvia miltiorrhiza extract, 2 parts of turmeric extract, 2 parts of schisandrin B, and 2 parts of apioin are mixed according to mass fraction. The preparation method of the ganoderma ergothioneine extract is as follows: (a) Ganoderma lucidum powder is mixed with acetic acid-sodium acetate buffer solution at a mass ratio of 1:6, enzyme solution is added, and enzyme hydrolysis reaction is carried out for 2 hours, and the enzyme is inactivated by heating at 85℃ for 5 minutes; the mass ratio of the buffer solution and the enzyme solution is 100:2.

[0029] The enzyme solution is: endo-β-1, 3-glucanase, endo-chitinase, β-glucosidase, and subtilisin, with a mass ratio of 2.5:1.5:1:1. (b) centrifuging the enzymatic reaction solution obtained in step (a) to collect the supernatant containing ergothioneine; (c) concentrating the supernatant containing ergothioneine and adding ethanol for alcohol precipitation treatment to collect the supernatant of alcohol precipitation; The alcohol precipitation treatment is performed as follows: the obtained supernatant is concentrated to 1 / 5 of the original volume at 50°C under reduced pressure, then transferred to an alcohol precipitation container, 3 times volume of 95% ethanol is slowly added under continuous stirring to make the final ethanol concentration reach 70% (v / v), and after the addition is completed, the mixture is continuously stirred for 15 minutes to make it fully mixed and uniform, then placed in a low temperature environment at 4°C for 8 hours of standing and precipitation, and finally the supernatant and precipitate are separated by centrifugation to collect the supernatant of alcohol precipitation rich in ergothioneine for subsequent treatment.

[0030] (d) passing the supernatant of alcohol precipitation through a macroporous adsorption resin column (type HP20) to elute with water first, then with 20% ethanol solution, and collecting the ethanol eluate; (e) concentrating and drying the collected ethanol eluate to obtain ganoderma ergothioneine extract.

[0031] Example 2

[0032] A preparation method of an oral liver-protecting composition containing ergothioneine, which comprises mixing 45 parts of ganoderma ergothioneine extract, 7 parts of silymarin, 5 parts of ginsenoside Rb1, 7 parts of pueraria extract, 7 parts of salvia miltiorrhiza extract, 5 parts of turmeric extract, 5 parts of schisandrin B, and 5 parts of apigenin. The preparation method of the ganoderma ergothioneine extract comprises the following steps: (a) mixing ganoderma lucidum powder with citric acid-sodium phosphate dibasic buffer solution at a mass ratio of 1:12, adding enzymatic solution, and performing enzymatic reaction for 3 hours, and then heating and inactivating the enzyme at 85°C for 5 minutes; the mass ratio of the buffer solution to the enzymatic solution is 100:3.

[0033] The enzymatic solution comprises endo-β-1,3-glucanase, endo-chitinase, β-glucosidase, and subtilisin protease at a mass ratio of 3.5:2.5:2:1. (b) centrifuging the enzymatic reaction solution obtained in step (a) to collect the supernatant containing ergothioneine; (c) concentrating the supernatant containing ergothioneine and adding ethanol for alcohol precipitation treatment to collect the supernatant of alcohol precipitation; The alcohol precipitation treatment is as follows: after the supernatant is concentrated to 1 / 10 of the original volume at 60°C under reduced pressure, it is transferred to an alcohol precipitation container, 5 times the volume of 95% ethanol is slowly added under continuous stirring, the final ethanol concentration is 80% (v / v), after the addition is completed, stirring is continued for 32 minutes to fully mix and uniform, then it is placed in a low temperature environment at 4°C and allowed to stand for 13 hours, finally the supernatant and precipitate are separated by centrifugation, and the alcohol precipitation supernatant rich in ergothioneine is collected for subsequent processing.

[0034] (d) The alcohol precipitation supernatant is passed through a macroporous adsorption resin column (type 816), first eluted with water, and then eluted with an ethanol solution with a concentration of 40%, and the ethanol eluate is collected; (e) The collected ethanol eluate is concentrated and dried to obtain ganoderma ergothioneine extract.

[0035] Example 3

[0036] A preparation method of an oral liver-protecting composition containing ergothioneine, 37 parts of ganoderma ergothioneine extract, 5 parts of silymarin, 3 parts of ginsenoside Rb1, 5 parts of pueraria extract, 5 parts of salvia miltiorrhiza extract, 3 parts of turmeric extract, 3 parts of schisandrin B, and 3 parts of apigenin are mixed. The preparation method of the ganoderma ergothioneine extract is as follows: (a) Ganoderma lucidum powder is mixed with acetic acid-sodium acetate buffer at a mass ratio of 1:7, an enzyme solution is added, and enzyme hydrolysis is carried out for 2 hours, and the enzyme is inactivated by heating at 83°C for 6 minutes; the mass ratio of the buffer and the enzyme solution is 100:2.

[0037] The enzyme solution is: endo-β-1, 3-glucanase, endo-chitinase, β-glucosidase, and subtilisin, with a mass ratio of 2.5:1.5:2:1; (b) The enzyme hydrolysis reaction solution obtained in step (a) is centrifuged, and the supernatant containing ergothioneine is collected; (c) The supernatant containing ergothioneine is concentrated, and ethanol is added for alcohol precipitation treatment, and the alcohol precipitation supernatant is collected; The alcohol precipitation treatment is as follows: after the supernatant is concentrated to 1 / 6 of the original volume at 53°C under reduced pressure, it is transferred to an alcohol precipitation container, 5 times the volume of 95% ethanol is slowly added under continuous stirring, the final ethanol concentration is 73% (v / v), after the addition is completed, stirring is continued for 19 minutes to fully mix and uniform, then it is placed in a low temperature environment at 4°C and allowed to stand for 10 hours, finally the supernatant and precipitate are separated by filtration, and the alcohol precipitation supernatant rich in ergothioneine is collected for subsequent processing.

[0038] (d) The alcohol precipitation supernatant is passed through a macroporous adsorption resin column (type HPD100), eluted with water first, and then eluted with an ethanol solution with a concentration of 25%, and the ethanol eluate is collected; (e) The collected ethanol eluate is concentrated and dried to obtain Ganoderma lucidum paxilline extract.

[0039] Example 4

[0040] A preparation method of an oral liver-protecting composition containing paxilline, in which 43 parts of Ganoderma lucidum paxilline extract, 6 parts of silymarin, 4 parts of ginsenoside Rb1, 6 parts of pueraria extract, 6 parts of salvia extract, 4 parts of turmeric extract, 4 parts of schisandrin B, and 4 parts of apigenin are mixed. The preparation method of the Ganoderma lucidum paxilline extract includes the following steps: (a) Ganoderma lucidum powder is mixed with acetic acid-sodium acetate buffer at a mass ratio of 1:11, and an enzyme solution is added for enzyme hydrolysis reaction for 3 hours, and the enzyme is inactivated by heating at 85°C for 5 minutes; the mass ratio of the buffer and the enzyme solution is 100:3.

[0041] The enzyme solution includes endo-β-1,3-glucanase, endo-chitinase, β-glucosidase, and subtilisin, and the mass ratio is 3:2:2:1. (b) The enzyme hydrolysis reaction solution obtained in step (a) is centrifuged, and the supernatant containing paxilline is collected. (c) The supernatant containing paxilline is concentrated, and ethanol is added for alcohol precipitation treatment, and the alcohol precipitation supernatant is collected. The alcohol precipitation treatment is performed as follows: the obtained supernatant is concentrated to 1 / 8 of the original volume at 57°C under reduced pressure, then transferred to an alcohol precipitation container, 5 times the volume of ethanol with a concentration of 95% is slowly added under continuous stirring, so that the final ethanol concentration reaches 76% (v / v), and after the addition is completed, the mixture is continuously stirred for 18 minutes to make it fully mixed and uniform, then placed in a low-temperature environment at 4°C for 9 hours of static precipitation, and finally separated by centrifugation to obtain the supernatant and the precipitate, and the alcohol precipitation supernatant rich in paxilline is collected for subsequent treatment.

[0042] (d) The alcohol precipitation supernatant is passed through a macroporous adsorption resin column (type D101), eluted with water first, and then eluted with an ethanol solution with a concentration of 26%, and the ethanol eluate is collected. (e) The collected ethanol eluate is concentrated and dried to obtain Ganoderma lucidum paxilline extract.

[0043] Example 5

[0044] A preparation method of an oral liver-protecting composition containing ergothioneine, which comprises mixing ganoderma lucidum ergothioneine extract 40 parts, silymarin 5 parts, ginsenoside Rb1 5 parts, pueraria extract 5 parts, salvia extract 5 parts, turmeric extract 5 parts, schisandrin B 5 parts, apigenin 5 parts. The preparation method of the ganoderma lucidum ergothioneine extract comprises the following steps: (a) Ganoderma lucidum powder is mixed with a citric acid-sodium phosphate dibasic buffer solution at a mass ratio of 1:9, and an enzymatic hydrolysis solution is added, and enzymatic hydrolysis reaction is carried out for 3 hours, and the enzyme is inactivated by heating at 85°C for 5 minutes; the mass ratio of the buffer solution and the enzymatic hydrolysis solution is 100:3.

[0045] The enzymatic hydrolysis solution comprises endo-β-1,3-glucanase, endo-chitinase, β-glucosidase and subtilisin, and the mass ratio is 3:2:1.5:1. (b) The supernatant containing ergothioneine is collected by centrifugation of the enzymatic hydrolysis reaction solution obtained in step (a); The alcohol precipitation treatment operation is as follows: the obtained supernatant is concentrated to 1 / 8 of the original volume at 55°C under reduced pressure, then transferred to an alcohol precipitation container, and 4 times the volume of 95% ethanol is slowly added under continuous stirring, so that the final ethanol concentration reaches 75% (v / v); after adding, continue to stir for 25 minutes to make it fully mixed and uniform, then place it in a low temperature environment at 4°C for 10 hours of static precipitation, and finally separate the supernatant and the precipitate by centrifugation, and collect the alcohol precipitation supernatant rich in ergothioneine for subsequent treatment.

[0046] (d) The alcohol precipitation supernatant is passed through a macroporous adsorption resin column (type HPD300), first eluted with water, and then eluted with an ethanol solution with a concentration of 30%, and the ethanol eluate is collected; (e) The collected ethanol eluate is concentrated and dried to obtain the ganoderma lucidum ergothioneine extract.

[0047] Control group 1

[0048] The other methods are the same as those in Example 3, except that the addition of ganoderma lucidum ergothioneine extract is abandoned.

[0049] Control group 2

[0050] The other methods are the same as those in Example 3, except that ginsenoside Rb1 is replaced by salidroside in equal amount.

[0051] Control group 3

[0052] The other methods are the same as in Example 3, except that the use of endo-β-1,3-glucanase and subtilisin is omitted in the preparation of Ganoderma lucidum p- acetyl coenzyme A synthetase extract.

[0053] Control group 4

[0054] The other methods are the same as in Example 3, except that subtilisin is replaced with an equal amount of papain.

[0055] Experimental Example 1

[0056] One hundred and five ICR mice were selected for the composition liver protection test, and the ICR mice were randomly divided into 7 groups, 15 mice in each group. Before modeling, the mice in each group could normally eat, had smooth fur, moved well, and had quick reactions. Experimental groups were set up. The details are as follows: Blank group: the mice were given distilled water by gavage according to 0.1 mL / 10 g of body weight.

[0057] Model group: the mice were given distilled water by gavage according to 0.1 mL / 10 g of body weight.

[0058] Experimental group: the mice were given the composition prepared in Example 3 at a standard of 0.3 g per 1 kg of body weight, the product was prepared into an aqueous solution with a concentration of 0.03 g / mL, and the mice were given the product by gavage according to 0.1 mL / 10 g of body weight.

[0059] During the experiment, the experimental group was given the composition prepared in Example 3 by gavage every day, and the blank group and the model group were given distilled water. The experiment lasted for 30 days, after 30 days, the animals in each experimental group were fasted for 16 h overnight, the model group and each experimental group were given 0.5% carbon tetrachloride solution by gavage once, the dose was 0.1 mL / 10 g of body weight, the blank group was given vegetable oil, and each experimental group was continuously given the composition (more than 4 h apart from the carbon tetrachloride gavage). The animals were sacrificed 24 hours after the carbon tetrachloride was given, the mouse blood was taken, and the serum was separated, and the values of alanine aminotransferase ALT and aspartate aminotransferase AST in the serum were determined. The determination results were all average values, as shown in Table 1.

[0060] Alanine aminotransferase and aspartate aminotransferase are two important enzymes present in hepatocytes. When the liver is damaged by various causes, hepatocytes will be destroyed, leading to the release of alanine aminotransferase and aspartate aminotransferase into the blood, causing the levels of alanine aminotransferase and aspartate aminotransferase in the blood to rise. By detecting the levels of alanine aminotransferase and aspartate aminotransferase in the blood, the damage to hepatocytes can be reflected, which is of great significance for the diagnosis, treatment and prognosis evaluation of liver diseases.

[0061] Table 1 Effect of 30-day feeding of each group on ALT and AST of mice

[0062] According to the experimental data in Table 1, the ergothioneine-containing oral liver-protecting composition of the present application showed a significant liver-protecting effect in the carbon tetrachloride-induced mouse liver injury model. The alanine aminotransferase (ALT) and aspartate aminotransferase (AST) values of the experimental group were 73.41 U / L and 79.52 U / L, respectively, which were significantly lower than the values of the model group (123.4 U / L and 136.2 U / L), indicating that the composition could effectively inhibit liver cell damage, reduce serum aminotransferase levels, and had a good liver-protecting effect.

[0063] Compared with the control groups, the ALT and AST values of the experimental group were the lowest, especially in control group 1 (without the addition of ganoderma ergothioneine extract) and control group 3 (incomplete enzymatic process), the aminotransferase levels were significantly increased, further verifying the key role of ganoderma ergothioneine extract and its complete enzymatic process in the composition. In addition, the aminotransferase levels of control group 2 (replaced with ginsenoside Rb1) and control group 4 (replaced with protease) were also higher than those of the experimental group, indicating that the components and process design of the original formula had an irreplaceable advantage in synergistic liver protection.

[0064] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, several improvements and refinements can be made, and these improvements and refinements should also be considered within the scope of protection of the present application.

Claims

1. A method for preparing a Ganoderma lucidum ergothioneine extract, characterized in that, Includes the following steps: (a) Mix Ganoderma lucidum powder with buffer solution, add enzymatic hydrolysate, carry out enzymatic hydrolysis reaction, heat to inactivate enzyme, and obtain enzymatic hydrolysis reaction solution; (b) Centrifuge the enzymatic hydrolysis solution obtained in step (a) and collect the supernatant containing ergothioneine; (c) After concentrating the supernatant containing ergothioneine, add ethanol for alcohol precipitation and collect the alcohol precipitation supernatant; (d) Pass the alcohol precipitation supernatant through a macroporous adsorption resin column, first elute with water, then elute with an ethanol solution of concentration of 20-40%, and collect the ethanol eluent; (e) The collected ethanol eluent was concentrated and dried to obtain Ganoderma lucidum ergothioneine extract.

2. The oral hepatoprotective composition containing ergothioneine according to claim 1, characterized in that, The enzymes contained in the enzymatic hydrolysate in step (a) are: endo-β-1,3-glucanase, endo-chitinase, β-glucosidase and subtilisin, in a mass ratio of 2.5~3.5:1.5~2.5:1~2:

1.

3. The oral hepatoprotective composition containing ergothioneine according to claim 2, characterized in that, The enzymatic hydrolysis time in step (a) is 2-3 hours.

4. The oral hepatoprotective composition containing ergothioneine according to claim 1, characterized in that, The buffer solution in step (a) is an acetic acid-sodium acetate buffer or a citrate-disodium hydrogen phosphate buffer.

5. The oral hepatoprotective composition containing ergothioneine according to claim 1, characterized in that, In step (a), the mass ratio of Ganoderma lucidum powder to buffer solution is 1:6~12; the mass ratio of buffer solution to enzyme hydrolysate is 100:2~3.

6. The oral hepatoprotective composition containing ergothioneine according to claim 1, characterized in that, The macroporous adsorption resin column in step (d) is of one type: HP20, HPD100, HPD300, D101, or 816.

7. An oral hepatoprotective composition containing ergothioneine, characterized in that, The product comprises the following components in parts by mass: 35-45 parts of Ganoderma lucidum ergothioneine extract prepared by any one of claims 1 to 6, 4-7 parts of silymarin, 2-5 parts of ginsenoside Rb1, 4-7 parts of kudzu root extract, 4-7 parts of salvia miltiorrhiza extract, 2-5 parts of turmeric extract, 2-5 parts of schisandrin B, and 2-5 parts of apigenin.

8. The method for preparing the oral hepatoprotective composition containing ergothioneine according to claim 7, characterized in that, Simply mix the following ingredients: Ganoderma lucidum ergothioneine extract, silymarin, ginsenoside Rb1, kudzu root extract, salvia miltiorrhiza extract, turmeric extract, schisandrin B, and apigenin.

9. The use of the oral hepatoprotective composition containing ergothioneine as described in claim 7 in the preparation of health products with hepatoprotective effects.

10. The application according to claim 9, characterized in that, The liver-protecting health products mentioned are one or more of the following: alcoholic liver damage care tablets, chemical liver damage protection capsules, hyperlipidemia and fatty liver conditioning pills, non-alcoholic fatty liver compound tablets, early intervention products for liver fibrosis, liver-protecting essence capsules for staying up late, hangover relief and liver protection pre-drink, liver repair granules after drinking, drug-assisted liver sustained-release capsules, liver and gallbladder comprehensive conditioning soft capsules, liver qi stagnation relief pills, liver fire clearing tea, liver and eye health care tablets, liver cirrhosis auxiliary nutritional powder, and liver detoxification and health preservation pills.

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