A Stropharia rugosoannulata glycoprotein extract, its preparation method and application

Abstract

The present invention belongs to the technical fields of glycoprotein preparation and biomedicine, and particularly relates to a Stropharia rugosoannulata glycoprotein extract, a preparation method thereof and an application thereof. The present invention firstly proposes a method for extracting Stropharia rugosoannulata glycoprotein. Compared with the conventional alkali solution-acid precipitation method for extracting plant glycoproteins, the extraction method of the present invention has a higher glycoprotein yield. The contents of protein and polysaccharide in the extracted Stropharia rugosoannulata glycoprotein extract are both relatively high, especially the polysaccharide content is significantly higher than that of the alkali solution-acid precipitation method. Furthermore, the extracted Stropharia rugosoannulata glycoprotein has excellent hypolipidemic and antioxidant activities and can be used for the treatment of hyperlipidemia. At the same time, the method for extracting Stropharia rugosoannulata glycoprotein of the present invention has a simple process, is easy to control, and is suitable for industrial production.

Description

Technical Field

[0001] The present invention belongs to the technical fields of glycoprotein preparation and biomedicine, and particularly relates to a Stropharia rugosoannulata glycoprotein extract, a preparation method thereof, and an application thereof. Background Art

[0002] Hyperlipidemia, also known as lipid metabolism disorder or abnormality, is caused by abnormal fat metabolism or transport, resulting in too high or too low levels of one or several lipids in plasma, such as triglyceride, total cholesterol, and low-density lipoprotein cholesterol levels. Hyperlipidemia mostly occurs due to excessive exogenous lipid intake or in vivo lipid metabolism disorders. Its harm lies in that excessive lipids deposit on the inner wall of arteries, forming atherosclerotic plaques, causing arterial stenosis, blood flow obstruction, reduced blood supply to target organs and subsequent damage, and forming various diseases such as coronary atherosclerotic heart disease, hypertension, cerebral thrombosis, diabetes, fatty liver, obesity, and cholelithiasis. Currently, hyperlipidemia is mainly treated with lipid-lowering chemical drugs, including statins, resins, niacin, bile acid sequestrants, etc. However, the treatment cost of these drugs is relatively high, and they may produce certain toxic and side effects. Recent studies have shown that food-derived natural active products, such as proteins, polypeptides, polysaccharides, etc., have a certain lipid-regulating effect and can be used as functional natural active factors for the adjuvant treatment of hyperlipidemia.

[0003] Stropharia rugosoannulata is one of the edible fungi recommended by the Food and Agriculture Organization of the United Nations for cultivation. It is rich in nutrients, including proteins, amino acids, vitamins, polysaccharides and other nutrients, and has significant effects in aspects such as antioxidant, anti-fatigue, anti-tumor, and hypoglycemic. It is an edible mushroom with both medicinal and edible properties and is deeply loved by consumers and researchers. The cap of Stropharia rugosoannulata is tender, the stipe is crispy, and the umami flavor is strong, enjoying the reputation of "the meat among vegetables". Its cultivation is simple, the raw materials are extensive, the input cost is low and the yield is high. These advantages have attracted much attention to Stropharia rugosoannulata in the promotion of science and technology for agriculture. In recent years, the research on edible mushroom proteins has also received increasing attention. Research shows that edible mushroom proteins have high antioxidant activity, can scavenge free radicals, slow down the process of cell oxidative aging; inhibit inflammatory reactions and reduce the progression of inflammatory lesions; inhibit the growth and spread of tumor cells and have a certain adjuvant effect on tumor treatment. However, there are few reports on the activity research of Stropharia rugosoannulata glycoprotein extract. Summary of the Invention

[0004] The purpose of the present invention is to provide a Stropharia rugosoannulata glycoprotein extract, a preparation method thereof, and an application thereof. The Stropharia rugosoannulata glycoprotein has lipid-lowering and antioxidant activities and can be used for preparing drugs for treating hyperlipidemia.

[0005] The present invention provides an extraction method of a Stropharia rugosoannulata glycoprotein extract, comprising the following steps:

[0006] Pre-treat the fresh Stropharia rugoso-annulata, mix the pre-treated Stropharia rugoso-annulata with water for wet homogenization to obtain a Stropharia rugoso-annulata slurry;

[0007] Perform solid-liquid separation on the Stropharia rugoso-annulata slurry, ultrafilter the obtained supernatant, and intercept the components with a molecular weight greater than 10,000 g / mol to obtain the Stropharia rugoso-annulata glycoprotein extract;

[0008] The pre-treatment includes: placing the fresh Stropharia rugoso-annulata at -20°C to 25°C for the first static placement for 4 to 6 hours, and then placing it at room temperature for the second static placement for 4 to 6 hours. The first static placement and the second static placement form one cycle, and a total of 2 cycles are performed.

[0009] Preferably, the temperature of the first static placement is -20°C to 5°C.

[0010] Preferably, the temperature of the first static placement is -20°C or 5°C to 25°C.

[0011] Preferably, the temperature of the first static placement is 5°C.

[0012] Preferably, the mass ratio of the fresh Stropharia rugoso-annulata to the volume of water is 1 to 2 g: 5 to 10 mL.

[0013] Preferably, the rotation speed of the wet homogenization is 1500 to 2500 rpm, and the time is 5 to 10 minutes.

[0014] Preferably, the method of solid-liquid separation includes centrifugation: the rotation speed of the centrifugation is 3000 to 4000 rpm, and the time is 5 to 15 minutes.

[0015] The present invention also provides a Stropharia rugoso-annulata glycoprotein extract obtained by the extraction method described in the above technical solution. The Stropharia rugoso-annulata glycoprotein extract includes proteins and polysaccharides. The content of the proteins is 40 to 55%, and the content of the polysaccharides is 20% to 25%.

[0016] The present invention also provides the application of the Stropharia rugoso-annulata glycoprotein extract described in the above technical solution in the preparation of drugs for preventing and treating hyperlipemia.

[0017] Preferably, the drug includes drugs with lipid-lowering and / or antioxidant activity functions.

[0018] Beneficial effects:

[0019] The present invention provides a method for extracting a glycoprotein extract from Stropharia rugosoannulata, comprising the following steps: pretreating fresh Stropharia rugosoannulata, mixing the pretreated Stropharia rugosoannulata with water for wet homogenization to obtain a Stropharia rugosoannulata slurry; performing solid-liquid separation on the Stropharia rugosoannulata slurry, and subjecting the obtained supernatant to ultrafiltration to intercept components with a molecular weight greater than 10,000 g / mol to obtain the Stropharia rugosoannulata glycoprotein extract; the pretreatment includes: placing fresh Stropharia rugosoannulata at -20°C to 25°C for the first static placement for 4 to 6 hours and then placing it at room temperature for the second static placement for 4 to 6 hours, and the first static placement and the second static placement form one cycle, with a total of 2 cycles. The present invention firstly proposes a method for extracting Stropharia rugosoannulata glycoprotein, and compared with the conventional alkali dissolution and acid precipitation method for extracting plant glycoprotein, the extraction method of the present invention has a higher glycoprotein yield, and the content of protein and polysaccharide in the extracted Stropharia rugosoannulata glycoprotein extract is relatively high, especially the polysaccharide content is significantly higher than that of the alkali dissolution and acid precipitation method; furthermore, the extracted Stropharia rugosoannulata glycoprotein has excellent hypolipidemic and antioxidant activities and can be used for the treatment of hyperlipidemia; at the same time, the method for extracting Stropharia rugosoannulata glycoprotein of the present invention has a simple process, is easy to control, and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments.

[0021] Figure 1 Results of the contents of protein and polysaccharide in the Stropharia rugosoannulata glycoprotein extract in Examples 1 to 3 and Comparative Example 1;

[0022] Figure 2 Lipid-lowering effect diagram of the Stropharia rugosoannulata glycoprotein extract on the high-fat L-O2 cell model in Examples 1 to 3 and Comparative Example 1 (Oil Red O staining × 25);

[0023] Figure 3 Results diagram of the change trends of TG, TC, HDL, and LDL indexes in different sample groups in Application Example 1;

[0024] Figure 4 Results diagram of the change trends of ALT, AST, T-AOC, and MDA indexes in different sample groups in Application Example 1;

[0025] Figure 5 Results diagram of the effects of different Stropharia rugosoannulata glycoprotein extracts on the sizes of white adipocytes (A-F) and epididymal adipocytes (a-f) of high-fat diet mice in Application Example 2 (HE staining × 25);

[0026] Among them, in Figures 3 - 4Among them, compared with the control group, #P<0.05, ##P<0.01, P<0.001, compared with the model group, *P<0.05, **P<0.01, ***P<0.001. Detailed implementation mode

[0027] The present invention provides a method for extracting a glycoprotein extract from Stropharia rugoso-annulata, comprising the following steps:

[0028] Pre-treat the fresh Stropharia rugoso-annulata, mix the pre-treated Stropharia rugoso-annulata with water for wet homogenization to obtain a Stropharia rugoso-annulata slurry;

[0029] Perform solid-liquid separation on the Stropharia rugoso-annulata slurry, ultrafilter the obtained supernatant, and intercept the components with a molecular weight greater than 10,000 g / mol to obtain the Stropharia rugoso-annulata glycoprotein extract;

[0030] The pre-treatment includes: placing the fresh Stropharia rugoso-annulata at -20°C to 25°C for the first static placement for 4 to 6 hours, and then placing it at room temperature for the second static placement for 4 to 6 hours. The first static placement and the second static placement form one cycle, and a total of 2 cycles are performed.

[0031] The present invention pre-treats the fresh Stropharia rugoso-annulata, and the steps of the pre-treatment include: placing the fresh Stropharia rugoso-annulata at -20°C to 25°C for the first static placement for 4 to 6 hours, and then placing it at room temperature for the second static placement for 4 to 6 hours. The first static placement and the second static placement form one cycle, and a total of 2 cycles are performed.

[0032] In the present invention, the temperature of the first static placement is preferably -20°C to 5°C or 25°C, more preferably -20°C, 5°C or 25°C, and even more preferably 5°C. That is, the present invention preferably pre-treats the fresh Stropharia rugoso-annulata by freezing and thawing, and this pre-treatment has the effect of promoting the dissolution of proteins and polysaccharides.

[0033] The time of the first static placement in the present invention is preferably 6 hours. The time of the second static placement in the present invention is preferably 4 hours. The number of cycles in the present invention is preferably 2 to 3 times, more preferably 2 times; this cycle helps the dissolution of the target substance.

[0034] After the pre-treatment, the present invention mixes the pre-treated Stropharia rugoso-annulata with water for wet homogenization to obtain a Stropharia rugoso-annulata slurry. Based on the fresh Stropharia rugoso-annulata, the mass ratio of the fresh Stropharia rugoso-annulata to the volume of water is preferably 1 to 2 g: 5 to 10 mL, more preferably 1 g: 5 mL. The rotation speed of the homogenization in the present invention is preferably 1500 to 2500 rpm, more preferably 2000 rpm; the time is preferably 5 to 10 minutes, more preferably 5 minutes.

[0035] After obtaining the Stropharia rugosoannulata slurry, the present invention separates the solid and liquid of the Stropharia rugosoannulata slurry to obtain a supernatant. The solid-liquid separation in the present invention preferably includes centrifugation. The rotation speed of the centrifugation is preferably 3000-4000 rpm, more preferably 4000 rpm, and the time is preferably 5-15 min, more preferably 10 min.

[0036] After obtaining the supernatant, the present invention ultrafilters the supernatant and intercepts the components with a molecular weight greater than 10000 g / mol to obtain a Stropharia rugosoannulata glycoprotein extract. The content, yield of protein and polysaccharide in the Stropharia rugosoannulata glycoprotein extract prepared by the method of the present invention are relatively high. Among them, the content of the protein is 40-55%, preferably 50.47%; the content of the polysaccharide is 20%-25%; the yield is preferably 45-55%. The extraction method of the present invention uses wet homogenization to physically destroy the tissue structure of the sample, thereby promoting the dissolution of protein and polysaccharide, and avoiding the problems that the protein obtained by conventional extraction methods such as alkali dissolution and acid precipitation is prone to denaturation, low activity efficiency and low yield of the extract. At the same time, the Stropharia rugosoannulata glycoprotein extracted by the extraction method of the present invention has excellent hypolipidemic and antioxidant activities and can be used to treat hyperlipidemia.

[0037] Based on the above advantages, the present invention also provides the application of the Stropharia rugosoannulata glycoprotein extract described in the above technical solution in the preparation of drugs for preventing and treating hyperlipemia. The drugs of the present invention include drugs with hypolipidemic and / or antioxidant activity functions, more preferably drugs with hypolipidemic and antioxidant activities. The hypolipidemic effect in the present invention preferably includes one or more of reducing triglyceride (TG), increasing high-density lipoprotein (HDL) and reducing low-density lipoprotein (LDL), more preferably reducing triglyceride, increasing high-density lipoprotein and reducing low-density lipoprotein. The antioxidant activity in the present invention preferably includes one or more of reducing the activity of alanine aminotransferase (ALT), reducing the activity of aspartate aminotransferase (AST), increasing the total antioxidant capacity (T-AOC) and reducing the content of malondialdehyde (MDA), more preferably reducing the activity of alanine aminotransferase, reducing the activity of aspartate aminotransferase, increasing the total antioxidant capacity and reducing the content of malondialdehyde.

[0038] In order to further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the drawings and embodiments, but they cannot be understood as limiting the protection scope of the present invention.

[0039] The fresh Stropharia rugosoannulata in this example was purchased from Shanghai Gulinyuan Mushroom Industry Professional Cooperative, and the strain was 'Stropharia rugosoannulata No. 5' [Shanghai Agricultural Product Recognition Edible Mushroom (2004) No. 062], which is a commercially available variety.

[0040] Reagents: PBS (self-prepared, pH 7.4, 0.1M), oleic acid, 1640, FBS, BSA: Sangon Biotech Co., Ltd.; Oil Red O: Beijing Solarbio Science & Technology Co., Ltd.; basal diet, high-fat diet, purified diet for the energy supply control group: Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd.; 4% paraformaldehyde: Shanghai Yuanye Bio-Technology Co., Ltd.; BCA protein concentration assay kit: Shanghai Beyotime Biotechnology Co., Ltd.; high-fat diet (product number XTHF60) was purchased from Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd.; total cholesterol (T-CHO), triglyceride (TG), aspartate aminotransferase (AST / GOT), alanine aminotransferase (ALT / GPT), high-density lipoprotein (HDL), low-density lipoprotein (LDL), total antioxidant capacity (T-AOC), total antioxidant malondialdehyde (MDA) test kits: Nanjing Jiancheng Bioengineering Institute; sodium hydroxide, hydrochloric acid, concentrated sulfuric acid, sodium carboxymethylcellulose: Sinopharm Chemical Reagent Co., Ltd., and all chemical reagents were of analytical grade.

[0041] Example 1

[0042] A preparation method of a Stropharia rugoso-annulata glycoprotein extract is as follows:

[0043] Weigh 200 g of fresh Stropharia rugoso-annulata, freeze it at -20 °C for 6 h, thaw it at room temperature for 4 h, and repeat this process twice; place the treated Stropharia rugoso-annulata in a homogenizer, add 1 L of distilled water for wet homogenization, the rotation speed of wet homogenization is 2000 rpm, and the time is 5 min; then centrifuge at 4000 rpm for 10 min, take the supernatant for ultrafiltration, intercept the component with a molecular weight greater than 10000 g / mol, rotary evaporate and concentrate the volume to 50 mL at 50 °C, and freeze-dry for later use.

[0044] Example 2

[0045] A preparation method of a Stropharia rugoso-annulata glycoprotein extract is as follows:

[0046] Weigh 200 g of fresh Stropharia rugoso-annulata, refrigerate it at 5 °C for 6 h, let it stand at room temperature for 4 h, and repeat this process twice; place the treated Stropharia rugoso-annulata in a homogenizer, add 1 L of distilled water for wet homogenization, the rotation speed of wet homogenization is 2000 rpm, and the time is 5 min; then centrifuge at 4000 rpm for 10 min, take the supernatant for ultrafiltration, intercept the component with a molecular weight greater than 10000 g / mol, rotary evaporate and concentrate the volume to 50 mL at 50 °C, and freeze-dry for later use.

[0047] Example 3

[0048] Weigh 200 g of fresh Stropharia rugosoannulata, place it at 25 °C for static placement for 6 h, then at room temperature for 4 h, and repeat this process twice. Place the treated Stropharia rugosoannulata in a homogenizer, add 1 L of distilled water for wet homogenization. The rotation speed of wet homogenization is 2000 rpm, and the time is 5 min. Then centrifuge at 4000 rpm for 10 min, take the supernatant for ultrafiltration, intercept the components with a molecular weight greater than 10000 g / mol, rotary evaporate and concentrate the volume to 50 mL at 50 °C, and freeze-dry for later use.

[0049] Comparative Example 1

[0050] Take 20 g of Stropharia rugosoannulata, beat it with a material-liquid (water) ratio of 1 g:10 mL, add 2 mol / L NaOH to adjust the pH value to 10.5, then stir and extract at 40 °C for 3 h, centrifuge at 6000 r / min for 10 min, and take the supernatant. Adjust the pH of the supernatant to 4.5 with 7% hydrochloric acid, let it stand and centrifuge at 6000 r / min or more for 10 min, collect the precipitate, dissolve the precipitate with a small amount of deionized water and transfer it to a container, and freeze-dry.

[0051] Adopt the Kjeldahl method, and refer to GB 5009.5-2016 "National Food Safety Standard Determination of Protein in Foods" to determine the protein content in the Stropharia rugosoannulata glycoprotein extracts and fresh Stropharia rugosoannulata in Examples 1-3 and Comparative Example 1;

[0052] Adopt the phenol-sulfuric acid method, and refer to NY / T1676--2008 "Determination of Crude Polysaccharide Content in Edible Fungi" to determine the polysaccharide content in the Stropharia rugosoannulata glycoprotein extracts and fresh Stropharia rugosoannulata in Examples 1-3 and Comparative Example 1;

[0053] Yield rate of extract (%) = (mass of extract / mass of Stropharia rugosoannulata after deducting water content (dry product)) × 100%

[0054] The measurement results are as Figure 1 shown.

[0055] It can be Figure 1 concluded that: the yield rate of the extract obtained from the sample treated at 5 °C in Example 2 is the highest (50.47%), followed by the sample treated with repeated freezing and thawing at -20 °C in Example 1 (41.18%). The protein content is the highest in the extract prepared by alkali dissolution and acid precipitation in Comparative Example 1 (72.56%), but its yield rate is only 13.55%. Secondly, the protein content of the sample treated at 5 °C in Example 2 is 50.47%. For the polysaccharide content, the values in the samples treated at -20 °C, 5 °C and 25 °C are 24.50%, 21.54% and 22.84% respectively, and there is no significant difference (P > 0.05). Considering the protein content, polysaccharide content and yield rate of each treatment method comprehensively, it can be known that the comprehensive evaluation of the content of each index of the extract obtained from the 5 °C sample in Example 2 is the best.

[0056] Application Example 1

[0057] Evaluation of the in vitro lipid-lowering activity of the Stropharia rugoso-annulata glycoprotein extract in Examples 1 to 3 and Comparative Example 1 was carried out as follows:

[0058] 1) Intervention effect of the Stropharia rugoso-annulata glycoprotein extract on the in vitro hyperlipidemia model

[0059] After culturing and adhering L-O2 cells (1.0×10 4 cells / well) in a 96-well plate, fresh serum-free medium containing 1 mmol / L oleic acid (OA) was added or not added. The culture wells without OA addition were used as the control group, and the culture wells with OA addition were divided into the model group and the Stropharia rugoso-annulata glycoprotein extract treatment group. Meanwhile, different concentrations of the Stropharia rugoso-annulata glycoprotein extract (50, 100, 200 μg / mL) were added to the Stropharia rugoso-annulata glycoprotein extract treatment group (as described in the figure below), and the cells were cultured for another 24 h. The changes in cell blood lipids were observed by Oil Red O staining method. Figure 2 as shown in the figure below), and the cells were cultured for another 24 h. The changes in cell blood lipids were observed by Oil Red O staining method.

[0060] 2) Oil Red O staining

[0061] After 24 h of OA and sample intervention, each culture well was washed once with PBS, and then fixed with 4% paraformaldehyde for 30 min; after fixation, it was rinsed once with 60% isopropanol. Then, 60% Oil Red O staining solution was added and incubated in the dark for 30 min, and 60% isopropanol was used to rinse off the excess staining solution. After the cells were rinsed twice with PBS, they were immersed in 100 μL PBS and photographed under an inverted microscope, and the positive occupancy ratio was calculated, where the positive occupancy ratio (%) = (total area of cell red regions / sum of cell areas / number of cells)×100%, and the results are as shown in Figure 2 shown.

[0062] It can be concluded from Figure 2 that the intracellular lipid deposition of each group of cells was characterized by Oil Red O staining. Compared with the control group, a large number of red-stained regions were presented in the L-O2 cells in the culture wells of the model group. After the intervention with 100 μg / mL Stropharia rugoso-annulata glycoprotein extract, the intracellular red-stained granules were significantly reduced ( Figure 2 A-F in the figure). Figure 2 The results in G in the figure showed that the three Stropharia rugoso-annulata glycoprotein extracts had an obvious improvement effect on the lipid metabolism of L-O2 cells. The average reduction rates of the positive occupancy ratios at 25 °C, 5 °C and -20 °C were 32.95%, 52.18% and 37.23% respectively, and the reduction rate of the positive occupancy ratio of the sample prepared by alkali dissolution and acid precipitation treatment was 36.78%. Among them, the sample prepared at 5 °C had the best effect.

[0063] Application Example 2

[0064] Intervention effect of the Stropharia rugoso-annulata protein extract on hyperlipidemic mice was carried out as follows:

[0065] 1) Grouping design of animal experiments

[0066] After being fed for 1 week under constant temperature and humidity conditions, ICR mice (purchased from Shanghai Bikai Keyi Biotechnology Co., Ltd.) were randomly divided into 5 groups (n = 6), including a blank control group fed with normal feed, a model control group fed with high-fat feed, a positive drug group based on the high-fat animal model (intragastric administration of simvastatin 0.8 mol / L daily), a high-dose intervention group of Stropharia rugosoannulata glycoprotein extract based on the high-fat animal model (intragastric administration of 300 mg / kg daily), and a low-dose intervention group of Stropharia rugosoannulata glycoprotein extract based on the high-fat animal model (intragastric administration of 150 mg / kg daily). The control group was intragastrically administered the same dose of water for 5 weeks. After the mice were fasted for 24 h, blood was collected from the orbital cavity, and samples such as the liver were collected for analysis.

[0067] 2) Detection of physiological and biochemical indexes

[0068] The contents of triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total antioxidant capacity (T-AOC), malondialdehyde (MDA), etc. in serum or liver were detected using kits. The experimental methods were determined according to the instructions of the kits (Nanjing Jiancheng), and the results are as Figure 3 and Figure 4 shown.

[0069] 3) Histopathological analysis

[0070] The cell samples were fixed with tissue fixative, embedded in paraffin, and cut into 5-μm tissue sections. After staining with Oil Red O and hematoxylin-eosin (H&E), the morphological changes of cells in the mouse liver, white adipose tissue, and epididymal adipose tissue were observed under a microscope. The results are as Figure 5 shown.

[0071] From Figure 3 and Figure 4 it can be concluded that: Through the analysis of the changing trends of the contents of triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL) in the serum of each group of animals and the contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) in the liver tissue, it is shown that there is a very significant difference in the TG content between the model group and the control group (P < 0.001), and there is a very significant difference between the experimental groups (including the positive sample and the high- and low-dose glycoprotein extract sample groups) and the model group (P < 0.001), reaching the level equivalent to that of the control group. It can be seen that the glycoprotein extract has a good effect on reducing triglyceride ( Figure 3 in A). In terms of TC, the effect of the glycoprotein extract on reducing total cholesterol is not significant (Figure 3 In B). For HDL and LDL, the samples in the high-dose group significantly increased the content of high-density lipoprotein and decreased the content of low-density lipoprotein ( Figure 3 in C and D in 3), thus reducing the risk of hyperlipidemia.

[0072] ALT and AST are key indicators for indicating liver function health. The experimental results are as Figure 4 shown in A in Figure 4 and B in. The results of the experimental group were comparable to those of the drug group, and there were very significant differences (P < 0.001) and extremely significant differences (P < 0.01) compared with the model group. It can be seen that the glycoprotein extract has a good effect on protecting the liver. In addition, compared with the control group, the T-AOC level in the liver tissue of the model group mice decreased extremely significantly (P < 0.01), and the MDA level increased very significantly (P < 0.001). After intervention with the glycoprotein of Stropharia rugosoannulata, the T-AOC level in the liver tissue of hyperlipidemic mice can be significantly increased, and its MDA level can be reduced ( Figure 4 in C and Figure 4 in D).

[0073] The results show that the glycoprotein extract of Stropharia rugosoannulata has good antioxidant activity on the liver tissue of hyperlipidemic mice, especially the high-dose group shows better effects in many indicators such as HDL and T-AOC.

[0074] It can be concluded from the above examples that the extraction method described in the present invention can improve the yield of the glycoprotein extract of Stropharia rugosoannulata, and the obtained glycoprotein extract of Stropharia rugosoannulata has high hypolipidemic and antioxidant activities.

[0075] Although the above examples have described the present invention in detail, they are only a part of the embodiments of the present invention, not all embodiments. People can also obtain other embodiments without creative efforts based on these embodiments, and these embodiments all belong to the protection scope of the present invention.

Claims

1. A method for extracting a glycoprotein extract from Stropharia rugosoannulata, characterized in that, it comprises the following steps: Pre-treat the fresh Stropharia rugosoannulata, mix the obtained Stropharia rugosoannulata after pre-treatment with water for wet homogenization to obtain a Stropharia rugosoannulata slurry; Perform solid-liquid separation on the Stropharia rugosoannulata slurry, ultrafilter the obtained supernatant, and intercept the components with a molecular weight greater than 10,000 g / mol to obtain the Stropharia rugosoannulata glycoprotein extract; The pre-treatment includes: placing the fresh Stropharia rugosoannulata at 5°C for the first static placement for 4 - 6 h, and then placing it at room temperature for the second static placement for 4 - 6 h. The first static placement and the second static placement form a cycle, and a total of 2 cycles are performed; The mass ratio of the fresh Stropharia rugosoannulata to the volume of water is 1 - 2 g: 5 - 10 mL; The rotation speed of the wet homogenization is 1500 - 2500 rpm, and the time is 5 - 10 min; The method of solid-liquid separation includes centrifugation: the rotation speed of the centrifugation is 3000 - 4000 rpm, and the time is 5 - 15 min.

2. The Stropharia rugosoannulata glycoprotein extract obtained by the extraction method described in claim 1, characterized in that, the Stropharia rugosoannulata glycoprotein extract contains protein and polysaccharide, the content of the protein is 40 - 55%, and the content of the polysaccharide is 20% - 25%.

3. Use of the Stropharia rugosoannulata glycoprotein extract described in claim 2 in the preparation of a drug for preventing and treating hyperlipidemia.

Images