Pretreatment method for increasing coriolus versicolor polysaccharide content

Abstract

The invention relates to the technical field of traditional Chinese medicine application and preparation, and provides a pretreatment method for improving the coriolus versicolor polysaccharide content, which comprises the following steps: carrying out hot-pressing treatment and rapid pressure relief on a thoroughly-moistened coriolus versicolor sporocarp, then carrying out ultrasonic treatment, and drying by adopting a microwave drying method to obtain the coriolus versicolor with high polysaccharide content. The invention also provides coriolus versicolor obtained by the method, wherein the polysaccharide content is not less than 6%. According to the method, the content of coriolus versicolor polysaccharide is remarkably increased through the combined technology of spraying moistening, hot-pressing steam heating, ultrasonic treatment and microwave drying on coriolus versicolor, the release rate and the release amount of the polysaccharide from the coriolus versicolor are increased, and the curative effect of the coriolus versicolor and the utilization rate of the medicinal material are improved.

Description

Technical Field

[0001] The invention relates to the technical field of application and preparation of traditional Chinese medicine, and relates to a pretreatment method of Coriolus versicolor. Background Art

[0002] The disclosure of this background information is intended to enhance understanding of the general background of the invention and should not necessarily be taken as an acknowledgment or any form of suggestion that this information constitutes the prior art already known to a person skilled in the art.

[0003] Coriolus versicolor Coriolus versicolor The dried fruiting body of (L. ex Fr.) Quel. It has the functions of strengthening the spleen and removing dampness, clearing away heat and detoxifying, and can be used for damp-heat jaundice, hypochondriasis, poor appetite, fatigue, etc. Modern research shows that enhancing immunity is the most important pharmacological effect of Yunzhi. It achieves immune enhancement by activating NK cells, T lymphocytes, B lymphocytes, promoting the production of cytokines, inducing IFN-r and other pathways. Yunzhi has a good inhibitory effect on digestive tract tumors. Yunzhi has a protective effect on liver damage, and the "Yunzhi Gantai" prepared from it is used as a drug for the treatment of hepatitis; Yunzhi also has a good anti-aging effect.

[0004] The active ingredients in Coriolus versicolor are mainly polysaccharides and Coriolus versicolor glycopeptides. Therefore, the content of Coriolus versicolor polysaccharides in Coriolus versicolor and the ease of releasing polysaccharides from Coriolus versicolor directly affect the effectiveness of Coriolus versicolor. Increasing the content of Coriolus versicolor polysaccharides and increasing the release of Coriolus versicolor polysaccharides are important means to improve the effectiveness of Coriolus versicolor. The fruiting body of Coriolus versicolor is leathery and hard. The hyphae wall is thick and thick, intertwined and entangled. The cell wall composed of glucan, chitin, cellulose, hemicellulose, lignin, protein, etc. is tough and has high mechanical strength. The polysaccharides contained in it are difficult to be fully extracted and utilized because they have a large adsorption force with other substances. Therefore, in order to improve the utilization rate of Coriolus versicolor, it is necessary to develop a method to improve the extraction rate of Coriolus versicolor polysaccharides. Summary of the invention

[0005] The present invention provides a pretreatment method for increasing the polysaccharide content of Coriolus versicolor. After the treatment of Coriolus versicolor by the present invention, the polysaccharide content of the active ingredient of Coriolus versicolor can be significantly increased, and the release rate of the polysaccharide of Coriolus versicolor can be increased, thereby improving the therapeutic effect of Coriolus versicolor.

[0006] To achieve the above purpose, the present invention adopts the following technical solution.

[0007] A pretreatment method for increasing the content of Coriolus versicolor polysaccharide, comprising the following steps: (1) Soak the dried fruiting bodies of Coriolus versicolor thoroughly; (2) subjecting the fruiting body in step (1) to heat-pressing treatment and rapidly releasing the pressure; (3) treating the fruiting body in step (2) with ultrasonic wave; (4) Drying the fruiting body in step (3) to obtain Versicolor versicolor with a high polysaccharide content.

[0008] Preferably, the specific operation of moistening is: cover the fruiting body with wet material, spray an appropriate amount of clean water frequently, and keep it moist for 12h-24h.

[0009] Preferably, the water vapor pressure of the hot pressing treatment is 1.2Mpa-2.0Mpa, the pressure treatment time is 1h-4h, and the pressure relief speed is 5min to normal pressure, such as 5min, 4min, 3min, 2min, 1min. More preferably, the water vapor pressure of the hot pressing treatment is 1.2Mpa-1.8Mpa, the pressure treatment time is 1h-3h, and the pressure relief speed is 2min-5min to normal pressure.

[0010] Preferably, the ultrasonic treatment conditions are: power 300 W, frequency 20 kHz, time 15 min-30 min; more preferably, the time is 15 min-20 min.

[0011] Preferably, the drying method is microwave drying. More preferably, the microwave frequency is 2450 MHz, the drying temperature is 60° C.-70° C., and the drying time is 20 min-30 min.

[0012] A kind of Coriolus versicolor obtained by the above method. The Coriolus versicolor can be a fruiting body, a slice of a fruiting body, or a crushed product.

[0013] The polysaccharide content in the above-mentioned Yunzhi is not less than 6%, or not less than 8%, or not less than 9%. The polysaccharide content is determined in accordance with the polysaccharide determination method under the "Content Determination" item of "Yunzhi" in the "Chinese Pharmacopoeia 2020 Edition".

[0014] The present invention has the following advantages: The present invention uses a combination of technologies of spraying and moistening, hot-pressing steam heating, ultrasonic treatment and microwave drying to transform related substances in the Coriolus versicolor into soluble polysaccharides on the one hand, and on the other hand, reduces the binding / adhesion between the polysaccharide and other substances, thereby significantly improving the content of Coriolus versicolor polysaccharides and increasing the release rate and amount of the polysaccharides from the Coriolus versicolor, which is beneficial to improving the therapeutic effect of Coriolus versicolor and the utilization rate of the medicinal materials. DETAILED DESCRIPTION

[0015] A method for processing Coriolus versicolor with high polysaccharide content, comprising the following steps: (1) Wash the dried fruiting bodies of Coriolus versicolor with water, cover the Coriolus versicolor with cotton cloth, and spray water on the cotton cloth to moisten the Coriolus versicolor for 12-24 hours, so that the Coriolus versicolor is fully moistened and there is no free water; (2) placing the moistened Coriolus versicolor in a hot-pressed steam cabinet, sealing it, evacuating it, and introducing hot-pressed steam to make the steam pressure in the hot-pressed steam cabinet 1.2Mpa-1.8Mpa, heating it at 188℃-208℃ for 1h-3h, and then quickly releasing the steam, completely releasing the hot steam in the hot-pressed steam cabinet within 2min-5min, and then cooling it; (3) placing the heated Coriolus versicolor in an ultrasonic processor for ultrasonic treatment, wherein the ultrasonic treatment conditions are 300 W of power, 20 kHz of frequency, and 15 min to 30 min of time; (4) Drying the Yunzhi after ultrasonic treatment in a microwave dryer. The microwave drying conditions are as follows: microwave frequency of 2450 MHz, drying temperature of 60° C.-70° C., and drying time of 20 min-30 min. Yunzhi with a high polysaccharide content is obtained. The polysaccharide content is not less than 6.0%. Under preferred conditions, the polysaccharide content is not less than 8%.

[0016] At present, in order to obtain Coriolus versicolor with high content of Coriolus versicolor polysaccharide, it is generally screened from germplasm. On the one hand, it is difficult to find germplasm resources, and on the other hand, the cultivation time of high-quality germplasm is long and the cost is high. The present invention uses a combination of hot pressing treatment, ultrasonic treatment and microwave drying to convert the relevant substances in Coriolus versicolor into soluble polysaccharides on the one hand, and on the other hand, reduce the binding / adhesion between polysaccharides and other substances, so that the content of Coriolus versicolor polysaccharide is significantly improved, and the utilization rate of Coriolus versicolor is improved. It can be verified by experimental results that a single method has limited effect on the improvement of the content of Coriolus versicolor polysaccharide. Only by combining the three treatments can the content of Coriolus versicolor polysaccharide be effectively improved.

[0017] The present invention will be further described below in conjunction with examples, but the present invention is not limited by the following examples.

[0018] Example 1 Preparation of Coriolus versicolor with high polysaccharide content Take 7 portions of Yunzhi fruiting bodies, wash them with water, cover them with cotton cloth, and spray water on the cotton cloth to moisten the Yunzhi for 24 hours, so that the Yunzhi is fully moistened and there is no free water; then process them according to the following methods: (1) Place the moistened Coriolus versicolor in a hot-pressed steam cabinet, seal it, evacuate it, and introduce hot-pressed steam to make the steam pressure in the hot-pressed steam cabinet 0.5Mpa-2Mpa, heat it at 151℃-212℃ for 2h, then quickly release the steam, completely release the hot steam in the hot-pressed steam cabinet within 5min, and then cool it; (2) placing the heated Coriolus versicolor in an ultrasonic processor for ultrasonic treatment, and the ultrasonic treatment conditions are as follows: power 300 W, frequency 20 kHz, and time 15 min; (3) The ultrasonically treated Coriolus versicolor was placed in a microwave dryer for drying. The microwave drying conditions were as follows: microwave frequency of 2450 MHz, drying temperature of 60°C, and drying time of 30 min to obtain Coriolus versicolor.

[0019] The total sugar and monosaccharide of Yunzhi treated under different pressures were determined respectively according to the determination method of total sugar and monosaccharide under Yunzhi [Content Determination] in Volume 1 of the 2020 edition of the "Chinese Pharmacopoeia", and the content of Yunzhi polysaccharide was calculated.

[0020] Table 1 Effects of different steam pressure treatments on the polysaccharide content of Coriolus versicolor The test results show that after the heat-pressing treatment, when the steam pressure exceeds 1.2MPa, the content of polysaccharides in Coriolus versicolor increases significantly, exceeding 1 times the content of polysaccharides in Coriolus versicolor without heat-pressing treatment or low-pressure treatment. When the pressure exceeds 1.8MPa, the increase in polysaccharide content is less.

[0021] Example 2 Preparation of Coriolus versicolor with high polysaccharide content Take 6 portions of soaked Yunzhi and process as follows: (1) Place the moistened Coriolus versicolor in a hot-pressed steam cabinet, seal it, evacuate it, and introduce hot-pressed steam to make the steam pressure in the hot-pressed steam cabinet reach 1.5 MPa. The heating treatment time is 0-4 hours, and then the steam is quickly released. The hot steam in the hot-pressed steam cabinet is completely released within 5 minutes, and then cooled; (2) placing the heated Coriolus versicolor in an ultrasonic processor for ultrasonic treatment, and the ultrasonic treatment conditions are as follows: power 300 W, frequency 20 kHz, and time 15 min; (3) The ultrasonically treated Coriolus versicolor was placed in a microwave dryer for drying. The microwave drying conditions were as follows: microwave frequency of 2450 MHz, drying temperature of 60°C, and drying time of 30 min to obtain Coriolus versicolor.

[0022] The total sugar and monosaccharide of Yunzhi treated with different heat pressing times were determined respectively according to the determination method of total sugar and monosaccharide under Yunzhi [Content Determination] in Volume 1 of the "Chinese Pharmacopoeia" 2020, and the content of Yunzhi polysaccharide was calculated.

[0023] Table 2 Effects of heat-pressing treatment of Coriolus versicolor at different times on Coriolus versicolor polysaccharides The test results show that different heat pressing times have a great influence on the content of Coriolus versicolor polysaccharides. When the heat pressing is 1 hour, the content of Coriolus versicolor polysaccharides increases significantly, and the content of polysaccharides gradually increases with the extension of time, but when it exceeds 3 hours, the content of polysaccharides changes less when the time is extended.

[0024] Example 3 Preparation of Coriolus versicolor with high polysaccharide content Take 5 portions of soaked Yunzhi and process as follows: (1) Place the moistened Coriolus versicolor in a hot-pressed steam cabinet, seal it, evacuate it, and introduce hot-pressed steam to make the steam pressure in the hot-pressed steam cabinet reach 1.5 MPa. The heating treatment time is 2 hours, and then the steam is quickly released. The hot steam in the hot-pressed steam cabinet is completely released within 2-20 minutes, and then cooled; (2) placing the heated Coriolus versicolor in an ultrasonic processor for ultrasonic treatment, and the ultrasonic treatment conditions are as follows: power 300 W, frequency 20 kHz, and time 15 min; (3) The ultrasonically treated Coriolus versicolor was placed in a microwave dryer for drying. The microwave drying conditions were as follows: microwave frequency of 2450 MHz, drying temperature of 60°C, and drying time of 30 min to obtain Coriolus versicolor.

[0025] Take 10g (dry weight) of Yunzhi with different steam release times, place it in a round-bottom flask, accurately add 100mL of water, weigh it, heat it to boiling, continue to heat and reflux for 10min, weigh it again, add water to make up for the lost water, shake it well, filter it, and determine the total sugar and monosaccharide in the extract according to the determination method of total sugar and monosaccharide under Yunzhi [Content Determination] in Volume 1 of the 2020 edition of the Chinese Pharmacopoeia, calculate the polysaccharide content, and use the following formula for extraction rate: Extraction rate = ×100%.

[0026] Table 3 Effects of different steam release times on Coriolus versicolor polysaccharides The test results show that different steam release times have a great influence on the extraction of Coriolus versicolor polysaccharides. The shorter the release time and the faster the speed, the easier it is to extract the polysaccharides in Coriolus versicolor. According to the test results, the steam release time should be less than 5 minutes.

[0027] Example 4 Preparation of Coriolus versicolor with high polysaccharide content Take 5 portions of soaked Yunzhi and process as follows: (1) Place the moistened Coriolus versicolor in a hot-pressed steam cabinet, seal it, evacuate it, and introduce hot-pressed steam to make the steam pressure in the hot-pressed steam cabinet reach 1.5 MPa. The heating treatment time is 2 hours, and then the steam is quickly released. The hot steam in the hot-pressed steam cabinet is completely released within 5 minutes, and then cooled; (2) placing the heated Coriolus versicolor in an ultrasonic processor for ultrasonic treatment, wherein the ultrasonic treatment conditions are 300 W of power, 20 kHz of frequency, and 0-30 min of time; (3) The ultrasonically treated Coriolus versicolor was placed in a microwave dryer for drying. The microwave drying conditions were as follows: microwave frequency of 2450 MHz, drying temperature of 60°C, and drying time of 30 min to obtain Coriolus versicolor.

[0028] Take 10g (dry weight) of Yunzhi with different ultrasonic times, place it in a round-bottom flask, accurately add 100mL of water, weigh it, heat it to boiling, continue to heat and reflux for 10min, weigh it again, add water to make up for the lost water, shake it well, filter it, and determine the total sugar and monosaccharide in the extract according to the determination method of total sugar and monosaccharide under Yunzhi [Content Determination] in Volume 1 of the 2020 edition of the Chinese Pharmacopoeia, calculate the polysaccharide content, and use the following formula for extraction rate: Extraction rate = ×100%.

[0029] Table 4 Effects of different ultrasound times on Coriolus versicolor polysaccharides The experimental results show that ultrasound has a great influence on the extraction and release of polysaccharides. The longer the ultrasound time, the higher the polysaccharide extraction rate. When the ultrasound time exceeds 15 minutes, the polysaccharide extraction rate increases significantly, but when it exceeds 20 minutes, the polysaccharide extraction rate does not increase much.

[0030] Example 5 Preparation of Coriolus versicolor with high polysaccharide content Take 5 portions of soaked Yunzhi and process as follows: (1) Place the moistened Coriolus versicolor in a hot-pressed steam cabinet, seal it, evacuate it, and introduce hot-pressed steam to make the steam pressure in the hot-pressed steam cabinet reach 1.5 MPa. The heating treatment time is 2 hours, and then the steam is quickly released. The hot steam in the hot-pressed steam cabinet is completely released within 5 minutes, and then cooled; (2) placing the heated Coriolus versicolor in an ultrasonic processor for ultrasonic treatment, and the ultrasonic treatment conditions are as follows: power 300 W, frequency 20 kHz, and time 15 min; (3) Drying the ultrasonically treated Coriolus versicolor in a microwave dryer or a hot air drying oven. The microwave drying conditions are as follows: microwave frequency of 2450 MHz, drying temperature of 60°C-70°C, and drying time of 20-30 min; the hot air drying temperature is 60°C-80°C; and Coriolus versicolor is obtained.

[0031] The total sugar and monosaccharide of Yunzhi treated with different drying methods were determined respectively according to the determination method of total sugar and monosaccharide under Yunzhi [Content Determination] in Volume 1 of the "Chinese Pharmacopoeia" 2020, and the content of Yunzhi polysaccharide was calculated.

[0032] Table 5 Effects of different drying methods and conditions on Coriolus versicolor polysaccharide The test results show that the drying method also affects the content of Coriolus versicolor polysaccharide, and the content of Coriolus versicolor polysaccharide obtained by microwave drying is high. Therefore, the drying conditions are determined to be microwave drying, with a microwave frequency of 2450MHz, a drying temperature of 60-70℃, and a drying time of 20min-30min.

[0033] Example 6 Preparation of Coriolus versicolor with high polysaccharide content This embodiment provides a pretreatment method for increasing the polysaccharide content of Coriolus versicolor, comprising the following steps: (1) Wash the dried fruiting bodies of Coriolus versicolor with water, cover the Coriolus versicolor with cotton cloth, and spray water on the cotton cloth to moisten the Coriolus versicolor for 12 hours; (2) placing the moistened Coriolus versicolor in a hot-pressed steam cabinet, sealing it, evacuating it, and introducing hot-pressed steam to make the steam pressure in the hot-pressed steam cabinet reach 1.6 MPa. The heating treatment time is 2 hours, and then the hot steam in the hot-pressed steam cabinet is completely released within 3 minutes and cooled; (3) placing the heated Coriolus versicolor in an ultrasonic processor for ultrasonic treatment, and the ultrasonic treatment conditions are 300 W of power, 20 kHz of frequency, and 20 min of time; (4) The ultrasonically treated Coriolus versicolor was placed in a microwave dryer for drying. The microwave drying conditions were as follows: microwave frequency of 2450 MHz, drying temperature of 60°C, and drying time of 30 min.

[0034] This embodiment also provides the above method for obtaining Coriolus versicolor with high polysaccharide content. The total sugar and monosaccharide are respectively determined according to the determination method of total sugar and monosaccharide under the item of Coriolus versicolor [Content Determination] in Volume 1 of the 2020 edition of the "Chinese Pharmacopoeia", and the Coriolus versicolor polysaccharide content is calculated to be 8.8%; the polysaccharide content of Coriolus versicolor without any treatment is 3.5%; the Coriolus versicolor with high polysaccharide content is cut into thick slices to obtain Coriolus versicolor decoction piece 1.

[0035] Example 7 Preparation of Coriolus versicolor with high polysaccharide content This embodiment provides a pretreatment method for increasing the polysaccharide content of Coriolus versicolor, comprising the following steps: (1) Wash the dried fruiting bodies of Coriolus versicolor with water, cover the Coriolus versicolor with cotton cloth, and spray water on the cotton cloth to moisten the Coriolus versicolor for 18 hours, so that the Coriolus versicolor is fully moistened and there is no free water; (2) placing the moistened Coriolus versicolor in a hot-pressed steam cabinet, sealing it, evacuating it, and introducing hot-pressed steam to make the steam pressure in the hot-pressed steam cabinet reach 1.3 MPa. The heating treatment time is 3 hours, and then the hot steam in the hot-pressed steam cabinet is completely released within 4 minutes and cooled; (3) placing the heated Coriolus versicolor in an ultrasonic processor for ultrasonic treatment, and the ultrasonic treatment conditions are as follows: power 300 W, frequency 20 kHz, and time 18 min; (4) The ultrasonically treated Coriolus versicolor was placed in a microwave dryer for drying. The microwave drying conditions were as follows: microwave frequency of 2450 MHz, drying temperature of 70°C, and drying time of 25 min.

[0036] This embodiment also provides the above method for obtaining Coriolus versicolor with high polysaccharide content. The total sugar and monosaccharide are respectively determined according to the determination method of total sugar and monosaccharide under the item of Coriolus versicolor [Content Determination] in Volume 1 of the 2020 edition of the "Chinese Pharmacopoeia", and the polysaccharide content of Coriolus versicolor is calculated to be 9.2%; the polysaccharide content of Coriolus versicolor without any treatment is 3.5%; the Coriolus versicolor with high polysaccharide content is cut into thick slices to obtain Coriolus versicolor decoction slices 2.

[0037] Comparative Example 1 Preparation of Yunzhi Decoction Pieces This comparative example pre-processes the fruiting bodies of Versicolor versicolor according to the following method: (1) Wash the dried fruiting bodies of Coriolus versicolor with water, cover the Coriolus versicolor with cotton cloth, and spray water on the cotton cloth to moisten the Coriolus versicolor for 18 hours, so that the Coriolus versicolor is fully moistened and there is no free water; (2) The ultrasonically treated Coriolus versicolor was placed in a microwave dryer for drying. The microwave drying conditions were as follows: microwave frequency of 2450 MHz, drying temperature of 70°C, and drying time of 25 min.

[0038] According to the determination method of total sugar and monosaccharide under the item of Yunzhi [Content Determination] in Volume 1 of the 2020 edition of the "Chinese Pharmacopoeia", the total sugar and monosaccharide of the above-mentioned Yunzhi were respectively determined, and the content of Yunzhi polysaccharide was calculated to be 4.2%; the polysaccharide content of Yunzhi without any treatment was 3.6%; the Yunzhi obtained by the above method was cut into thick slices to obtain Yunzhi decoction slice 3.

[0039] Application Example 1 Extraction rate of Coriolus versicolor polysaccharide from Coriolus versicolor Take 50g of each of the Yunzhi decoction pieces prepared in Examples 6, 7 and Comparative Example 1, add water and decoct twice, add 8 times the amount of water for the first time, and add 6 times the amount of water for the second time, decoct for 1 hour each time, filter, combine the decoctions, concentrate, and set to 100mL. Determine the content of total sugar and monosaccharide in the decoction according to the determination method of polysaccharides under Yunzhi [Content Determination] in the 2020 edition of the Chinese Pharmacopoeia, and calculate the content of Yunzhi polysaccharide.

[0040] Table 6 The content of polysaccharides in the decoction of Coriolus versicolor treated with and without the treatment of the present invention The results in Table 6 show that the content of polysaccharides and the extraction rate of polysaccharides in the decoction of Coriolus versicolor treated by the present invention are significantly higher than those in the decoction of Coriolus versicolor not treated by the present invention.

[0041] Application Example 2 Effect of Coriolus versicolor extract on immune enhancement The decoctions of the Coriolus versicolor slices 1 and 3 in Application Example 1 were concentrated to obtain extracts 1 and 3, each 1 mL of the concentrate being equivalent to 1 g of Coriolus versicolor.

[0042] 150 healthy adult mice were randomly divided into extract group 1, extract group 3 and blank control group, with 50 mice in each group. Each group was intragastrically gavaged with extract or saline 0.1mL / 10g every day, and various immune function indicators were measured after 30 days of gavage.

[0043] Specific immune function index: 30 days after mice were given the extract, 2% sheep red blood cell suspension was prepared, and 0.2 mL of sheep red blood cell suspension was injected intraperitoneally into each mouse for immunization. Blood was collected 4 days later, the mice were killed, and the spleen was removed. The spleen was prepared with 5×10 6 Cell suspension. Mix RPMI-1640 medium with an equal amount of Hank's solution of 2 times concentration, dispense into small test tubes, 0.5 mL per tube, add 50 μL 10% sheep red blood cells and 25 μL spleen cell suspension, pour onto a glass slide with a thin layer of agarose, incubate in a 37°C, 5% CO2 incubator for 1.5 hours, then add complement, continue incubation for 1.5 hours, centrifuge, take serum and dilute 200 times, and measure the absorbance value of the test tube and sheep red blood cells when half of the hemolysis occurs. The hemolysin content is expressed as the half hemolysis value (HC 50 )express.

[0044] Nonspecific immune function index Mouse carbon clearance experiment: 30 days after the mice were given the extract, 0.1mL / 10g of 4-fold diluted Indian ink was injected into the tail vein. 20μL of blood was collected from the medial canthal venous plexus 2 and 10 minutes after the injection of ink, and added to 2mL of 0.1% Na2CO3 solution. The absorbance value was measured at a wavelength of 600nm using an enzyme marker, and the thymus, liver, and spleen were weighed. The phagocytic index was calculated using the absorbance value, liver weight, and spleen weight.

[0045] Non-specific immune function index Peritoneal macrophage phagocytosis of chicken red blood cells: 30 days after mice were given the extract, 0.2 mL of 2% packed sheep red blood cells were injected into the peritoneum of each mouse. The mice were killed, 4 mL of Hank's solution was added, 0.5 mL of peritoneal wash fluid was added to 0.5 mL of 1% chicken red blood cell suspension, added to the agar circle of the slide, incubated at 37°C for 20 minutes, fixed, and stained with Giemsa solution. The number of macrophages, the number of macrophages phagocytizing chicken red blood cells, and the number of chicken red blood cells phagocytosed were observed under an optical microscope, and the phagocytic percentage and phagocytic index were calculated.

[0046] Determination of NK cell activity, an indicator of nonspecific immune function: After 30 days of administration of the extract, the mice were killed. The spleen was removed aseptically, and spleen cell suspension was prepared. After lysing red blood cells, the cell concentration was adjusted to 2×10 per ml. 7Take YAC-1 cells (4×10 per ml) 5 100 μL of each of 100 μL of culture medium (cells) and 100 μL of spleen cells (effector-target ratio 50:1) were added to a 96-well culture plate and cultured in a 37°C, 5% CO2 incubator for 4 h. Then, the cells were centrifuged for 5 min. 100 μL of supernatant was aspirated from each well and placed in a flat-bottom 96-well culture plate. 100 μL of lactate dehydrogenase matrix solution was added to react for 10 min. 30 μL of HCl (1 mol / L) was added to each well, and the absorbance was measured at 490 nm on a microplate reader.

[0047] In the above experiments, the number of animals used in each group of each experiment was 10.

[0048] Table 7 Effects of Coriolus versicolor extract on immunity of mice Note: ✱ Compared with the control group P <0.05.

[0049] The above test results show that Yunzhi has the effect of improving the immunity of mice, and the effect of Yunzhi obtained by the method of the present invention is more obvious, with obvious differences compared with the control group ( P <0.05).

[0050] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. A pretreatment method for increasing the polysaccharide content of Coriolus versicolor, characterized in that: The following steps are involved: (1) Soak the dried fruiting bodies of Coriolus versicolor thoroughly; (2) subjecting the fruiting body in step (1) to heat-pressing treatment and rapidly releasing the pressure; (3) treating the fruiting body in step (2) with ultrasonic wave; (4) Drying the fruiting body in step (3) to obtain Versicolor versicolor with a high polysaccharide content.

2. The pretreatment method according to claim 1, characterized in that The specific operation of moistening is: cover the fruiting body with wet objects, spray an appropriate amount of clean water, and keep it moist for 12h-24h.

3. The pretreatment method according to claim 1, characterized in that: The water vapor pressure of the hot pressing treatment is 1.2Mpa-2.0Mpa, and the pressurization treatment time is 1h-4h; the pressure relief speed is to release the pressure to normal pressure within 5min, such as 5min, 4min, 3min, 2min, 1min.

4. The pretreatment method according to claim 1, characterized in that: The water vapor pressure of the hot pressing treatment is 1.2Mpa-1.8Mpa, the pressurization treatment time is 1h-3h; the pressure relief speed is to release the pressure to normal pressure within 2min-5min.

5. The pretreatment method according to claim 1, characterized in that: The conditions of ultrasonic treatment are: power 300W, frequency 20kHz, time 15min-30min; more preferably, time 15min-20min.

6. The pretreatment method according to claim 1, characterized in that: The drying method is microwave drying.

7. The pretreatment method according to claim 6, characterized in that: The microwave frequency is 2450 MHz, the drying temperature is 60°C-70°C, and the drying time is 20min-30min.

8. A Coriolus versicolor obtained by the pretreatment method according to any one of claims 1 to 7.

9. The Coriolus versicolor according to claim 8, characterized in that The Versicolor coriolus can be a fruiting body, a slice of a fruiting body, or a crushed product.

10. The Coriolus versicolor according to claim 8, characterized in that The polysaccharide content in the versicolor is not less than 6%, or not less than 8%, or not less than 9%.

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