Chinese herbal medicine composition for improving immunity as well as preparation method and application of Chinese herbal medicine composition

Abstract

The invention provides a Chinese herbal medicine composition for improving immunity as well as a preparation method and application thereof, and belongs to the technical field of medicines. The preparation method comprises the following steps: fermenting rhizoma anemarrhenae, poria cocos, rhizoma polygonati and fructus lycii to prepare Chinese herbal medicine selenium / germanium-rich polysaccharide, carrying out carboxylation modification on the Chinese herbal medicine selenium / germanium-rich polysaccharide and phycobiliprotein to prepare a Chinese herbal medicine selenium / germanium-rich polysaccharide-phycobiliprotein compound, and dissolving bacterial sludge in the fermentation process with helicase to prepare a bacteria-dissolving product; and uniformly stirring and mixing with the Chinese herbal medicine selenium / germanium-rich polysaccharide-phycobiliprotein compound, berberine and camptothecin to prepare the Chinese herbal medicine composition for improving immunity. The prepared Chinese herbal medicine composition capable of improving immunity contains rich glycoprotein and organic selenium / germanium compounds, has the effects of consolidating body resistance and strengthening yang, has the advantages of good sterilization, oxidation resistance, inflammation resistance, virus resistance and tumor resistance, no toxicity, high bioavailability, easiness in absorption and the like, is high in patient compliance and has a wide application prospect.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a Chinese herbal medicine composition for improving immunity, a preparation method and application thereof. Background Art

[0002] With the continuous development of social economy and the improvement of people's living standards, people are paying more and more attention to health. The health of each of us is threatened by many external factors, including air pollution, environmental pollution, work pressure, economic pressure, bacteria and viruses, improper diet, etc. Moreover, due to the changes in the pace of work and life in the city, it is easy for people to enter a sub-healthy state. All these unhealthy factors usually make people unable to concentrate, memory decline, often feel lazy, and age prematurely. Therefore, it is very important to improve one's immunity and enhance the ability to fight against various unfavorable health factors.

[0003] There is no such thing as "low immunity" in traditional Chinese medicine, but there are similar concepts such as qi deficiency and body weakness. Patients are prone to colds, pharyngitis, pneumonia and other infection symptoms because the body lacks positive energy and external pathogens can easily invade repeatedly. Western medicine explains it as low immunity, which means the same thing, so improving immunity is actually a common goal pursued by both Chinese and Western medicine. Summary of the invention

[0004] The purpose of the present invention is to propose a Chinese herbal medicine composition for improving immunity, a preparation method and an application thereof, which is rich in glycoprotein and organic selenium / germanium complex, can strengthen the body and support yang, has good bactericidal, antioxidant, anti-inflammatory, antiviral and anti-tumor effects, is non-toxic, has high bioavailability, is easy to absorb, has high patient compliance, and has broad application prospects.

[0005] The technical solution of the present invention is achieved in this way:

[0006] The invention provides a method for preparing a Chinese herbal medicine composition for improving immunity. The method comprises the following steps: fermenting rhizoma anemarrhenae, poria cocos, polygonatum sibiricum and wolfberry to obtain Chinese herbal medicine selenium / germanium-rich polysaccharide, and reacting the polysaccharide with phycobiliprotein after carboxyl modification to obtain a Chinese herbal medicine selenium / germanium-rich polysaccharide-phycobiliprotein complex; subjecting bacterial sludge in the fermentation process to snail enzyme lysis to obtain a lysate, and mixing the lysate with the Chinese herbal medicine selenium / germanium-rich polysaccharide-phycobiliprotein complex, berberine and camptothecin to obtain the Chinese herbal medicine composition for improving immunity.

[0007] As a further improvement of the present invention, the following steps are included:

[0008] S1. Wash Rhizoma Anemarrhenae, Poria, Polygonatum sibiricum and Lycium barbarum, dry, crush, add to water, inoculate with selenium yeast and germanium yeast seed solution, ferment, filter, wash solid, collect bacterial solution, freeze-dry to obtain bacterial mud, add ethanol to the filtrate for precipitation, filter, wash solid, dry, and obtain Chinese herbal selenium / germanium-rich polysaccharide;

[0009] S2. Dissolve the selenium-rich / germanium-rich Chinese herbal medicine polysaccharide in water, add alkali solution and monochloroacetic acid, heat and stir the reaction, add ethanol to precipitate, filter, wash the solid, and dry to obtain a carboxylated Chinese herbal medicine polysaccharide with selenium / germanium;

[0010] S3. The rice and Spirulina were dried, crushed, added to water, frozen with liquid nitrogen, thawed at room temperature, filtered, the filtrate was ultrafiltered, and freeze-dried to obtain phycobiliprotein;

[0011] S4. Add the carboxylated Chinese herbal selenium-rich / germanium polysaccharide to water, add NHS and EDC, activate the reaction, add phycobiliprotein, stir the reaction, and prepare the Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex;

[0012] S5. Add the bacterial sludge in step S1 to water, add snail enzyme, enzymolyze, inactivate the enzyme, filter, and freeze-dry the filtrate to obtain a lysate;

[0013] S6. Evenly mix the Chinese herbal selenium / germanium polysaccharide-phycobiliprotein complex and the lysate, add berberine and camptothecin, stir and mix evenly to prepare a Chinese herbal medicine composition for improving immunity.

[0014] As a further improvement of the present invention, the mass ratio of Anemarrhena asphodeloides, Poria cocos, Polygonatum sibiricum and Lycium barbarum in step S1 is 2-4:7-9:3-5:1-3, and the bacterial content of the selenium yeast and germanium yeast seed liquid is 10 8 -10 9 cfu / mL, the inoculation amounts are 2-4v / v% and 1-3v / v%, respectively. The fermentation conditions are 38-42°C, 50-150r / min, and the fermentation is carried out for 36-48h. Ethanol is added until the ethanol content of the system is 70-80wt%, and the precipitation time is 1-2h.

[0015] As a further improvement of the present invention, the alkali solution in step S2 is a 10-15wt% NaOH or KOH solution, the mass ratio of the Chinese herbal selenium / germanium polysaccharide and monochloroacetic acid is 10:1-2, the temperature of the heating and stirring reaction is 50-60°C, the time is 3-4h, ethanol is added until the ethanol content of the system is 70-80wt%, and the precipitation time is 1-2h.

[0016] As a further improvement of the present invention, in step S3, the mass ratio of the radix schizonepetae to the spirulina is 3-5:10, and the ultrafiltration has a protein cut-off molecular weight of 10K-25K.

[0017] As a further improvement of the present invention, the mass ratio of the carboxylated Chinese herbal selenium / germanium polysaccharide, NHS, EDC, and phycobiliprotein in step S4 is 10:2-4:1-2:3-5, the activation reaction time is 20-30 minutes, and the stirring reaction time is 10-15 hours.

[0018] As a further improvement of the present invention, in step S5, the mass ratio of the bacterial sludge to the snail enzyme is 10:1-2, the temperature of the enzymatic hydrolysis is 40-50°C, and the time is 1-2h.

[0019] As a further improvement of the present invention, the mass ratio of the Chinese herbal selenium / germanium polysaccharide-phycobiliprotein complex, lysate, berberine and camptothecin in step S6 is 12-17:4-8:0.5-1:0.1-0.2.

[0020] The present invention further protects a Chinese herbal medicine composition for improving immunity prepared by the above preparation method.

[0021] The present invention further protects the use of the above-mentioned Chinese herbal medicine composition for improving immunity in the preparation of a medicine for improving immunity.

[0022] The present invention has the following beneficial effects:

[0023] There are many polysaccharides in Chinese herbal medicines, some of which are biologically inactive, such as starch, gum and mucilage, and are usually removed as impurities. Most polysaccharides with pharmacological effects are active polysaccharides, which are mainly found in fungi, algae, and rhizomes. The polysaccharides extracted from Rhizoma Anemarrhenae, Poria, Polygonatum sibiricum and Lycium barbarum in the present invention can act on IL-2 secreting cells, promote cell secretion, increase the number or enhance the activity of IL-2, promote the conversion rate of T lymphocytes and B cells, increase the synthesis and secretion of IL-2 by T cells and B cells, thereby improving the function of the immune system, and stimulate various immune cells to increase their activity, increase their number or produce corresponding antibodies, activate the reticuloendothelial system and complement system, and produce interferon.

[0024] In selenium-rich and germanium-rich polysaccharides, selenium and germanium are combined on macromolecular polysaccharides to form polysaccharide-selenium / germanium complexes, which can significantly inhibit the proliferation of tumor cells, maintain normal immune function, and have good antioxidant capacity. In addition, organic selenium / germanium complexes have the advantages of being non-toxic, highly bioavailable, and easily absorbed, which are significantly superior to the direct addition of inorganic selenium and inorganic germanium.

[0025] Pueraria lobata and Spirulina are rich in phycobiliprotein, a type of light-harvesting pigment protein that can stimulate the body's immune system response, increase the activity of lymphocytes and macrophages, and enhance the body's ability to kill cancer cells. It can also form a complex by coupling with carboxylated Chinese herbal selenium / germanium polysaccharides, which greatly enhances the effect of promoting immunity.

[0026] In addition, carboxyl-modified Chinese herbal selenium / germanium-rich polysaccharides also improve biological activity by changing the polysaccharide structure. They have the advantages of low cost, few side effects, and easy absorption by the body. They significantly improve the antioxidant and anti-inflammatory activities of polysaccharides, and improve their anti-tumor and immunomodulatory activities.

[0027] Selenium yeast and germanium yeast themselves are rich in active ingredients such as β-glucan and antimicrobial peptides. The selenium and germanium elements are rich in organic form and have good immunomodulatory effects. They also reduce waste generation and improve the resource utilization of by-products.

[0028] Berberine and camptothecin have good bactericidal, antiviral and anti-tumor effects, and have a synergistic effect.

[0029] The immunity-enhancing Chinese herbal medicine composition prepared by the present invention contains rich glycoproteins and organic selenium / germanium complexes, strengthens the body and supports yang, has good bactericidal, antioxidant, anti-inflammatory, antiviral and anti-tumor effects, is non-toxic, has high bioavailability, is easy to absorb, and has the advantages of high patient compliance and broad application prospects. DETAILED DESCRIPTION

[0030] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0031] Selenium yeast, 10 billion cfu / g, was purchased from Angel Yeast Co., Ltd.; germanium yeast, 10 billion cfu / g, was purchased from Guangzhou Bosiao Biochemical Technology Research Co., Ltd.

[0032] Preparation method of bacterial seed liquid: inoculate the bacterial strain into yeast extract peptone glucose medium, at 40°C, 100r / min, activate and culture for 18-24h to obtain the bacterial seed liquid.

[0033] Snail enzyme, with a cell wall breaking rate of 90%, was purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0034] Example 1

[0035] This embodiment provides a method for preparing a Chinese herbal medicine composition for improving immunity, and the preparation method comprises the following steps:

[0036] S1. Wash 2g of Anemarrhena, 7g of Poria, 3g of Polygonatum, and 1g of Lycium barbarum, dry them, crush them, add them to 5 times the mass of water, and inoculate them with a bacterial content of 10 8 -10 9 cfu / mL of selenium yeast and germanium yeast seed liquid, inoculation amounts of 2v / v% and 1v / v%, respectively, at 38°C, 50r / min, fermentation culture for 36h, filtration, solid washing, collection of bacterial liquid, freeze drying, preparation of bacterial mud, addition of ethanol to the filtrate until the ethanol content of the system is 70wt%, precipitation for 1h, filtration, solid washing, drying, preparation of Chinese herbal selenium / germanium-rich polysaccharides;

[0037] S2. Dissolve 10g of Chinese herbal selenium / germanium polysaccharide in 300mL of water, add 100mL of 10wt% KOH solution and 1g of monochloroacetic acid, heat to 50°C, stir and react for 3h, add ethanol until the ethanol content of the system is 70wt%, precipitate for 1h, filter, wash the solid, and dry to obtain carboxylated Chinese herbal selenium / germanium polysaccharide;

[0038] S3. 3g of Gexianmi and 10g of Spirulina were dried, crushed, added to 5 times the mass of water, frozen with liquid nitrogen, thawed at room temperature, filtered, and the filtrate was ultrafiltered to a protein with a molecular weight cutoff of 10K-25K, and freeze-dried to obtain phycobiliprotein;

[0039] S4. 10 g of carboxylated Chinese herbal selenium-rich / germanium polysaccharide was added to 500 mL of water, 2 g of NHS and 1 g of EDC were added, the reaction was activated for 20 min, 3 g of phycobiliprotein was added, and the reaction was stirred for 10 h to obtain a Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex;

[0040] S5. Add 10 g of the bacterial sludge in step S1 to 200 mL of water, add 1 g of snail enzyme, enzymolyze at 40 ° C for 1 h, inactivate the enzyme, filter, and freeze-dry the filtrate to obtain a lysate;

[0041] S6. Evenly mix 12 g of the Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex and 4 g of the lysate, add 0.5 g of berberine and 0.1 g of camptothecin, and stir for 20 minutes to prepare a Chinese herbal composition for improving immunity.

[0042] Example 2

[0043] This embodiment provides a method for preparing a Chinese herbal medicine composition for improving immunity, and the preparation method comprises the following steps:

[0044] S1. Wash 4g of Rhizoma Anemarrhenae, 9g of Poria, 5g of Polygonatum, and 3g of Lycium barbarum, dry them, crush them, add them to 10 times the mass of water, and inoculate them with a bacterial content of 10 8 -10 9cfu / mL of selenium yeast and germanium yeast seed liquid, inoculation amounts of 4v / v% and 3v / v%, respectively, at 42°C, 150r / min, fermentation culture for 48h, filtration, solid washing, collection of bacterial liquid, freeze-drying to obtain bacterial mud, adding ethanol to the filtrate until the ethanol content of the system is 80wt%, precipitation for 2h, filtration, solid washing, drying, to obtain Chinese herbal selenium / germanium-rich polysaccharides;

[0045] S2. Dissolve 10 g of Chinese herbal selenium / germanium polysaccharide in 300 mL of water, add 100 mL of 15 wt% NaOH solution and 2 g of monochloroacetic acid, heat to 60 ° C, stir and react for 4 h, add ethanol to the ethanol content of the system to 80 wt%, precipitate for 2 h, filter, wash the solid, and dry to obtain carboxylated Chinese herbal selenium / germanium polysaccharide;

[0046] S3. 5 g of Gexianmi and 10 g of Spirulina were dried, crushed, added to 10 times the mass of water, frozen with liquid nitrogen, thawed at room temperature, filtered, and the filtrate was ultrafiltered to cut off the protein with a molecular weight of 10K-25K, and freeze-dried to obtain phycobiliprotein;

[0047] S4. 10 g of carboxylated Chinese herbal selenium-rich / germanium polysaccharide was added to 500 mL of water, 4 g of NHS and 2 g of EDC were added, the activation reaction was carried out for 30 min, 5 g of phycobiliprotein was added, and the reaction was stirred for 15 h to obtain a Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex;

[0048] S5. Add 10 g of the bacterial sludge in step S1 to 200 mL of water, add 2 g of snail enzyme, enzymolyze at 50 ° C for 2 h, inactivate the enzyme, filter, and freeze-dry the filtrate to obtain a lysate;

[0049] S6. Evenly mix 17 g of Chinese herbal selenium / germanium polysaccharide-phycobiliprotein complex and 8 g of lysate, add 1 g of berberine and 0.2 g of camptothecin, and stir for 20 minutes to prepare a Chinese herbal composition for improving immunity.

[0050] Example 3

[0051] This embodiment provides a method for preparing a Chinese herbal medicine composition for improving immunity, and the preparation method comprises the following steps:

[0052] S1. Wash 3g of Anemarrhena, 8g of Poria, 4g of Polygonatum, and 2g of Lycium barbarum, dry them, crush them, add them to 7 times the mass of water, and inoculate them with a bacterial content of 10 8 -10 9cfu / mL of selenium yeast and germanium yeast seed liquid, inoculation amount of 3v / v% and 2v / v%, respectively, 40°C, 100r / min, fermentation culture for 42h, filtration, solid washing, collection of bacterial liquid, freeze drying, preparation of bacterial mud, ethanol added to the filtrate until the ethanol content of the system is 75wt%, precipitation for 1.5h, filtration, solid washing, drying, preparation of Chinese herbal selenium / germanium-rich polysaccharides;

[0053] S2. Dissolve 10 g of Chinese herbal selenium / germanium polysaccharide in 300 mL of water, add 100 mL of 12 wt% NaOH solution and 1.5 g of monochloroacetic acid, heat to 55 ° C, stir and react for 3.5 h, add ethanol to the ethanol content of the system to 75 wt%, precipitate for 1.5 h, filter, wash the solid, and dry to obtain carboxylated Chinese herbal selenium / germanium polysaccharide;

[0054] S3. 4 g of Gexianmi and 10 g of Spirulina were dried, crushed, added to 7 times the mass of water, frozen with liquid nitrogen, thawed at room temperature, filtered, and the filtrate was ultrafiltered to a protein with a molecular weight cutoff of 10K-25K, and freeze-dried to obtain phycobiliprotein;

[0055] S4. 10 g of carboxylated Chinese herbal selenium-rich / germanium polysaccharide was added to 500 mL of water, 3 g of NHS and 1.5 g of EDC were added, the activation reaction was carried out for 25 min, 4 g of phycobiliprotein was added, and the reaction was stirred for 12 h to obtain a Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex;

[0056] S5. Add 10 g of the bacterial sludge in step S1 to 200 mL of water, add 1.5 g of snail enzyme, enzymolysis at 45 ° C for 1.5 h, inactivate the enzyme, filter, and freeze-dry the filtrate to obtain a lysate;

[0057] S6. Evenly mix 15 g of Chinese herbal selenium / germanium polysaccharide-phycobiliprotein complex and 6 g of lysate, add 0.7 g of berberine and 0.15 g of camptothecin, and stir for 20 minutes to prepare a Chinese herbal composition for improving immunity.

[0058] Comparative Example 1

[0059] Compared with Example 3, the difference is that no selenium yeast seed liquid is inoculated in step S1.

[0060] The details are as follows:

[0061] S1. Wash 3g of Anemarrhena, 8g of Poria, 4g of Polygonatum, and 2g of Lycium barbarum, dry them, crush them, add them to 7 times the mass of water, and inoculate them with a bacterial content of 10 8 -10 9cfu / mL of germanium yeast seed liquid, inoculation amount of 5v / v%, 40℃, 100r / min, fermentation culture for 42h, filtration, solid washing, collection of bacterial liquid, freeze-drying to obtain bacterial mud, addition of ethanol to the filtrate until the ethanol content of the system is 75wt%, precipitation for 1.5h, filtration, solid washing, drying to obtain Chinese herbal selenium / germanium rich polysaccharide.

[0062] Comparative Example 2

[0063] Compared with Example 3, the difference is that the germanium yeast seed liquid is not inoculated in step S1.

[0064] The details are as follows:

[0065] S1. Wash 3g of Anemarrhena, 8g of Poria, 4g of Polygonatum, and 2g of Lycium barbarum, dry them, crush them, add them to 7 times the mass of water, and inoculate them with a bacterial content of 10 8 -10 9 cfu / mL of selenium yeast seed liquid, inoculation amount of 5v / v%, 40℃, 100r / min, fermentation culture for 42h, filtration, solid washing, collection of bacterial liquid, freeze-drying to obtain bacterial sludge, addition of ethanol to the filtrate until the ethanol content of the system is 75wt%, precipitation for 1.5h, filtration, solid washing, drying to obtain Chinese herbal selenium / germanium-rich polysaccharides.

[0066] Comparative Example 3

[0067] Compared with Example 3, the difference is that fermentation is not performed in step S1, and heating and boiling extraction is adopted.

[0068] The details are as follows:

[0069] S1. Wash 3g of Anemarrhena asphodeloides, 8g of Poria cocos, 4g of Polygonatum sibiricum and 2g of Lycium barbarum, dry and crush them, add them into 7 times the mass of water, heat and reflux for 4h, filter, add ethanol to the filtrate until the ethanol content of the system is 75wt%, precipitate for 1.5h, filter, wash the solid and dry it to obtain Chinese herbal polysaccharide.

[0070] Comparative Example 4

[0071] Compared with Example 3, the difference is that step S2 is not performed.

[0072] The details are as follows:

[0073] S1. Wash 3g of Anemarrhena, 8g of Poria, 4g of Polygonatum, and 2g of Lycium barbarum, dry them, crush them, add them to 7 times the mass of water, and inoculate them with a bacterial content of 10 8 -10 9cfu / mL of selenium yeast and germanium yeast seed liquid, inoculation amount of 3v / v% and 2v / v%, respectively, 40°C, 100r / min, fermentation culture for 42h, filtration, solid washing, collection of bacterial liquid, freeze drying, preparation of bacterial mud, ethanol added to the filtrate until the ethanol content of the system is 75wt%, precipitation for 1.5h, filtration, solid washing, drying, preparation of Chinese herbal selenium / germanium-rich polysaccharides;

[0074] S2. 4 g of Gexianmi and 10 g of Spirulina were dried, crushed, added to 7 times the mass of water, frozen with liquid nitrogen, thawed at room temperature, filtered, and the filtrate was ultrafiltered to a protein with a molecular weight cutoff of 10K-25K, and freeze-dried to obtain phycobiliprotein;

[0075] S3. 10 g of Chinese herbal selenium-rich / germanium polysaccharide was added to 500 mL of water, 3 g of NHS and 1.5 g of EDC were added, the reaction was activated for 25 min, 4 g of phycobiliprotein was added, and the reaction was stirred for 12 h to obtain a Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex;

[0076] S4. Add 10 g of the bacterial sludge in step S1 to 200 mL of water, add 1.5 g of snail enzyme, enzymolysis at 45 ° C for 1.5 h, inactivate the enzyme, filter, and freeze-dry the filtrate to obtain a lysate;

[0077] S5. Evenly mix 15 g of Chinese herbal selenium / germanium polysaccharide-phycobiliprotein complex and 6 g of lysate, add 0.7 g of berberine and 0.15 g of camptothecin, and stir for 20 minutes to prepare a Chinese herbal composition for improving immunity.

[0078] Comparative Example 5

[0079] Compared with Example 3, the difference is that no reaction is performed in step S4, and the carboxylated Chinese herbal selenium / germanium-rich polysaccharide and phycobiliprotein are mixed.

[0080] The details are as follows:

[0081] S4. 10 g of carboxylated Chinese herbal selenium / germanium-rich polysaccharide and 4 g of phycobiliprotein were stirred and mixed for 10 min to prepare a mixture.

[0082] Comparative Example 6

[0083] Compared with Example 3, the difference is that no lysate is added in step S6.

[0084] The details are as follows:

[0085] S6. Add 0.7 g of berberine and 0.15 g of camptothecin to 21 g of the Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex, and stir and mix for 20 minutes to prepare a Chinese herbal composition for improving immunity.

[0086] Comparative Example 7

[0087] Compared with Example 3, the difference is that no Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex is added in step S6.

[0088] The details are as follows:

[0089] S6. Add 0.7 g of berberine and 0.15 g of camptothecin to 21 g of the lysate, stir and mix for 20 minutes to prepare a Chinese herbal medicine composition for improving immunity.

[0090] Test Example 1

[0091] 120 mice were randomly divided into 7 groups, blank group, model group (cyclophosphamide), Example 1-3 and Comparative Example 1-7 groups, 10 mice in each group. Adaptive feeding was performed for 7 days, the temperature of the feeding room was 22±3°C, the humidity was 50±5%, the light / dark alternation was 12h, and drinking water and food were free.

[0092] The mice were maintained on medication for 15 days, twice a day, with a daily dose of 5 g / kg of the corresponding Chinese herbal composition for improving immunity. The blank group and the model group mice were given the same dose of distilled water. Except for the blank group, the other groups were intraperitoneally injected with cyclophosphamide at a dose of 60 mg / kg·d on the 7th to 15th day of medication to construct an immunosuppressive mouse model, once a day, while the blank group mice were given the same amount of normal saline.

[0093] On the 16th day, the eyeballs of the mice were removed, blood was collected, centrifuged, and the contents of IL-4 and IL-2 in the blood were detected according to the instructions of the mouse interleukin-4 (IL-4) ELISA kit and the mouse interleukin-2 (IL-2) ELISA kit. The results are shown in Table 1.

[0094] Table 1

[0095] Group IL-4 content (pg / mL) IL-2 content (pg / mL) Blank group 11.75±0.39 16.92±0.52 Model Group 8.04±0.71* 13.19±0.89* Example 1 10.92±0.44# 16.10±0.61# Example 2 11.10±0.38# 16.04±0.58# Example 3 11.15±0.48# 16.19±0.64# Comparative Example 1 10.16±0.67 15.03±0.77 Comparative Example 2 10.03±0.74 14.87±0.89 Comparative Example 3 9.47±0.69 13.95±0.82 Comparative Example 4 10.84±0.71 15.72±0.80 Comparative Example 5 10.29±0.68 15.31±0.84 Comparative Example 6 9.21±0.59 14.23±0.69 Comparative Example 7 8.92±0.64 14.02±0.74

[0096] Note: * compared with the blank group, P < 0.05; # compared with the model group, P < 0.05.

[0097] It can be seen from the above table that the Chinese herbal medicine composition for improving immunity prepared in Examples 1-3 of the present invention can significantly promote the secretion of IL-4 and IL-2, thereby improving the body's immune function.

[0098] The mice were weighed, cervical dislocation was performed and then the mice were killed. The mice were dissected, the spleen and thymus were taken out, the wet weight data were measured, and the spleen index and thymus index were calculated.

[0099] Organ index = organ mass (mg) / mouse weight (g)

[0100] The results are shown in Table 2.

[0101] Table 2

[0102] Group Thymus index (mg / g) Spleen index (mg / g) Blank group 5.12±0.75 2.71±0.39 Model Group 1.90±0.42* 2.01±0.33* Example 1 3.89±0.58# 2.91±0.43# Example 2 3.92±0.61# 2.90±0.38# Example 3 3.97±0.44# 2.95±0.47# Comparative Example 1 3.91±0.69 2.85±0.52 Comparative Example 2 3.88±0.72 2.81±0.49 Comparative Example 3 3.72±0.85 2.72±0.56 Comparative Example 4 3.93±0.67 2.86±0.55 Comparative Example 5 3.90±0.73 2.82±0.64 Comparative Example 6 3.70±0.74 2.69±0.62 Comparative Example 7 3.52±0.87 2.45±0.77

[0103] Note: * compared with the blank group, P < 0.05; # compared with the model group, P < 0.05.

[0104] As can be seen from the above table, the immunity-enhancing Chinese herbal medicine compositions prepared in Examples 1-3 of the present invention can significantly improve the organ index of immune organs, indicating that they have a good immune-promoting effect.

[0105] Test Example 2 DPPH free radical scavenging rate determination

[0106] Prepare a 0.2mmol / L DPPH solution, and prepare the Chinese herbal medicine composition for improving immunity prepared in Examples 1-3 or Comparative Examples 1-7 (concentration of 0.5wt%) with PBS at pH=7.4. Prepare a vitamin C solution with a mass concentration of 0.5wt% with PBS at pH=7.4, prepare the reaction system according to the formula in Table 3, react for 30min at 25℃ in the dark, centrifuge, measure the absorbance at a wavelength of 517nm, measure 3 times in parallel for each group, and take the average value.

[0107] Clearance rate (%) = (A 2 +A 3 -A 1 ) / A 2 ×100%

[0108] In the formula, A 2 : absorbance of negative control group; A 3 : absorbance of blank control group; A 1 : Absorbance of sample group / positive control group.

[0109] Table 3

[0110] Test liquid Test sample (mL) PBS solution with pH = 7.4 (mL) DPPH solution (mL) Sample set 1 0 1 Positive control group 1 0 1 Negative control group 0 1 1 Blank control group 1 1 0

[0111] The results are shown in Table 4.

[0112] Table 4

[0113] Group DPPH free radical scavenging rate (%) Vitamin C Group 70.2 Example 1 91.5 Example 2 92.2 Example 3 93.0 Comparative Example 1 88.5 Comparative Example 2 85.6 Comparative Example 3 80.4 Comparative Example 4 82.2 Comparative Example 5 81.9 Comparative Example 6 78.7 Comparative Example 7 72.4

[0114] It can be seen from the above table that the Chinese herbal medicine compositions for improving immunity prepared in Examples 1-3 of the present invention have good antioxidant effects.

[0115] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a Chinese herbal medicine composition for improving immunity, characterized in that: The Chinese herbal selenium / germanium polysaccharide is prepared by fermenting Anemarrhena asphodeloides, Poria cocos, Polygonatum sibiricum and Lycium barbarum, which is then carboxylated and reacted with phycobiliprotein to prepare a Chinese herbal selenium / germanium polysaccharide-phycobiliprotein complex. The bacterial sludge in the fermentation process is lysed by snail enzyme to obtain a lysate, which is then stirred and evenly mixed with the Chinese herbal selenium / germanium polysaccharide-phycobiliprotein complex, berberine and camptothecin to prepare a Chinese herbal medicine composition for improving immunity.

2. The preparation method according to claim 1, characterized in that: The following steps are involved: S1. Wash Rhizoma Anemarrhenae, Poria, Polygonatum sibiricum and Lycium barbarum, dry, crush, add to water, inoculate with selenium yeast and germanium yeast seed solution, ferment, filter, wash solid, collect bacterial solution, freeze-dry to obtain bacterial mud, add ethanol to the filtrate for precipitation, filter, wash solid, dry, and obtain Chinese herbal selenium / germanium-rich polysaccharide; S2. Dissolve the selenium-rich / germanium-rich Chinese herbal medicine polysaccharide in water, add alkali solution and monochloroacetic acid, heat and stir the reaction, add ethanol to precipitate, filter, wash the solid, and dry to obtain a carboxylated Chinese herbal medicine polysaccharide with selenium / germanium; S3. The rice and Spirulina were dried, crushed, added to water, frozen with liquid nitrogen, thawed at room temperature, filtered, the filtrate was ultrafiltered, and freeze-dried to obtain phycobiliprotein; S4. Add the carboxylated Chinese herbal selenium-rich / germanium polysaccharide to water, add NHS and EDC, activate the reaction, add phycobiliprotein, stir the reaction, and prepare the Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex; S5. Add the bacterial sludge in step S1 to water, add snail enzyme, enzymolyze, inactivate the enzyme, filter, and freeze-dry the filtrate to obtain a lysate; S6. Evenly mix the Chinese herbal selenium / germanium polysaccharide-phycobiliprotein complex and the lysate, add berberine and camptothecin, stir and mix evenly to prepare a Chinese herbal medicine composition for improving immunity.

3. The preparation method according to claim 2, characterized in that: The mass ratio of Anemarrhena asphodeloides, Poria cocos, Polygonatum sibiricum and Lycium barbarum in step S1 is 2-4:7-9:3-5:1-3, and the bacterial content of the selenium yeast and germanium yeast seed liquid is 10 8 -10 9 cfu / mL, the inoculation amounts are 2-4v / v% and 1-3v / v%, respectively. The fermentation conditions are 38-42°C, 50-150r / min, and the fermentation is carried out for 36-48h. Ethanol is added until the ethanol content of the system is 70-80wt%, and the precipitation time is 1-2h.

4. The preparation method according to claim 2, characterized in that: The alkali solution in step S2 is a 10-15wt% NaOH or KOH solution, the mass ratio of the Chinese herbal selenium / germanium polysaccharide and monochloroacetic acid is 10:1-2, the temperature of the heating and stirring reaction is 50-60°C, the time is 3-4h, ethanol is added until the ethanol content of the system is 70-80wt%, and the precipitation time is 1-2h.

5. The preparation method according to claim 2, characterized in that: In step S3, the mass ratio of radix puerariae to spirulina is 3-5:10, and the ultrafiltration has a protein cut-off molecular weight of 10K-25K.

6. The preparation method according to claim 2, characterized in that: In step S4, the mass ratio of the carboxylated Chinese herbal selenium / germanium-rich polysaccharide, NHS, EDC, and phycobiliprotein is 10:2-4:1-2:3-5, the activation reaction time is 20-30 minutes, and the stirring reaction time is 10-15 hours.

7. The preparation method according to claim 2, characterized in that: In step S5, the mass ratio of bacterial sludge to snail enzyme is 10:1-2, the temperature of enzymolysis is 40-50°C, and the time is 1-2h.

8. The preparation method according to claim 2, characterized in that: In step S6, the mass ratio of the Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex, the lysate, berberine and camptothecin is 12-17:4-8:0.5-1:0.1-0.

2.

9. A Chinese herbal medicine composition for improving immunity obtained by the preparation method according to any one of claims 1 to 8.

10. Use of the Chinese herbal medicine composition for improving immunity as claimed in claim 9 in the preparation of a medicine for improving immunity.