A Chinese herbal medicine composition for improving immunity and its preparation method and application

Abstract

The present invention proposes a Chinese herbal medicine composition for improving immunity and its preparation method and application, which belong to the field of medical technology. Rhizoma Anemarrhenae, Poria, Polygonatum, and Lycium barbarum are fermented to obtain Chinese herbal medicine selenium-rich / germanium polysaccharide, which is reacted with phycobiliprotein after carboxyl modification to obtain Chinese herbal medicine selenium-rich / germanium polysaccharide-phycobiliprotein complex. After the bacterial mud in the fermentation process is lysed by snail enzyme, a lysate is obtained, which is stirred and mixed with Chinese herbal medicine selenium-rich / germanium polysaccharide-phycobiliprotein complex, berberine, and camptothecin to obtain a Chinese herbal medicine composition for improving immunity. The Chinese herbal medicine composition for improving immunity prepared by the present invention contains rich glycoprotein, organic selenium / germanium complex, strengthens the foundation and supports yang, has good bactericidal, antioxidant, anti-inflammatory, antiviral, antitumor effects, is non-toxic, has high bioavailability, and is easily absorbed and other advantages, and has high patient compliance and broad application prospects.

Description

Technical Field

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

[0002] With the continuous development of society and the improvement of people's living standards, people are paying more and more attention to their health. Each of us is exposed to many external threats to our health, including air pollution, environmental pollution, work pressure, financial pressure, bacteria and viruses, and improper diet. Furthermore, the changing pace of work and life in cities can easily lead to sub-health. All these adverse health factors often lead to poor concentration, memory loss, frequent fatigue, and premature aging. Therefore, it is crucial to improve our immunity and enhance our ability to fight these adverse health factors.

[0003] Traditional Chinese Medicine doesn't have the term "low immunity," but it does have similar concepts like qi deficiency and body weakness. Patients are susceptible to infections like colds, sore throats, and pneumonia because of a lack of vital energy, which allows for repeated invasions of external pathogens. Western medicine explains this as low immunity, conveying the same concept. Therefore, improving immunity is 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, as well as its preparation method and application. The composition 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, and is easy to absorb. It has high patient compliance and has broad application prospects.

[0005] The technical solution of the present invention is achieved as follows:

[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 a Chinese herbal medicine selenium / germanium-rich polysaccharide, which is then carboxylated and modified and reacted with phycobiliprotein to obtain a Chinese herbal medicine selenium / germanium-rich polysaccharide-phycobiliprotein complex; lysing bacterial sludge produced during the fermentation process with snail enzyme to obtain a lysate, which is then stirred and uniformly mixed 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. Rhizoma Anemarrhenae, Poria, Polygonatum, and Lycium barbarum were washed, dried, crushed, added to water, inoculated with selenium yeast and germanium yeast seed liquid, fermented, filtered, washed, and the liquid was collected and freeze-dried to obtain a slurry. The filtrate was added with ethanol for precipitation, filtered, washed, and dried to obtain a selenium / germanium herbal polysaccharide.

[0009] S2. The herbal selenium-rich / germanium polysaccharide was dissolved in water, alkali solution and monochloroacetic acid were added, the reaction was heated with stirring, ethanol was added for precipitation, filtered, the solid was washed, and dried to obtain a carboxylated herbal selenium-rich / germanium polysaccharide;

[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. The carboxylated herbal selenium-rich / germanium polysaccharide was added to water, NHS and EDC were added, the reaction was activated, phycobiliprotein was added, and the reaction was stirred to obtain a herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex;

[0012] S5. The bacterial sludge in step S1 is added to water, snail enzyme is added, enzymatically hydrolyzed, the enzyme is inactivated, filtered, and the filtrate is freeze-dried to obtain a lysate;

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

[0014] As a further improvement of the present invention, the mass ratio of the Rhizoma Anemarrhenae, Poria, Polygonatum 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 were 2-4v / v% and 1-3v / v%, respectively. The fermentation conditions were 38-42°C, 50-150r / min, and the fermentation was carried out for 36-48h. Ethanol was added until the ethanol content of the system was 70-80wt%, and the precipitation time was 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, the mass ratio of the kudzu vine and spirulina in step S3 is 3-5:10, and the ultrafiltration has a molecular weight cut-off of proteins in the range 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 1-2 h.

[0019] As a further improvement of the present invention, the mass ratio of the Chinese herbal selenium-rich / 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. Polysaccharides with pharmacological effects are mostly active polysaccharides, which are mainly found in fungi, algae and rhizomes. The polysaccharides extracted from Rhizoma Anemarrhenae, Poria, Polygonatum and Lycium barbarum of the present invention can act on IL-2 secreting cells, promote cell secretion, increase the number of IL-2 or enhance its activity, 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 make them 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] Kudzu vine 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 form a complex by coupling with carboxylated selenium / germanium-rich polysaccharides in Chinese herbal medicines, greatly enhancing the effect of promoting immunity.

[0026] In addition, carboxyl-modified Chinese herbal selenium / germanium 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 and exist in organic form. They have good immunomodulatory effects, 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, and is easily absorbed. It has high patient compliance and has broad application prospects. DETAILED DESCRIPTION

[0030] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts 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 Boshiao Biochemical Technology Research Co., Ltd.

[0032] Preparation method of bacterial seed liquid: inoculate the bacterial strain into yeast extract peptone glucose medium, activate and culture at 40°C, 100r / min for 18-24h to obtain 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, the preparation method comprising 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 weight of water, and inoculate them with a bacterial count of 10 8 -10 9 cfu / mL of selenium yeast and germanium yeast seed liquid, inoculated at 2v / v% and 1v / v%, respectively, at 38°C, 50r / min, fermented for 36h, filtered, washed with solids, collected the bacterial liquid, freeze-dried to obtain bacterial sludge, added ethanol to the filtrate until the ethanol content of the system was 70wt%, precipitated for 1h, filtered, washed with solids, and dried to obtain Chinese herbal selenium / germanium-rich polysaccharides;

[0037] S2. 10 g of Chinese herbal selenium-rich / germanium polysaccharide was dissolved in 300 mL of water, 100 mL of 10 wt% KOH solution and 1 g of monochloroacetic acid were added, heated to 50 ° C, stirred for 3 h, ethanol was added to the system ethanol content of 70 wt%, precipitated for 1 h, filtered, the solid was washed, and dried to obtain carboxylated Chinese herbal selenium-rich / germanium polysaccharide;

[0038] S3. The 3g Gexianmi and 10g 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 molecular weight cutoff of 10K-25K protein, 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 activation reaction was carried out 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. 10 g of the bacterial sludge from step S1 was added to 200 mL of water, 1 g of snail enzyme was added, and enzymatic hydrolysis was carried out at 40 ° C for 1 h, the enzyme was inactivated, filtered, and the filtrate was freeze-dried 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, the preparation method comprising the following steps:

[0044] S1. Wash 4g of Anemarrhena, 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 count of 10. 8 -10 9cfu / mL of selenium yeast and germanium yeast seed liquid, inoculated at 4v / v% and 3v / v%, respectively, at 42°C, 150r / min, fermented for 48h, filtered, washed with solids, collected the bacterial liquid, freeze-dried to obtain bacterial sludge, added ethanol to the filtrate until the ethanol content of the system was 80wt%, precipitated for 2h, filtered, washed with solids, and dried to obtain Chinese herbal selenium / germanium-rich polysaccharides;

[0045] S2. 10 g of Chinese herbal selenium-rich / germanium polysaccharide was dissolved in 300 mL of water, 100 mL of 15 wt% NaOH solution and 2 g of monochloroacetic acid were added, heated to 60 ° C, stirred for 4 h, ethanol was added to the system ethanol content of 80 wt%, precipitated for 2 h, filtered, the solid was washed, and dried to obtain carboxylated Chinese herbal selenium-rich / germanium polysaccharide;

[0046] S3. 5g of Gexianmi and 10g 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 a molecular weight cutoff of 10K-25K protein, freeze-dried, and phycobiliprotein was obtained;

[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 reaction was activated 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. 10 g of the bacterial sludge from step S1 was added to 200 mL of water, 2 g of snail enzyme was added, and enzymatic hydrolysis was carried out at 50 ° C for 2 h, the enzyme was inactivated, filtered, and the filtrate was freeze-dried to obtain a lysate;

[0049] S6. Evenly mix 17 g of the Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex and 8 g of the 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, the preparation method comprising 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 count of 10 8 -10 9cfu / mL of selenium yeast and germanium yeast seed liquid, inoculation amounts of 3v / v% and 2v / v%, respectively, at 40°C, 100r / min, fermentation culture for 42h, filtration, solid washing, collection of bacterial liquid, freeze-drying to obtain bacterial sludge, ethanol was added to the filtrate until the ethanol content of the system was 75wt%, precipitation for 1.5h, filtration, solid washing, and drying to obtain Chinese herbal selenium / germanium-rich polysaccharides;

[0053] S2. 10 g of Chinese herbal selenium-rich / germanium polysaccharide was dissolved in 300 mL of water, 100 mL of 12 wt% NaOH solution and 1.5 g of monochloroacetic acid were added, heated to 55 ° C, stirred for 3.5 h, ethanol was added to the system ethanol content of 75 wt%, precipitated for 1.5 h, filtered, the solid was washed, and dried to obtain carboxylated Chinese herbal selenium-rich / germanium polysaccharide;

[0054] S3. 4g of Gexianmi and 10g 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 molecular weight cutoff of 10K-25K protein, 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 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;

[0056] S5. 10 g of the bacterial sludge from step S1 was added to 200 mL of water, 1.5 g of snail enzyme was added, and enzymatic hydrolysis was carried out at 45 ° C for 1.5 h, the enzyme was inactivated, filtered, and the filtrate was freeze-dried to obtain a lysate;

[0057] S6. Evenly mix 15 g of the Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex and 6 g of the 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 the selenium yeast seed liquid is not 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 count 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 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 count 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 mud, ethanol was added to the filtrate until the ethanol content of the system was 75wt%, precipitation for 1.5h, filtration, solid washing, and drying to obtain Chinese herbal selenium / germanium rich polysaccharide.

[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 them, crush them, add them into 7 times the weight 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 count of 10 8 -10 9cfu / mL of selenium yeast and germanium yeast seed liquid, inoculation amounts of 3v / v% and 2v / v%, respectively, at 40°C, 100r / min, fermentation culture for 42h, filtration, solid washing, collection of bacterial liquid, freeze-drying to obtain bacterial sludge, ethanol was added to the filtrate until the ethanol content of the system was 75wt%, precipitation for 1.5h, filtration, solid washing, and drying to obtain Chinese herbal selenium / germanium-rich polysaccharides;

[0074] S2. 4g of Gexianmi and 10g 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 molecular weight cutoff of 10K-25K protein, 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. 10 g of the bacterial sludge in step S1 was added to 200 mL of water, 1.5 g of snail enzyme was added, and enzymatic hydrolysis was carried out at 45 ° C for 1.5 h, the enzyme was inactivated, filtered, and the filtrate was freeze-dried to obtain a lysate;

[0077] S5. Evenly mix 15 g of the Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex and 6 g of the 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 minutes 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, and stir and mix for 20 minutes to prepare a Chinese herbal 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 group, and Comparative Example 1-7 group, with 10 mice in each group. The mice were adaptively fed for 7 days in a feeding room with a temperature of 22±3°C, a humidity of 50±5%, and a 12-h light / dark cycle. They had free access to water and food.

[0092] The mice were maintained on medication for 15 days, twice a day, with a daily dose of 5g / kg of the corresponding immune-enhancing herbal composition. 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 60mg / 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 day 16, the mice's eyes were enucleated, and blood was collected and centrifuged. Blood levels of IL-4 and IL-2 were measured using mouse interleukin-4 (IL-4) ELISA kits and mouse interleukin-2 (IL-2) ELISA kits, as described in the manufacturer's instructions. 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] 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 promote the secretion of IL-4 and IL-2, thereby improving the body's immune function.

[0098] After weighing the mice, the mice were sacrificed by cervical dislocation and autopsied to remove the spleen and thymus. The wet weight data were measured, and the spleen index and thymus index were calculated.

[0099] Organ index = organ mass (mg) / mouse body 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: * indicates P < 0.05 compared with the blank group; # indicates P < 0.05 compared with the model group.

[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 indexes of immune organs, indicating that they have a good immune-promoting effect.

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

[0106] A 0.2 mmol / L DPPH solution was prepared, and the immunity-enhancing Chinese herbal medicine composition prepared in Examples 1-3 or Comparative Examples 1-7 (at a concentration of 0.5 wt%) was prepared in PBS at pH 7.4. A 0.5 wt% vitamin C solution was prepared in PBS at pH 7.4. The reaction system was prepared according to the formula in Table 3. The reaction was carried out at 25° C. in the dark for 30 min, followed by centrifugation. The absorbance at a wavelength of 517 nm was measured. Three replicate measurements were performed for each group, and the average value was calculated.

[0107] Clearance (%) = (A2 + A3 - A1) / A2 × 100%

[0108] Wherein, A2: absorbance of negative control group; A3: absorbance of blank control group; A1: 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 Group 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] As can be seen from the above table, the immunity-enhancing Chinese herbal medicine compositions 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 principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing a Chinese herbal medicine composition for improving immunity, characterized in that: 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 count of 10 8 -10 9 cfu / mL of selenium yeast and germanium yeast seed liquid, inoculation amounts of 3v / v% and 2v / v%, respectively, at 40°C, 100r / min, fermentation culture for 42h, filtration, solid washing, collection of bacterial liquid, freeze-drying to obtain bacterial sludge, ethanol was added to the filtrate until the ethanol content of the system was 75wt%, precipitation for 1.5h, filtration, solid washing, and drying to obtain Chinese herbal selenium / germanium-rich polysaccharides; S2. 10 g of Chinese herbal selenium-rich / germanium polysaccharide was dissolved in 300 mL of water, 100 mL of 12 wt% NaOH solution and 1.5 g of monochloroacetic acid were added, heated to 55 ° C, stirred for 3.5 h, ethanol was added to the system ethanol content of 75 wt%, precipitated for 1.5 h, filtered, the solid was washed, and dried to obtain carboxylated Chinese herbal selenium-rich / germanium polysaccharide; S3. 4g of Gexianmi and 10g 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 molecular weight cutoff of 10K-25K protein, and freeze-dried to obtain phycobiliprotein; 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; S5. 10 g of the bacterial sludge from step S1 was added to 200 mL of water, 1.5 g of snail enzyme was added, and enzymatic hydrolysis was carried out at 45 ° C for 1.5 h, the enzyme was inactivated, filtered, and the filtrate was freeze-dried to obtain a lysate; S6. Evenly mix 15 g of the Chinese herbal selenium-rich / germanium polysaccharide-phycobiliprotein complex and 6 g of the 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.

2. A Chinese herbal medicine composition for improving immunity obtained by the preparation method according to claim 1.

3. Use of the Chinese herbal medicine composition for improving immunity as claimed in claim 2 in the preparation of a medicine for improving immunity.