Composition containing ginseng extract and preparation method and application thereof

Abstract

The present invention provides a composition containing ginseng extract, a preparation method, and an application thereof, belonging to the field of biomedicine. The composition containing ginseng extract provided by the present invention is composed of the following raw materials, measured by weight: 6-10 parts of Lobelia chinensis, 2-4 parts of Bogudan, 1-3 parts of wax fern, 3-6 parts of lantern grass, 5-8 parts of hibiscus leaves, 7-9 parts of lotus leaves, 8-10 parts of cassia seeds, and 10-15 parts of ginseng extract. The various medicinal ingredients in the composition containing ginseng extract provided by the present invention are used in combination, achieving synergistic effects and exhibiting a favorable therapeutic effect for both lung cancer and systemic lupus erythematosus.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a composition containing ginseng extract, a preparation method and an application thereof. Background Art

[0002] Ginseng's active ingredients (such as ginsenosides, polysaccharides, and polyphenols) have been extensively studied, particularly in the field of anti-tumor efficacy. Existing studies have shown that ginsenosides (such as Rg3 and Rh2) can exert anti-cancer effects through mechanisms such as inhibiting tumor cell proliferation, inducing apoptosis, and inhibiting angiogenesis. However, existing technologies suffer from the following drawbacks: low bioavailability: Components such as ginsenosides have limited in vivo efficacy due to poor water solubility and low intestinal absorption; single-ingredient limitations: Most studies have focused on single saponins, ignoring the potential for synergistic effects among multiple components; and crude preparation processes: Traditional extraction methods (such as decoction and ethanol reflux) easily destroy heat-sensitive components and fail to effectively enrich highly active components, resulting in poor formulation stability.

[0003] Lung cancer is a highly prevalent cancer worldwide. While existing targeted drugs are partially effective, they are associated with significant drug resistance and toxic side effects. Lung cancer is in urgent need of new, low-toxic, multi-target therapies. Systemic lupus erythematosus is an autoimmune disease affecting multiple organs. Current treatments primarily focus on glucocorticoids, immunosuppressants, and biologics (such as belimumab). However, long-term use can lead to infection, cardiovascular risks, and cumulative organ damage. Although some traditional Chinese medicine formulas (such as tripterygium wilfordii) are used as adjuvant therapy, their hepato-renal toxicity and unclear mechanisms of action limit their application. Currently, there are no studies in this field on ginseng extracts and their compositions that can simultaneously treat lung cancer and systemic lupus erythematosus. Summary of the Invention

[0004] In view of this, one of the objects of the present invention is to provide a composition containing ginseng extract and a preparation method thereof, which has a more significant therapeutic effect on lung cancer and systemic lupus erythematosus.

[0005] The second object of the present invention is to provide the use of the above composition in the preparation of drugs for treating lung cancer or systemic lupus erythematosus.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a composition containing ginseng extract, which is composed of the following raw materials in parts by weight: 6-10 parts of lobelia chinensis, 2-4 parts of bogudan, 1-3 parts of wax fern, 3-6 parts of lantern grass, 5-8 parts of hibiscus leaves, 7-9 parts of lotus leaves, 8-10 parts of cassia seeds, and 10-15 parts of ginseng extract;

[0008] The preparation method of the ginseng extract comprises: crushing ginseng, mixing the mixture with a complex enzyme and water in a weight ratio of 100:(6-8):100, and performing enzymolysis to obtain an enzymatic hydrolyzate; mixing the enzymatic hydrolyzate, corn flour, soybean meal powder, sucrose, and water in a weight-to-volume ratio of (30-50) g:(8-10) g:(6-10) g:(5-8) g:0.8 L to obtain a fermentation medium; mixing the fermentation medium with a Ganoderma lucidum liquid in a weight ratio of 100:4-6, fermenting at 30°C-35°C for 20 h-24 h, and drying to obtain the ginseng extract; the complex enzyme comprises cellulase, pectinase, and α-amylase; and the weight of Ganoderma lucidum mycelium in the Ganoderma lucidum liquid is 8 g / L-14 g / L.

[0009] Preferably, the weight ratio of the cellulase, pectinase and α-amylase is (1~2):(2~3):(0.5~1); the enzymatic activity of the cellulase is greater than 100,000 U / g; the enzymatic activity of the pectinase is greater than 30,000 U / g; and the enzymatic activity of the α-amylase is greater than 10,000 U / g.

[0010] Preferably, the initial pH of the enzymatic hydrolysis is 5.2-6.0, the temperature of the enzymatic hydrolysis is 40° C.-50° C., and the time of the enzymatic hydrolysis is 12 h-16 h.

[0011] Preferably, the Ganoderma lucidum liquid is obtained by inoculating Ganoderma lucidum into a liquid culture medium and culturing at 25°C~28°C; the composition of the liquid culture medium is 20g~30g glucose, 5g~10g peptone, 3g~5g yeast extract, 1g~2g potassium dihydrogen phosphate, 0.5g~1g magnesium sulfate, 10mg~20mg vitamin B1 and 1L water; the pH value of the liquid culture medium is 5~6.

[0012] The present invention also provides a preparation method of the above-mentioned composition, comprising the following steps: mixing lobelia chinensis, radix scutellariae, radix scutellariae, hibiscus leaves, lotus leaves and cassia seeds to obtain a mixture, mixing the mixture with water and soaking it, heating it to boiling and then decocting it for 1-2 hours, respectively collecting the decoction and the medicinal residue; concentrating the decoction to obtain a water extract; mixing the medicinal residue with an ethanol solution with a volume fraction of 70%-80% and soaking it, heating and refluxing it for extraction for 1.5-2.5 hours, collecting the extract and concentrating it to obtain an alcohol extract; mixing the alcohol extract, the water extract, wax fern and ginseng extract, and drying it to obtain the composition.

[0013] Preferably, the soaking time is 30 min to 60 min; and the concentration is to concentrate to a relative density of 0.9 to 1.0.

[0014] Preferably, the weight ratio of the mixture to water is 1:8-10; the weight ratio of the medicinal residue to the ethanol solution with a volume fraction of 70%-80% is 1:7-9.

[0015] The present invention also provides the use of the above composition in preparing medicine for preventing and treating lung cancer.

[0016] The present invention also provides the use of the above composition in preparing a medicine for treating systemic lupus erythematosus.

[0017] Beneficial effects of the present invention:

[0018] The present invention provides a composition containing ginseng extract, wherein the ginseng extract is enzymatically hydrolyzed using a composite enzyme, thereby releasing more of the originally encapsulated active ingredients in the ginseng, reducing the molecular weight of polysaccharides, and generating more easily absorbed oligosaccharides or oligosaccharides, thereby enhancing immunomodulatory activity. Furthermore, the enzymatic hydrolysis can convert ginsenosides in the ginseng into rare ginsenosides, thereby enhancing anti-tumor activity. Furthermore, the present invention enzymatically hydrolyzes the ginseng using a composite enzyme and then inoculates it with Ganoderma lucidum for fermentation. The fermentation process allows for biotransformation (converting the components in the ginseng into new active substances, such as ganoderic acid and a complex of Ganoderma lucidum polysaccharides and saponins), thereby enhancing the activity of the original components. Furthermore, fermentation with Ganoderma lucidum generates secondary metabolites (such as antioxidant peptides and organic acids), while fermentation also degrades potentially irritating components in ginseng (such as certain saponin precursors), thereby improving safety. In summary, the present invention utilizes a combined enzymatic hydrolysis and Ganoderma lucidum fermentation method to prepare a ginseng extract, achieving a synergistic effect and ultimately producing a ginseng extract with multi-target effects, which has excellent therapeutic effects on both lung cancer and systemic lupus erythematosus.

[0019] The present invention combines ginseng extract with lobelia chinensis, radix scutellariae, wax fern, hibiscus sibiricum, hibiscus rosa-sinensis, lotus leaf and cassia seed for use. The medicinal flavors synergistically enhance the efficacy, thereby further enhancing the therapeutic effect of ginseng extract on lung cancer and systemic lupus erythematosus. Specifically: Lobelia chinensis helps inhibit tumor cell proliferation and induce tumor cell apoptosis, can synergize with ginseng extract to activate immune response, and reduce systemic lupus erythematosus-related inflammation; Bogudan has anti-tumor and antioxidant effects; Baibai wax can promote mucosal and skin healing, reduce skin ulcers and lung damage in patients with systemic lupus erythematosus, and can inhibit excessive autoimmune reactions; Lantern grass can induce tumor cell autophagy, inhibit lung cancer metastasis, and can regulate metabolism, and improve abnormal liver and kidney function in patients with systemic lupus erythematosus; Hibiscus leaf can resist angiogenesis, block the vascular supply of lung cancer, and can reduce systemic lupus erythematosus-related pulmonary fibrosis and renal fibrosis; lotus leaf has antioxidant, anti-inflammatory and lipid metabolism regulating effects; Cassia seed exerts a bidirectional immune regulatory effect, inhibiting over-activated B cells (reducing autoantibodies), enhancing T cell anti-tumor activity, and promoting bowel movements and detoxification, promoting the excretion of metabolic waste, and reducing toxin accumulation in patients with tumors and systemic lupus erythematosus. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1The cell scratch results of Example 3 group, where the left picture is the initial scratch picture, and the right picture is the 24h scratch picture;

[0021] Figure 2 The cell scratch results of the comparative example 1 group, where the left picture is the initial scratch picture and the right picture is the 24h scratch picture;

[0022] Figure 3 This is the cell migration result of the Example 3 group;

[0023] Figure 4 This is the cell migration result of Comparative Example 1. DETAILED DESCRIPTION

[0024] The present invention provides a composition containing ginseng extract, which is composed of the following raw materials in parts by weight: 6-10 parts of lobelia chinensis, 2-4 parts of bogudan, 1-3 parts of wax fern, 3-6 parts of lantern grass, 5-8 parts of hibiscus leaves, 7-9 parts of lotus leaves, 8-10 parts of cassia seeds, and 10-15 parts of ginseng extract;

[0025] The preparation method of the ginseng extract comprises: crushing ginseng, mixing the mixture with a complex enzyme and water in a weight ratio of 100:(6-8):100, and performing enzymolysis to obtain an enzymatic hydrolyzate; mixing the enzymatic hydrolyzate, corn flour, soybean meal powder, sucrose, and water in a weight-to-volume ratio of (30-50) g:(8-10) g:(6-10) g:(5-8) g:0.8 L to obtain a fermentation medium; mixing the fermentation medium with a Ganoderma lucidum liquid in a weight ratio of 100:4-6, fermenting at 30°C-35°C for 20 h-24 h, and drying to obtain the ginseng extract; the complex enzyme comprises cellulase, pectinase, and α-amylase; and the weight of Ganoderma lucidum mycelium in the Ganoderma lucidum liquid is 8 g / L-14 g / L.

[0026] The present invention does not specifically limit the specific sources of the raw materials used in the preparation of ginseng extract; any commercially available products in the art can be used. In the present invention, the mesh size of the pulverization is preferably 100-150 mesh, and the specific method of pulverization is not specifically limited. In the present invention, the weight ratio of cellulase, pectinase, and α-amylase in the complex enzyme is preferably (1-2):(2-3):(0.5-1), more preferably (1.2-1.8):(2.2-2.5):(0.7-0.8). The cellulase activity is preferably greater than 100,000 U / g; the pectinase activity is preferably greater than 30,000 U / g; and the α-amylase activity is preferably greater than 10,000 U / g. In the present invention, the weight ratio of pulverized ginseng to the complex enzyme and water is preferably 100:(6.5-7.5):100, more preferably 100:(6.8-7.2):100. In the present invention, after pulverizing ginseng and mixing it with a complex enzyme and water, the pH is preferably adjusted to 5.2-6.0 before enzymatic hydrolysis, and the pH is more preferably 5.4-5.8. The enzymatic hydrolysis temperature is preferably 40-50°C, more preferably 42-48°C, and the enzymatic hydrolysis time is preferably 12-16 hours, more preferably 13-15 hours. After the enzymatic hydrolysis is completed, the enzyme is preferably inactivated, and the inactivation method is preferably 80-90°C for 5-8 minutes.

[0027] In the present invention, the weight-to-volume ratio of the enzymatic hydrolysate, corn flour, soybean meal, sucrose, and water is preferably (35-45) g:9 g:(7-9) g:(6-7) g:0.8 L, and more preferably (38-40) g:9 g:8 g:6 g:0.8 L. In the present invention, the fermentation medium is preferably sterilized, and the sterilization method is preferably 121°C for 2-4 minutes. In the present invention, the Ganoderma lucidum liquid is preferably obtained by inoculating Ganoderma lucidum into a liquid culture medium and culturing at 25°C~28°C; the culturing method is preferably shaking culture, and the shaking culture speed is preferably 120r / min~140r / min, more preferably 130r / min; the composition of the liquid culture medium is preferably 20g~30g glucose, 5g~10g peptone, 3g~5g yeast extract, 1g~2g potassium dihydrogen phosphate, 0.5g~1g magnesium sulfate, 10mg~20mg vitamin B1 and 1L water, more preferably 22g~28g glucose, 6g~8g peptone, 4g yeast extract, 1.5g potassium dihydrogen phosphate, 0.7g~0.8g magnesium sulfate, 12mg~16mg vitamin B1 and 1L water; the pH value of the liquid culture medium is preferably 5~6. In the present invention, the weight of the mycelia of the Ganoderma lucidum culture liquid is preferably 10 g / L to 12 g / L, and the weight of the mycelia of Ganoderma lucidum refers to the wet weight of the mycelia per unit volume of the culture liquid.

[0028] In the present invention, when the fermentation medium is mixed with the Ganoderma lucidum liquid, the weight ratio of the fermentation medium to the Ganoderma lucidum liquid is preferably 100:5. In the present invention, the fermentation temperature is preferably 31°C to 34°C, more preferably 32°C to 33°C; the fermentation time is preferably 21 hours to 23 hours, more preferably 22 hours. After fermentation, the entire fermentation system is dried. The specific drying method is not particularly limited in the present invention. Drying to a moisture content of 5% to 10% yields a ginseng extract.

[0029] The present invention does not particularly limit the specific sources of Lobelia chinensis, Bogudan, Baiwax, Lanterna sinensis, Hibiscus sangreae leaves, Lotus leaf, and Cassia seed; conventional commercial products in this field can be used. In the present invention, the weight of Lobelia chinensis is preferably 7 to 9 parts, more preferably 8 parts; the weight of Bogudan is preferably 3 parts; the weight of Baiwax is preferably 2 parts; the weight of Lanterna sinensis is preferably 4 to 5 parts; the weight of Hibiscus sangreae leaves is preferably 6 to 7 parts; the weight of Lotus leaf is preferably 8 parts; the weight of Cassia seed is preferably 9 parts; and the weight of the ginseng extract is preferably 11 to 14 parts, more preferably 12 to 13 parts.

[0030] The composition of the present invention uses ginseng extract as the main ingredient, and has the effects of greatly replenishing vital energy, restoring the pulse and strengthening the body, tonifying the spleen and lungs, promoting the production of body fluids and nourishing blood, and calming the mind and improving intelligence. It enhances the body's immunity and strengthens the body's vital energy to resist pathogenic factors, playing a key role in strengthening the body's vital energy and eliminating pathogenic factors. Lobelia chinensis and Cassia seed are both auxiliary ingredients. Lobelia chinensis clears heat and detoxifies, promotes diuresis and reduces swelling, and assists ginseng extract in enhancing its heat-clearing and detoxifying effects in the formula, helping to eliminate heat and toxins in the body. Cassia seed clears heat and improves eyesight, and moistens the intestines and promotes bowel movements. On the one hand, it assists Lobelia chinensis in clearing heat, and on the other hand, by moistening the intestines and promoting bowel movements, it excretes toxins from the intestines, reducing the body's burden and assisting the main ingredient in treating diseases. Bogudan, white wax, lantern grass, hibiscus leaf and lotus leaf are adjuvants. Bogudan can dispel rheumatism, dredge meridians, assist the main and auxiliary drugs, enhance the effects of dredging meridians and relieving pain, and is an adjuvant drug; white wax can stop bleeding, promote tissue regeneration, and relieve pain, and help repair body damage. In the process of treating diseases, it has a certain therapeutic effect on possible tissue damage and bleeding. As an adjuvant drug, lantern grass clears away heat and detoxifies, promotes diuresis and stops bleeding, assists the clearing of heat and detoxification of lobelia and other herbs, and can also promote diuresis to expel dampness from the body. It can relieve the damp-heat symptoms that may accompany systemic lupus erythematosus, and is an adjuvant drug; hibiscus leaves clear away heat and detoxify, reduce swelling and relieve pain, and assist the main and auxiliary drugs in clearing heat and detoxifying, reducing swelling and dispersing nodules, and have an improving effect on local swelling and pain that may be caused by the disease, and serve as an adjuvant drug; lotus leaves clear away heat and dampness, promote the development of clear yang, cool blood and stop bleeding, and prevent ginseng and other drugs from being too warm and promoting heat. At the same time, its cooling blood and hemostasis effect is helpful for the bleeding symptoms that may occur in the disease, and it also has the function of an adjuvant drug.

[0031] The present invention also provides a preparation method of the above-mentioned composition, comprising the following steps: mixing lobelia chinensis, radix scutellariae, radix scutellariae, hibiscus leaves, lotus leaves and cassia seeds to obtain a mixture, mixing the mixture with water and soaking it, heating it to boiling and then decocting it for 1-2 hours, respectively collecting the decoction and the medicinal residue; concentrating the decoction to obtain a water extract; mixing the medicinal residue with an ethanol solution with a volume fraction of 70%-80% and soaking it, heating and refluxing it for extraction for 1.5-2.5 hours, collecting the extract and concentrating it to obtain an alcohol extract; mixing the alcohol extract, the water extract, wax fern and the above-mentioned ginseng extract, and drying it to obtain the composition.

[0032] In the present invention, the weight ratio of the mixture to water is preferably 1:8-10, more preferably 1:9. When the mixture is mixed with water and soaked, the soaking time is preferably 30-60 minutes, and in specific embodiments, it can be 30, 35, 40, 45, 50, 55, and 60 minutes. In the present invention, the boiling time is preferably 1.2-1.8 hours, more preferably 1.4-1.6 hours, and the boiling is preferably maintained at a slight boil. The present invention does not particularly limit the specific method of concentration. In one embodiment, a reduced pressure concentration method is used, specifically concentration at 50°C-60°C. The concentration is preferably concentrated to a relative density of 0.9-1.0. In the present invention, relative density refers to the ratio of the density of the substance to the density of water at 50°C-60°C.

[0033] In the present invention, when the medicinal residue is soaked in an ethanol solution, the volume fraction of the ethanol solution is preferably 72% to 78%, more preferably 74% to 76%; the weight ratio of the medicinal residue to the 70% to 80% ethanol solution is preferably 1:7 to 9, more preferably 1:8. When the medicinal residue is mixed and soaked in a 70% to 80% ethanol solution, the soaking time is preferably 30 minutes to 60 minutes. In specific embodiments, it can be 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, and 60 minutes. In the present invention, the heating and reflux extraction time is preferably 1.8 to 2.4 hours, more preferably 2.0 to 2.2 hours. The present invention does not specifically limit the specific method for concentrating the ethanol extract. In one embodiment, a reduced pressure concentration method is used, specifically concentration at 50°C to 60°C. The concentration is preferably concentrated to a relative density of 0.9 to 1.0.

[0034] The present invention also provides the use of the above composition in the preparation of a drug for preventing and treating lung cancer. In the pharmaceutical application of the present invention, the composition can exert an anti-tumor effect as the sole active ingredient.

[0035] The present invention also provides the use of the above composition in the preparation of a drug for treating systemic lupus erythematosus. In the pharmaceutical application of the present invention, the composition can be used as the sole active ingredient to treat systemic lupus erythematosus.

[0036] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0037] In the following examples, unless otherwise specified, all methods are conventional.

[0038] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0039] In the following examples, cellulase was purchased from Shanghai Xiangqi Biotechnology Co., Ltd. with an enzyme activity of more than 100,000 U / g; pectinase was purchased from Anhui Weimao Biotechnology Co., Ltd. with an enzyme activity of more than 30,000 U / g; α-amylase was purchased from Beina Biotechnology with a number of BNCC382029. The resource name is Ganoderma lucidum (red ganoderma), which is Latin for Ganoderma lucidum .

[0040] Example 1

[0041] A ginseng extract is prepared by the following preparation method:

[0042] The ginseng is crushed until it can completely pass through a 100-mesh sieve, and then mixed with a complex enzyme and water in a weight ratio of 100 g ginseng: 6 g complex enzyme: 100 g water, and the pH is adjusted to 5.2. The mixture is enzymatically hydrolyzed at 40° C. for 16 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated at 80° C. for 8 min to obtain an enzymolyzate; the complex enzyme is composed of cellulase, pectinase and α-amylase, and the weight ratio of cellulase, pectinase and α-amylase is 1:2:0.5.

[0043] The Ganoderma lucidum was inoculated into a liquid culture medium and cultured at 25° C. and 120 r / min until the weight of the Ganoderma lucidum mycelium reached 8 g / L, thereby obtaining a Ganoderma lucidum liquid; the liquid culture medium consisted of 20 g of glucose, 5 g of peptone, 3 g of yeast extract, 1 g of potassium dihydrogen phosphate, 0.5 g of magnesium sulfate, 10 mg of vitamin B1, and 1 L of water, and was sterilized at 121° C. for 10 min.

[0044] The enzymatic hydrolysate, corn flour, soybean meal powder, sucrose and water were mixed in a weight-to-volume ratio of 30 g enzymatic hydrolysate: 8 g corn flour: 6 g soybean meal powder: 5 g sucrose: 0.8 L water, and sterilized at 121° C. for 2 min to obtain a fermentation medium; the fermentation medium was mixed with Ganoderma lucidum liquid in a weight ratio of 100:4, and fermented at 30° C. and 130 r / min for 20 h. The entire fermentation system was dried with hot air at 50° C. to a moisture content of 5% to obtain a ginseng extract.

[0045] Example 2

[0046] A ginseng extract is prepared by the following preparation method:

[0047] The ginseng is crushed until it can completely pass through a 150-mesh sieve, and then mixed with a complex enzyme and water in a weight ratio of 100 g ginseng: 8 g complex enzyme: 100 g water, and the pH is adjusted to 6.0. The mixture is enzymatically hydrolyzed at 50° C. for 12 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated at 90° C. for 5 min to obtain an enzymolyzate; the complex enzyme is composed of cellulase, pectinase and α-amylase, and the weight ratio of cellulase, pectinase and α-amylase is 2:3:1.

[0048] The Ganoderma lucidum was inoculated into a liquid culture medium and cultured at 28° C. and 120 r / min until the weight of the Ganoderma lucidum mycelium reached 14 g / L, thereby obtaining a Ganoderma lucidum liquid; the liquid culture medium consisted of 30 g of glucose, 10 g of peptone, 5 g of yeast extract, 2 g of potassium dihydrogen phosphate, 1 g of magnesium sulfate, 20 mg of vitamin B1, and 1 L of water, and was sterilized at 121° C. for 10 min.

[0049] The enzymatic hydrolysate, corn flour, soybean meal powder, sucrose and water were mixed in a weight-to-volume ratio of 50 g enzymatic hydrolysate: 10 g corn flour: 10 g soybean meal powder: 8 g sucrose: 0.8 L water, and sterilized at 121° C. for 4 min to obtain a fermentation medium; the fermentation medium was mixed with Ganoderma lucidum liquid in a weight ratio of 100:6, and fermented at 35° C. and 120 r / min for 24 h. The entire fermentation system was dried with hot air at 50° C. to a moisture content of 10%, to obtain a ginseng extract.

[0050] Example 3

[0051] A ginseng extract is prepared by the following preparation method:

[0052] The ginseng is crushed until it can completely pass through a 120-mesh sieve, and then mixed with a complex enzyme and water in a weight ratio of 100 g ginseng: 7 g complex enzyme: 100 g water, and the pH is adjusted to 5.6. The mixture is enzymatically hydrolyzed at 45° C. for 14 h. After the enzymatic hydrolysis is completed, the enzyme is inactivated at 85° C. for 6 min to obtain an enzymolyzate; the complex enzyme is composed of cellulase, pectinase and α-amylase, and the weight ratio of cellulase, pectinase and α-amylase is 1.8:2.5:0.8.

[0053] The Ganoderma lucidum was inoculated into a liquid culture medium and cultured at 26° C. and 130 r / min until the weight of the Ganoderma lucidum mycelium reached 12 g / L, thereby obtaining a Ganoderma lucidum liquid; the liquid culture medium consisted of 25 g of glucose, 7 g of peptone, 4 g of yeast extract, 2 g of potassium dihydrogen phosphate, 0.8 g of magnesium sulfate, 15 mg of vitamin B1, and 1 L of water, and was sterilized at 121° C. for 10 min.

[0054] The enzymatic hydrolysate, corn flour, soybean meal powder, sucrose and water were mixed in a weight-to-volume ratio of 40 g enzymatic hydrolysate: 9 g corn flour: 8 g soybean meal powder: 7 g sucrose: 0.8 L water, and sterilized at 121° C. for 4 min to obtain a fermentation medium; the fermentation medium was mixed with Ganoderma lucidum liquid in a weight ratio of 100:5, and fermented at 32° C. and 120 r / min for 22 h. The entire fermentation system was dried with hot air at 50° C. to a moisture content of 8% to obtain a ginseng extract.

[0055] Example 4

[0056] A composition is prepared from the following raw materials in parts by weight: 6 parts of Lobelia chinensis, 2 parts of Bogudan, 1 part of wax fern, 3 parts of lantern grass, 5 parts of hibiscus leaves, 7 parts of lotus leaves, 8 parts of cassia seeds, and 10 parts of the ginseng extract obtained in Example 1;

[0057] The preparation method is:

[0058] Lobelia chinensis, Psoralea corylifolia, Herba Lycopodii, Hibiscus sinensis leaves, Lotus leaves and Cassia seed are mixed to obtain a mixture, the mixture is mixed with water in a weight ratio of 1:8 and soaked for 30 minutes, heated to boiling, and decocted in a boiling state for 1 hour, and the decoction and the medicinal residue are collected separately; the decoction is concentrated under reduced pressure at 50° C. to a relative density of 0.9 to obtain a water extract.

[0059] The medicinal residue was mixed with 70% ethanol solution in a weight ratio of 1:7 and soaked for 30 minutes, then heated under reflux for extraction for 1.5 hours. The extract was collected and concentrated under reduced pressure at 50°C to a relative density of 0.9 to obtain an alcohol extract.

[0060] The alcohol extract and the water extract were mixed, stirred evenly, heated to 70°C, and paraffin wax was added, and stirred continuously to completely dissolve. After cooling to room temperature, the ginseng extract obtained in Example 1 was added and mixed, and dried with hot air at 50°C to a moisture content of 5% to obtain a composition.

[0061] Example 5

[0062] A composition is prepared from the following raw materials in parts by weight: 10 parts of Lobelia chinensis, 4 parts of Bogudan, 3 parts of wax fern, 6 parts of lantern grass, 8 parts of hibiscus leaves, 9 parts of lotus leaves, 10 parts of cassia seeds, and 15 parts of the ginseng extract obtained in Example 2;

[0063] The preparation method is:

[0064] Lobelia chinensis, Psoralea corylifolia, Herba Lycopodii, Hibiscus sinensis leaves, Lotus leaves and Cassia seed are mixed to obtain a mixture, the mixture is mixed with water in a weight ratio of 1:10 and soaked for 60 minutes, heated to boiling, and decocted in a boiling state for 2 hours, and the decoction and the medicinal residue are collected separately; the decoction is concentrated under reduced pressure at 60°C to a relative density of 1.0 to obtain a water extract.

[0065] The medicinal residue was mixed with 80% ethanol solution in a weight ratio of 1:9 and soaked for 60 minutes, heated under reflux for extraction for 2.5 hours, and the extract was collected and concentrated under reduced pressure at 60°C to a relative density of 1.0 to obtain an alcohol extract.

[0066] The alcohol extract and the water extract were mixed, stirred evenly, heated to 70°C, and paraffin wax was added, and stirred continuously to completely dissolve. After cooling to room temperature, the ginseng extract obtained in Example 2 was added and mixed, and dried with hot air at 50°C to a moisture content of 6% to obtain a composition.

[0067] Example 6

[0068] A composition is prepared from the following raw materials in parts by weight: 8 parts of Lobelia chinensis, 3 parts of Bogudan, 2 parts of wax wax, 4 parts of lantern grass, 7 parts of hibiscus leaves, 8 parts of lotus leaves, 9 parts of cassia seeds, and 12 parts of the ginseng extract obtained in Example 3;

[0069] The preparation method is:

[0070] (1) Mix Lobelia chinensis, Herba Bulbosae, Herba Lycopodii, Hibiscus sangreae, Lotus leaf and Semen Cassiae to obtain a mixture, mix the mixture with water in a weight ratio of 1:9 and soak for 40 minutes, heat to boiling, and keep boiling for 1.5 hours, collect the decoction and the residue respectively; concentrate the decoction under reduced pressure at 55°C to a relative density of 0.9 to obtain a water extract.

[0071] (2) The medicinal residue was mixed with 75% ethanol solution in a weight ratio of 1:8 and soaked for 40 min. The mixture was heated under reflux for 2 h. The extract was collected and concentrated under reduced pressure at 55 °C to a relative density of 0.9 to obtain the alcohol extract.

[0072] (3) The alcohol extract and the water extract were mixed, stirred evenly, and heated to 70°C. White wax was added and stirred continuously to completely dissolve the mixture. After cooling to room temperature, the ginseng extract obtained in Example 3 was added and mixed. The mixture was dried with hot air at 50°C until the moisture content was 5%, thereby obtaining a composition.

[0073] Comparative Example 1

[0074] The difference from Example 3 is that Ganoderma lucidum was not inoculated, and the rest was the same as Example 3.

[0075] Comparative Example 2

[0076] The difference from Example 3 is that α-amylase is not added to the complex enzyme, and the rest is the same as Example 3.

[0077] Comparative Example 3

[0078] The difference from Example 3 is that the weight ratio of cellulase, pectinase and α-amylase in the complex enzyme is 3:1:1, and the rest is the same as Example 3.

[0079] Comparative Example 4

[0080] The difference from Example 6 is that the ginseng extract is replaced by the ginseng extract obtained in Comparative Example 1, and the rest is the same as Example 6.

[0081] Comparative Example 5

[0082] The difference from Example 6 is that Cassia seed is not contained, and the rest is the same as Example 6.

[0083] Comparative Example 6

[0084] The difference from Example 6 is that the insect wax is not contained, and the rest is the same as Example 6.

[0085] Comparative Example 7

[0086] The difference from Example 6 is that the volume fraction of the ethanol solution is 90%, and the rest is the same as Example 6.

[0087] Comparative Example 8

[0088] The difference from Example 6 is that the ethanol solution in step (2) is replaced by water, and the reflux extraction is changed to heating to boiling and then maintaining the boiling state for 1.5 hours. The rest is the same as Example 6.

[0089] Example 7

[0090] (1) Culture of human non-small cell lung cancer A549 cells

[0091] After the A549 cells were taken out from the liquid nitrogen tank, they were quickly thawed in a 37°C water bath.

[0092] Centrifuge at a speed of rpm / min, remove and discard the supernatant after 3 minutes, add 1mL of DMEM culture medium containing 10% fetal bovine serum, and gently pipette to suspend. After pipetting several times to mix evenly, transfer the suspension into a 25mL cell culture flask, add DMEM complete medium to 5mL, pipette to mix evenly, and place in a cell culture incubator (5% CO2, 37℃) for culture. When it is observed under a microscope that the cells have covered 80%~90% of the bottom of the culture flask, start the cell passaging step. First, discard the original culture medium, add physiological saline to the culture flask, lay the culture flask flat and shake gently to wash the cells, 2mL each time, wash 3 times to ensure that the floating objects and metabolites produced during the culture are cleaned. Then add 2mL of trypsin and shake gently to allow the trypsin to infiltrate the cell surface. Observe under a microscope. When the cells are observed to change from the original state to a round state, it proves that the digestion is complete, and add an appropriate amount of complete culture medium to stop the digestion. Collect the digested cell suspension, centrifuge, take a certain amount of cells and place them in a new culture flask, add complete culture medium and place it in the culture flask.

[0093] Continue to culture in a culture box (5% CO2, 37℃).

[0094] (2) Effects of the ginseng extracts or compositions obtained in Examples 1 to 6 and Comparative Examples 1 to 8 on the proliferation inhibition rate of A549 cells

[0095] The ginseng extracts or compositions obtained in Examples 1 to 6 and Comparative Examples 1 to 8 were prepared into ginseng extract solutions or composition solutions with a concentration of 50 μg / mL using complete culture medium, and the following tests were performed:

[0096] Collect A549 cells in the logarithmic growth phase, which is the best growth phase, for plating experiments. Discard the culture medium and add physiological saline to the culture flask. Lay the flask flat and gently shake to wash the cells, using 2 mL each time. Wash three times to ensure that floating objects and metabolites produced during the culture are completely removed. Discard the physiological saline, trypsinize, and centrifuge. Collect the centrifuged cells and dilute them with complete culture medium to 4 × 10 cells per well. 3 Cells were seeded into 96-well plates using a dispenser at a density of 100 μL per well. The plates were incubated in an incubator for 24 hours. The supernatant was discarded. A blank control group received 100 μL of complete culture medium, while the sample groups received 100 μL of either a 50 μg / mL ginseng extract solution or a combination solution. Six replicates were added to each well and incubated for a further 24 hours at 37°C in a 5% CO2 incubator. 10 μL of 5 mg / mL MTT was added to each well. After 4 hours, the supernatant was discarded, and 100 μL of DMSO was added. The plates were shaken for 5 minutes on a microplate reader and measured at 490 nm. The absorbance was measured and the cell proliferation inhibition rate for each group was calculated according to the following formula. The experiment was repeated three times. Data were analyzed using SPSS 24.0 software. Data are presented as mean ± standard deviation (x ± s). One-way analysis of variance (ANOVA) was used to compare the groups. A p < 0.05 indicated statistical significance. The results are shown in Table 1.

[0097] Cell proliferation inhibition rate (%) = [A 空白组 -A 给药组 / A 空白组 ]×100%

[0098] Table 1 Effects of different groups on the inhibition rate of A549 cell proliferation

[0099]

[0100] Note: The letters in the table represent the differences between the groups. Different lowercase letters in the same column represent significant differences. p＜0.05。

[0101] (3) Effects of the ginseng extracts or compositions obtained in Examples 3 and 6 and Comparative Examples 1 to 8 on the migration ability of A549 cells

[0102] Cell scratch assay

[0103] A549 cells in the logarithmic growth phase were seeded in a 6-well plate, scratched using a 200 μL sterilized pipette tip, and the floating broken cells were washed away with PBS. The cells were then observed and photographed under a fluorescent inverted microscope. The ginseng extracts or compositions obtained in Examples 3 and 6 and Comparative Examples 1 to 8 were prepared in DMEM medium containing 1% FBS to a concentration of 100 μg / mL. Three replicates were set up for each group. The control group was added with DMEM medium containing 1% FBS.

[0104] 2.5 mL of DMEM culture medium was placed in an environment of 37°C and 5% carbon dioxide for 24 h, and then the cell migration was observed and photographed under a microscope. The cell migration results of Example 3 and Comparative Example 1 were as follows: Figure 1 and Figure 2 As shown ( Figure 1 and Figure 2 The magnification of the scratches was the same), and the scratch repair rate was calculated using ImageJ software. The initial area of the scratch area (A0) and the remaining area at the observation time point (A t ), and finally calculate the scratch repair rate according to the formula: The experiment was repeated three times, and data were analyzed using SPSS 24.0 software. Data are presented as mean ± standard deviation (x ± s). Intergroup comparisons were performed using one-way analysis of variance (ANOVA). The results are shown in Table 2.

[0105] Table 2 Scratch repair rate of different groups

[0106]

[0107] Note: ** in the table indicates that compared with the blank group, p＜0.01。

[0108] Transwell cell migration assay

[0109] A549 cells in the logarithmic growth phase were digested and prepared into suspensions and then counted. The Transwell chamber was placed in a 24-well plate and 800 μL of complete medium was added to the lower chamber. 100 μL of cell suspension was taken and added to the upper chamber. The cell count was 5×10 5 / mL, the ginseng extracts or compositions obtained in Examples 3 and 6 and Comparative Examples 1 to 8 were used as drugs at a dosage concentration of 100μg / mL, and only complete culture medium was added to the control group. After culturing for 24 hours, the chamber was taken out, the cells were fixed with 4% paraformaldehyde, and stained with 1% crystal violet. After washing three times with PBS, three fields of view were randomly selected under a microscope to take pictures, record and count, and the average value was taken as the number of migrating cells. The relative migration rate (%) of each group was calculated according to the following formula. The independent experiment was repeated three times, and the data were analyzed using SPSS 24.0 data processing software. The experimental data were expressed as mean ± standard deviation (x ± s). ANOVA single-factor analysis of variance was used for inter-group comparison. The results are shown in Table 3. The field of view photography results of Example 3 and Comparative Example 1 groups are shown in Table 3. Figure 3 and Figure 4 As shown ( Figure 3 and Figure 4 The magnification is the same).

[0110] Relative migration rate (%) = (number of migrating cells in the experimental group ÷ number of migrating cells in the control group) × 100%

[0111] Table 3 Relative migration rates of different groups

[0112]

[0113] Note: *** in the table means compared with the blank group, p＜0.001。

[0114] Example 8

[0115] Eight-week-old wild-type female BALB / c mice served as the normal group. Sixty MRL / lpr female mice were randomly divided into a positive drug group (glucocorticoid prednisone administered orally, 1.0 mg / kg body weight / day), a blank group (normal saline administered orally, 1.0 mg / kg body weight / day), and Example 3, Example 6, and Comparative Examples 1 to 8 (gavage, 1.0 mg / kg body weight / day). Animal care adhered to the 3Rs principle. Surgical procedures and data analysis were performed in a single-blind fashion. Animals were maintained in a 12-hour light / 12-hour dark cycle with free access to food and water. They were acclimated for 7 days prior to the experiment. Mice in each group were gavage administered at 12 weeks of age, with dosing every other day for 2 weeks. At 14 weeks of age, 24-hour urine samples were collected for protein and creatinine analysis. Skin lesions and erythema on the face and back of the mice were also observed. Serum was collected for anti-nucleosome antibodies (AnuA). The results are expressed as mean.

[0116] The results showed that compared with the blank group, the positive drug group and Example 3, Example 6, and Comparative Examples 1 to 8 groups had significantly reduced facial and back lesions, and the number of erythema was significantly reduced. The degree of reduction in facial and back lesions in Example 3 and Example 6 groups was significantly higher than that in Comparative Examples 1 to 8 groups, and even better than that in the positive drug group. The results of urine protein, urine creatinine, and AnuA are shown in Table 4.

[0117] Table 4 Results of urine protein, urine creatinine and AnuA in different groups

[0118]

[0119] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A composition containing ginseng extract, characterized in that: The invention is composed of the following raw materials in parts by weight: 6-10 parts of Lobelia chinensis, 2-4 parts of Bogudan, 1-3 parts of wax wax, 3-6 parts of lantern grass, 5-8 parts of hibiscus leaves, 7-9 parts of lotus leaves, 8-10 parts of cassia seeds and 10-15 parts of ginseng extract; The preparation method of the ginseng extract comprises: crushing ginseng, mixing the mixture with a complex enzyme and water in a weight ratio of 100:(6-8):100, and performing enzymolysis to obtain an enzymatic hydrolyzate; mixing the enzymatic hydrolyzate, corn flour, soybean meal powder, sucrose, and water in a weight-to-volume ratio of (30-50) g:(8-10) g:(6-10) g:(5-8) g:0.8 L to obtain a fermentation medium; mixing the fermentation medium with a Ganoderma lucidum liquid in a weight ratio of 100:4-6, fermenting at 30°C-35°C for 20 h-24 h, and drying to obtain the ginseng extract; the complex enzyme comprises cellulase, pectinase, and α-amylase; and the weight of Ganoderma lucidum mycelium in the Ganoderma lucidum liquid is 8 g / L-14 g / L.

2. The composition according to claim 1, characterized in that The weight ratio of the cellulase, pectinase and α-amylase is (1-2):(2-3):(0.5-1); the enzymatic activity of the cellulase is greater than 100,000 U / g; the enzymatic activity of the pectinase is greater than 30,000 U / g; and the enzymatic activity of the α-amylase is greater than 10,000 U / g.

3. The composition according to claim 1, characterized in that The initial pH of the enzymatic hydrolysis is 5.2~6.0, the enzymatic hydrolysis temperature is 40℃~50℃, and the enzymatic hydrolysis time is 12h~16h.

4. The composition according to claim 1, characterized in that The Ganoderma lucidum liquid is obtained by inoculating Ganoderma lucidum into a liquid culture medium and culturing at 25°C to 28°C; the composition of the liquid culture medium is 20g to 30g of glucose, 5g to 10g of peptone, 3g to 5g of yeast extract, 1g to 2g of potassium dihydrogen phosphate, 0.5g to 1g of magnesium sulfate, 10mg to 20mg of vitamin B1 and 1L of water; the pH value of the liquid culture medium is 5 to 6.

5. The method for preparing the composition according to any one of claims 1 to 4, characterized in that: The method comprises the following steps: mixing lobelia chinensis, radix scutellariae, radix scutellariae, hibiscus sangreae leaves, lotus leaves and cassia seeds to obtain a mixture, mixing the mixture with water and soaking it, heating it to boiling and then decocting it for 1-2 hours, collecting the decoction and the medicinal residue respectively; concentrating the decoction to obtain a water extract; mixing the medicinal residue with an ethanol solution with a volume fraction of 70%-80% and soaking it, heating and refluxing it for extraction for 1.5-2.5 hours, collecting the extract and concentrating it to obtain an alcohol extract; and mixing the alcohol extract, the water extract, wax fern and ginseng extract, and drying it to obtain a composition.

6. The preparation method according to claim 5, characterized in that The soaking time is 30 min to 60 min; and the concentration is to concentrate to a relative density of 0.9 to 1.

0.

7. The preparation method according to claim 5, characterized in that The weight ratio of the mixture to water is 1:8-10; the weight ratio of the medicinal residue to the ethanol solution with a volume fraction of 70%-80% is 1:7-9.

8. Use of the composition according to any one of claims 1 to 4 in the preparation of a drug for preventing and treating lung cancer.

9. Use of the composition according to any one of claims 1 to 4 in the preparation of a medicament for treating systemic lupus erythematosus.

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