Composition containing ginseng extract as well as preparation method and application of composition

Abstract

The invention provides a composition containing a ginseng extract as well as a preparation method and application of the composition, and belongs to the technical field of biological medicines. The composition containing the ginseng extract is prepared from the following raw materials in parts by weight: 6-10 parts of Chinese lobelia, 2-4 parts of gendarussa vulgaris, 1-3 parts of Chinese insect wax, 3-6 parts of peruvian groundcherry herb, 5-8 parts of Chinese hibiscus leaves, 7-9 parts of lotus leaves, 8-10 parts of cassia seeds and 10-15 parts of the ginseng extract. According to the composition containing the ginseng extract, all the medicines are combined and matched for use, all the medicines have a synergistic effect, and the composition has a good treatment effect on lung cancer and systemic lupus erythematosus.

Description

Technical Field

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

[0002] The active substances of ginseng (such as ginsenosides, polysaccharides, polyphenols, etc.) have been widely studied, especially in the field of anti-tumor. Existing studies have shown that ginsenosides (such as Rg3, Rh2) can play an anti-cancer role through mechanisms such as inhibiting tumor cell proliferation, inducing apoptosis, and inhibiting angiogenesis. However, the existing technology has the following defects: low bioavailability: components such as ginsenosides have poor water solubility and low intestinal absorption rate, resulting in limited efficacy in vivo; limitations of single components: most studies focus on monomeric saponins, ignoring the potential of synergistic effects of multiple components; crude preparation process: traditional extraction methods (such as decoction with water, ethanol reflux) are easy to damage thermosensitive components and fail to effectively enrich highly active components, resulting in poor stability of the preparation.

[0003] Lung cancer is a globally high-incidence cancer type. Although existing targeted drugs are partially effective, the problems of drug resistance and side effects are prominent. Currently, new low-toxic and multi-target therapies are urgently needed for lung cancer. Systemic lupus erythematosus is an autoimmune disease involving multiple organs. Existing therapies mainly include glucocorticoids, immunosuppressants, and biological agents (such as belimumab), but long-term use is prone to cause infections, cardiovascular risks, and cumulative organ damage. Although some traditional Chinese medicine compound prescriptions (such as Tripterygium wilfordii) are used for adjuvant treatment, their liver and kidney toxicity and unclear action mechanisms limit their application. Currently, there is no research in this field on ginseng extract and its composition that can simultaneously treat lung cancer and systemic lupus erythematosus. Summary of the Invention

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

[0005] Another purpose of the present invention is to provide the application of the above composition in the preparation of drugs for treating lung cancer or systemic lupus erythematosus.

[0006] In order to achieve the above invention purposes, the present invention provides the following technical solutions: 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 Lour., 2-4 parts of Justicia gendarussa Burm. f., 1-3 parts of Chinese insect wax, 3-6 parts of Physalis angulata L., 5-8 parts of Abutilon indicum (L.) Sweet, 7-9 parts of lotus leaf, 8-10 parts of Cassia obtusifolia L., and 10-15 parts of ginseng extract; The preparation method of the ginseng extract is as follows: Ginseng is crushed and mixed with a complex enzyme and water in a weight ratio of 100:(6 - 8):100 to obtain an enzymolysate through enzymolysis; The enzymolysate, corn flour, soybean meal powder, sucrose and water are mixed in a weight - volume ratio of (30 - 50) g:(8 - 10) g:(6 - 10) g:(5 - 8) g:0.8 L to obtain a fermentation medium; The fermentation medium and Ganoderma lucidum bacterial liquid are mixed in a weight ratio of 100:4 - 6, fermented at 30°C - 35°C for 20 h - 24 h, and dried to obtain the ginseng extract; The complex enzyme is composed of cellulase, pectinase and α - amylase; The weight of Ganoderma lucidum mycelium in the Ganoderma lucidum bacterial liquid is 8 g / L - 14 g / L.

[0007] Preferably, the weight ratio of the cellulase, pectinase and α - amylase is (1 - 2):(2 - 3):(0.5 - 1); The enzyme activity of the cellulase is above 100,000 U / g; The enzyme activity of the pectinase is above 30,000 U / g; The enzyme activity of the α - amylase is above 10,000 U / g.

[0008] Preferably, the initial pH of enzymolysis is 5.2 - 6.0, the temperature of enzymolysis is 40°C - 50°C, and the time of enzymolysis is 12 h - 16 h.

[0009] Preferably, the Ganoderma lucidum bacterial liquid is obtained by inoculating Ganoderma lucidum into a liquid medium and culturing at 25°C - 28°C; The composition of the liquid medium is 20 g - 30 g of glucose, 5 g - 10 g of peptone, 3 g - 5 g of yeast extract, 1 g - 2 g of potassium dihydrogen phosphate, 0.5 g - 1 g of magnesium sulfate, 10 mg - 20 mg of vitamin B1 and 1 L of water; The pH value of the liquid medium is 5 - 6.

[0010] The present invention also provides a preparation method of the above - mentioned composition, which consists of the following steps: Lobelia chinensis, Justicia gendarussa, Physalis angulata, Flacourtia indica leaves, lotus leaves and Cassia obtusifolia are mixed to obtain a mixture, the mixture is mixed with water and soaked, heated to boiling and decocted for 1 - 2 h, and the decoction liquid and medicinal residues are collected respectively; The decoction liquid is concentrated to obtain a water extract; The medicinal residues are mixed with an ethanol solution with a volume fraction of 70% - 80% and soaked, heated under reflux for 1.5 - 2.5 h, and the extract is collected and concentrated to obtain an ethanol extract; The ethanol extract, water extract, Chinese insect wax and ginseng extract are mixed and dried to obtain the composition.

[0011] Preferably, the soaking time is 30 min - 60 min; The concentration is to concentrate to a relative density of 0.9 - 1.0.

[0012] Preferably, the weight ratio of the mixture to water is 1:8 - 10; The weight ratio of the medicinal residues to the ethanol solution with a volume fraction of 70% - 80% is 1:7 - 9.

[0013] The present invention also provides the application of the above - mentioned composition in the preparation of drugs for preventing and treating lung cancer.

[0014] The present invention also provides the use of the above composition in the preparation of a medicament for treating systemic lupus erythematosus.

[0015] Advantages of the present invention: The composition containing ginseng extract provided by the present invention, in which the ginseng extract is enzymatically hydrolyzed with a complex enzyme, can release more active ingredients originally encapsulated in ginseng, and can reduce the molecular weight of polysaccharides to generate oligosaccharides or oligosaccharides that are more easily absorbed, enhancing immunomodulatory activity. Moreover, enzymatic hydrolysis can convert ginsenosides in ginseng into rare ginsenosides, enhancing anti-tumor activity. In addition, after enzymatically hydrolyzing ginseng with a complex enzyme, the present invention inoculates Ganoderma lucidum for fermentation, and carries out biotransformation through the fermentation process (converting the components in ginseng into new active substances such as complexes of ganoderic acid, ganoderma polysaccharide and saponins), enhancing the activity of the original components. Moreover, fermenting with Ganoderma lucidum can generate secondary metabolites (such as antioxidant peptides and organic acids), and at the same time, fermentation can degrade potential irritant components in ginseng (such as certain saponin precursors), improving safety. In short, the present invention prepares ginseng extract by means of the linkage of complex enzymatic hydrolysis and Ganoderma lucidum fermentation, playing a synergistic effect, and finally obtaining a ginseng extract with multi-target effects, which has good therapeutic effects on lung cancer and systemic lupus erythematosus.

[0016] The present invention combines and uses ginseng extract with Lobelia chinensis, Justicia gendarussa, Chinese insect wax, Physalis angulata, Mulberry leaf, Lotus leaf and Cassia obtusifolia. The various medicinal flavors synergistically enhance the effect, and can further improve the therapeutic effect of ginseng extract on lung cancer and systemic lupus erythematosus. Specifically: Lobelia chinensis helps to inhibit the proliferation of tumor cells and induce apoptosis of tumor cells, can synergistically activate the immune response with ginseng extract, and alleviate the inflammation related to systemic lupus erythematosus; Justicia gendarussa has anti-tumor and antioxidant effects; Chinese insect wax can promote the healing of mucous membranes and skin, alleviate skin ulcers and lung injuries in patients with systemic lupus erythematosus, and can inhibit excessive autoimmune reactions; Physalis angulata can induce autophagy of tumor cells, inhibit the metastasis of lung cancer, and can regulate metabolism and improve abnormal liver and kidney functions in patients with systemic lupus erythematosus; Mulberry leaf can anti-angiogenesis, block the blood supply of lung cancer, and can alleviate the pulmonary fibrosis and renal fibrosis related to systemic lupus erythematosus; Lotus leaf has antioxidant, anti-inflammatory and lipid metabolism regulating effects; Cassia obtusifolia plays a dual-directional immunomodulatory role, inhibits over-activated B cells (reducing autoantibodies), enhances the anti-tumor activity of T cells, and promotes defecation and detoxification, promotes the excretion of metabolic wastes, and reduces the accumulation of toxins in the bodies of patients with tumors and systemic lupus erythematosus. Description of the drawings

[0017] Figure 1 The results of cell scratch for the third group of examples, where the left figure is the starting scratch picture and the right figure is the scratch picture after 24 h; Figure 2 The cell scratch results of Comparative Example 1 group, where the left figure is the starting scratch picture and the right figure is the scratch picture after 24 h; Figure 3 The cell migration results of Example 3 group; Figure 4 The cell migration results of Comparative Example 1 group. Detailed implementation mode

[0018] 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 Lour., 2-4 parts of Justicia gendarussa Burm. f., 1-3 parts of Chinese insect wax, 3-6 parts of Physalis angulata L., 5-8 parts of Abutilon indicum (L.) Sweet, 7-9 parts of lotus leaf, 8-10 parts of Cassia obtusifolia L. and 10-15 parts of ginseng extract; The preparation method of the ginseng extract is as follows: ginseng is pulverized and mixed with a composite enzyme and water in a weight ratio of 100: (6-8): 100 to obtain an enzymolyzed product by enzymatic hydrolysis; the enzymolyzed product, corn flour, soybean meal powder, sucrose and water are mixed 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; the fermentation medium and Ganoderma lucidum spore liquid are mixed in a weight ratio of 100: 4-6, fermented at 30 °C-35 °C for 20 h-24 h, and dried to obtain the ginseng extract; the composite enzyme is composed of cellulase, pectinase and α-amylase; the weight of Ganoderma lucidum mycelium in the Ganoderma lucidum spore liquid is 8 g / L-14 g / L.

[0019] The present invention does not have special limitations on the specific sources of each raw material in the preparation process of ginseng extract, and conventional commercially available products in the art can be used. In the present invention, the mesh number of the pulverization is preferably 100 to 150 meshes, and the present invention does not have special limitations on the specific pulverization method. In the present invention, the weight ratio of cellulase, pectinase and α-amylase in the complex enzyme is preferably (1 to 2):(2 to 3):(0.5 to 1), more preferably (1.2 to 1.8):(2.2 to 2.5):(0.7 to 0.8); the enzyme activity of the cellulase is preferably above 100,000 U / g; the enzyme activity of the pectinase is preferably above 30,000 U / g; the enzyme activity of the α-amylase is preferably above 10,000 U / g. In the present invention, the weight ratio of ginseng after pulverization, complex enzyme and water is preferably 100:(6.5 to 7.5):100, more preferably 100:(6.8 to 7.2):100. In the present invention, after ginseng is pulverized and mixed with the complex enzyme and water, the pH is preferably adjusted to 5.2 to 6.0, and then enzymatic hydrolysis is carried out. The pH is more preferably 5.4 to 5.8; the temperature of the enzymatic hydrolysis is preferably 40°C to 50°C, more preferably 42°C to 48°C; the time of the enzymatic hydrolysis is preferably 12 h to 16 h, more preferably 13 h to 15 h. After the enzymatic hydrolysis is completed, it is preferably necessary to carry out enzyme inactivation treatment, and the enzyme inactivation method is preferably enzyme inactivation at 80°C to 90°C for 5 min to 8 min.

[0020] In the present invention, the weight-to-volume ratio of the enzymatic hydrolysate, corn flour, soybean meal powder, sucrose and water is preferably (35 to 45) g:9 g:(7 to 9) g:(6 to 7) g:0.8 L, more preferably (38 to 40) g:9 g:8 g:6 g:0.8 L. In the present invention, the fermentation medium is preferably a sterilized fermentation medium, and the sterilization method is preferably sterilization at 121°C for 2 min to 4 min. In the present invention, the Ganoderma lucidum bacterial liquid is preferably obtained by inoculating Ganoderma lucidum into a liquid medium and culturing at 25°C to 28°C; the culture method is preferably shaking culture, and the rotation speed of the shaking culture is preferably 120 r / min to 140 r / min, more preferably 130 r / min; the composition of the liquid medium is preferably 20 g to 30 g of glucose, 5 g to 10 g of peptone, 3 g to 5 g of yeast extract, 1 g to 2 g of potassium dihydrogen phosphate, 0.5 g to 1 g of magnesium sulfate, 10 mg to 20 mg of vitamin B1 and 1 L of water, more preferably 22 g to 28 g of glucose, 6 g to 8 g of peptone, 4 g of yeast extract, 1.5 g of potassium dihydrogen phosphate, 0.7 g to 0.8 g of magnesium sulfate, 12 mg to 16 mg of vitamin B1 and 1 L of water; the pH value of the liquid medium is preferably 5 to 6. In the present invention, the weight of Ganoderma lucidum mycelium in the Ganoderma lucidum bacterial liquid is preferably 10 g / L to 12 g / L, and the weight of Ganoderma lucidum mycelium refers to the wet weight of the mycelium in the bacterial liquid per unit volume.

[0021] In the present invention, when the fermentation medium is mixed with Ganoderma lucidum broth, the weight ratio of the fermentation medium to Ganoderma lucidum broth 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 h to 23 h, more preferably 22 h. After fermentation, the entire fermentation system obtained is dried. The present invention has no special limitation on the specific drying method, and it is dried to a moisture content of 5% to 10% to obtain ginseng extract.

[0022] The present invention has no special limitation on the specific sources of Lobelia chinensis Lour., Justicia gendarussa Burm. f., Chinese insect wax, Physalis angulata L., Hibiscus rosa-sinensis L. leaves, lotus leaf and Cassia obtusifolia L. Conventional commercially available products in the art can be used. In the present invention, the weight parts of Lobelia chinensis Lour. are preferably 7 to 9 parts, more preferably 8 parts; the weight parts of Justicia gendarussa Burm. f. are preferably 3 parts; the weight parts of Chinese insect wax are preferably 2 parts; the weight parts of Physalis angulata L. are preferably 4 to 5 parts; the weight parts of Hibiscus rosa-sinensis L. leaves are preferably 6 to 7 parts; the weight parts of lotus leaf are preferably 8 parts; the weight parts of Cassia obtusifolia L. are preferably 9 parts; the weight parts of the above-mentioned ginseng extract are preferably 11 to 14 parts, more preferably 12 to 13 parts.

[0023] The composition of the present invention uses ginseng extract as the monarch drug, which exerts the effects of greatly tonifying primordial qi, restoring pulse and arresting collapse, invigorating the spleen and benefiting the lung, promoting fluid production and nourishing blood, calming the mind and improving intelligence, etc. By improving the body's immunity and enhancing vital qi, it can resist pathogenic factors and play a key role in strengthening the healthy qi and eliminating pathogenic factors. Lobelia chinensis Lour. and Cassia obtusifolia L. together serve as ministerial drugs. Lobelia chinensis Lour. clears heat and detoxifies, promotes diuresis and reduces swelling. In the formula, it assists ginseng extract to enhance the power of clearing heat and detoxifying, and helps to eliminate heat toxins in the body; Cassia obtusifolia L. clears heat and improves eyesight, promotes defecation by moistening the intestines. On the one hand, it can assist Lobelia chinensis Lour. in clearing heat, and on the other hand, by promoting defecation, it enables the body's toxins to be excreted from the intestines, reducing the body burden and assisting the monarch drug in treating diseases. Justicia gendarussa Burm. f., Chinese insect wax, Physalis angulata L., Hibiscus rosa-sinensis L. leaves and lotus leaf are adjuvant drugs. Justicia gendarussa Burm. f. dispels wind-dampness and dredges meridians, assisting the monarch drug and ministerial drugs to enhance the effects of dredging meridians and relieving pain, and is an adjuvant drug for assistance; Chinese insect wax stops bleeding, promotes granulation and relieves pain, helps to repair body injuries, and has a certain therapeutic effect on possible tissue injuries, bleeding, etc. during the treatment of diseases, and serves as an adjuvant drug for assistance; Physalis angulata L. clears heat and detoxifies, promotes diuresis and stops bleeding, assists Lobelia chinensis Lour. etc. in clearing heat and detoxifying, and can also promote diuresis to excrete dampness in the body, and has a relieving effect on possible damp-heat symptoms such as systemic lupus erythematosus, and is an adjuvant drug for assistance; Hibiscus rosa-sinensis L. leaves clears heat and detoxifies, reduces swelling and relieves pain, assists the monarch and ministerial drugs in clearing heat and detoxifying, reducing swelling and dissipating nodules, and has an improving effect on possible local swelling and pain symptoms caused by diseases, and serves as an adjuvant drug for assistance; lotus leaf clears summer heat and resolves dampness, promotes the ascending of clear yang, cools blood and stops bleeding, prevents drugs such as ginseng from being too warm and nourishing to generate heat, and at the same time its effect of cooling blood and stopping bleeding is helpful for possible bleeding symptoms during the disease, and has the role of an adjuvant drug for assistance.

[0024] The present invention also provides a method for preparing the above composition, which comprises the following steps: mixing Lobelia chinensis, Justicia gendarussa, Physalis angulata, Mulberry leaves, lotus leaves and Cassia obtusifolia to obtain a mixture, mixing the mixture with water and soaking, heating to boiling and decocting for 1-2 h, and separately collecting the decoction and the medicinal residues; concentrating the decoction to obtain a water extract; mixing the medicinal residues with an ethanol solution with a volume fraction of 70%-80% and soaking, heating under reflux for 1.5-2.5 h, and collecting the extract and concentrating to obtain an ethanol extract; mixing the ethanol extract, the water extract, Chinese wax and the above-mentioned ginseng extract, and drying to obtain the composition.

[0025] 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 min-60 min, and in specific embodiments, it can be 30 min, 35 min, 40 min, 45 min, 50 min, 55 min and 60 min. In the present invention, the decocting time is preferably 1.2 h-1.8 h, more preferably 1.4 h-1.6 h, and the decocting is preferably maintained at a slightly boiling state. The present invention has no special limitation on the specific method of concentration. In one embodiment, the vacuum concentration method is adopted, specifically, concentrating at 50°C-60°C, and the concentration is preferably concentrated to a relative density of 0.9-1.0. In the present invention, the relative density mentioned refers to the ratio of the density of the substance to the density of water at 50°C-60°C.

[0026] In the present invention, when the medicinal residues are soaked with an ethanol solution, the volume fraction of the ethanol solution is preferably 72%-78%, more preferably 74%-76%; the weight ratio of the medicinal residues to the ethanol solution with a volume fraction of 70%-80% is preferably 1:7-9, more preferably 1:8. When the medicinal residues are mixed with an ethanol solution with a volume fraction of 70%-80% and soaked, the soaking time is preferably 30 min-60 min, and in specific embodiments, it can be 30 min, 35 min, 40 min, 45 min, 50 min, 55 min and 60 min. In the present invention, the heating reflux extraction time is preferably 1.8-2.4 h, more preferably 2.0-2.2 h. The present invention has no special limitation on the specific method of concentration for obtaining the ethanol extract. In one embodiment, the vacuum concentration method is adopted, specifically, concentrating at 50°C-60°C, and the concentration is preferably concentrated to a relative density of 0.9-1.0.

[0027] The present invention also provides the application of the above composition in the preparation of drugs for preventing and treating lung cancer. In the pharmaceutical application of the present invention, the composition can play an anti-tumor role as the only active ingredient.

[0028] The present invention also provides the use of the above composition in the preparation of a medicament for treating systemic lupus erythematosus. In the pharmaceutical use of the present invention, the composition can act as the sole active ingredient to treat systemic lupus erythematosus.

[0029] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0030] In the following embodiments, unless otherwise specified, all are conventional methods.

[0031] In the following embodiments, the materials, reagents, etc. used, unless otherwise specified, can be obtained from commercial channels.

[0032] In the following embodiments, the cellulase is purchased from Shanghai Xiangqi Biotechnology Co., Ltd., and the enzyme activity is above 100,000 U / g; the pectinase is purchased from Anhui Weimao Biotechnology Co., Ltd., and the enzyme activity is above 30,000 U / g; the enzyme activity of α-amylase is above 10,000 U / g; the Ganoderma lucidum is purchased from Beina Biology, with the number BNCC382029, the resource name is Ganoderma lucidum (Ganoderma rubra), and the Latin name is Ganoderma lucidum .

[0033] Example 1 A ginseng extract is prepared by the following preparation method: The ginseng is pulverized to pass through a 100-mesh sieve, and then mixed with a complex enzyme and water according to a weight ratio of 100 g of ginseng: 6 g of complex enzyme: 100 g of water, the pH is adjusted to 5.2, and enzymolysis is carried out at 40 °C for 16 h. After the enzymolysis is completed, the enzyme is inactivated at 80 °C for 8 min to obtain an enzymolysate; 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.

[0034] The Ganoderma lucidum is inoculated into a liquid medium and cultured at 25 °C and 120 r / min until the weight of Ganoderma lucidum mycelium is 8 g / L to obtain a Ganoderma lucidum liquid; the liquid medium is composed 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 is sterilized at 121 °C for 10 min.

[0035] The enzymolysate, corn flour, soybean meal powder, sucrose and water are mixed according to a weight-to-volume ratio of 30 g of enzymolysate: 8 g of corn flour: 6 g of soybean meal powder: 5 g of sucrose: 0.8 L of water, and sterilized at 121 °C for 2 min to obtain a fermentation medium; the fermentation medium and the Ganoderma lucidum liquid are mixed according to a weight ratio of 100:4, fermented at 30 °C and 130 r / min for 20 h, and the whole fermentation system is dried at 50 °C by hot air until the water content is 5% to obtain the ginseng extract.

[0036] Example 2 A ginseng extract, prepared by the following preparation method: The ginseng is crushed to be able to pass through a 150-mesh sieve completely, and then mixed with a complex enzyme and water according to the weight ratio of 100 g of ginseng: 8 g of complex enzyme: 100 g of water, the pH is adjusted to 6.0, and enzymatic hydrolysis is carried out 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 enzymolysate; 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.

[0037] The Ganoderma lucidum fungus is inoculated into a liquid medium and cultured at 28 °C and 120 r / min until the weight of Ganoderma lucidum mycelium is 14 g / L to obtain a Ganoderma lucidum fungal liquid; the liquid medium is composed 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 is sterilized at 121 °C for 10 min.

[0038] The enzymolysate, corn flour, soybean meal powder, sucrose and water are mixed according to the weight-to-volume ratio of 50 g of enzymolysate: 10 g of corn flour: 10 g of soybean meal powder: 8 g of sucrose: 0.8 L of water, and sterilized at 121 °C for 4 min to obtain a fermentation medium; the fermentation medium and the Ganoderma lucidum fungal liquid are mixed according to the weight ratio of 100:6, and fermented at 35 °C and 120 r / min for 24 h. The entire fermentation system is dried with hot air at 50 °C until the water content is 10% to obtain the ginseng extract.

[0039] Example 3 A ginseng extract, prepared by the following preparation method: The ginseng is crushed to be able to pass through a 120-mesh sieve completely, and then mixed with a complex enzyme and water according to the weight ratio of 100 g of ginseng: 7 g of complex enzyme: 100 g of water, the pH is adjusted to 5.6, and enzymatic hydrolysis is carried out 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 enzymolysate; 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.

[0040] The Ganoderma lucidum fungus is inoculated into a liquid medium and cultured at 26 °C and 130 r / min until the weight of Ganoderma lucidum mycelium is 12 g / L to obtain a Ganoderma lucidum fungal liquid; the liquid medium is composed 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 is sterilized at 121 °C for 10 min.

[0041] Mix the enzymatic hydrolysate, corn flour, soybean meal powder, sucrose and water according to the weight-to-volume ratio of 40 g of enzymatic hydrolysate: 9 g of corn flour: 8 g of soybean meal powder: 7 g of sucrose: 0.8 L of water, sterilize at 121 °C for 4 min to obtain a fermentation medium; mix the fermentation medium and Ganoderma lucidum liquid according to the weight ratio of 100:5, ferment at 32 °C and 120 r / min for 22 h, and dry the whole fermentation system at 50 °C with hot air until the moisture content is 8% to obtain ginseng extract.

[0042] Example 4 A composition is made from the following raw materials by weight: 6 parts of Lobelia chinensis Lour., 2 parts of Justicia gendarussa Burm. f., 1 part of Chinese insect wax, 3 parts of Physalis minima L., 5 parts of Abutilon indicum (L.) Sweet, 7 parts of lotus leaf, 8 parts of Cassia obtusifolia L. and 10 parts of the ginseng extract obtained in Example 1; The preparation method is as follows: Mix Lobelia chinensis Lour., Justicia gendarussa Burm. f., Physalis minima L., Abutilon indicum (L.) Sweet, lotus leaf and Cassia obtusifolia L. to obtain a mixture, mix the mixture and water according to the weight ratio of 1:8, soak for 30 min, heat to boiling, and keep boiling for 1 h, and collect the decoction and the medicinal residues respectively; concentrate the decoction at 50 °C under reduced pressure to a relative density of 0.9 to obtain a water extract.

[0043] Mix the medicinal residues and ethanol solution with a volume fraction of 70% according to the weight ratio of 1:7, soak for 30 min, heat under reflux for 1.5 h, collect the extract, and concentrate it at 50 °C under reduced pressure to a relative density of 0.9 to obtain an ethanol extract.

[0044] Mix the ethanol extract and the water extract, stir evenly, heat to 70 °C, add Chinese insect wax, continuously stir until it is completely dissolved, wait until it cools to room temperature, add the ginseng extract obtained in Example 1 and mix, dry at 50 °C with hot air until the moisture content is 5% to obtain the composition.

[0045] Example 5 A composition is made from the following raw materials by weight: 10 parts of Lobelia chinensis Lour., 4 parts of Justicia gendarussa Burm. f., 3 parts of Chinese insect wax, 6 parts of Physalis minima L., 8 parts of Abutilon indicum (L.) Sweet, 9 parts of lotus leaf, 10 parts of Cassia obtusifolia L. and 15 parts of the ginseng extract obtained in Example 2; The preparation method is as follows: Mix Lobelia chinensis Lour., Justicia gendarussa Burm. f., Physalis minima L., Abutilon indicum (L.) Sweet, lotus leaf and Cassia obtusifolia L. to obtain a mixture, mix the mixture and water according to the weight ratio of 1:10, soak for 60 min, heat to boiling, and keep boiling for 2 h, and collect the decoction and the medicinal residues respectively; concentrate the decoction at 60 °C under reduced pressure to a relative density of 1.0 to obtain a water extract.

[0046] Mix the medicinal residues and ethanol solution with a volume fraction of 80% according to the weight ratio of 1:9, soak for 60 min, heat under reflux for 2.5 h, collect the extract, and concentrate it at 60 °C under reduced pressure to a relative density of 1.0 to obtain an ethanol extract.

[0047] Mix the ethanol extract and the water extract, stir evenly, heat to 70 °C, add Chinese insect wax, continuously stir until it is completely dissolved, after cooling to room temperature, add the ginseng extract obtained in Example 2, and dry at 50 °C with hot air until the moisture content is 6% to obtain the composition.

[0048] Example 6 A composition is made from the following raw materials by weight: 8 parts of Lobelia chinensis, 3 parts of Justicia gendarussa, 2 parts of Chinese insect wax, 4 parts of Physalis angulata, 7 parts of Mulberry leaf, 8 parts of Lotus leaf, 9 parts of Cassia obtusifolia and 12 parts of the ginseng extract obtained in Example 3; The preparation method is as follows: (1) Mix Lobelia chinensis, Justicia gendarussa, Physalis angulata, Mulberry leaf, Lotus leaf and Cassia obtusifolia to obtain a mixture, mix the mixture with water at a weight ratio of 1:9 and soak for 40 min, heat to boiling, and keep boiling for 1.5 h, and collect the decoction and the medicinal residues respectively; Concentrate the decoction under reduced pressure at 55 °C to a relative density of 0.9 to obtain the water extract.

[0049] (2) Mix the medicinal residues with an ethanol solution with a volume fraction of 75% at a weight ratio of 1:8 and soak for 40 min, heat under reflux for 2.0 h, collect the extract and concentrate it under reduced pressure at 55 °C to a relative density of 0.9 to obtain the ethanol extract.

[0050] (3) Mix the ethanol extract and the water extract, stir evenly, heat to 70 °C, add Chinese insect wax, continuously stir until it is completely dissolved, after cooling to room temperature, add the ginseng extract obtained in Example 3, and dry at 50 °C with hot air until the moisture content is 5% to obtain the composition.

[0051] Comparative Example 1 The difference from Example 3 is that Ganoderma lucidum was not inoculated, and the rest are the same as Example 3.

[0052] Comparative Example 2 The difference from Example 3 is that α-amylase was not added to the complex enzyme, and the rest are the same as Example 3.

[0053] Comparative Example 3 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 are the same as Example 3.

[0054] Comparative Example 4 The difference from Example 6 is that the ginseng extract is replaced with the ginseng extract obtained in Comparative Example 1, and the rest are the same as Example 6.

[0055] Comparative Example 5 The difference from Example 6 is that it does not contain Cassia obtusifolia, and the rest are the same as Example 6.

[0056] Comparative Example 6 It is different from Example 6 in that it does not contain Chinese insect wax, and the rest is the same as Example 6.

[0057] Comparative Example 7 It is different from Example 6 in that the volume fraction of the ethanol solution is 90%, and the rest is the same as Example 6.

[0058] Comparative Example 8 It is different from Example 6 in that the ethanol solution in step (2) is replaced with water, and the reflux extraction is changed to boiling and kept boiling for 1.5 h, and the rest is the same as Example 6.

[0059] Example 7 (1) Cultivation of human non-small cell lung cancer cell line A549 After taking out the A549 cells frozen in the liquid nitrogen tank, place them in a 37 °C water bath for rapid thawing, and centrifuge at a speed of 1000 rpm / min for 3 min, then take out and discard the supernatant. Add 1 mL of DMEM medium containing 10% fetal bovine serum and gently pipette to suspend. After pipetting several times to mix evenly, transfer the suspension into a 25 mL cell culture flask, supplement the DMEM complete medium to 5 mL, pipette and mix well, and then place it in a cell culture incubator (5% CO2, 37 °C) for cultivation. When it is observed under the microscope that the cells have covered 80% - 90% of the bottom of the culture flask, start the cell passage procedure. First, discard the original medium, add physiological saline to the culture flask, place the culture flask flat and gently shake to wash the cells, 2 mL each time, wash 3 times to ensure that the floating matter and metabolites produced during the culture are washed away. Then add 2 mL of trypsin and gently shake to make the trypsin moisten the cell surface. Observe under the microscope. When it is observed that the cells change from the original state to a round state, it proves that the digestion is complete, and add an appropriate amount of complete medium to terminate the digestion. Collect the digested cell suspension, centrifuge, take a certain amount of cells and put them into a new culture flask, supplement the complete medium and then place it in the culture incubator (5% CO2, 37 °C) for continuous cultivation.

[0060] (2) Effects of ginseng extracts or compositions obtained in Examples 1 - 6 and Comparative Examples 1 - 8 on the proliferation inhibition rate of A549 cells The ginseng extracts or compositions obtained in Examples 1 - 6 and Comparative Examples 1 - 8 were respectively formulated into ginseng extract solutions or composition solutions with a concentration of 50 μg / mL using complete medium, and the following tests were carried out: Collect A549 cells in the best logarithmic growth phase of the growth state for the plating experiment. Discard the culture medium, add physiological saline to the culture flask, lay the culture flask flat and gently shake to wash the cells, 2 mL each time, wash 3 times to ensure that the floating substances and metabolites produced during the culture are washed clean. Discard the physiological saline, digest with trypsin and centrifuge. Collect the centrifuged cells and dilute them with complete medium to a density of 4×10 3 cells per well. Use a multi-channel pipette to inoculate the cells into a 96-well plate, 100 μL per well. Culture in an incubator for 24 h. Discard the supernatant. Add 100 μL of complete medium to the blank control group, and add 100 μL of ginseng extract solution or composition solution with a concentration of 50 μg / mL to the sample groups respectively. There are 6 replicate wells in each group. Continue to culture in a 37 °C, 5% CO2 incubator for 24 h. Add 10 μL of 5 mg / mL MTT to each well. After 4 h, discard the supernatant, add 100 μL of DMSO, place it on an enzyme-linked immunosorbent assay (ELISA) reader and shake for 5 min. Detect at a wavelength of 490 nm. According to the measured absorbance value, calculate the cell proliferation inhibition rate of each group according to the following formula. The independent experiment is repeated three times. Use SPSS 24.0 data processing software to analyze the data. The experimental data are expressed as mean ± standard deviation (x±s). ANOVA one-way analysis of variance is used for comparison between groups. p<0.05 represents a significant difference between the two groups of data, and the data results are statistically significant. The results are shown in Table 1.

[0061] Cell proliferation inhibition rate (%) = [A 空白组 -A 给药组 / A 空白组 ×100% Table 1 Effects of different groups on the cell proliferation inhibition rate of A549 cells

[0062] Note: The superscript letters used in the table represent the differences between groups. Different lowercase letters in the same column indicate significant differences. p＜0.05。

[0063] (3) Effects of the ginseng extracts or compositions obtained in Examples 3 and 6 and Comparative Examples 1-8 on the migration ability of A549 cells Cell scratch assay Inoculate A549 cells in the logarithmic growth phase into a 6-well plate. Use a 200 μL sterile pipette tip to make a scratch. After rinsing off the floating broken cells with PBS, observe and take pictures under a fluorescence inverted microscope. Prepare the ginseng extracts or compositions obtained in Examples 3 and 6 and Comparative Examples 1-8 into a concentration of 100 μg / mL with DMEM medium containing 1% fetal bovine serum (FBS). Set 3 replicate wells in each group. The control group is added with 2.5 mL of DMEM medium was placed in an environment of 37 °C and 5% carbon dioxide and continued to be cultured for 24 h, and then the cell migration was observed and photographed under a microscope. The cell migration results of the experimental group 3 and the control group 1 were respectively as Figure 1 and Figure 2 shown ( Figure 1 and Figure 2 had the same magnification), and the scratch repair rate was analyzed and calculated using ImageJ software. The initial area (A0) of the scratch area and the remaining area (A t ) at the observation time point were measured. Finally, the scratch repair rate was calculated according to the formula: The independent experiment was repeated three times. The data were analyzed using SPSS 24.0 data processing software. The experimental data were expressed as mean ± standard deviation (x ± s). One-way ANOVA was used for inter-group comparison. The results are shown in Table 2.

[0064] Table 2 Scratch repair rates of different groups

[0065] Note: ** in the table indicates compared with the blank group, p＜0.01。

[0066] Transwell cell migration experiment The A549 cells in the logarithmic growth phase were digested and prepared into a suspension 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 the cell suspension was taken and added to the upper chamber, and the cell number was 5×10 5 cells / mL. The ginseng extracts or compositions obtained in Examples 3 and 6 and Comparative Examples 1-8 were used as drugs, and the administration concentration was 100 μg / mL. The control group only added complete medium. After culturing for 24 h, the chamber was taken out, and the cells were fixed with 4% paraformaldehyde and stained with 1% crystal violet. After washing three times with PBS, three random fields of view were photographed and counted under a microscope, and the average value was taken as the number of migrated cells. The relative migration rate (%) of each group was calculated according to the following formula. The independent experiment was repeated three times. The data were analyzed using SPSS 24.0 data processing software. The experimental data were expressed as mean ± standard deviation (x ± s). One-way ANOVA was used for inter-group comparison. The results are shown in Table 3. The field-of-view photographing results of the experimental group 3 and the control group 1 were respectively as Figure 3 and Figure 4 shown ( Figure 3 and Figure 4 had the same magnification).

[0067] Relative migration rate (%) = (number of migrated cells in the experimental group ÷ number of migrated cells in the control group) × 100% Table 3 Relative migration rates of different groups

[0068] Note: *** in the table indicates that compared with the blank group, p＜0.001。

[0069] Example 8 Eight-week-old wild-type female BALB / c mice were selected as the normal group (Normal group). Sixty MRL / lpr female mice were randomly divided into a positive drug group (intragastric administration of glucocorticoid prednisone, 1.0 mg / kg body weight / day), a blank group (intragastric administration of normal saline, 1.0 mg / kg body weight / day), Example 3, Example 6, and Comparative Example 1 to Comparative Example 8 groups (intragastric administration, 1.0 mg / kg body weight / day). The handling of animals in the experiment complied with the 3R rules. The surgical procedures and experimental data analysis for each group were performed in a single-blind mode. The animals were maintained in an environment with a 12h light / 12h dark cycle and had free access to food and water, and were acclimated to the conditions for 7 days before the experiment. Each group of mice was intragastrically administered at 12 weeks of age, once every other day for a total of 2 weeks. At 14 weeks of age, 24-hour urine of the mice was collected for the detection of urinary protein and creatinine; and the skin lesions and erythema on the faces and backs of the mice were observed; and the serum of the mice was collected for the detection of anti-nucleosome antibody (AnuA). The results were expressed as averages.

[0070] The results showed that compared with the blank group, the skin lesions on the faces and backs of the positive drug group, Example 3, Example 6, and Comparative Example 1 to Comparative Example 8 groups were significantly reduced, and the number of erythema was significantly decreased. Among them, the degree of reduction of skin lesions on the faces and backs of Example 3 and Example 6 groups was significantly higher than that of Comparative Example 1 to Comparative Example 8 groups, and even better than that of the positive drug group. The detection results of urinary protein, creatinine, and AnuA are shown in Table 4.

[0071] Table 4 Detection results of urinary protein, creatinine, and AnuA in different groups

[0072] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A composition containing ginseng extract, characterized in that, It consists of the following raw materials by weight parts: 6 - 10 parts of Lobelia chinensis Lour., 2 - 4 parts of Justicia gendarussa Burm. f., 1 - 3 parts of Chinese insect wax, 3 - 6 parts of Physalis angulata L., 5 - 8 parts of Morus alba L. var. rotundata W. T. Wang, 7 - 9 parts of lotus leaf, 8 - 10 parts of Cassia obtusifolia L. seeds and 10 - 15 parts of ginseng extract; The preparation method of the ginseng extract is as follows: Ginseng is crushed and mixed with a complex enzyme and water in a weight ratio of 100:(6 - 8):100 for enzymatic hydrolysis to obtain an enzymatic hydrolysate; The enzymatic hydrolysate, corn flour, soybean meal powder, sucrose and water are mixed in a weight - volume ratio of (30 - 50) g:(8 - 10) g:(6 - 10) g:(5 - 8) g:0.8 L to obtain a fermentation medium; The fermentation medium and Ganoderma lucidum spore suspension are mixed in a weight ratio of 100:4 - 6, fermented at 30°C - 35°C for 20 h - 24 h, and dried to obtain the ginseng extract; The complex enzyme is composed of cellulase, pectinase and α - amylase; The weight of Ganoderma lucidum mycelium in the Ganoderma lucidum spore suspension is 8 g / L - 14 g / L.

2. The composition according to claim 1, wherein The weight ratio of the cellulase, pectinase and α - amylase is (1 - 2):(2 - 3):(0.5 - 1); The enzyme activity of the cellulase is above 100,000 U / g; The enzyme activity of the pectinase is above 30,000 U / g; The enzyme activity of the α - amylase is above 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 temperature of the enzymatic hydrolysis is 40°C - 50°C, and the time of the enzymatic hydrolysis is 12 h - 16 h.

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

5. A method for preparing the composition according to any one of claims 1 to 4, characterized in that, It consists of the following steps: Mix Lobelia chinensis Lour., Justicia gendarussa Burm. f., Physalis angulata L., Morus alba L. var. rotundata W. T. Wang, lotus leaf and Cassia obtusifolia L. seeds to obtain a mixture, mix the mixture with water and soak it, heat to boiling and decoct for 1 - 2 h, and collect the decoction and the medicinal residues respectively; Concentrate the decoction to obtain a water extract; Mix the medicinal residues with an ethanol solution with a volume fraction of 70% - 80% and soak it, heat under reflux for 1.5 - 2.5 h, collect the extract and concentrate it to obtain an ethanol extract; Mix the ethanol extract, the water extract, Chinese insect wax and ginseng extract, and dry to obtain a composition.

6. The preparation method according to claim 5, wherein, The soaking time is 30 min - 60 min; The concentration is to concentrate to a relative density of 0.9 - 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 residues 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 - 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 - 4 in the preparation of a drug for treating systemic lupus erythematosus.

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