CN111643649A - Chinese medicinal materials for preventing and treating immune related diseases and composition thereof - Google Patents

Abstract

The invention relates to the technical field of traditional Chinese medicine, in particular to traditional Chinese medicinal materials and compositions for preventing and treating immune-related diseases. The present invention provides at least one of Angelica dahuricae, Goosebum, Inula, Ginger, Artemisia annua, Chuan Shegan, Gardenia, L. japonica, Wild Chrysanthemum, Hibiscus Leaf, Artemisia argyi, Agrimony, Honeysuckle and Arranthaceae Application in the preparation of immunomodulators; any of the above single drugs can significantly act on one or more key genes regulating immune function, such as genes related to viral infection, chronic inflammation and/or autoimmune diseases. These traditional Chinese medicine compositions can inhibit a number of key genes and thus play a role in regulating immune function.

Description

Chinese medicinal materials and compositions for preventing and treating immune-related diseases

technical field

The invention relates to the technical field of traditional Chinese medicine, in particular to traditional Chinese medicinal materials and compositions for preventing and treating immune-related diseases.

Background technique

Monocytes-macrophages are an important immune cell group in the body. They can non-specifically phagocytose a variety of pathogenic microorganisms. They are an important part of the body's non-specific immune defense and play an important role in anti-viral infection. At the same time, in the specific immune response, most TD antigens need to be phagocytosed and processed by macrophages to form antigen peptide-MHC complexes, which are expressed on the surface of the cell membrane, and there are many adhesion molecules on the surface of macrophages, which can interact with T cells. The co-stimulatory molecule receptors on the cell surface bind to generate a co-stimulatory signal, induce the activation of T cells, and initiate an immune response. In the specific immune response, macrophages can also secrete and release a variety of cytokines, which are involved in a variety of immune regulation. In addition, macrophages can effectively kill tumor cells after being activated by certain cytokines such as IFN-γ, and are important effector cells involved in immune surveillance. Therefore, in addition to fighting viral infections, macrophages are involved in a variety of chronic inflammatory and autoimmune diseases such as rheumatoid arthritis, atherosclerosis, fibrosis, asthma, allergic rhinitis, viral hepatitis, multiple sclerosis and inflammatory bowel disease.

In the prior art, there are many reports on immune regulation, and a large number of reports show that traditional Chinese medicine has a good effect on the regulation of the immune system, but the current research on the target of Chinese medicine to regulate the immune system is far from enough.

SUMMARY OF THE INVENTION

In view of this, the technical problem to be solved by the present invention is to provide Chinese medicinal materials and compositions for the prevention and treatment of immune-related diseases. Studies have shown that a variety of traditional Chinese medicine extracts have significant effects on inflammation and cytokine storm-related genes, apoptosis-related genes and/or non-inflammatory effects. The key genes of fibrosis have inhibitory effects, and the traditional Chinese medicine composition prepared by combining them has significant advantages.

The invention provides the application of traditional Chinese medicine in the preparation of medicines for preventing and treating immune-related diseases; the traditional Chinese medicine is selected from inula flower, wild chrysanthemum, angelica, hibiscus leaf, gooseberry, mugwort, ginger, artemisia, honeysuckle, gardenia At least one of the seeds, two-sided needles, arthropods, Chuanshegan, and Agrimony.

The prevention and treatment include: regulating the expression of genes related to viral infection and/or regulating the expression of genes related to respiratory failure;

The viral infection-related genes include CXCL1, IL1B, IL6, NFKB1, NFKB1A, PTGS2, IL1A, TNF, CCL2, RELA and/or NFKB2; the respiratory failure-related genes include ITGA1, ITGB3, ITGB5, ITGB1, INHBA, VEGFB , VEGFA, TGFB1, MMP9, ACTB, FN1 and/or BMP2.

The immune-related diseases include: viral infections, chronic inflammation, allergic diseases and/or autoimmune diseases;

The viruses include: coronavirus, adenovirus, herpes virus, enterovirus, measles virus, hepatitis B virus, hepatitis C virus, human papilloma virus, influenza A (H1N1) virus, and human T cell leukemia virus.

The autoimmune diseases include: rheumatoid arthritis, atherosclerosis, pulmonary fibrosis, viral hepatitis, multiple sclerosis and/or inflammatory bowel disease;

The allergic diseases include allergic asthma and/or allergic rhinitis.

The respiratory failure includes respiratory failure resulting from acute lung injury manifested as alveolar epithelial damage, uncontrolled inflammatory response and/or pulmonary fibrosis.

In the embodiment of the present invention, the traditional Chinese medicine with the effect of regulating immunity is the ethanolic extract of the traditional Chinese medicinal material. The preparation method is as follows: standard Soxhlet reflux extraction is performed on the medicinal material with 90 vol% ethanol aqueous solution, and the obtained extract is dried to obtain the ethanol extract of the traditional Chinese medicinal material.

Chinese medicine believes that: Artemisia annua, bitter in taste, pungent, cold in nature, returns to the liver and gallbladder meridians, clears heat from deficiency, eliminates Gu steaming, is used for warming pathogens and injuring yin, fever due to yin deficiency, fever caused by steaming of bones, heat caused by summer heat, cold and heat caused by malaria , Damp heat jaundice. Honeysuckle, sweet in taste, cold in nature, returns to the lung, heart, and stomach meridians, clears heat and detoxifies, disperses wind-heat, and is used for carbuncle swollen furuncle, throat arthralgia, erysipelas, heat toxin and blood dysentery, wind-heat cold, and fever due to febrile diseases. Gardenia, slightly bitter and pungent in taste, warm in nature, returns to the heart, spleen, and stomach meridians, regulates qi and relieves pain, eliminates food and stops diarrhea, dispels wind and removes dampness, and calms the nerves. Goose does not eat herbs, is acrid in taste, warm in nature, returns to the lung meridian, dissipates wind-cold, clears the nose orifices, and stops constipation. Inula flower, bitter, pungent, salty, slightly warm, enters the lung, spleen, stomach, and large intestine meridians, lowers qi, eliminates phlegm, promotes water, and stops vomiting. It is used for wind-cold cough, accumulation of phlegm and phlegm, chest and diaphragm suffocation, dyspnea and expectoration with excessive phlegm, vomiting and suffocation, and hard heart. Chuanshegan, bitter in taste, cold in nature, returns to the lung meridian, clears heat and detoxifies, removes phlegm, and relieves throat. For heat-toxic phlegm-fire stagnation, sore throat, phlegm and saliva, cough and asthma. Angelica sinensis, spicy, sweet, warm in nature, expelling wind and dispelling cold, relieving cough and wheezing, promoting qi and relieving pain, main wind-cold cold, cough, asthma, headache, toothache, etc. Ginger, spicy, slightly warm, returns to the lung, spleen and stomach meridians, relieves the exterior and disperses cold, warms and stops vomiting, resolves phlegm and relieves cough, and resolves fish and crab poison. For wind-cold cold, stomach-cold vomiting, cough with cold phlegm, fish and crab poisoning. Two-sided acupuncture, bitter in taste, pungent, flat in nature, returns to the liver and stomach meridians, can activate blood and remove blood stasis, promote qi and relieve pain, dispel wind and dredge collaterals, detoxify and reduce swelling. Wild chrysanthemum, bitter, acrid, slightly cold in nature, returns to the liver and heart meridians, clears heat and detoxifies, purifies fire and calms the liver. Hibiscus leaf, pungent in taste, flat in nature, returns to the lung and liver meridians, cools blood, detoxifies, reduces swelling, and relieves pain. Artemisia argyi, pungent and bitter in taste, warm in nature, returns to the liver, spleen and kidney meridians, warms the meridians to stop bleeding, disperses cold and relieves pain. Agrimony, bitter in taste, astringent, flat in nature, return to the heart and liver meridians, can astringe and stop bleeding, cut malaria, stop dysentery, detoxify, and tonic. System Tian Nanxing, bitter in taste, pungent, warm in nature, return to lung, liver and spleen meridians to dry dampness and resolve phlegm, dispel wind and stop spasm, disperse knot and reduce swelling.

The present invention proves that:

The ethanolic extract of Inula flower can inhibit the expression of PTGS2, IL1A, CXCL1, NFKB1A, NFKB1, IL6, INHBA, ITGB1, ITGA1, TGFB1, ITGB5 and/or ITGB3 genes.

The ethanolic extract of wild chrysanthemum can inhibit the expression of PTGS2, IL1A, CXCL1, NFKB1A, NFKB1, FN1, INHBA, ITGB1, ITGA1, TGFB1 and/or ITGB3 genes.

The ethanol extract of Angelica dahurica can inhibit the expression of CCL2, TNF, IL1A, CXCL1, NFKB1A, NFKB1, IL1B, RELA, VEGFB, VEGFA, INHBA, ITGB1, ITGA1, TGFB1, ITGB5 and/or ITGB3 genes.

The ethanolic extract of Hibiscus hibiscus leaves can inhibit the expression of TNF, IL1A, CXCL1, NFKB1, RELA, VEGFA, INHBA, ITGA1 and/or TGFB1 genes.

The ethanolic extract of geese can inhibit the expression of PTGS2, TNF, IL1A, CXCL1, NFKB1A, NFKB1, IL6, RELA, VEGFA, INHBA, ITGB1 and/or ITGA1 genes.

The ethanolic extract of Artemisia argyi can inhibit the expression of PTGS2, IL1A, CXCL1, NFKB1A, NFKB1, INHBA, ITGB1, ITGA1 and/or TGFB1 genes.

The ethanolic extract of ginger can inhibit the expression of CCL2, PTGS2, TNF, IL1A, CXCL1, NFKB1A, ACTB, VEGFA and/or INHBA genes.

The ethanolic extract of Artemisia annua can inhibit the expression of NFKB2, CCL2, PTGS2, TNF, CXCL1, MMP9, ACTB, INHBA and/or ITGB1 genes.

The ethanol extract of honeysuckle can inhibit the expression of BMP2 gene.

The ethanolic extract of Gardenia can inhibit the expression of NFKB2 and/or CCL2 gene.

The ethanolic extract of L. japonicus can inhibit the expression of CCL2 and/or IL1A gene.

The ethanolic extract of Arata radix can inhibit the expression of NFKB2, CCL2, TNF, IL1A, CXCL1, NFKB1A, NFKB1, IL6, FN1, MMP9, ACTB, VEGFA, ITGB1 and/or ITGA1 genes.

The ethanol extract of Chuanshegan can inhibit the expression of CCL2, TNF, IL1A, CXCL1, IL6, FN1, VEGFA, INHBA, ITGA1 genes.

The ethanolic extract of Agrimony can inhibit the expression of CCL2, TNF, CXCL1, NFKB1A, FN1, MMP9, ACTB, VEGFA and/or TGFB1 genes.

The present invention also provides a traditional Chinese medicine composition, which is composed of Artemisia annua, honeysuckle, gardenia, goosebum, inula, Chuanshegan, Angelica dahurica, ginger, two-sided needle, wild chrysanthemum, hibiscus leaf, mugwort, and crane. Composition of grass and Arranthaceae.

In some embodiments, the traditional Chinese medicine composition is composed of the following components by mass: 6-15 parts of Artemisia annua, 3-30 parts of honeysuckle, 6-25 parts of gardenia, 6-15 parts of goosebum 3 to 9 parts of Inula, 6 to 10 parts of Chuan Shegan, 2 to 12 parts of Angelica Radix, 3 to 30 parts of ginger, 5 to 10 parts of two-sided needles, 9 to 15 parts of wild chrysanthemum, 10 to 30 parts of Hibiscus hibiscus, 3 to 15 parts of Artemisia argyi, 6 to 12 parts of Agrimony, and 3 to 9 parts of Aronia star.

In some specific embodiments, the traditional Chinese medicine composition is made up of the following components by mass: 10 parts of Artemisia annua, 10 parts of honeysuckle, 10 parts of Gardenia, 10 parts of goosebum, 10 parts of Inula flower, 10 parts of Chuan Shegan. parts, 10 parts of Angelica dahurica, 10 parts of ginger, 10 parts of two-sided needles, 10 parts of wild chrysanthemum, 10 parts of hibiscus leaves, 10 parts of mugwort leaves, 10 parts of Agrimony, and 10 parts of Tiannanxing.

In some specific embodiments, the traditional Chinese medicine composition is made up of the following components by mass: 6 parts of Artemisia annua, 3 parts of honeysuckle, 6 parts of Gardenia, 6 parts of Goosebum, 3 parts of Inula vulgaris, 6 parts of Chuan Shegan. parts, 2 parts of Angelica dahurica, 3 parts of ginger, 5 parts of two-sided needle, 9 parts of wild chrysanthemum, 10 parts of hibiscus leaves, 3 parts of mugwort leaves, 6 parts of Agrimony, and 3 parts of Arranthaceae.

In some specific embodiments, the traditional Chinese medicine composition is made up of the following components by mass: 15 parts of Artemisia annua, 30 parts of honeysuckle, 25 parts of Gardenia, 15 parts of goosebum, 9 parts of Inula chinensis, 10 parts of Chuan Shegan. parts, 12 parts of Angelica, 30 parts of ginger, 10 parts of two-sided needles, 15 parts of wild chrysanthemum, 30 parts of Hibiscus, 15 parts of Artemisia argyi, 12 parts of Agrimony, and 9 parts of Tiannanxing.

In some specific embodiments, the traditional Chinese medicine composition is made up of the following components by mass: 9 parts of Artemisia annua, 9 parts of honeysuckle, 6 parts of Gardenia, 6 parts of goosebum, 6 parts of Inula chinensis, 6 parts of Chuan Shegan. parts, 6 parts of Angelica dahurica, 9 parts of ginger, 6 parts of two needles, 12 parts of wild chrysanthemum, 15 parts of hibiscus leaves, 6 parts of mugwort leaves, 9 parts of Agrimony, and 6 parts of Arranthaceae.

The present invention provides a traditional Chinese medicine extract, which is obtained by extracting the traditional Chinese medicine composition of the present invention.

The preparation method of the traditional Chinese medicine extract of the present invention comprises the steps of mixing Artemisia annua, honeysuckle, gardenia, goosebum, inula, Chuanshegan, Angelica dahurica, ginger, two-sided needle, wild chrysanthemum, hibiscus leaf, mugwort, and crane. The traditional Chinese medicine extracts were prepared by standard Soxhlet reflux extraction of the herbs from the herbal medicine Tiannanxing with 90 vol% ethanol aqueous solution, and the obtained extract was dried.

The standard Soxhlet reflux extraction time of the 90 vol% ethanol aqueous solution is 3 hours, and every 10 g of medicinal materials is extracted with 150 mL of 90 vol% ethanol aqueous solution. The standard Soxhlet reflux extraction time of the water is 2h, and every 10g of medicinal materials is extracted with 150mL of water.

In the preparation method of the traditional Chinese medicine extract of the present invention, each component can be mixed with the extract, or the extracts can be extracted successively and then mixed, which is not limited in the present invention.

The application of the traditional Chinese medicine composition, the traditional Chinese medicine extract or the extract prepared by the preparation method of the present invention in the preparation of medicines for the treatment of immune-related diseases.

The prevention and treatment includes: regulating the expression of genes related to viral infection and/or regulating the expression of genes related to respiratory failure.

That is, the traditional Chinese medicine composition of the present invention can regulate the expression of viral infection-related genes and/or regulate the expression of respiratory failure-related genes.

The viral infection-related genes include inflammation-related genes, cytokine storm-related genes, and apoptosis-related genes; the respiratory failure-related genes include pulmonary fibrosis key genes;

The immunomodulation includes inhibition of NFKB2, CCL2, PTGS2, TNF, IL1A, CXCL1, NFKB1A, NFKB1, IL6, IL1B, RELA, BMP2, VEGFB, FN1, MMP9, ACTB, VEGFA, INHBA, ITGB1, ITGA1, TGFB1, ITGB3 and / or ITGB5.

Wherein, the viral infection-related genes include CXCL1, IL1B, IL6, NFKB1, NFKB1A, PTGS2, IL1A, TNF, CCL2, RELA and/or NFKB2; the respiratory failure-related genes include ITGA1, ITGB3, ITGB5, ITGB1, INHBA , VEGFB, VEGFA, TGFB1, MMP9, ACTB, FN1 and/or BMP2.

The traditional Chinese medicine composition, the traditional Chinese medicine extract or the extract prepared by the preparation method of the present invention are used in the preparation and treatment of immune-related diseases.

The immune-related diseases include: viral infections, chronic inflammation, allergic diseases and/or autoimmune diseases.

The viruses include: coronavirus, adenovirus, herpes virus, enterovirus, measles virus, hepatitis B virus, hepatitis C virus, human papilloma virus, influenza A (H1N1) virus, and human T cell leukemia virus.

The autoimmune diseases include: rheumatoid arthritis, atherosclerosis, pulmonary fibrosis, viral hepatitis, multiple sclerosis and/or inflammatory bowel disease.

The allergic diseases include allergic asthma and/or allergic rhinitis.

The occurrence of pulmonary fibrosis affects breathing, causing dyspnea, and in severe cases, respiratory failure, and the treatment also includes the treatment of respiratory failure caused by pulmonary fibrosis.

The present invention also provides a medicine, which comprises the traditional Chinese medicine composition, the traditional Chinese medicine extract or the extract prepared by the preparation method of the present invention.

The medicine also includes pharmaceutically acceptable excipients;

The pharmaceutically acceptable adjuvants are one or more of flavoring agents, fillers, lubricants, preservatives, suspending agents, food colorings, diluents, emulsifiers, disintegrating agents or plasticizers mixture;

The dosage form of the medicine is injection, tablet, pill, oral liquid, capsule, syrup, drop pill or granule.

The present invention also provides a method for treating immune-related diseases, which is to administer the medicament of the present invention. The mode of administration is injection or oral.

The present invention provides at least one of Angelica dahuricae, Goosebum, Inula, Ginger, Artemisia annua, Chuan Shegan, Gardenia, L. japonica, Wild Chrysanthemum, Hibiscus Leaf, Artemisia argyi, Agrimony, Honeysuckle and Arranthaceae Application in the preparation of immunomodulators; any of the above single drugs can significantly act on one or more key genes regulating immune function, such as genes related to viral infection, chronic inflammation and/or autoimmune diseases. These traditional Chinese medicine compositions can inhibit a number of key genes and thus play a role in regulating immune function, and their effects are significantly better than any single medicinal material, and are better than compositions lacking a prescription.

Description of drawings

Figure 1 shows the expression profiles of 14 kinds of traditional Chinese medicines in THP-1 cells of viral infection-related genes and respiratory failure-related genes;

Figure 2 shows the expression profiles of the viral infection-related genes and respiratory failure-related genes of the traditional Chinese medicine compositions of Examples 2, 3, 4, and 5 in THP-1 cells;

Figure 3 shows the expression profiles of viral infection-related genes and respiratory failure-related genes of the traditional Chinese medicine compositions of Example 2 and Comparative Examples 1 to 3 in THP-1 cells;

Figure 4 shows the expression profiles of the viral infection-related genes and respiratory failure-related genes of the traditional Chinese medicine compositions of Example 2 and Comparative Examples 4 to 6 in THP-1 cells.

Detailed ways

The present invention provides traditional Chinese medicinal materials and compositions for preventing and treating immune-related diseases, and those skilled in the art can learn from the content of this article and appropriately improve process parameters to achieve. It should be particularly pointed out that all similar substitutions and modifications are obvious to those skilled in the art, and they are deemed to be included in the present invention. The method and application of the present invention have been described through the preferred embodiments, and it is obvious that relevant persons can make changes or appropriate changes and combinations of the methods and applications herein without departing from the content, spirit and scope of the present invention, so as to realize and apply the present invention. Invention technology.

The test materials used in the present invention are all common commercial products and can be purchased in the market.

The detection gene of the present invention includes:

NFKB2, nuclear factor of kappa light polypeptide gene enhancer in Bcells 2, nuclear factor activated B cell κ light chain enhancer 2;

CCL2, chemokine (C-C motif) ligand2, chemokine ligand 2;

PTGS2, prostaglandin-endoperoxide synthase 2, prostaglandin-endoperoxide synthase 2;

TNF, tumor necrosis factor, tumor necrosis factor;

IL1A, interleukin 1, alpha, interleukin 1, alpha;

CXCL1, chemokine (C-X-C motif) ligand 1, chemokine (C-X-C motif) ligand 1;

NFKB1A, NFKB inhibitor alpha, NFKB inhibitor alpha;

NFKB1, nuclear factor of kappa light polypeptide gene enhancer in B-cells 1, nuclear factor activated B cell κ light chain enhancer 1;

IL6, Interleukin-6, Interleukin-6;

IL1B, interleukin 1, beta, interleukin 1, beta

RELA, V-Rel reticuloendotheliosis viral oncogene homolog A, V-Rel reticuloendotheliosis virus oncogene homolog A;

BMP2, bone morphogenetic protein 2, bone morphogenetic protein 2;

VEGFB, vascular endothelial growth factor B, vascular endothelial growth factor B.

FN1, fibronectin 1, fibronectin 1;

MMP9, matrix metallopeptidase 9, matrix metallopeptidase 9;

ACTB, actin, beta, actin, beta;

VEGFA, vascular endothelial growth factor A, vascular endothelial growth factor A;

INHBA, inhibin, beta A, arrestin, beta A;

ITGB1, integrin, beta 1, integrin, beta 1;

ITGA1, integrin, alpha 1, integrin, alpha 1;

ITGB3, integrin, beta 3, integrin, beta 3;

ITGB5, integrin, beta 5, integrin, beta 5.

Below in conjunction with embodiment, the present invention is further elaborated:

Example 1

The preparation of traditional Chinese medicine extract, the specific process is:

1. Weigh the traditional Chinese medicines Angelica dahurica, Goosebukki, Inula flower, Ginger, Artemisia annua, Chuan Shegan, Gardenia, L. japonica, Wild Chrysanthemum, Hibiscus Leaf, Artemisia argyi, Agrimony, Honeysuckle and Arranthaceae.

2. Preparation method: After the above medicines are crushed, 10 g are weighed respectively, and each medicine is extracted with 150 mL of 90% ethanol, standard Soxhlet reflux for 3 hours, the extract is concentrated for 20 minutes to a volume of 2 mL, and then lyophilized under reduced pressure using a freeze dryer for 24 hours. , were made into the freeze-dried powder of the above 13 kinds of various traditional Chinese medicine extracts.

A certain mass of lyophilized powder was weighed and dissolved in DMSO to prepare an extract mother solution. The concentration of the mother solution was 20 mg/mL. The Chinese herbal extract with a final concentration of 100 μg/mL was prepared and stored at -20°C.

Examples 2 to 5

The preparation of the traditional Chinese medicine composition, the specific process is:

1. Weigh the traditional Chinese medicines Angelica dahurica, Goosebukki, Inula flower, Ginger, Artemisia annua, Chuan Shegan, Gardenia, L. japonica, Wild Chrysanthemum, Hibiscus Leaf, Artemisia argyi, Agrimony, Honeysuckle and Arranthaceae.

2. Preparation method: The above medicines are all prepared separately.

Weigh 10g of each drug respectively, extract with 150mL of 90% ethanol, standard Soxhlet reflux for 3h, concentrate the extract for 20min to a volume of 2mL, and freeze-dried under reduced pressure in a lyophilizer for 24h to prepare the above 13 kinds of traditional Chinese medicine extraction. lyophilized powder. The lyophilized powder of each extract was weighed according to the proportion shown in Table 1, and dissolved in DMSO to prepare the mother liquor of each traditional Chinese medicine extract. save.

The proportioning (g) of the compositions of Table 1 Examples 2-5

Example 2 Example 3 Example 4 Example 5 Artemisia annua 10 15 6 9 honeysuckle 10 30 3 9 Gardenia 10 25 6 6 geese do not eat grass 10 15 6 6 Inula 10 9 3 6 Kawasaki 10 10 6 6 Mountain Angelica 10 12 2 6 ginger 10 30 3 9 double needle 10 10 5 6 wild chrysanthemum 10 15 9 12 wood hibiscus leaves 10 30 10 15 Ai Ye 10 15 3 6 Agrimony 10 12 6 9 Arranus star 10 9 3 6

Comparative Example 1

Taking Zhenwu Decoction in the prior art as positive control, the preparation method is:

Poria cocos, white peony root, Atractylodes macrocephala, ginger and aconite are respectively crushed and prepared separately. Weigh 10g of each medicine, respectively use 150mL of 90% ethanol, extract with standard Soxhlet reflux for 3h, concentrate the extract for 20min to a volume of 2mL, and use a freeze dryer to freeze-dry under reduced pressure for 24h to prepare the above five kinds of traditional Chinese medicines. Lyophilized powder of the extract. A certain mass of lyophilized powder was weighed and dissolved in DMSO to prepare an extract mother solution. The concentration of the mother solution was 20 mg/mL. The components were proportioned in equal proportions, and the cells were treated with a final concentration of 100 μg/mL Chinese medicine extract.

Comparative Example 2

Taking Liujunzi Decoction in the prior art as positive control, the preparation method is:

Ginseng, Atractylodes, Poria, Licorice, dried tangerine peel, and Pinellia are respectively crushed and prepared separately. Weigh 10g of each medicine, respectively use 150mL of 90% ethanol, extract with standard Soxhlet reflux for 3h, concentrate the extract for 20min to a volume of 2mL, and use a freeze dryer to freeze-dry under reduced pressure for 24h to prepare the above 6 kinds of traditional Chinese medicines. Lyophilized powder of the extract. A certain mass of lyophilized powder was weighed and dissolved in DMSO to prepare an extract mother solution. The concentration of the mother solution was 20 mg/mL. The components were proportioned in equal proportions, and the cells were treated with a final concentration of 100 μg/mL Chinese medicine extract.

Comparative Example 3

Taking Bufei Decoction in the prior art as positive control, the preparation method is:

Morus alba, Rehmannia glutinosa, ginseng, aster, astragalus and schisandra are respectively crushed and prepared separately. Weigh 10g of each medicine, respectively use 150mL of 90% ethanol, extract with standard Soxhlet reflux for 3h, concentrate the extract for 20min to a volume of 2mL, and use a freeze dryer to freeze-dry under reduced pressure for 24h to prepare the above 6 kinds of traditional Chinese medicines. Lyophilized powder of the extract. A certain mass of lyophilized powder was weighed and dissolved in DMSO to prepare an extract mother solution. The concentration of the mother solution was 20 mg/mL. The components were proportioned in equal proportions, and the cells were treated with a final concentration of 100 μg/mL Chinese medicine extract.

Comparative Example 4

Part of the Chinese medicinal materials in Example 1 were used to make part of the prescription, as a control, the preparation method was:

Angelica chinensis, goosebum, honeysuckle, mugwort, and Agrimony are separately crushed and prepared separately. Weigh 10g of each medicine, respectively use 150mL of 90% ethanol, extract with standard Soxhlet reflux for 3h, concentrate the extract for 20min to a volume of 2mL, and use a freeze dryer to freeze-dry under reduced pressure for 24h to prepare the above five kinds of traditional Chinese medicines. Lyophilized powder of the extract. A certain mass of lyophilized powder was weighed and dissolved in DMSO to prepare an extract mother solution. The concentration of the mother solution was 20 mg/mL. The components were proportioned in equal proportions, and the cells were treated with a final concentration of 100 μg/mL Chinese medicine extract.

Comparative Example 5

Part of the Chinese medicinal materials in Example 1 were used to make part of the prescription, as a control, the preparation method was:

Inula flower, ginger, Artemisia annua, Chuan Shegan, and Gardenia are crushed and prepared separately. Weigh 10g of each medicine, respectively use 150mL of 90% ethanol, extract with standard Soxhlet reflux for 3h, concentrate the extract for 20min to a volume of 2mL, and use a freeze dryer to freeze-dry under reduced pressure for 24h to prepare the above five kinds of traditional Chinese medicines. Lyophilized powder of the extract. A certain mass of lyophilized powder was weighed and dissolved in DMSO to prepare an extract mother solution. The concentration of the mother solution was 20 mg/mL. The components were proportioned in equal proportions, and the cells were treated with a final concentration of 100 μg/mL Chinese medicine extract.

Comparative Example 6

Part of the Chinese medicinal materials in Example 1 were used to make part of the prescription, as a control, the preparation method was:

Two-sided needles, wild chrysanthemum, hibiscus leaves, mugwort leaves, and Tiannanxing were crushed and prepared separately. Weigh 10g of each medicine, respectively use 150mL of 90% ethanol, extract with standard Soxhlet reflux for 3h, concentrate the extract for 20min to a volume of 2mL, and use a freeze dryer to freeze-dry under reduced pressure for 24h to prepare the above five kinds of traditional Chinese medicines. Lyophilized powder of the extract. A certain mass of lyophilized powder was weighed and dissolved in DMSO to prepare an extract mother solution. The concentration of the mother solution was 20 mg/mL. The components were proportioned in equal proportions, and the cells were treated with a final concentration of 100 μg/mL Chinese medicine extract.

Efficacy verification

1. The influence of single drug on key genes

Using HTS2 (high-throughput screening based on high-throughput sequencing) technology, the effects of several traditional Chinese medicine extracts on the expression of multiple key genes in macrophages induced by THP-1 human acute monocytic leukemia cells were detected. The experimental process is:

Using HTS2 (high-throughput screening based on high-throughput sequencing) technology, the effects of several traditional Chinese medicine extracts on the expression of multiple key genes in macrophages induced by THP-1 human acute monocytic leukemia cells were detected. The experimental process is:

1. Using 14 kinds of Chinese herbal extracts, namely Artemisia annua, Honeysuckle, Gardenia, Goosebum, Inula, Chuan Shegan, Angelica, Ginger, Radix, Wild Chrysanthemum, Hibiscus, Artemisia, Agrimony, Tian Nanxing, 100 μg/mL concentration of cells were treated for 24 hours.

2. Cells are lysed, gene-specific probes are added to bind mRNA, and DNA ligase is used to connect the successfully bound pair of probes.

3. Incubate at 65°C for 10 min, elute the successfully ligated probe, use PCR technology to amplify and build a library for sequencing.

4. Data processing: The solvent control DMSO treatment group was used as the control group, the gene change fold of the drug treatment group was ≥1.5 times, and the P value <0.05 was defined as significantly up-regulated or down-regulated gene expression.

The results are shown in Figure 1, in which the data shows the effect of 14 traditional Chinese medicines on the expression of 23 genes on THP-1 cells. Red indicates that the gene expression is up-regulated, blue indicates down-regulation, and white indicates no significant effect.

Gene expression results showed that: Angelica dahurica, Goosebum, Inula flower, Ginger, Artemisia annua, Chuan Shegan, Gardenia, L. japonica, Wild Chrysanthemum, Hibiscus leaf, Artemisia argyi, Agrimony, Arranthaceae can inhibit inflammation and cytokines Storm-related genes CXCL1, IL1B, NFKB1, IL6, NFKB1, NFKB1A, PTGS2, IL1A, TNF, CCL2, RELA, NFKB2, Angelica sinensis, Goosebum, Inula, Ginger, Artemisia annua, Chuanshegan, Honeysuckle, Wild Chrysanthemum, Hibiscus leaf, Artemisia argyi, Agrimony, Arranthaceae can inhibit pulmonary fibrosis-related genes ITGBA1, ITGB3, ITGB5, ITGB1, INHBA, VEGFB, VEGFA, TGFB1, MMP9, ACTB, FN1, BMP2. The traditional Chinese medicine composition can improve the symptoms of respiratory failure by inhibiting multiple key genes regulating viral infection and respiratory failure.

2. Effects of Examples 2 to 5 on key genes

Using the same method as (1), the effects of the traditional Chinese medicine compositions of Examples 2 to 5 on key genes were detected, and the induced THP1 cells were treated with the same final concentration (100 μg/mL) for each composition for 24 hours. The results are shown in Figure 2, red indicates up-regulation of the gene expression, blue indicates down-regulation, and white indicates no significant effect. The results show that the compositions of Examples 2, 3, 4, and 5 all have regulatory effects on 13 key genes. Among them, the regulatory effects of Examples 2 to 3 are stronger than those of Examples 4 to 5. The composition has a more significant therapeutic effect on immune-related diseases.

3. Effects of the drugs of Example 2 and Comparative Examples 1 to 3 on key genes

The method was the same as (1), the drugs of Example 2 (Shanfang) and Comparative Examples 1-3 were treated with the induced THP1 cells at a concentration of 100 μg/mL for 24 hours. The results are shown in Figure 3, red indicates up-regulation of the gene expression, blue indicates down-regulation, and white indicates no significant effect. The results show that compared with the drugs shown in Example 2, the drugs in Comparative Examples 1 to 3 have fewer targets, such as:

Among the 23 targets, Zhenwu Decoction only had inhibitory effects on 13 of them, but also promoted the expression of ITGA1 and ITGB3. Liujunzi decoction only had inhibitory effects on 6 of them, but promoted many targets. Bufei Decoction only has inhibitory effects on two targets, and has a promoting effect on INHBA at the same time.

IV. Effects of the drugs of Example 2 and Comparative Examples 4 to 6 on key genes

The method was the same as (1), the drugs of Example 2 and Comparative Examples 4-6 were treated with the induced THP1 cells at a concentration of 100 μg/mL for 24 hours respectively. The results are shown in Figure 4, red indicates up-regulation of the gene expression, blue indicates down-regulation, and white indicates no significant effect. The results showed that the drugs of Comparative Examples 4-6 had fewer targets. Compared with Shanfang (the drug of Example 2), the target of Comparative Example 4 was 2 less, and the composition of Comparative Example 5 had fewer targets. 4, and has a promoting effect on VEGFB; the composition of Comparative Example 6 has 3 less targets, and has a promoting effect on BMP2. Explain that in the traditional Chinese medicine composition provided by the present invention, each component is indispensable.

The above are only the preferred embodiments of the present invention. It should be pointed out that for those skilled in the art, some improvements and modifications can be made without departing from the principles of the present invention, and these improvements and modifications should also be regarded as It is the protection scope of the present invention.

Claims (12)

1. The application of traditional Chinese medicine in preparing medicine for preventing and treating immune related diseases; the Chinese medicinal materials are at least one selected from Inulae flos, flos Chrysanthemi Indici, radix Angelicae Dahuricae, folium Hibisci Mutabilis, herba Centipedae, folium Artemisiae Argyi, rhizoma Zingiberis recens, herba Artemisiae Annuae, flos Lonicerae, fructus Gardeniae, radix Zanthoxyli, rhizoma arisaematis preparata, rhizoma Belamcandae, and herba et Gemma Agrimoniae.

2. Use according to claim 1, characterized in that the control comprises: regulating the expression of a gene associated with viral infection and/or regulating the expression of a gene associated with respiratory failure;

the viral infection-associated genes include CXCL1, IL1B, IL6, NFKB1, NFKB1A, PTGS2, IL1A, TNF, CCL2, RELA, and/or NFKB 2; the respiratory failure related genes comprise ITGA1, ITGB3, ITGB5, ITGB1, INHBA, VEGFB, VEGFA, TGFB1, MMP9, ACTB, FN1 and/or BMP 2.

3. The use of claim 2, wherein the immune-related disorder comprises: viral infections, chronic inflammation, allergic diseases and/or autoimmune diseases;

the virus comprises: coronavirus, adenovirus, herpesvirus, enterovirus, measles virus, hepatitis b virus, hepatitis c virus, human papilloma virus, influenza a H1N1 virus, human T cell leukemia virus;

the autoimmune diseases include: rheumatoid arthritis, atherosclerosis, pulmonary fibrosis, viral hepatitis, multiple sclerosis and/or inflammatory bowel disease;

the allergic disease comprises allergic asthma and/or allergic rhinitis.

4. A Chinese medicinal composition is prepared from herba Artemisiae Annuae, flos Lonicerae, fructus Gardeniae, herba Centipedae, Inulae flos, rhizoma Belamcandae, radix Angelicae Dahuricae, rhizoma Zingiberis recens, radix Zanthoxyli, flos Chrysanthemi Indici, folium Hibisci Mutabilis, folium Artemisiae Argyi, herba et Gemma Agrimoniae and rhizoma arisaematis preparata.

5. The traditional Chinese medicine composition according to claim 4, which is characterized by comprising the following components in parts by mass: 9 to 15 parts of sweet wormwood, 3 to 30 parts of honeysuckle, 9 to 25 parts of gardenia, 6 to 15 parts of centipeda minima, 3 to 9 parts of inula flower, 6 to 10 parts of rhizoma ligustici wallichii, 2 to 12 parts of dahurian angelica root, 6 to 30 parts of ginger, 5 to 10 parts of radix zanthoxyli, 9 to 15 parts of wild chrysanthemum flower, 10 to 30 parts of cotton rose leaf, 3 to 15 parts of argy wormwood leaf, 6 to 12 parts of hairyvein agrimony and 3 to 9 parts of arisaema cum bile.

6. A Chinese medicinal extract obtained by extracting the Chinese medicinal composition of claim 4 or 5.

7. The method for preparing a herbal extract of claim 6, comprising: performing standard Soxhlet reflux extraction on sweet wormwood, honeysuckle, cape jasmine, centipeda minima, inula flower, Sichuan blackberry lily, dahurian angelica root, ginger, shinyleaf pricklyash root, wild chrysanthemum, cotton rose leaf, argyi leaf, hairyvein agrimony and prepared arisaema tuber by using 90 vol% ethanol water solution, drying the obtained extract, and preparing the traditional Chinese medicine extract.

8. Use of the Chinese medicinal composition of claim 4 or 5, the Chinese medicinal extract of claim 6 or the extract prepared by the preparation method of claim 7 in the preparation of medicaments for treating immune-related diseases.

9. Use according to claim 8, wherein the control comprises: regulating the expression of a gene associated with viral infection and/or regulating the expression of a gene associated with respiratory failure.

10. The use of claim 8, wherein said immunomodulation comprises inhibition of NFKB2, CCL2, PTGS2, TNF, IL1A, CXCL1, NFKB1A, NFKB1, IL6, IL1B, RELA, BMP2, VEGFB, FN1, MMP9, ACTB, VEGFA, INHBA, ITGB1, ITGA1, TGFB1, ITGB3, and/or ITGB 5.

11. The use of the Chinese medicinal composition of claim 4 or 5, the Chinese medicinal extract of claim 6 or the extract prepared by the preparation method of claim 7 in the preparation of medicaments for treating immune-related diseases, wherein the immune-related diseases comprise: viral infections, chronic inflammation, allergic diseases and/or autoimmune diseases;

the virus comprises: coronavirus, adenovirus, herpesvirus, enterovirus, measles virus, hepatitis b virus, hepatitis c virus, human papilloma virus, influenza a H1N1 virus, human T cell leukemia virus; the autoimmune diseases include: rheumatoid arthritis, atherosclerosis, pulmonary fibrosis, viral hepatitis, multiple sclerosis and/or inflammatory bowel disease; the allergic disease comprises allergic asthma and/or allergic rhinitis.

12. A medicament, which is characterized by comprising the traditional Chinese medicine composition as claimed in claim 4 or 5, the traditional Chinese medicine extract as claimed in claim 6 or the extract prepared by the preparation method as claimed in claim 7.

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