A traditional Chinese medicine composition for preventing and / or treating interstitial lung disease, a traditional Chinese medicine decoction, and its preparation method and application

Abstract

The present invention provides a traditional Chinese medicine composition for preventing and / or treating interstitial lung disease, a traditional Chinese medicine decoction, and a preparation method and application thereof, belonging to the technical field of traditional Chinese medicine. The Chinese medicine composition comprises: 25-35 parts of astragalus, 15-25 parts of codonopsis, 10-15 parts of atractylodes, 10-20 parts of tuckahoe, 10-15 parts of bitter almond, 10-15 parts of platycodon, 10-20 parts of salvia, 10-20 parts of chuanxiong, 10-15 parts of angelica, 4-10 parts of schisandra, and 5-12 parts of liquorice. The astragalus, codonopsis, atractylodes and tuckahoe are combined as the monarch medicine to exert the effect of invigorating qi, the salvia, angelica and chuanxiong are combined as the assistant medicine to exert the effect of promoting blood circulation and removing blood stasis, the platycodon, bitter almond and schisandra are combined as the auxiliary medicine to exert the effect of relieving cough and asthma, and the liquorice is used as the guiding medicine to harmonize the properties of the medicines, and the combined medicine has the effects of invigorating qi (lung and spleen), promoting blood circulation and removing blood stasis, and relieving cough and asthma, thereby preventing and / or treating interstitial lung disease.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and in particular relates to a traditional Chinese medicine composition for preventing and / or treating interstitial lung disease, a traditional Chinese medicine decoction, and a preparation method and application thereof. Background Art

[0002] Interstitial lung disease (ILD) is a general term for a group of heterogeneous, non-tumor, and non-infectious lung diseases characterized by inflammation and interstitial fibrosis of alveolar units. It is also known as diffuse parenchymal lung disease (DPLD). It often manifests as decreased respiratory function in patients, with clinical symptoms such as dyspnea, long-term cough and wheezing, and shortness of breath.

[0003] The high morbidity and mortality of ILD are becoming increasingly prominent. To date, ideal interventions for the treatment of ILD are still lacking. Western medicine clinical treatments primarily rely on oxygen therapy and anti-fibrosis. Pirfenidone and nintedanib can slow the decline in lung function in patients with pulmonary fibrosis. However, in actual clinical use, these drugs have a slow onset of efficacy in treating idiopathic pulmonary fibrosis and are associated with difficult-to-tolerate side effects. In addition, some new treatments have been developed, such as bone marrow stem cell transplantation, which can promote the repair of lung tissue structure and function. However, stem cell transplantation is still in the experimental research stage and is still a long way from clinical practice.

[0004] In Traditional Chinese Medicine (TCM), there's no specific syndrome corresponding to ILD. Most consider ILD to fall under the category of "lung bi," with the disease located in the lungs and spleen. The primary pathogenesis is a combination of deficiency of the underlying qi and excess of the superficial qi, based on lung and spleen qi deficiency. The principle of "cultivating the earth and generating metal" is the primary treatment. The Yellow Emperor's Classic of Internal Medicine states, "When wind, cold, and dampness converge, they cause bi," "If skin bi persists, it is further affected by pathogenic factors, which then lodge internally in the lungs," and "Lung bi causes restlessness, fullness, shortness of breath, and vomiting." These are the earliest records of the etiology and pathogenesis of lung bi. Later generations of physicians have also largely adopted the Huangdi Neijing's theory of "lung bi," and its discussion suggests that internal organ deficiency is the key to lung bi. Currently, reports on the effectiveness of traditional Chinese medicine in preventing and treating ILD primarily focus on clinical efficacy and preclinical studies of related mechanisms, including mitigation of oxidative stress, improvement of inflammation, inhibition of epithelial-mesenchymal transition and myofibroblast activation, and regulation of autophagy and apoptosis. Summary of the Invention

[0005] In view of this, the object of the present invention is to provide a traditional Chinese medicine composition, a traditional Chinese medicine decoction, and a preparation method and use thereof for preventing and / or treating interstitial lung disease. The traditional Chinese medicine composition or traditional Chinese medicine decoction can improve lung function, inhibit pulmonary fibrosis and inflammation, and regulate intestinal flora, thereby preventing and / or treating interstitial lung disease.

[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 traditional Chinese medicine composition for preventing and / or treating interstitial lung disease, comprising the following raw materials in parts by weight:

[0008] 25-35 parts of Astragalus, 15-25 parts of Codonopsis, 10-15 parts of Atractylodes, 10-20 parts of Poria, 10-15 parts of Apricot Kernel, 10-15 parts of Platycodon, 10-20 parts of Salvia miltiorrhiza, 10-20 parts of Chuanxiong, 10-15 parts of Chinese Angelica, 4-10 parts of Schisandra, and 5-12 parts of Licorice.

[0009] As a preferred embodiment, the Chinese medicine composition comprises the following raw materials in parts by weight:

[0010] 28-32 parts of Astragalus, 16-23 parts of Codonopsis, 11-14 parts of Atractylodes, 12-17 parts of Poria, 11-13 parts of Apricot Kernel, 11-13 parts of Platycodon, 12-18 parts of Salvia miltiorrhiza, 13-17 parts of Chuanxiong, 11-14 parts of Angelica sinensis, 5-7 parts of Schisandra chinensis, and 8-10 parts of Licorice.

[0011] The Chinese medicine composition used for treating ILD in the present invention, this side takes "Qi, deficiency, stasis" as the core pathogenesis of ILD, Qi deficiency and blood stasis are carried out throughout, the main deficiency is excess in the superficial, its source is lung and spleen, viscera qi weakness causes blood vessels obstruction, produces pathological products such as blood stasis, phlegm turbidity and further damages vital energy, and ultimately the accumulated damage is difficult to recover the lung and lose its function. Therefore, the treatment method should take into account both strengthening the body's health (strengthening the spleen and benefiting the lungs) and dispelling pathogenic factors (activating blood circulation, removing blood stasis and eliminating phlegm).

[0012] The formula of the Chinese medicine composition for preventing and / or treating interstitial lung disease of the present invention is as follows:

[0013] Astragalus membranaceus was first mentioned in Shennong's Classic of Materia Medica. It is sweet and slightly warm, and enters the spleen and lung meridians. It is often used to strengthen the body's defenses and consolidate the body's foundation. It contains numerous active ingredients, including astragaloside IV, astragaloside polysaccharides, astragaloglycoproteins, and astragaloflavonoids. These ingredients exert varying degrees of immune regulation, anti-inflammatory effects, and inhibition of epithelial-mesenchymal transition (EMT), and are widely used in the treatment of pulmonary fibrosis. Codonopsis pilosula was first mentioned as a medicinal herb in the Qing Dynasty's Bencao Congxin (Compendium of Materia Medica). Codonopsis pilosula is sweet and neutral, and enters the spleen and lung meridians. It has the effects of strengthening the spleen and lungs, nourishing blood, and promoting the production of body fluids. It can inhibit EMT and inflammatory responses. Astragalus membranaceus, Codonopsis pilosula, Atractylodes macrocephala, and Poria cocos are combined as the monarch herb, exerting qi-tonifying effects, particularly strengthening the spleen and lungs.

[0014] Minister: Salvia miltiorrhiza, first mentioned in Shennong's Herbal Classic and listed as a top-grade herb, is a commonly used herb for promoting blood circulation and removing blood stasis. Modern pharmacological research indicates that the tanshinones and salvianolic acids in Salvia miltiorrhiza have multiple effects, including anti-inflammatory, inhibiting collagen production, promoting fibrin degradation, and inhibiting cell proliferation. This herb holds great promise for preventing and treating tissue and organ fibrosis. Salvia miltiorrhiza, along with Angelica sinensis and Chuanxiong, serve as ministerial herbs, synergistically promoting blood circulation and removing blood stasis.

[0015] Adjuvants: Platycodon grandiflorum, as recorded in the Shennong's Herbal Classic, is "pungent, slightly warm, and enters the Lung Meridian," boasting lung-clearing and expectorant properties. Bitter almond, as recorded in the Shennong's Herbal Classic, is bitter, slightly warm, and slightly toxic. It enters the Lung and Large Intestine Meridians, promoting the descent of Qi, relieving cough and asthma. Schisandra chinensis, as recorded in the Bencao Classic, is warm in nature, sour and sweet in flavor. It enters the Lung, Heart, and Kidney Meridians, invigorating Qi, relieving cough and shortness of breath, astringing and firming, relieving cough and expectorant, and treating chronic cough and asthma. It also promotes immune function. Platycodon grandiflorum, bitter almond, and Schisandra chinensis work together as adjuvants, achieving a synergistic effect of relieving cough and asthma.

[0016] Licorice: Licorice is sweet and mild in nature. It has the effects of removing phlegm and relieving cough, replenishing the middle and invigorating qi, and harmonizing the properties of medicines. Modern pharmacological research shows that licorice contains a variety of chemical components, which have anti-inflammatory, immune regulation, anti-fibrosis and other effects.

[0017] The whole formula: Astragalus, Codonopsis, Atractylodes, Poria, Platycodon, Salvia, Chuanxiong, Chinese Angelica, Bitter Apricot, Schisandra and Licorice, which work together to strengthen Qi (lungs and spleen), promote blood circulation and remove blood stasis, and relieve cough and asthma.

[0018] The present invention also provides a method for preparing the above-mentioned Chinese medicine composition, comprising the following steps:

[0019] The astragalus, codonopsis, atractylodes, poria, bitter almond, platycodon, salvia miltiorrhiza, ligusticum, angelica, schisandra chinensis and liquorice are mixed to obtain a mixed raw material; the mixed raw material is mixed with water and soaked, decocted and extracted, and filtered to obtain a filtrate.

[0020] In the present invention, the astragalus, codonopsis, atractylodes, poria, bitter almond, platycodon, salvia miltiorrhiza, ligusticum chuanxiong, angelica, schisandra chinensis and liquorice are all prepared as decoction pieces of each raw material commercially available in the art, and mixed to obtain a mixed raw material. After obtaining the mixed raw material, the mixed raw material is mixed with water and soaked, decocted and extracted, and filtered to obtain a filtrate. The mass volume ratio of the mixed raw material to water is preferably 1:6 to 11, more preferably 1:7 to 10, more preferably 1:8 to 9, such as 1:8 or 1:9, and the soaking time is preferably 0.5 to 1.5 hours, such as 0.5 hours, 1 hour or 1.5 hours; the number of extractions is 1 to 3 times, such as 1 time or 2 times, and the time of each extraction is preferably 0.5 to 1.5 hours, such as 0.5 hours, 1 hour or 1.5 hours.

[0021] As a preferred embodiment, the method for preparing the Chinese medicine composition further comprises the following steps:

[0022] The filtrate is subjected to a first concentration to obtain concentrated solution 1, which is then cooled, allowed to stand, and filtered to obtain filtrate a. Filtrate a is then subjected to a second concentration and filtration to obtain concentrated solution 2, thereby obtaining the traditional Chinese medicine composition. The first or second concentration method is preferably performed under reduced pressure at 60-90°C and -0.04--0.1 MPa, more preferably at 70-80°C and -0.05--0.08 MPa. The relative density of concentrated solution 1 is preferably 1.04-1.06, such as 1.04, 1.05, or 1.06; the relative density of concentrated solution 2 is preferably 1.02-1.04, such as 1.02, 1.03, or 1.04. The cooling temperature is below 10°C. The standing time is preferably 8-24 hours, more preferably 10-20 hours, and more preferably 14-18 hours, such as 14, 15, 16, 17, or 18 hours. The filtration method can be performed using 300-mesh nylon filter cloth.

[0023] The present invention also provides a traditional Chinese medicine decoction for treating interstitial lung disease, comprising the traditional Chinese medicine composition or the traditional Chinese medicine composition obtained by the preparation method.

[0024] Preferably, the Chinese herbal decoction further comprises 2 to 4 parts of potassium sorbate.

[0025] The present invention also provides a method for preparing the above-mentioned Chinese medicine decoction, comprising the following steps:

[0026] The Chinese medicine composition obtained by the above preparation method, potassium sorbate, and water are mixed, boiled, and cooled to obtain a Chinese medicine decoction. The cooling temperature is below 50°C. Based on a total volume of 1000 mL of the above Chinese medicine composition, potassium sorbate, and water, the amount of water used is the amount of water added to 1000 mL after adding the Chinese medicine composition and potassium sorbate.

[0027] The present invention is set up by modern preparation process research, multi-index quality control system, makes Chinese medicine decoction, forms the Chinese medicine compound preparation process and quality standard that process is stable, quality controllable, patient compliance is good, and lays the foundation for the main pharmacodynamics research of its Chinese medicine composition, clinical data collection.Simultaneously, obtain active pharmaceutical ingredient and target information by network pharmacology, Chinese medicine decoction has been carried out the main pharmacodynamics verification inside and outside the body, and has tentatively excavated the molecular mechanism of action of this side treatment ILD.Clinically, improving clinical symptoms, Chinese medicine syndrome, improving quality of life, is well received by patients.Above-mentioned research, all confirmed the validity of Chinese medicine decoction treatment ILD.For next step, Chinese medicine decoction is transformed into new Chinese medicine and laid a solid foundation.

[0028] The present invention also provides a use of the above-mentioned traditional Chinese medicine composition, the traditional Chinese medicine composition obtained by the preparation method, the traditional Chinese medicine decoction, or the traditional Chinese medicine decoction obtained by the preparation method, comprising at least one of the following:

[0029] (1) Use in the preparation of drugs for preventing and / or treating interstitial lung diseases;

[0030] (2) Application in the preparation of drugs for regulating intestinal flora.

[0031] In the present invention, at the phylum level, the intestinal flora regulation includes reducing the Firmicutes flora, increasing the Bacteroidetes flora, and increasing the ratio of Proteobacteria and Actinobacteria. At the genus level, the intestinal flora regulation includes short-chain fatty acid producing flora, such as Pseudomonas and unclassified_Muribaculaceae.

[0032] Based on this, the present invention also provides an application of the above-mentioned Chinese medicine composition, the Chinese medicine composition obtained by the preparation method, the Chinese medicine decoction or the Chinese medicine decoction obtained by the preparation method in preparing a product of a short-chain fatty acid-producing bacterial community.

[0033] In the present invention, the short-chain fatty acid-producing bacterial community includes Pseudomonas and unclassified_Muribaculaceae. The product includes a reagent or a medicine.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] The present invention provides a traditional Chinese medicine composition and a traditional Chinese medicine decoction for preventing and / or treating interstitial lung disease, as well as their preparation method and use. The traditional Chinese medicine composition comprises four herbs: Astragalus, Codonopsis, Atractylodes, and Poria, as the main ingredients to invigorate qi. Salvia miltiorrhiza, Angelica sinensis, and Chuanxiong are used as the auxiliary ingredients to promote blood circulation and remove blood stasis. Platycodon grandiflorum, Apricot kernel, and Schisandra chinensis are used as the auxiliary ingredients to relieve cough and asthma. Licorice is used as the guiding ingredient to eliminate phlegm and relieve cough, invigorate the middle qi, and harmonize the medicinal properties. Together, these herbs strengthen qi (lungs and spleen), promote blood circulation and remove blood stasis, and relieve cough and asthma. Results demonstrate that the traditional Chinese medicine composition or decoction can improve lung function, inhibit pulmonary fibrosis and inflammation, and regulate intestinal flora, thereby preventing and / or treating interstitial lung disease. The preparation method of the traditional Chinese medicine composition or decoction is simple and easy to operate, making it suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 Effects of serum containing Chinese herbal medicines on TGF-β1-induced EMT in A549 cells;

[0037] Figure 2 The regulatory effect of serum prepared from Chinese herbal decoction on the transformation of lung fibroblasts into myofibroblasts induced by TGF-β1;

[0038] Figure 3Western blot was used to detect the effect of serum containing Chinese herbal medicine decoction on the levels of E-cadherin and vimentin in A549 cells.

[0039] Figure 4 The changes of lung coefficients of mice after treatment in different groups;

[0040] Figure 5 is the tidal volume of mice after treatment in different groups;

[0041] Figure 6 The minute ventilation of mice after treatment in different groups;

[0042] Figure 7 The corresponding expiratory flow rate of 50% of the exhaled volume of mice after treatment in different groups;

[0043] Figure 8 HE staining results of lung tissues of mice after treatment in different groups;

[0044] Figure 9 The results of Masson, picrosirius red, and VG staining of mouse lung tissues after treatment in different groups;

[0045] Figure 10 is the IL-1β content in the serum of mice after treatment in different groups;

[0046] Figure 11 is the IL-6 content in the serum of mice after treatment in different groups;

[0047] Figure 12 is the TNF-α content in the serum of mice after treatment in different groups;

[0048] Figure 13 is the TGF-β1 content in the serum of mice after treatment in different groups;

[0049] Figure 14 The figure shows the Venn diagram of different groups after treatment, where CON represents the blank control group, MOD represents the model group, Nib represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0050] Figure 15 The distribution diagram of the number of OTUs after treatment in different groups, where CON represents the blank control group, MOD represents the model group, NIB represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0051] Figure 16This is the Alpha diversity index analysis chart after treatment in different groups, where CON represents the blank control group, MOD represents the model group, NIB represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0052] Figure 17 Beta diversity analysis diagram after treatment of different groups, where CON represents the blank control group, MOD represents the model group, NIB represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0053] Figure 18 To analyze the bacterial flora differences between different groups of samples based on phylum, CON represents the blank control group, MOD represents the model group, NIB represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0054] Figure 19 represents the difference in Bacteroidota between samples in different groups, where CON represents the blank control group, MOD represents the model group, NIB represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0055] Figure 20 represents the difference in Firmicutes between samples in different groups, where CON represents the blank control group, MOD represents the model group, NIB represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0056] Figure 21 is the ratio of Firmicutes to Bacteroidota between samples in different groups, where CON represents the blank control group, MOD represents the model group, NIB represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0057] Figure 22 To analyze the differences in bacterial flora between different groups of samples based on genus, CON represents the blank control group, MOD represents the model group, NIB represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0058] Figure 23 The difference in the bacterial flora of the genus Alloprevotella that can increase the production of short-chain fatty acids between different groups of samples, where CON represents the blank control group, MOD represents the model group, NIB represents the nintedanib group, FJD represents the low-dose group of the prescription, and FJG represents the high-dose group of the prescription;

[0059] Figure 24 The differences in the unclassified_Muribaculaceae bacterial community that can increase the production of short-chain fatty acids between different groups of samples. DETAILED DESCRIPTION

[0060] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0061] 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.

[0062] In the following examples, all the raw materials used are medicinal pieces.

[0063] Example 1

[0064] A traditional Chinese medicine composition for preventing and / or treating interstitial lung disease, comprising the following raw materials:

[0065] Astragalus 30g, Codonopsis pilosula 20g, Atractylodes macrocephala 12g, Poria 15g, Bitter Apricot 12g, Platycodon grandiflorum 12g, Salvia miltiorrhiza 15g, Chuanxiong 15g, Angelica sinensis 12g, Schisandra chinensis 6g, Licorice root 9g.

[0066] The preparation method of the Chinese medicine composition comprises the following steps:

[0067] (1) Take the prescription medicinal materials that meet the standards of the first volume of the Chinese Pharmacopoeia (2020 edition) and pass the inspection, and weigh astragalus, codonopsis, atractylodes, poria, bitter almond, platycodon, salvia miltiorrhiza, ligusticum, angelica, schisandra chinensis and liquorice according to the above prescription amount to obtain mixed medicinal pieces.

[0068] (2) Soak the mixed slices in 1442 mL (9 times the amount) of water for 1 hour, decoct for 1 hour, and filter through 300 mesh nylon cloth to obtain a first filtrate and a medicinal residue. Add 1264 mL (8 times the amount) of water to the medicinal residue, decoct for 1 hour, filter through 300 mesh nylon cloth, discard the medicinal residue, and obtain a second filtrate. The obtained first filtrate and second filtrate are respectively concentrated under reduced pressure at 60-90°C and -0.04--0.1 MPa to a relative density of the concentrated solution at 50-60°C of 1.04-1.06. Combine the first concentrated solution and the second concentrated solution to obtain a concentrated solution 1;

[0069] (3) The concentrated solution 1 is cooled to below 10°C and allowed to stand for 16 hours. The supernatant 1 is filtered through 300-mesh nylon cloth. The precipitate is allowed to stand for another 16 hours. The supernatant 2 is filtered through 300-mesh nylon cloth. The supernatant 1 and the supernatant 2 are combined to obtain a filtrate a. The filtrate a is concentrated under reduced pressure at 60-90°C and -0.04--0.1 MPa to a relative density of 1.02-1.04 at 50-60°C. The concentrated solution is filtered through 300-mesh nylon cloth to obtain a concentrated solution 2, thereby obtaining a traditional Chinese medicine composition.

[0070] Example 2

[0071] A traditional Chinese medicine decoction for preventing and / or treating interstitial lung disease, comprising the following ingredients:

[0072] Astragalus 30g, Codonopsis pilosula 20g, Atractylodes macrocephala 12g, Poria 15g, Bitter Apricot 12g, Platycodon grandiflorum 12g, Salvia miltiorrhiza 15g, Chuanxiong 15g, Angelica 12g, Schisandra chinensis 6g, Licorice root 9g and 3g potassium sorbate and purified water.

[0073] The preparation method of the Chinese medicine decoction comprises the following steps:

[0074] The preparation method of the Chinese medicine composition is the same as that described in Example 1.

[0075] The above-mentioned amount of potassium sorbate was added to the concentrated solution 2, and the volume was made up to 1000 mL with water. The mixture was stirred and mixed, boiled for 20 minutes, and cooled to below 50° C. to obtain a Chinese medicine decoction.

[0076] [Properties]: The Chinese herbal decoction is a brown liquid with a slight fragrance, bitter taste and slightly sweet taste.

[0077]

Identification

[0078] (1) TLC identification of Atractylodes macrocephala

[0079] Take 20 mL of the traditional Chinese medicine decoction and extract it twice with n-hexane, 25 mL each time. Combine the n-hexane extracts and evaporate to dryness. Dissolve the residue in 2 mL of methanol to prepare the test solution. Separately, extract 1 g of Atractylodes macrocephala control material with 10 mL of n-hexane and ultrasonicate for 30 minutes. Filter, evaporate the filtrate to dryness, and dissolve the residue in 2 mL of methanol to prepare the Atractylodes macrocephala control solution. According to thin-layer chromatography (General Chapter 0502 of the 2020 edition of the Chinese Pharmacopoeia), spot 10 μL of each of the two solutions on a single silica gel G thin-layer plate using petroleum ether (60-90°C)-ethyl acetate (50:1) as the developing solvent. Develop the plate, remove the plate, air-dry, spray with 5% vanillin-sulfuric acid solution, heat at 105°C until the spots are clearly colored, and inspect under sunlight.

[0080] Result: In the chromatogram of the test sample, spots of the same color appeared at the corresponding positions in the chromatogram of the control medicinal material.

[0081] (2) TLC identification of Poria cocos

[0082] Take 20 mL of the traditional Chinese medicine decoction and extract it twice with 30 mL of ether each time. Combine the ether solutions and evaporate to dryness. Dissolve the residue in 2 mL of n-hexane to prepare the test solution. Separately, extract 1 g of Poria cocos control material with 10 mL of ether and sonicate for 30 minutes. Filter, evaporate the filtrate to dryness, and dissolve the residue in 2 mL of n-hexane to prepare the Poria cocos control solution. According to thin-layer chromatography (General Chapter 0502 of the 2020 edition of the Chinese Pharmacopoeia), spot 5 μL of each of the two solutions on the same silica gel G thin-layer plate using petroleum ether (30-60°C)-ethyl acetate-acetone (84:3:15) as the developing solvent. Develop the plate, remove the plate, air dry, spray with 5% vanillin in sulfuric acid, heat at 105°C until the spots are clearly colored, and inspect under ultraviolet light (365 nm).

[0083] Result: In the chromatogram of the test sample, spots of the same color appeared at the corresponding positions in the chromatogram of the control medicinal material.

[0084] (3) TLC identification of Platycodon grandiflorum

[0085] Take 30 mL of the traditional Chinese medicine decoction and evaporate it to dryness. Add 50 mL of 7% sulfuric acid ethanol-water (1:3) to the residue and heat under reflux for 3 hours. Extract with ether twice, 30 mL each time. Combine the ether solutions and evaporate to dryness. Dissolve the residue in 2 mL of n-hexane to prepare the test solution. Separately, take 1 g of Poria cocos control material and add 10 mL of ether. Ultrasonicate for 30 minutes, filter, evaporate the filtrate to dryness, and dissolve the residue in 2 mL of n-hexane to prepare the Poria cocos control solution. According to thin-layer chromatography (General Chapter 0502 of the 2020 edition of the Chinese Pharmacopoeia), 5 μL of each solution was spotted onto the same silica gel G thin-layer plate using petroleum ether (30-60°C)-ethyl acetate-acetone (84:3:15) as the developing solvent. Develop the plate, remove the plate, air-dry, spray with 5% vanillin in sulfuric acid, heat at 105°C until the spots are clearly colored, and inspect under ultraviolet light (365 nm).

[0086] Result: In the chromatogram of the test sample, spots of the same color appeared at the corresponding positions in the chromatogram of the control medicinal material.

[0087]

Assay

[0088] Astragaloside IV was determined by high performance liquid chromatography (General Chapter 0512 of the 2020 edition of the Chinese Pharmacopoeia).

[0089] The chromatographic conditions and system suitability test used octadecylsilane bonded silica as the filler, acetonitrile-water (36:64) as the mobile phase, and evaporative light scattering detection. The theoretical plate number, calculated based on the astragaloside IV peak, should be no less than 4000.

[0090] Preparation of reference solution: Take an appropriate amount of astragaloside IV reference substance, accurately weigh it, and add methanol to make a solution containing 0.2 mg per 1 mL.

[0091] Preparation of test solution: After mixing the Chinese herbal medicine decoction, take 20 mL and shake and extract it 4 times with water-saturated n-butanol, 20 mL each time, combine the n-butanol solutions, wash with ammonia test solution 2 times, 40 mL each time, discard the ammonia solution, evaporate the n-butanol solution to dryness, dissolve the residue in methanol, transfer it to a 5 mL volumetric flask, add methanol to the scale, shake well, filter, and take the filtrate to obtain the product.

[0092] Determination method: Densely aspirate 10 μL of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0093] Results: 1mL of this mixture contains astragaloside IV (C 41 H 68 O 14 ) is above 0.15 mg.

[0094] [Usage and Dosage] Oral administration, 30mL / time, 3 times a day. Shake well before use.

[0095] [Effects and Indications] This Chinese herbal decoction strengthens the spleen and lungs, activates blood circulation, removes blood stasis, and resolves phlegm and dredges the meridians. It is used for patients with lung bi syndrome characterized by qi deficiency, blood stasis, and phlegm stagnation. Common clinical symptoms include dyspnea, wheezing, fatigue, cough with or without phlegm, dark red tongue, thin white fur, and a weak pulse. This condition is known in Western medicine as interstitial pneumonia or interstitial lung disease.

[0096] Example 3

[0097] Pharmacodynamics research on Chinese herbal decoctions

[0098] 3.1 Cell experiment: Effects of Chinese herbal decoction on TGF-β1-induced mesenchymal transition in A549 cells

[0099] A549 cells were cultured in vitro and induced with TGF-β1. Then, a Chinese herbal decoction was used for intervention. The expression of epithelial cell transdifferentiation markers was detected to determine the fibrosis process. The expression of fibrosis markers, oxidative stress indicators, and inflammatory cytokines was detected to elucidate the effect of the Chinese herbal decoction in inhibiting TGF-β1-induced A549 cell transdifferentiation.

[0100] Research methods

[0101] (1) Cell selection

[0102] A549 is a human lung adenocarcinoma epithelial cell line that exhibits the characteristics of type II alveolar epithelial cells during in vitro culture and is currently a commonly used cell model for evaluating lung epithelial-mesenchymal transition (EMT).

[0103] (2) Preparation of Chinese herbal decoctions

[0104] According to the method for preparing the Chinese medicinal decoction in Example 2, a Chinese medicinal decoction is obtained for later use.

[0105] (3) Preparation of drug-containing serum in rats

[0106] Six-week-old SD rats were fed adaptively for one week and then orally administered 8.1 mL of the Chinese herbal decoction once a day for 7 days. After the last dose, rats were fasted but not watered for 9 hours. They were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (50 mg / kg). Blood was collected from the abdominal aorta, allowed to stand for 1 hour, and then centrifuged at 4000×g for 10 minutes. The serum was collected, aliquoted, labeled, and stored at -80°C until use.

[0107] Another 10 SD rats were injected with an equal amount of physiological saline solution, and blank serum was prepared in the same way and stored for later use.

[0108] Both drug-containing serum and blank serum were inactivated at 56°C for 30 min and filter-sterilized before use.

[0109] (4) Model preparation and drug treatment

[0110] Current research has found that alveolar epithelial-mesenchymal transition (EMT) is the key to fibrosis formation, and transforming growth factor-1β (TGF-1β) is the master switch of the EMT process. It can promote the proliferation and differentiation of mesenchymal cells and promote the deposition of EMT. Therefore, it is considered to be the most important fibrogenic factor and can be used to induce cells to establish a fibrosis model.

[0111] Human alveolar epithelial carcinoma cell line A549 was cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and maintained in a CO2 incubator at 37°C and 5% CO2.

[0112] Establishment of the fibrosis model and treatment with traditional Chinese medicine decoctions: A549 cells were trypsinized and seeded into 6-well plates. The cells were incubated overnight in DMEM containing 10% FBS at 37°C in a 5% CO2 incubator. The cells were then treated with 0, 5, or 10 ng / mL TGF-1β for 24 hours. To determine the effects of traditional Chinese medicine decoctions on A549 cells, the blank serum control group was cultured with 10% rat blank serum for 48 hours. The drug-containing serum group was cultured with 10% rat drug-containing serum for 24 hours, followed by the addition of medium containing 10% rat drug-containing serum at a total concentration of 10 ng / mL TGF-1β for an additional 24 hours. The model group was cultured with medium containing 10% rat blank serum for 24 hours, followed by the addition of medium containing 10% rat blank serum at a total concentration of 10 ng / mL TGF-1β for an additional 24 hours. After treatment, the morphology of A549 cells was observed under an inverted light microscope, and the cell culture supernatant and cells were collected.

[0113] Figure 1 The results showed that compared with the control group, after 24 hours of stimulation with 10 ng / mL TGF-β1, the morphology of A549 cells changed from cobblestone-like polygonal epithelial cells to spindle-like long spindle-shaped mesenchymal cells, with larger intercellular spaces and loose connections between adjacent cells, showing a fibroblast-like phenotype. At the same time, compared with the model group and the blank serum control group, the drug-containing serum group was able to alleviate the cell morphological transformation induced by TGF-β1.

[0114] (5) Sirius red staining

[0115] Collagen fibers are the most widespread and abundant type of connective tissue, found throughout various organs. Type I collagen fibers are primarily found in bone, skin, and tendon; type II collagen fibers are primarily found in cartilage; type III collagen fibers are primarily found in embryonic tissue, adult blood vessels, and the gastrointestinal tract; and type IV collagen fibers are primarily found in the basement membrane. Sirius red and its contrasting dye are both strongly acidic dyes that readily bind to the basic groups in collagen molecules, resulting in strong adsorption. Polarized light microscopy reveals that collagen fibers exhibit positive uniaxial birefringence. Combining this with a Sirius red stain enhances birefringence, improving resolution and enabling differentiation between the two types of collagen fibers.

[0116] This example uses a modified Sirius Red staining solution (for collagen fiber staining), G1472-Reagent B, purchased from Beijing Solebo Technology Co., Ltd. Collagen fibers in tissues such as cardiovascular tissues stain red under a standard optical microscope, which is helpful for the classification and grading of various fibrotic lesions under a polarized microscope. Changes in Sirius Red staining concentration can be used to preliminarily assess intracellular collagen expression.

[0117] The present invention uses an inverted microscope to observe the morphological changes of A549 cells after the action of different concentrations of TGF-β1, and observes the collagen fibers by Sirius red staining. An in vitro pulmonary fibrosis model is constructed, and the inhibitory effect of drug-containing serum on TGF-β1-induced EMT of A549 cells is detected.

[0118] Figure 2 The results showed that after 24 hours of TGF-β1 stimulation, the expression of collagen fibers in A549 cells increased significantly. At the same time, compared with the model group and the blank serum control group, the drug-containing serum group was able to inhibit TGF-β1-induced cell morphological transformation and reduce the content of collagen fibers in cells.

[0119] (6) Western blot detection

[0120] The cell samples of each group were lysed with RIPA solution, and the total cell protein was extracted. The protein samples were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to PVDF membrane. The membrane was blocked in Tween-20-tris saline buffer (TBST) containing 5% skim milk for 2 h at 37°C and rinsed three times with Tris-buffered saline (TBST) containing 0.1% Tween-20 for 10 min each. The membrane was incubated with primary antibodies against GAPDH (1:2500 dilution), fibrosis-related proteins E-cadherin, and vimentin for 2 h. The membrane was rinsed three times with TBST for 10 min each. The membrane was incubated with horseradish peroxidase-conjugated secondary antibody (1:2000 dilution) at 37°C for 1-2 h and washed three times with TBST for 10 min each. Protein bands were visualized using an enhanced chemiluminescence substrate kit and detected using the BIO-RAD ChemiDocXRS+ chemiluminescence gel imaging system. The grayscale values ​​of the internal reference GAPDH and fibrosis-related protein bands were analyzed with Image J software, and the expression level of the target protein was expressed as the ratio of the target protein to the internal reference GAPDH.

[0121] Figure 3 The results showed that compared with the blank control group cells, TGF-β1 stimulation caused the expression level of E-cadherin protein in A549 cells to decrease and the expression of Vimentin protein to increase, while the serum containing Chinese herbal decoction could upregulate the expression of E-cadherin protein and downregulate the expression of Vimentin protein.

[0122] The above results indicate that Yifei Tongbi Decoction has the effect of reversing EMT of A549 cells.

[0123] 3.2 Animal Experiment: Protective Effects of Traditional Chinese Medicine Decoction on Bleomycin-Induced Pulmonary Fibrosis in Mice

[0124] In this example, a bleomycin-induced pulmonary fibrosis mouse model was established, and intervention studies were conducted using a traditional Chinese medicine decoction and nintedanib to observe the effects of the traditional Chinese medicine decoction on bleomycin-induced lung inflammation and fibrosis in mice. The mechanism of action was explored from the perspectives of inflammation, macrophage polarization, epithelial-mesenchymal transition, and intestinal flora.

[0125] 3.2.1 Research Methods

[0126] (1) Selection of inducible model drugs

[0127] Bleomycin is an aminoglycoside drug commonly used in clinical practice to treat various malignant tumors. However, its most serious side effect is pulmonary fibrosis. Due to its side effects, it is often used to induce animal pulmonary fibrosis models to explore the mechanisms of pulmonary fibrosis and screen anti-fibrotic drugs. Common routes of administration include a single intratracheal administration, intravenous injection, nasal drops, and intraperitoneal injection. The most common and classic method for pulmonary fibrosis models is a single intratracheal injection of bleomycin. This method replicates the pathological process of human pulmonary interstitial fibrosis in an animal model with a short development time, with fibrosis typically occurring within 14 to 28 days.

[0128] (2) Experimental mice

[0129] Thirty C57BL / 6 male mice, weighing 18–20 g, aged 6–8 weeks, were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd., with animal production license number: SCXK (Xiang) 2019-0004.

[0130] Mice were maintained in a suitable breeding environment with an ambient temperature of 20-25°C, a relative humidity of 40%-70%, a noise level below 85 decibels, and an ammonia concentration below 20 PPm. The mice were provided with SPF-grade bedding and feed, and had free access to water and food daily. All animal experimental procedures were performed in accordance with standard procedures and approved by the Experimental Animal Welfare and Ethics Committee of Chongqing Institute of Traditional Chinese Medicine.

[0131] (3) Establishment of a bleomycin-induced pulmonary fibrosis mouse model

[0132] After one week of adaptive feeding, the mice were randomly divided into: blank control (Control) group (n=10), model (BLM) group (n=12), nintedanib (BLM+Nib) group (n=12), low-dose prescription (BLM+prescription) group (n=12), and high-dose prescription (BLM+prescription*3) group (n=12). Mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (50 mg / kg). The model group, nintedanib group, low-dose prescription group, or high-dose prescription group were respectively slowly injected with 3 mg / kg bleomycin solution using a microsyringe, and then the mice were immediately rotated upright to evenly distribute the drug solution in the lungs. The mice in the blank control group were slowly injected with normal saline using a microsyringe. 48 hours after modeling, the drugs were administered. The blank control group and the model group were gavaged with an equal amount of normal saline. The nintedanib (BLM+Nib) group was gavaged with nintedanib ethanesulfonate at 22.75 mg / kg / time, twice a day, with an interval of 12 hours between each dose. The low-dose prescription group was gavaged with the traditional Chinese medicine decoction prepared in Example 2 at 20.54 g / kg / d, once a day. The high-dose prescription group was gavaged with the traditional Chinese medicine decoction prepared in Example 2 at 61.62 g / kg / d, once a day, which are equivalent to 1 times and 3 times the recommended daily dose for humans, respectively.

[0133] The mice were administered for 28 consecutive days, and their coat color and diet were observed daily, and their body weights were measured. On day 28, mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (50 mg / kg). The mice were exsanguinated from the abdominal aorta, and lung tissue was immediately removed. The lung tissue was washed with cold saline, dried with filter paper, and the wet weight of the lung was measured. The same lobe of the right lung was placed in 4% paraformaldehyde solution, and the remaining lung tissue was collected and stored at -80°C for subsequent experimental studies.

[0134] After one week of adaptive culture, male C57BL / 6 mice showed good spirits, normal breathing, smooth fur, well-developed muscles, and vigorous movements. Their eyes were bright and alert, their reactions were agile, their limbs were well-proportioned, their bodies were free of deformities and scars, and their natural orifices showed no abnormal secretions. The blank control group appeared normal throughout the experiment. Mice in the model group developed matted fur and lethargy after modeling, while the symptoms of mice in the drug-treated group improved.

[0135] (4) Calculation of lung coefficient

[0136] The lung coefficient of each group of mice was calculated according to the following formula.

[0137] Lung coefficient = lung mass (g) / body mass (kg).

[0138] The lung coefficient results of each group are shown in Figure 4 . Figure 4 The results showed that the lung coefficient of the model mice was significantly reduced, indicating that the model was successfully constructed. The lung coefficient of the mice in the drug-treated group increased, indicating that the drug may have the effect of improving the lung function of mice.

[0139] (5) Mouse lung function test

[0140] Three days before the end of the experiment, lung function in each group of mice was assessed using an unrestrained whole-body plethysmography (WBP) system from Shanghai Tawa Intelligent Technology Co., Ltd. The indicators and their meanings are described in the Results Analysis section. Airway pressure, lung volume, and airflow rate are the three key elements of lung function measurement. The basic working principle of WBP is based on Boyle's law, which states that the pressure and volume of a fixed amount of gas in a closed container at constant temperature are inversely proportional.

[0141] The WBP plethysmographic chamber is like a sealed container. When an animal inhales, the temperature and humidity of the respiratory system are higher than those of the inhaled gas. As the inhaled gas mixes with the air in the lungs, its temperature and relative humidity increase, causing the inhaled gas to expand. This expansion through the animal's chest cavity increases the pressure within the chamber. The greater the volume of inhaled gas (tidal volume), the greater the increase in intraplethysmographic pressure. Conversely, during exhalation, the exhaled gas mixes with air, which, due to the decrease in air temperature and humidity, compresses the gas and reduces intraplethysmographic pressure. The changes in intraplethysmographic pressure (ΔP) caused by respiration are recorded by a screen-mounted respiratory sensor within the chamber and amplified by an amplifier. These pressure changes can be used to derive other indicators of lung function using specialized algorithms in analysis software.

[0142] The following parameters were collected using WBP: tidal volume (the volume of air inhaled or exhaled with each breath during quiet breathing), minute ventilation (the total volume of air moving in and out of the lungs per minute), and mid-expiratory flow (the expiratory flow rate at which 50% of the tidal volume is exhaled).

[0143] Figures 5 to 7 The results showed that the drug could increase the tidal volume of mice, improve minute ventilation, and increase the expiratory flow rate when 50% of the tidal volume was exhaled, indicating that the drug could improve the lung ventilation function of mice and reduce the degree of airway obstruction.

[0144] (6) Lung tissue pathological observation

[0145] The lung tissues of each group of mice were fixed in 4% paraformaldehyde overnight, and then different concentrations of alcohol were used as dehydrating agents, from low concentration to high concentration, to gradually remove the water in the lung tissues. The lung tissues were then transparentized with xylene; the lung tissues were embedded in paraffin, and after the wax blocks solidified, the lung tissues were cut into 5μM thin slices using a microtome; the mouse lung tissues were stained with hematoxylin-eosin (HE), and the results were observed under an optical microscope to evaluate lung inflammation.

[0146] The lung tissues of mice were stained with Masson's, picrosirius red, and VG, and the results were observed under light microscopy to evaluate the level of lung and collagen deposition.

[0147] Figure 8 The results showed that the alveolar structure of mice in the blank control group was clear, and there was no obvious alveolar inflammatory exudation and fibrosis in the lung tissue; the lung structure of mice in the model group was distorted, the number of pulmonary interstitial cells increased, the fibrous tissue proliferated and fibrosed, and the alveolar cavity enlarged and fused; after intervention with Nib and Chinese herbal decoction, the degree of lung tissue pathology in the mice with pulmonary fibrosis in the nintedanib group, the low-dose prescription group, and the high-dose prescription group was alleviated, the integrity of the lung tissue structure was improved, the thickness of the alveolar septum was reduced, and the infiltration of inflammatory cells was reduced.

[0148] Figure 9 The results showed that the lung tissue structure and morphology of the mice in the blank control group were normal; there was obvious collagen deposition in the lung interstitium of the mice in the model group, which was distributed in large bundles and sheets, and the degree of lung fibrosis was severe; after intervention with Nib and Chinese herbal decoction, the collagen deposition in the lung tissue of the mice with pulmonary fibrosis in the nintedanib group, the low-dose prescription group and the high-dose prescription group was reduced, and the degree of lung fibrosis was significantly alleviated.

[0149] (7) Cytokine detection

[0150] Blood was collected to separate serum, and the ELISA kits provided by Shanghai ELISA Biotechnology Co., Ltd. were used to detect the levels of TNF-α, IL-1β, IL-6 and TGF-β1 in mouse serum.

[0151] Figures 10 to 13 The results showed that after intervention with Chinese herbal decoction, the inflammation levels in mice in the low-dose and high-dose groups of the prescription were reduced, and the drug had the effect of regulating the immune response.

[0152] (8) 16S sequencing of intestinal flora

[0153] Currently, microbial diversity research primarily focuses on conserved regions of ribosomal RNA (ribosomal RNA) encoding nucleic acid sequences. For bacteria, this is primarily based on 16S rDNA, the DNA sequence encoding the small subunit rRNA (rRNA) of the prokaryotic ribosome. By assembling and filtering reads, clustering or denoising them, and performing species annotation and abundance analysis, the species composition of a sample can be revealed. Further analysis, such as alpha diversity, beta diversity, significant species differences, correlation analysis, and functional prediction, can be used to uncover differences between samples. In this study, 16S rRNA gene sequencing was used to analyze fecal microbial diversity. The workflow involved fecal DNA extraction using the CTAB method, amplicon generation, PCR product pooling and purification, library construction, and sequencing. Microbial diversity was determined using paired-end sequencing on the Illumina Novaseq platform, with small fragment libraries constructed for sequencing.

[0154] The steps for extracting fecal DNA are as follows: (1) Lysis: Pipette 1 mL of CTAB lysis buffer into a 2.0 mL EP tube, add lysozyme, add an appropriate amount of fecal sample to the lysis buffer, incubate at 65°C, mix thoroughly, and centrifuge for 10 min; take the supernatant, add chloroform:isoamyl alcohol (24:1), mix thoroughly, and centrifuge for 10 min; (3) Precipitation: Pipette the supernatant into a 1.5 mL centrifuge tube, add isopropanol, shake up and down, and precipitate at -20°C; (4) Wash: Centrifuge for 10 min, pour out the liquid, be careful not to pour out the precipitate, wash twice with 1 mL of 75% ethanol, and the remaining small amount of liquid can be collected by centrifugation again and aspirated with a pipette tip; (5) Drying: Dry at room temperature; (6) Add DEPC water to dissolve the DNA sample. If necessary, incubate at 55-60°C for 10 min to aid dissolution;

[0155] (7) Digestion: Add 1 μL RNase A to digest RNA and incubate at 37°C for 15 min.

[0156] Library construction and sequencing: After total DNA was extracted from the samples, primers were designed based on conserved regions. The upstream primer 338F: 5'-ACTCCTACGGGAGGCAGCAG-3' (SEQ ID NO. 1) and the downstream primer 806R: 5'-GGACTACHVGGGTWTCTAAT-3' (SEQ ID NO. 2) were used to amplify the bacterial 16S V3-V4 region. Sequencing adapters were added to the primer ends, and PCR amplification was performed. The products were purified, quantified, and normalized to form sequencing libraries. The constructed libraries were quality-checked, and qualified libraries were sequenced using the Illumina Novaseq 6000. Raw image data files generated by high-throughput sequencing (such as those on Illumina Novaseq sequencing platforms) were converted into raw sequenced reads through base calling analysis. The results were stored in the FASTQ (fq) file format, which contains sequence information and corresponding sequencing quality information for the sequenced reads. In the sequence shown in SEQ ID NO. 2, H, V and W are degenerate sequences, H = A / C / T; V = A / C / G; W = A / T.

[0157] Data preprocessing: (1) Quality filtering: First, use Trimmomatic v0.33 software to filter the raw reads obtained by sequencing; then use cutadapt 1.9.1 software to identify and remove primer sequences to obtain clean reads that do not contain primer sequences; (2) Double-end sequence splicing: Use Usearch v10 software to splice the clean reads of each sample through overlap, and then filter the length of the spliced ​​data according to the length range of different regions; (3) Remove chimeras: Use UCHIME v4.2 software to identify and remove chimera sequences to obtain the final effective data (Effective Reads). In this project, the Biomarker cloud platform tool was used to perform microbial diversity analysis.

[0158] The feces of mice in each group were collected on the 27th day of continuous administration. The feces samples were collected and sequenced. The sequencing and analysis results are shown in the figure. Figures 14 to 24 As shown, the quality of all stool samples was qualified, and 5 samples were selected from each group for sequencing.

[0159] Figures 14 to 24The results showed that the traditional Chinese medicine decoction had a certain regulatory effect on the intestinal flora. The differences in the microbial flora between the different groups of samples were analyzed based on phylum. At the phylum level, the relative abundance of Firmicutes, Bacteroidota, Proteobacteria, and Actinobacteriota changed significantly. The traditional Chinese medicine decoction of the present invention was able to reduce the Firmicutes flora, increase Bacteroidota, reduce the Firmicutes / Bacteroidota ratio, and increase the Proteobacteria and Actinobacteriota ratios. The differences in the microbial flora between the different groups of samples were analyzed based on genus. At the genus level, the traditional Chinese medicine decoction of the present invention was able to increase the proportion of Alloprevotella and Unclassified_Muribaculaceae, which are bacteria that produce short-chain fatty acids.

[0160] Chinese herbal decoctions have a protective effect on the lungs of mice with bleomycin-induced pulmonary fibrosis, can improve lung coefficient and lung function, improve pathological changes in lung tissue, inhibit the level of inflammation in the body, and have a certain regulatory effect on intestinal flora.

[0161] 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 Chinese medicine composition for treating pulmonary fibrosis, characterized in that: Made from the following raw materials in parts by weight: 28-32 parts of Astragalus, 16-23 parts of Codonopsis, 11-14 parts of Atractylodes, 12-17 parts of Poria, 11-13 parts of Apricot Kernel, 11-13 parts of Platycodon, 12-18 parts of Salvia miltiorrhiza, 13-17 parts of Chuanxiong, 11-14 parts of Angelica sinensis, 5-7 parts of Schisandra chinensis, and 8-10 parts of Licorice.

2. A method for preparing the Chinese medicine composition according to claim 1, characterized in that: The following steps are involved: The astragalus, codonopsis, atractylodes, poria, bitter almond, platycodon, salvia miltiorrhiza, ligusticum, angelica, schisandra chinensis and liquorice are mixed to obtain a mixed raw material; the mixed raw material is mixed with water and soaked, decocted and extracted, and filtered to obtain a filtrate.

3. The preparation method according to claim 2, characterized in that The following steps are also included: The filtrate is subjected to a first concentration to obtain concentrated solution 1, which is then cooled, allowed to stand, and filtered to obtain filtrate a. The filtrate a is then subjected to a second concentration and filtered to obtain concentrated solution 2, thereby obtaining a traditional Chinese medicine composition.

4. The preparation method according to claim 2, characterized in that The mass volume ratio of the mixed raw material to water is 1:6-11; the soaking time is 0.5-1.5 hours; the number of extractions is 1-3 times, and the time for each extraction is 0.5-1.5 hours.

5. The preparation method according to claim 3, characterized in that The first concentration or the second concentration method is reduced pressure concentration at 60-90° C. and -0.04--0.1 MPa; the relative density of the concentrated solution 1 is 1.04-1.06; the relative density of the concentrated solution 2 is 1.02-1.

04.

6. A Chinese medicinal decoction for treating pulmonary interstitial fibrosis, characterized in that: The invention comprises the Chinese medicine composition according to claim 1 or the Chinese medicine composition obtained by the preparation method according to any one of claims 2 to 5.

7. The Chinese medicinal decoction according to claim 6, characterized in that It also includes 2 to 4 parts of potassium sorbate.

8. A method for preparing the Chinese medicinal decoction according to claim 6 or 7, characterized in that: The following steps are involved: The Chinese medicine composition obtained by the preparation method according to any one of claims 2 to 5, potassium sorbate and water are mixed, boiled and cooled to obtain a Chinese medicine decoction.

9. Use of the traditional Chinese medicine composition according to claim 1, the traditional Chinese medicine composition obtained by the preparation method according to any one of claims 2 to 5, the traditional Chinese medicine decoction according to claim 6 or 7, or the traditional Chinese medicine decoction obtained by the preparation method according to claim 8 in the preparation of a medicament for treating pulmonary fibrosis.

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