Traditional Chinese medicine composition and traditional Chinese medicine decoction for preventing and / or treating interstitial lung diseases as well as preparation method and application of traditional Chinese medicine composition and traditional Chinese medicine decoction

Abstract

The invention provides a traditional Chinese medicine composition and traditional Chinese medicine decoction for preventing and / or treating interstitial lung diseases as well as a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicines. The traditional Chinese medicine composition is prepared from 25-35 parts of radix astragali, 15-25 parts of radix codonopsis, 10-15 parts of rhizoma atractylodis macrocephalae, 10-20 parts of poria cocos, 10-15 parts of semen armeniacae amarae, 10-15 parts of radix platycodonis, 10-20 parts of radix salviae miltiorrhizae, 10-20 parts of rhizoma chuanxiong, 10-15 parts of radix angelicae sinensis, 4-10 parts of fructus schizandrae and 5-12 parts of radix glycyrrhizae, the radix astragali, the radix codonopsis, the rhizoma atractylodis macrocephalae and the poria cocos are combined to serve as monarch drugs to achieve the effect of tonifying qi, and the radix salviae miltiorrhizae, the radix angelicae sinensis and the rhizoma chuanxiong are combined to serve as ministerial drugs to jointly achieve the effect of promoting blood circulation to remove blood stasis. The radix platycodonis, the semen armeniacae amarae and the fructus schizandrae are combined to serve as adjuvant drugs and jointly achieve the effects of relieving cough and asthma, and the liquorice serves as conductant drugs, has the effect of regulating drug properties and jointly achieves the effects of tonifying qi (lung and spleen), promoting blood circulation to remove blood stasis and relieving cough and asthma, so that the interstitial lung diseases are prevented and / or treated.

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 with inflammation and interstitial fibrosis of alveolar units as basic lesions. It is also called diffuse parenchymal lung disease (DPLD). It is often manifested by decreased respiratory function of patients, and 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. So far, there is still a lack of ideal intervention measures for the treatment of ILD. In Western medicine clinical treatment, oxygen therapy and anti-fibrosis are the main treatments. Pirfenidone and nintedanib can delay the decline of lung function in patients with pulmonary fibrosis, but in actual clinical use, the above drugs have a slow onset of efficacy in the treatment of idiopathic pulmonary fibrosis and the side effects are difficult to tolerate. In addition, some new treatments have also been developed, such as bone marrow stem cell transplantation, which can promote the repair of lung tissue structure and function. However, stem cell transplantation treatment is still in the experimental research stage and is still a long way from clinical practice.

[0004] In the theoretical system of traditional Chinese medicine, there is no specific disease corresponding to ILD. Most people believe that ILD belongs to the category of "lung arthralgia", and its location is in the lungs and spleen. The main pathogenesis is the deficiency of the lung and spleen qi, and the method of cultivating soil to produce gold is the main treatment principle. The Yellow Emperor's Internal Classic says that "the three qi of wind, cold and dampness are mixed together and become arthralgia", "the skin arthralgia continues, and the evil is affected again, and the lungs are lodged inside", and "the lung arthralgia patient is irritable, full, panting and vomiting", which is the earliest record of the etiology and pathogenesis of lung arthralgia. Later generations of doctors also followed the theory of "lung arthralgia" in the Yellow Emperor's Internal Classic, and from its discussion, it can be seen that the internal deficiency of the internal organs is the key to the occurrence of lung arthralgia. At present, reports on the effective prevention and treatment of ILD by traditional Chinese medicine mainly focus on clinical efficacy and preclinical research related mechanisms, including reducing oxidative stress, improving inflammation, inhibiting epithelial-mesenchymal transition and myofibroblast activation, and regulating autophagy and apoptosis. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a Chinese medicine composition, a Chinese medicine decoction and a preparation method and use thereof for preventing and / or treating interstitial lung disease. The Chinese medicine composition or 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 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 Bitter Apricot, 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 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 Bitter Apricot, 11-13 parts of Platycodon, 12-18 parts of Salvia, 13-17 parts of Chuanxiong, 11-14 parts of Angelica, 5-7 parts of Schisandra, 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, and Qi deficiency and blood stasis run through the whole process, the deficiency in the main body and the excess in the superficial part, its origin lies in the lung and spleen two internal organs, the weakness of internal Qi leads to the obstruction of the meridians, the pathological products such as blood stasis and turbid phlegm are retained and further damage the healthy Qi, and the final accumulation of damage is difficult to restore the lung and lose its use. Therefore, the method of treatment should take into account both strengthening the body (strengthening the spleen and benefiting the lung) and dispelling evil (activating blood circulation, removing blood stasis and removing phlegm).

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

[0013] Sovereign: Astragalus was first recorded in Shennong's Herbal Classic. It is sweet and slightly warm. It enters the spleen and lung meridians. It is often used to strengthen the body. There are many active ingredients in the medicine. Among them, astragalus total saponins include astragaloside A, astragalus polysaccharide, astragaloglycoprotein, astragalus flavonoids and other effective ingredients. They play different degrees of regulating immunity, anti-inflammatory, and inhibiting epithelial-mesenchymal transformation. They are widely used in the treatment of pulmonary fibrosis. Codonopsis was first recorded as a medicine in the Qing Dynasty's Bencao Congxin. Codonopsis is sweet and neutral in nature. It enters the spleen and lung meridians. It has the effects of strengthening the spleen and lungs, nourishing blood and promoting body fluid. It can inhibit high epithelial-mesenchymal transformation and inflammatory response. Astragalus, Codonopsis, Atractylodes and Poria are combined as the sovereign, which plays the role of replenishing qi, especially strengthening the spleen and lungs.

[0014] Minister: Danshen was first recorded in Shennong's Herbal Classic and was listed as a top-grade medicine. It is a commonly used blood-activating and stasis-removing medicine. Modern pharmacological studies have shown that tanshinones and salvianolic acids in Danshen have anti-inflammatory effects, inhibit collagen fiber production, promote fibrin degradation, inhibit cell proliferation, and have good application prospects in preventing and treating tissue and organ fibrosis. Danshen, Angelica sinensis, and Chuanxiong are combined as ministers to achieve the effect of promoting blood circulation and removing stasis.

[0015] Auxiliary: Platycodon grandiflorum is recorded in Shennong's Herbal Classic as "spicy, slightly warm, and enters the lung meridian", which has the effect of clearing the lung and removing phlegm; Bitter almond is recorded in Shennong's Herbal Classic as bitter, slightly warm, and slightly toxic. It enters the lung and large intestine meridians, descends qi, relieves cough and asthma; Schisandra chinensis is recorded in Bencao as warm in nature, sour and sweet in taste. It enters the lung, heart, and kidney meridians, mainly replenishes qi, relieves cough and reverses breathing, astringes and consolidates, relieves cough and removes phlegm, treats chronic cough and asthma, and has the effect of promoting the body's immune function. Platycodon grandiflorum, bitter almond and Schisandra chinensis are combined as adjuvants to achieve the 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, tonifying the middle and replenishing 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, Ligusticum, 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, chuanxiong, angelica, schisandra and liquorice are mixed to obtain a mixed raw material; the mixed raw material is mixed with water and soaked, then boiled and extracted, filtered to obtain a filtrate.

[0020] In the present invention, the astragalus, codonopsis, atractylodes, poria, bitter almond, platycodon, salvia miltiorrhiza, chuanxiong, angelica, schisandra chinensis and liquorice are all used as medicines in the form of 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 first concentrated to obtain a concentrated solution 1, cooled, allowed to stand, and filtered to obtain a filtrate a, and then the filtrate a is second concentrated and filtered to obtain a concentrated solution 2 to obtain a Chinese medicine composition. The first or second concentration method is preferably concentrated under reduced pressure at 60 to 90°C and -0.04 to -0.1MPa, and more preferably concentrated under reduced pressure at 70 to 80°C and -0.05 to -0.08MPa; the relative density of the concentrated solution 1 is preferably 1.04 to 1.06, such as 1.04, 1.05 or 1.06; the relative density of the concentrated solution 2 is preferably 1.02 to 1.04, such as 1.02, 1.03 or 1.04. The cooling temperature is below 10°C; the standing time is preferably 8 to 24h, more preferably 10 to 20h, and more preferably 14 to 18h, such as 14, 15, 16, 17 or 18h. The filtering method can be filtered using a 300-mesh nylon filter cloth.

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

[0024] Preferably, the Chinese medicine decoction also includes 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 the total volume of the above Chinese medicine composition, potassium sorbate and water being 1000 mL, the amount of water used is to add the Chinese medicine composition and potassium sorbate and then make up to 1000 mL with water.

[0027] The present invention is established through modern preparation process research and multi-index quality control system, and Chinese medicine decoction is made, forming a Chinese medicine compound preparation process and quality standard with stable process, controllable quality and good patient compliance, which lays the foundation for the main pharmacodynamics research and clinical data collection of its Chinese medicine composition. At the same time, the main pharmacodynamics of Chinese medicine decoction are verified in vivo and in vitro by obtaining drug active ingredients and target information through network pharmacology, and the molecular mechanism of action of the prescription for treating ILD is preliminarily excavated. Clinically, in improving clinical symptoms, Chinese medicine syndrome, improving quality of life, it is well received by patients. The above-mentioned studies have all confirmed the effectiveness of Chinese medicine decoction for treating ILD. For the next step, Chinese medicine decoction is transformed into a new Chinese medicine and laid a solid foundation.

[0028] 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, 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 is regulated to reduce the Firmicutes flora, increase the Bacteroidetes flora, and increase the ratio of Proteobacteria and Actinobacteria. At the genus level, the intestinal flora is regulated to include flora producing short-chain fatty acids, such as Pseudo-Prevotella 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 the preparation of a product of a short-chain fatty acid-producing bacterial community.

[0033] In the present invention, the short-chain fatty acid producing bacterial flora 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 Chinese medicine composition, Chinese medicine decoction and its preparation method and application for preventing and / or treating interstitial lung disease. The Chinese medicine composition is combined with four medicines of Astragalus, Codonopsis, Atractylodes and Poria as the monarch, exerting the effect of invigorating qi, with Salvia miltiorrhiza, Angelica sinensis and Chuanxiong as the minister, playing the effect of promoting blood circulation and removing blood stasis, with Platycodon grandiflorum, Apricot kernel and Schisandra chinensis as the assistant, playing the effect of relieving cough and relieving asthma, and Licorice as the guide, having the effects of expectorant and relieving cough, invigorating the middle and replenishing qi, and harmonizing the properties of the medicine, playing the effect of strengthening and replenishing qi (lung and spleen), promoting blood circulation and removing blood stasis, relieving cough and relieving asthma. The results show that the Chinese medicine composition or Chinese medicine decoction can improve lung function, inhibit pulmonary fibrosis and inflammation, and can also regulate intestinal flora, thereby preventing and / or treating interstitial lung disease. The preparation method of the Chinese medicine composition or Chinese medicine decoction of the present invention is simple and easy to operate, and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 Effects of serum containing Chinese herbal medicine prepared from decoction on EMT of A549 cells induced by TGF-β1;

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

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

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

[0040] Figure 5 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 air volume of mice after treatment in different groups;

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

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

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

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

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

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

[0049] Fig.14 It is the Venn 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;

[0050] Fig.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] Fig.16This is the Alpha diversity index analysis chart after different groups were treated, 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] Fig.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] Fig.18 To analyze the differences in bacterial flora between samples in different groups 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] Fig.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] Fig. 20 represents the difference of 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] Fig.21 is the ratio of Firmicutes / 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] Fig. 22 To analyze the differences in bacterial flora between samples of different groups 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] Fig.23 The difference of alloprevotella in the bacterial flora 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] Fig.24 The differences in the unclassified_Muribaculaceae bacteria 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 in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

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

[0063] Example 1

[0064] A 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 kernel 12g, Platycodon grandiflorum 12g, Salvia miltiorrhiza 15g, Ligusticum 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 decoction pieces in Part I of the Chinese Pharmacopoeia (2020 edition) and have passed the inspection, and weigh astragalus, codonopsis, atractylodes, poria, bitter almond, platycodon, salvia miltiorrhiza, ligusticum chuanxiong, angelica, schisandra chinensis and liquorice according to the above prescription amount to obtain mixed decoction pieces.

[0068] (2) Soaking the mixed slices in 1442 mL (9 times the amount) of water for 1 hour, boiling for 1 hour, filtering through 300 mesh nylon cloth to obtain a first filtrate and a medicinal residue, adding 1264 mL (8 times the amount) of water to the medicinal residue, boiling for 1 hour, filtering through 300 mesh nylon cloth, discarding the medicinal residue to obtain a second filtrate, and concentrating the first filtrate and the second filtrate under reduced pressure at 60 to 90° C. and -0.04 to -0.1 MPa to a relative density of 1.04 to 1.06 at 50 to 60° C., combining to obtain a first concentrated solution and a second concentrated solution, thereby obtaining 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 a 300-mesh nylon cloth. The precipitate is allowed to stand for another time and allowed to stand for 16 hours. The supernatant 2 is filtered through a 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 to 90° C. and -0.04 to -0.1 MPa to a relative density of 1.02 to 1.04 at 50 to 60° C., and filtered through a 300-mesh nylon cloth to obtain a concentrated solution 2, thereby obtaining a traditional Chinese medicine composition.

[0070] Example 2

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

[0072] 30g of Astragalus, 20g of Codonopsis, 12g of Atractylodes, 15g of Poria, 12g of Bitter Apricot, 12g of Platycodon, 15g of Salvia miltiorrhiza, 15g of Chuanxiong, 12g of Angelica sinensis, 6g of Schisandra chinensis, 9g of Licorice and 3g of potassium sorbate and pure 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 the preparation method described in Example 1.

[0075] Add the above-mentioned amount of potassium sorbate to the concentrated solution 2, add water to make up to 1000 mL, stir and mix, boil for 20 minutes, and cool to below 50° C. to obtain a Chinese medicine decoction.

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

[0077]

Identification

[0078] (1) TLC identification of Atractylodes macrocephala

[0079] Take 20mL of Chinese medicine decoction, extract twice with n-hexane, 25mL each time, combine the n-hexane solution, evaporate to dryness, add 2mL of methanol to the residue to dissolve it as the test solution. Take 1g of Atractylodes macrocephala control medicinal material, add 10mL of n-hexane, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and add 2mL of methanol to the residue to dissolve it as the Atractylodes macrocephala control medicinal material solution. According to the thin layer chromatography method (General Rules 0502 of the 2020 Edition of the Chinese Pharmacopoeia), take 10μL of each of the above two solutions and spot them on the same silica gel G thin layer plate, use petroleum ether (60-90℃)-ethyl acetate (50:1) as the developing solvent, develop, take out, dry, spray with 5% vanillin sulfuric acid solution, heat at 105℃ until the spots are clearly colored, and place them under sunlight for inspection.

[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

[0082] Take 20mL of Chinese medicine decoction, extract twice with ether, 30mL each time, combine the ether solution, evaporate to dryness, add 2mL of n-hexane to the residue to dissolve it as the test solution. Take another 1g of Poria cocos control medicinal material, add 10mL of ether, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and add 2mL of n-hexane to the residue to dissolve it as the Poria cocos control medicinal material solution. According to the thin layer chromatography method (General Rules 0502 of the 2020 Edition of the Chinese Pharmacopoeia), take 5μL of each of the above two solutions and spot them on the same silica gel G thin layer plate, use petroleum ether (30-60℃)-ethyl acetate-acetone (84:3:15) as the developing agent, develop, take out, dry, spray with 5% vanillin sulfuric acid solution, heat at 105℃ until the spots are clearly colored, and place under ultraviolet light (365nm) for inspection.

[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 30mL of Chinese medicine decoction, evaporate to dryness, add 50mL of 7% sulfuric acid ethanol-water (1:3) to the residue, heat and reflux for 3 hours, extract with ether twice, 30mL each time, combine the ether solution, evaporate to dryness, add 2mL of n-hexane to the residue to dissolve it as the test solution. Take 1g of Poria cocos control medicinal material, add 10mL of ether, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, add 2mL of n-hexane to the residue to dissolve it as Poria cocos control medicinal material solution. According to the thin layer chromatography method (General Rules 0502 of the 2020 Edition of the Chinese Pharmacopoeia), take 5μL of each of the above two solutions and spot them on the same silica gel G thin layer plate, use petroleum ether (30-60℃)-ethyl acetate-acetone (84:3:15) as the developing solvent, develop, take out, dry, spray with 5% vanillin sulfuric acid solution, heat at 105℃ until the spots are clearly colored, and place under ultraviolet light (365nm) for inspection.

[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]

Content determination

[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 gel as filler, acetonitrile-water (36:64) as mobile phase, and evaporative light scattering detector for 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, weigh it accurately, 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 20mL, shake and extract 4 times with water-saturated n-butanol, 20mL each time, combine the n-butanol solutions, wash twice with ammonia test solution, 40mL each time, discard the ammonia solution, evaporate the n-butanol solution, dissolve the residue in methanol, transfer it to a 5mL volumetric flask, add methanol to the scale, shake well, filter, and take the filtrate to obtain the solution.

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

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

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

[0095] [Effects and indications] Chinese herbal decoction has the effects of strengthening the spleen and lungs, promoting blood circulation and removing blood stasis, and removing phlegm and unblocking collaterals. It is used for patients with lung arthritis syndrome, which is caused by qi deficiency, blood stasis and phlegm obstruction. Common clinical symptoms include dyspnea, wheezing, fatigue, cough with or without phlegm, dark red tongue, thin white fur, and weak pulse; the Western medical name for this disease is interstitial pneumonia or interstitial lung disease.

[0096] Example 3

[0097] Pharmacodynamics study of Chinese herbal medicine decoction

[0098] 3.1 Cell experiment: Study on the effect of Chinese herbal decoction on TGF-β1-induced mesenchymal transformation of A549 cells

[0099] A549 cells were cultured in vitro and induced by TGF-β1. Then, 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 were detected to elucidate the effect of 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 decoction

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

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

[0106] After 1 week of adaptive feeding, 6-week-old SD rats were continuously gavaged with Chinese herbal decoction for 7 days, once a day, 8.1 mL each time. After the last dose, they 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, packaged, labeled, and stored at -80°C for 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 filtered for sterilization before use.

[0109] (4) Model preparation and drug treatment

[0110] Current studies have 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, which 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] The human alveolar epithelial carcinoma cell line A549 was maintained in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin in a CO2 incubator at 37°C and 5% CO2.

[0112] Establishment of fibrosis model and treatment with Chinese herbal decoction: A549 cells were digested with trypsin and inoculated in 6-well plates. They were incubated in DMEM containing 10% FBS at 37°C and 5% CO2 overnight, and then treated with 0, 5, and 10 ng / mL TGF-1β for 24 hours. At the same time, in order to determine the effect of Chinese herbal decoction 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, and then a medium with a total concentration of 10 ng / mL TGF-1β (containing 10% rat drug-containing serum) was added for a continuous action of 24 hours. The model group was cultured with a medium containing 10% rat blank serum for 24 hours, and then a medium with a total concentration of 10 ng / mL TGF-1β (containing 10% rat blank serum) was added for a continuous action of 24 hours. After the treatment, the morphology of A549 cells was observed under an inverted optical microscope, and the cell culture medium 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 interstitial cells, the intercellular gap became larger, and the connection between adjacent cells became loose, 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 fiber is the most widely distributed and abundant fiber in connective tissue, and is widely distributed in various organs. Type I collagen fibers are mainly bones, skin, and tendons; type II collagen fibers are mainly cartilage collagen; type III collagen fibers are mainly found in embryonic tissues, adult blood vessels, and the gastrointestinal tract; and type IV collagen fibers are mainly found in the basement membrane. Sirius red and its contrast dye are both strong acid dyes, which are easy to combine with the basic groups in the collagen molecules and adsorb firmly. Under polarized light microscopy, collagen fibers have the property of positive uniaxial birefringence light. When combined with Sirius red composite staining solution, birefringence can be enhanced, and resolution can be improved, thereby distinguishing the two types of collagen fibers.

[0116] This example uses a modified Sirius red staining solution (collagen fiber staining) purchased from Beijing Solebow Technology Co., Ltd., which is G1472-reagent B. Collagen fibers in tissues such as heart and blood vessels are dyed red under an ordinary optical microscope, which is helpful for the classification and grading of various fibrotic lesions under a polarized microscope. The expression of collagen in cells can be preliminarily determined by observing the changes in Sirius red staining concentration.

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

[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; the membrane was rinsed three times with Tris buffer (TBST) containing 0.1% Tween-20 for 10 min each time; the membrane was incubated with primary antibodies such as GAPDH (1:2500 dilution), fibrosis-related protein E-cadherin and vimentin for 2 h; the membrane was rinsed three times with TBST for 10 min each time; the membrane was incubated with horseradish peroxidase-conjugated secondary antibody (1:2000 dilution) at 37°C for 1-2 h; the membrane was washed three times with TBST for 10 min each time; the protein bands were developed using an enhanced chemiluminescence substrate kit and detected by the BIO-RAD ChemiDocXRS+ chemiluminescence gel imaging system; the grayscale values ​​of the internal reference GAPDH and fibrosis-related protein bands were analyzed using 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 a decrease in the expression level of E-cadherin protein and an increase in the expression of Vimentin protein in A549 cells, 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 show that Yifei Tongbi Decoction has the effect of reversing EMT of A549 cells.

[0123] 3.2 Animal experiment: Protective effect of Chinese herbal decoction on mice with bleomycin-induced pulmonary fibrosis

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

[0125] 3.2.1 Research Methods

[0126] (1) Selection of induction model drugs

[0127] Bleomycin is an aminoglycoside drug that is commonly used in the clinic to treat a variety of malignant tumors, but its most serious side effect can cause pulmonary fibrosis. Based on its side effects, it is often used to induce animal pulmonary fibrosis models to explore the mechanism of pulmonary fibrosis and screen anti-fibrotic drugs. The commonly used administration methods are single intratracheal administration, or intravenous injection, nasal drops, intraperitoneal injection, etc. The most commonly used and classic model administration method for pulmonary fibrosis models is a one-time injection of bleomycin into the trachea, which can replicate an animal model similar to the pathological process of human pulmonary interstitial fibrosis, and the modeling time is short. Generally, the time range of fibrosis occurrence is 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] The mice were maintained in a suitable breeding environment with an ambient temperature of 20-25℃, a relative humidity of 40%-70%, a noise level below 85 decibels, an ammonia concentration below 20PPm, SPF grade bedding and feed for mice, and free access to water and food every day. All animal experimental operations were performed in accordance with standard procedures and approved by the Experimental Animal Welfare Ethics Committee of Chongqing Institute of Traditional Chinese Medicine.

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

[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 group or high-dose group were respectively slowly injected with 3 mg / kg of 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, and 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 22.75 mg / kg / time, twice a day, with an interval of 12 hours each time; the low-dose group was gavaged with the Chinese medicine decoction prepared in Example 2 20.54 g / kg / d, once a day; the high-dose group was gavaged with the Chinese medicine decoction prepared in Example 2 61.62 g / kg / d, once a day, which were equivalent to 1 times and 3 times the recommended daily dose for humans, respectively.

[0133] The drug was administered for 28 consecutive days, and the coat color and diet of the mice were observed and their body weight was measured every day. On the 28th day, the mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (50 mg / kg); the mice were killed by bleeding from the abdominal aorta, and the lung tissue was immediately dissected and washed with cold saline. The wet weight of the lung was weighed after the water was absorbed by filter paper; the same lobe of the right lung was placed in a 4% paraformaldehyde solution, and the remaining lung tissues were collected and placed at -80°C for subsequent experimental studies.

[0134] After one week of adaptive culture, the male C57BL / 6 mice were in good spirits, had normal breathing, smooth fur, plump muscles, and strong movements; their eyes were bright and energetic, and their reactions were quick; their limbs were well-proportioned, their bodies were not deformed, and there were no scars; there was no abnormal secretion in the natural pores. The blank control group performed normally during the experiment, the model group mice had messy fur and were listless after modeling, and the symptoms of the mice in the drug treatment 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, and 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] The lung function of mice in each group was detected 3 days before the end of the experiment using the unrestrained whole body plethysmography system (WBP) of Shanghai Tower Wang Intelligent Technology Co., Ltd. The indicators and their meanings are shown in the result analysis section. Airway pressure, lung volume, and flow rate are the three elements of lung function measurement. The basic working principle of WBP is based on Boyle's law, that is, the pressure and volume of a quantitative gas in a closed container are inversely proportional at constant temperature.

[0141] The volumetric volume cavity of WBP is equivalent to a closed container. When the animal is placed in the cavity, because the temperature and humidity of the respiratory system are higher than the inhaled gas, the temperature and relative humidity of the inhaled gas increase after mixing with the gas in the lungs, causing the inhaled gas to expand, and the pressure in the cavity increases through the expansion of the animal's chest cavity. The larger the amount of inhaled gas (tidal volume), the more obvious the increase in cavity pressure. Conversely, when exhaling, the exhaled gas mixes with the air, and because the air temperature and humidity decrease, the gas is compressed and the cavity pressure decreases. The change in the pressure in the volumetric volume cavity caused by the animal's breathing (ΔP) can be recorded by the screen-type respiratory sensor in the cavity and amplified by the amplifier. Based on these pressure changes, other indicators of lung function can be derived using special algorithms of the analysis software.

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

[0143] Figure 5 to Figure 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 pulmonary ventilation function of mice and reduce the degree of airway obstruction.

[0144] (6) Pathological observation of lung tissue

[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, and then the lung tissues were 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 with a slicer; 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, 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 fibrosized, 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] Fig. 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 tissue fibrosis was serious; after the intervention of 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 group of the prescription and the high-dose group of the prescription 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] Figure 10 to Figure 13 The results showed that after intervention with Chinese herbal decoction, the levels of inflammation 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] At present, the study of microbial diversity is mainly carried out on the conserved regions of nucleic acid sequences encoding ribosomal RNA. Bacteria are mainly based on 16SrDNA, that is, the DNA sequence encoding the small subunit rRNA (16S rRNA) of the prokaryotic ribosome. By splicing and filtering Reads, clustering or denoising, and performing species annotation and abundance analysis, the species composition of the sample can be revealed; further analysis of alpha diversity (Alpha Diversity), beta diversity (Beta Diversity), significant species difference analysis, correlation analysis, functional prediction analysis, etc. can be carried out to explore the differences between samples. In this study, 16S rRNA gene sequencing was used to detect and analyze the diversity of fecal intestinal microecology. The process was to extract fecal DNA by CTAB method-amplicon generation-PCR product mixing and purification-library construction and sequencing on the machine. Microbial diversity is based on the IlluminaNovaseq sequencing platform, and a small fragment library is constructed for sequencing using the paired-end sequencing method.

[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 by inversion, and fully lyse the sample; (2) Centrifugation: Add phenol (pH 8.0): chloroform: isoamyl alcohol (25:24:1), mix by inversion, and centrifuge for 10 min; take the supernatant, add chloroform: isoamyl alcohol (24:1), mix by inversion, 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) Washing: 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 sucked out 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 extracting the total DNA of the sample, primers were designed according to the conserved region. 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 as the primer corresponding region. Sequencing adapters were added to the ends of the primers, and PCR amplification was performed. The products were purified, quantified and normalized to form a sequencing library. The constructed library was first subjected to library quality inspection, and the qualified library was sequenced using Illumina Novasek 6000. The original image data file obtained by high-throughput sequencing (such as Illumina Novasek sequencing platform) was converted into original sequencing sequences (Sequenced Reads) by base calling analysis, and the results were stored in FASTQ (abbreviated as fq) file format, which contains the sequence information of the sequencing sequence (Reads) and its corresponding sequencing quality information. 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 RawReads obtained by sequencing; then use cutadapt 1.9.1 software to identify and remove primer sequences to obtain CleanReads 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 were collected from each group on the 27th day of continuous administration. The feces samples were collected and sequenced. The sequencing and analysis results are shown in Figure 14 to Figure 24 As shown, the quality of all stool samples was qualified, and 5 samples were selected from each group for sequencing.

[0159] Figure 14 to Figure 24The results showed that the Chinese herbal decoction had a certain regulatory effect on intestinal flora. The differences in flora between 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 Chinese herbal decoction of the present invention can reduce the Firmicutes flora, increase Bacteroidota, reduce the ratio of Firmicutes / Bacteroidota, and increase the ratio of Proteobacteria and Actinobacteriota. The differences in flora between different groups of samples were analyzed based on genus. At the genus level, the Chinese herbal decoction of the present invention can increase the proportion of alloprevotella and unclassified_Muribaculaceae, which produce short-chain fatty acids.

[0160] Chinese herbal decoction has 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 has 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 principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicine composition for preventing and / or treating interstitial lung disease, characterized in that: The invention comprises the following raw materials in parts by weight: 25-35 parts of Astragalus, 15-25 parts of Codonopsis, 10-15 parts of Atractylodes, 10-20 parts of Poria, 10-15 parts of Bitter Apricot, 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 Licorice.

2. The Chinese medicine composition according to claim 1, characterized in that: The invention comprises 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 Bitter Apricot, 11-13 parts of Platycodon, 12-18 parts of Salvia, 13-17 parts of Chuanxiong, 11-14 parts of Angelica, 5-7 parts of Schisandra, and 8-10 parts of Licorice.

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

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

5. The preparation method according to claim 3, 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 of each extraction is 0.5-1.5 hours.

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

04.

7. A Chinese medicinal decoction for preventing and / or treating interstitial lung disease, characterized in that: The invention comprises the Chinese medicine composition according to claim 1 or 2 or the Chinese medicine composition obtained by the preparation method according to any one of claims 3 to 6.

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

9. A method for preparing the Chinese medicine decoction according to claim 7 or 8, characterized in that: The following steps are involved: The Chinese medicine composition obtained by the preparation method according to any one of claims 3 to 6, potassium sorbate and water are mixed, boiled and cooled to obtain a Chinese medicine decoction.

10. Use of the Chinese medicine composition according to claim 1 or 2, the Chinese medicine composition obtained by the preparation method according to any one of claims 3 to 6, the Chinese medicine decoction according to claim 7 or 8, or the Chinese medicine decoction obtained by the preparation method according to claim 9, characterized in that: Include at least one of the following: (1) Use in the preparation of drugs for preventing and / or treating interstitial lung diseases; (2) Application in the preparation of drugs for regulating intestinal flora.

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