CN116726086A - A Tibetan medicinal composition for treating chronic obstructive pneumonia and its preparation method and application - Google Patents

Abstract

The invention discloses a Tibetan medicine composition for treating chronic obstructive pneumonia and its preparation method and application. The Tibetan medicine composition is obtained by repeatedly optimizing the drug composition and content through clinical and basic research, and is made of seven medicinal materials: gentian flower, chinensis bark, rabbit ear grass, nutmeg, licorice, travertine and slag paste. The Tibetan medicine composition of the present invention can reduce the expression levels of inflammatory factors in the serum and lung tissue of chronic obstructive pneumonia model mice and improve lung function indicators, induce autophagy by activating the AMPK/mTOR pathway to reduce respiratory tract inflammation, and inhibit MAPK Activation of pathway proteins reduces inflammatory protein expression. This prescription is clinically suitable for the treatment of chronic obstructive pulmonary disease. It is safe, effective, and of controllable quality. Its efficacy is better than that of chemical drugs such as glucocorticoids that are already on the market. It is an ethnic medicine that is urgently needed in clinical practice.

Description

A Tibetan medicinal composition for treating chronic obstructive pneumonia and its preparation method and application

Technical field

The invention belongs to the technical field of Tibetan medicine, and specifically relates to a Tibetan medicine composition for treating chronic obstructive pneumonia and its preparation method and application.

Background technique

Chronic Obstructive Pulmonary Disease (COPD, referred to as COPD) is a disease state characterized by incompletely reversible airflow limitation. The airflow limitation develops repeatedly and progressively, which is related to the lung disease. It is related to the abnormal inflammatory response of the body to harmful gases or harmful particles; the exact cause of COPD is unknown and is similar to emphysema. Clinically, COPD is diagnosed when patients with chronic bronchitis or/and emphysema show airflow limitation on pulmonary function tests that cannot be fully reversible. Chronic bronchitis refers to chronic, non-specific inflammation of the bronchial wall; emphysema refers to the abnormal and persistent expansion of the air spaces distal to the terminal bronchioles of the lungs, accompanied by the destruction of the alveolar walls and bronchioles. In addition, COPD is a drain on the body, can damage immunity, and make patients prone to infectious diseases. Severe cases can lead to complications in other organs, such as pulmonary heart disease, pulmonary encephalopathy, etc., leading to severe damage to the heart and brain, seriously threatening people's health and quality of life. With the development of society, air pollution has increased, haze environment has increased, smoking and other lung diseases have increased, which has caused the incidence of COPD to increase year by year. Currently, there is no effective clinical treatment drug and patients cannot receive timely prevention and treatment. It results in low disease control rate and high patient mortality rate, making it one of the diseases that endangers the health of people in society.

The pathogenesis of COPD is complex and is related to chronic inflammation, oxidative stress, protease/anti-protease imbalance and immune imbalance. Its clinical manifestations include chronic cough, sputum production, shortness of breath, dyspnea and other symptoms. Conventional treatment includes antitussive, expectorant and anti-inflammatory treatments. Mainly, the main measure is to use bronchodilators. When the condition is severe, they are often combined with glucocorticoids and low-concentration oxygen inhalation. Bronchodilators can better relieve the symptoms of COPD, but they have no clear anti-inflammatory effect and treat the symptoms but not the root cause. Although glucocorticoids can It has good anti-inflammatory effect, but long-term use has serious side effects, and the body will develop glucocorticoid resistance, making the condition more serious. The traditional medicine of the motherland has accumulated rich experience in the treatment of lung diseases. It is of great scientific significance to find COPD treatment drugs from traditional Chinese medicine and classic ethnic medicine prescriptions.

Contents of the invention

The purpose of this invention is to search for suitable drugs for treating COPD from classic Tibetan medicine prescriptions, perform chemical tailoring and optimization, discover new clinical indications for traditional Tibetan medicine to treat COPD, and develop suitable medicinal dosage forms to achieve clear efficacy and increase the number of patients. Compliance, medication safety purposes.

In order to achieve the above object, the preparation scheme of the Tibetan medicine composition provided by the present invention is as follows:

A Tibetan medicine composition for treating chronic obstructive pneumonia. The composition is made of the following raw materials by weight: 120-160 parts of gentian flower, 50-55 parts of chinensis bark, 50-55 parts of rabbit ear grass, 19-25 parts of nutmeg, 50-60 parts of travertine, 20-30 parts of slag paste, and 75-85 parts of licorice.

Preferably, the above composition is made of the following raw materials by weight: 140-160 parts of gentian flower, 50-55 parts of chinensis bark, 50-55 parts of rabbit ear grass, 19-23 parts of nutmeg, and 50-55 parts of travertine. 26-30 parts of slag paste and 75-79 parts of licorice.

The above composition is added with pharmaceutically acceptable auxiliary materials to prepare a Tibetan medicine compound preparation. The preparation may be one of granules, tablets, and capsules.

The preparation method of the above-mentioned Tibetan medicine composition for treating chronic obstructive pulmonary disease includes the following steps: taking gentian flower, chinensis bark, rabbit ear grass, nutmeg, and licorice in proportion, soaking in water, decoction or heating and refluxing to extract, filtering, and concentrating the filtrate to obtain Concentrate the extract, then add the residue and extract, mix and then alcohol precipitate. Take the supernatant and concentrate it, then combine it with travertine, dry, crush and sieve to obtain extract powder.

The above-mentioned water soaking time is 30-60min, and the preferred soaking time is 30-40min.

The above-mentioned decoction or heating and reflux extraction is performed 2-3 times, and the time for each decoction or extraction is 1-2 hours. The water consumption for each decoction or extraction is 8-14 times the weight of the medicinal material, preferably 8-12 times the weight.

The above-mentioned alcohol precipitation involves adding 3-4 times the volume of 95% ethanol, and the alcohol precipitation time is 24 hours.

The Tibetan medicine compound provided by the invention can significantly improve the symptoms of chronic obstructive pulmonary disease, and the Tibetan medicine composition has significant curative effect when used in the preparation of medicines for the treatment of chronic obstructive pneumonia.

The specific functions of the pharmaceutical components used in this invention are as follows:

Gentian flower: This product is the dry flowering aerial part of Gentian veitchiorum Hemsl., a plant of the Gentianaceae family. Basis of identification: It complies with the provisions of Gentian double flower on page 98 of the 2004 edition of the "Standards for Tibetan Medicinal Materials of the Tibet Autonomous Region". "Jingzhu Materia Medica" records that Tibetan medicine uses gentian flower as a "bangjian warm and protective medicine" to treat poisonous diseases, various fevers, and sore throat.

Qinpi: This product is the dried branch bark or dried bark of Fraxinus rhynchophylla Hance, Fraxinus chinensis Roxb, Fraxinus szaboana Lingelsh. or Fraxinus stylosa Lingelsh. of Oleaceae plants. It complies with the regulations under Qinpi on page 282 of the 2020 edition of the Chinese Pharmacopoeia. "Chinese Materia Medica" records that the main functions of Qinpi are clearing away heat and drying dampness; clearing the liver and improving eyesight; relieving cough and asthma. Mainly caused by damp-heat and diarrhea; vaginal discharge, red and swollen eyes; sores and nebula on the eyes; lung-heat, asthma and cough.

Lagotis brevituba: This product is the dried whole plant of Scrophulariaceae plant Lagotisbrevituba Maxim. or Lagotis integra W.Smith. It is recorded in the Flora of China that rabbit ear grass can treat lung abscess, cough, tuberculosis, chest fullness and vomiting pus and blood, blood-heat purulent cavity, yellow water disease and other diseases. It has the effects of clearing away heat, removing fire, reducing inflammation and detoxifying.

Nutmeg: This product is the dried seed kernel of Myristica fragrans Houtt., a plant of the Myristicaceae family. According to the "Chinese Pharmacopoeia", the main functions of nutmeg are to warm the body and promote qi, astringent intestines and relieve diarrhea. It is used for spleen and stomach deficiency, persistent diarrhea, abdominal distension and pain, and vomiting due to lack of food.

Licorice: This product is the dried root and rhizome of the leguminous plant Glycyrrhizauralensis Fisch., Glycyrrhiza inflata Bat. or Glycyrrhiza glabra L. "Chinese Pharmacopoeia" records that the main functions of licorice include nourishing the spleen and replenishing qi, clearing heat and detoxifying, eliminating phlegm and relieving cough, relieving pain and relieving pain, and reconciling various medicines. It is used for weak spleen and stomach, fatigue and fatigue, palpitation and shortness of breath, cough with excessive phlegm, acute pain in the epigastrium and limbs, carbuncle and sore, and relieves the toxicity and potency of drugs.

Travertine: This product is a carbonate mineral, mainly containing calcium carbonate (CaCO3). It is recorded in the "National Collection of Chinese Herbal Medicine" that travertine has the effect of clearing away heat and nourishing the lungs, and is suitable for treating various lung-heat diseases.

Zhajiu ointment: This product is a concentrated extract made by decoction and concentration of Zhajiu with water. The Tibetan medical work "Four Medical Classics" records that Zha Tie Ointment can treat all types of fever, and is particularly effective in clearing stomach heat, liver heat, and kidney heat.

In the "Four Tantras", the foundational classic of Tibetan medicine, it is pointed out that the bodies of all living beings are formed by five elements (earth, water, fire, wind, and space). The lungs belong to the bacon area, and the five elements are earth and water. They are cold and have the function of regulating water and secreting mucus. Due to the characteristics of bacon being cold and damp and spreading in the skin and orifices of the body, the respiratory tract is susceptible to infection by external factors, causing symptoms of cough, thick phlegm, fever and inflammation. If bacon is disturbed, it will become bacon evil, which will cause the heat, sharpness, lightness, greasiness and other properties of Chiba to change, causing Chiba disease, resulting in heat syndrome, plague and various inflammations. If it does not heal for a long time or has repeated attacks, it will evolve. Into chronic bronchitis. When a five-element substance disorder in the human body is diagnosed clinically, drugs with different properties and effects will be used to treat it. Cold and cooling drugs are generally used to treat this disease. There are many famous prescriptions for treating respiratory diseases clearly recorded in the "Four Medical Classics", such as the fifteen-bit gentian flower pills, the nine-flavored travertine powder, the three-flavored gentian flower pills, the six-flavored clove pills, etc. The above famous Tibetan medicine prescriptions It is clinically suitable for the treatment of bronchitis, senile asthma and other diseases. The medicine has a complex taste and has no clinical use in the treatment of chronic obstructive pneumonia.

The present invention uses gentian flower as the main medicine to make a reasonable prescription, adds cold and cool Tibetan medicine that can cure lung heat to enhance the efficacy of gentian flower, and is compatible with corresponding medicines that can prevent bacon syndrome or long syndrome caused by cold and cold medicine, so as to To achieve the goal of the three factors of Long, Chiba and Bacon being in a coordinated state. The specific formula is that gentian flower is the main medicine, and Qinpi helps gentian flower clear away heat and dampness; travertine helps gentian flower clear away heat and nourish the lungs; rabbit ear grass and slag paste help gentian flower clear away heat and detoxify, among which Rabbit ear grass can not only prevent bacon syndrome, but also clear lung and stomach congestion and expel pus; nutmeg, a pungent and warming medicine, prevents long syndrome, warms the middle and promotes qi, and regulates the symptoms of spleen and stomach deficiency and cold; and is combined with licorice to reconcile various drugs to nourish the spleen and replenish qi. , relieve pain and relieve pain. Optimization of clinical trials of Tibetan medicine and modern pharmacological research have proven that this prescription is suitable for the treatment of chronic obstructive pneumonia.

Compared with the prior art, the beneficial effects of the present invention are:

1. The Tibetan medicine composition of the present invention has reasonable compatibility and is an empirical formula composed by the inventor based on the theoretical knowledge of Tibetan medicine combined with many years of clinical practice.

2. The Tibetan medicine composition of the present invention has a small number of medicinal flavors and a simple process, which is beneficial to pharmaceutical production and quality control.

3. The Tibetan medicine composition of the present invention is mainly made of seven medicinal materials: gentian flower, chinensis bark, rabbit ear grass, nutmeg, licorice, travertine and slag paste. It is obtained by repeatedly optimizing the drug composition and ratio through clinical and basic research. . The Tibetan medicine composition can reduce the expression levels of inflammatory factors in the serum and lung tissue of chronic obstructive pneumonia model mice and improve lung function indicators, induce autophagy by activating the AMPK/mTOR pathway to reduce respiratory inflammation, and inhibit the MAPK pathway Protein activation reduces inflammatory protein expression. Therefore, based on a large number of animal experimental research data, this Tibetan medicine composition can improve lung respiratory function, exert anti-inflammatory and immune-enhancing effects, and has no obvious toxicity or adverse reactions. This prescription is clinically suitable for the treatment of respiratory inflammatory diseases such as chronic obstructive pulmonary disease. It is safe and effective, has controllable quality, and is more effective than chemical drugs such as glucocorticoids that are already on the market. It is an ethnic drug that is urgently needed in clinical practice.

Description of drawings

The drawings forming a part of the present invention are used to provide a further understanding of the present invention. The illustrative embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the attached picture:

Figure 1 is a picture of the HE-stained pathological section of the lung tissue of COPD mice using the Tibetan medicine composition of the present invention (×100);

Figure 2 is a picture of the HE-stained pathological section of the lung tissue of COPD mice using the Tibetan medicine composition of the present invention (×200);

Figure 3 is a diagram showing the mechanism of the Tibetan medicine composition of the present invention exerting a protective effect on respiratory tract inflammation on macrophages Raw264.7 by activating the AMPK/mTOR autophagy pathway; in the figure, (A) Different concentrations of LDZK on Raw264.7 cells Vitality effect bar chart; (B) Release amount of inflammatory factor IL-1β from cells in each group; (C) Release amount of inflammatory factor IL-6 from cells in each group; (D) Release amount of inflammatory factor TNF-α from cells in each group ; (E) Original band diagram of AMPK/mTOR pathway proteins;

Figure 4 is a diagram showing the mechanism of the Tibetan medicine composition of the present invention exerting protective effects on respiratory tract inflammation on alveolar epithelial cells A549 by inhibiting activation of the MAPK pathway; in the figure, (A) Toxicity study of different concentrations of LPS on A549 cells; (B) Bar chart of the effect of different concentrations of LDZK on A549 cell viability; (C) Protective effect of LDZK on LPS-induced cytotoxicity at different time points (12, 24, 48h); (D) Inflammatory proteins INOS, COX2 and MAPK pathway proteins of the original strip chart.

Specific implementation methods

The present invention will be further elucidated below in conjunction with specific examples. The following examples are only used to explain the present invention in detail and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the content of the present invention, those skilled in the art will not understand the present invention. Modifications in various equivalent forms fall within the scope defined by the appended claims of this application.

Example 1

Take 55g of travertine, crush it, sieve it (80 mesh), and set aside. Weigh 120g of the raw materials gentian flower, 55g of bark, 55g of rabbit ear grass, 23g of nutmeg, and 80g of licorice. Remove impurities, cut them into pieces and place them in the extraction equipment. Add 10 times the amount of water of the raw materials, soak for 30 minutes, and heat and fry. Boil and extract for 1 hour, filter with 100 mesh filter cloth to obtain the first extraction liquid, then add 8 times the amount of water of the raw material, heat and decoct for 1 hour, filter with 100 mesh filter cloth to obtain the second extraction liquid, combine the two extraction liquids, and reduce Press and concentrate to obtain the concentrated extract, add 26g of the residue extract, mix evenly, and add 3 times the volume of 95% ethanol to the extract mixture, stir evenly and let it stand for 24 hours, take the supernatant and concentrate, combine with travertine, dry, and pulverize Sieve to obtain extract powder. Then add conventional doses of microcrystalline cellulose and lactose, mix with 75% ethanol, perform granulation by conventional methods, and sieve to prepare granules.

Example 2

Take 50g of travertine, crush it, sieve it (80 mesh), and set aside. Weigh 160g of the raw materials gentian flower, 50g of bark, 50g of rabbit ear grass, 19g of nutmeg, and 75g of licorice. Remove impurities, cut them into pieces and place them in the extraction equipment. Add 12 times the weight of the raw material in water, soak for 30 minutes, and heat and fry. Boil and extract for 2 hours, filter with 100 mesh filter cloth to obtain the first extraction liquid, then add 10 times the amount of water of the raw material, heat and decoct for 1 hour, filter with 100 mesh filter cloth to obtain the second extraction liquid, combine the two extraction liquids, and reduce Concentrate under pressure to obtain a concentrated extract, add 30g of residue extract, mix evenly, add 3 times the volume of 95% ethanol to the extract mixture, stir evenly and let it stand for 24 hours, concentrate the supernatant, combine with travertine, dry, and grind Sift the extract powder. Then add conventional dosages of starch, powdered sugar, and dextrin, then adopt conventional wet granulation, mix with magnesium stearate, and then press into tablets to prepare tablets.

Example 3

Take 52.5g of travertine, crush it, sieve it (80 mesh), and set aside. Weigh 140g of the raw materials gentian flower, 50g of bark, 52.5g of rabbit ear grass, 21g of nutmeg, and 79g of licorice. Remove impurities, cut them into pieces and place them in the extraction equipment. Add 10 times the weight of the raw material in water, soak for 30 minutes, and then heat Decoct and extract for 1 hour, filter with 100 mesh filter cloth to obtain the first extraction liquid, then add 8 times the weight of the raw material in water, heat and decoct for 1 hour, filter with 100 mesh filter cloth to obtain the second extraction liquid, combine the two extraction liquids, Concentrate under reduced pressure to obtain a concentrated extract, add 28g of residue extract, mix evenly, add 3 times the volume of 95% ethanol to the extract mixture, stir evenly and let it stand for 24 hours, take the supernatant and concentrate, combine with travertine, dry and pulverize Sieve the extract powder to obtain the extract powder, add a conventional dose of starch, and fill hollow capsules to prepare capsules.

Example 4

Take 52.5g of travertine, crush it, sieve it (80 mesh), and set aside. Weigh 140g of the raw materials gentian flower, 52.5g of bark, 52.5g of rabbit ear grass, 21g of nutmeg, and 77g of licorice, remove impurities, cut into pieces and place in the extraction equipment, add 10 times the amount of water of the raw material, and soak for 30 minutes. Heat, decoct and extract for 1 hour. Filter with 100 mesh filter cloth to obtain the first extraction solution. Add 10 times the amount of water of the API. Heat, decoct and extract for 2 hours. Filter with 100 mesh filter cloth to obtain the second extraction solution. Combine the two extracts. , concentrate under reduced pressure to obtain a concentrated extract, add 28g of slag extract, mix evenly, add 4 times the volume of 95% ethanol to the extract mixture, stir evenly and let it stand for 24 hours, take the supernatant and concentrate, combine with travertine and dry. Crush and sieve the extract powder to obtain the extract powder, add a conventional dose of concentrated sucrose aqueous solution to prepare a syrup.

Example 5

Take 55g of travertine, crush it, sieve it (80 mesh), and set aside. Weigh 140g of the raw materials gentian flower, 52.5g of bark, 52.5g of rabbit ear grass, 21g of nutmeg, and 77g of licorice. Remove impurities, cut them into pieces and place them in the extraction equipment. Add 12 times the amount of water as the raw material and soak for 30 minutes. Heat, decoct and extract for 1 hour. Filter with 100 mesh filter cloth to obtain the first extraction solution. Add 10 times the amount of water of the raw material. Heat, decoct and extract for 1 hour. Filter with 100 mesh filter cloth to obtain the second extraction solution. Combine the two extracts. , concentrate under reduced pressure to obtain a concentrated extract, add 28g of slag extract, mix evenly, add 3 times the volume of 95% ethanol to the extract mixture, stir evenly and let it stand for 24 hours, concentrate the supernatant, combine with travertine, and dry , crush and sieve to obtain the extract powder, add a pharmaceutically acceptable wetting agent to obtain pills, and round into pills using a pill making machine to prepare pills.

The above-mentioned slag paste is a concentrated extract made by adding 5-10 times the weight of the slag to water and boiling it 1-2 times, and then concentrating the filtrate.

Example 6

In order to study the efficacy of this Tibetan medicine compound on COPD, the present invention uses the method of instilling LPS into the respiratory tract combined with atomized papain to establish a mouse model of chronic obstructive pulmonary disease, and uses this as the research object to conduct drug efficacy tests. The feasibility of the modeling method was initially judged by observing the mouse body weight and blood inflammation indicators during the modeling period. After the experiment, the lung tissue pathological sections, lung respiratory function, and bronchoalveolar perfusion fluid of the model group mice and the control group mice were measured. (BALF), blood inflammatory cell indicators and immune organ indexes to make a final judgment on the feasibility of the COPD modeling method, and determine the therapeutic effect of this Tibetan medicine prescription on COPD by comparing various indicators between each drug group and the model mice.

The present invention is verified and explained below through drug efficacy test examples:

Test drug (drug extract powder prepared by the method of Example 2), control drug (dexamethasone), lipopolysaccharide LPS, papain, 60 C57 male mice, isoflurane, 25% urethane, mist Chemical analyzer, blood analyzer, (EMMS) FM forced lung function testing system, tissue homogenizer, lipid oxidation (MDA) detection kit, Mouse IL-6 ELISA Kit, MouseTNF-α ELISA Kit.

6.2 Animal grouping

A total of 60 C57 mice were quarantined for 7 days. They were weighed on the last day of the quarantine period, and 9 mice were randomly selected as a blank group according to body weight. The remaining mice were modeled together. After modeling, the number of deaths was eliminated and randomly divided. They are the COPD model group, the dexamethasone (DEX) group, the low-concentration Tibetan medicine group, and the high-concentration Tibetan medicine group. 8-9 animals per group.

6.3 Model establishment and drug administration

Except for the normal control group, the other groups were instilled with 50 μl of LPS (1 mg/ml) into the nasal cavity on the first day of the experiment, and 25 μl of LPS (1 mg/ml) on the 15th and 27th days, and the LPS recovery period was 1 -After 3 days, atomize papain (1mg/ml), 1min/animal, atomize every other day, and build the model for 4 weeks.

After the modeling was completed, the drugs were given. The model group was given 10ml/kg of normal saline, the DEX group was given 0.2mg/kg of DEX solution, and the low-concentration Tibetan medicine group was given 102.75mg/kg of drug extract powder suspension, which is equivalent to the crude drug dose. The amount of crude drug/kg was 0.9g. In the high-concentration Tibetan medicine group, the drug extract powder suspension was administered orally at 205.5 mg/kg. The equivalent crude drug amount was 1.8g crude drug/kg. A total of 4 weeks of intragastric administration. For the first 2 weeks of the dosing period, atomized papain (1 mg/ml) was maintained every other day, 1 min/animal. The total experimental period is 8 weeks.

After the end of the 8th week of intragastric administration, 5 animals from each group were selected for lung function measurement. After the measurement, the chest cavity was opened to remove lung tissue. The left lung was fixed in 4% paraformaldehyde, and the right lung was weighed and stored at -80°C. Later, the kit indicators were determined using it; in addition, 3-4 mice in each group were bled from their eyeballs and then underwent bronchoalveolar lavage to collect BALF fluid and measure the differential count of white blood cells.

6.4 Experimental indicators

(1)Weight testing

The weight of the mice was measured every week, and their activity status, diet, etc. were observed.

(2) Blood inflammatory cell indicators

Blood was collected from the mouse orbits at weeks 4 and 8, and an automatic blood analyzer was used to perform statistical analysis on the number of white blood cells, neutrophils, and lymphocytes in the blood.

(3) Inflammatory cell indicators in bronchoalveolar perfusion fluid BALF

Cut the skin of the mouse's throat, use forceps to separate the tissue around the trachea, and expose the trachea. Use the No. 7 needle of the syringe, cut off the needle tip, then grind it flat, and fix the animal in the supine position. First, thread two surgical sutures between the trachea and esophagus. Use small scissors to make a T-shaped incision across the thyroid. Insert a homemade needle into the trachea. Tighten the place where the casing enters the trachea and the distal end of the casing. Connect the homemade needle Lavage 0.5ml normal saline with a 1ml syringe, stay for 10 seconds and withdraw slowly, repeat twice, until milky white foamy liquid is visible. Stop when you feel that the piston cannot move and there is resistance, transfer to the EP tube, repeat the above lavage process 3 times, and combine The extracted liquid has a recovery rate of more than 80% and is regarded as qualified perfusate. Use an automatic hematology analyzer to count the total number of white blood cells, neutrophils, lymphocytes, and eosinophils.

(4) Measurement of IL-6 and TNF-α in lung tissue

Take the lower lobe of the right lung that has not been perfused, weigh about 30 mg, add PBS at a ratio of 1:9 to homogenize the lung tissue, centrifuge at 12000r, 4°C for 10 minutes, take the supernatant, and strictly follow the steps of the kit to perform IL- 6. TNF-α detection.

(5) Lung function indicators

After the eighth week, the lung function of the mice was measured. The limbs and head of the anesthetized mouse were fixed, and the neck was disinfected. The subcutaneous tissue of the neck was separated layer by layer with blunt force until the trachea was exposed and separated. A small transverse incision was made in the upper part of the tracheal ring. The tracheal tube was inserted and ligated and fixed with surgical thread. , then put the mouse into the mouse chamber of the pulmonary function instrument, connect the small animal pulmonary function instrument for mechanical ventilation and measure. The measurement results of each mouse are repeated three times. The lung function indicators include 50ms forced expiratory volume (FEV50) and forced vital capacity. (FVC), functional residual capacity FRC, maximum expiratory flow PEF, maximum mid-expiratory flow MMEF.

(6) Pathological sections of lung tissue

Take the fixed left lung and process it according to the steps of dehydration and wax dipping, embedding, sectioning, dewaxing to water, hematoxylin staining, eosin staining, dehydration and sealing, and finally microscopic examination under 100x and 200x visual fields. Image acquisition and analysis.

(7)Immune organ index

After the mice were subjected to bronchoalveolar lavage and the respiratory function of the mice was measured, the thymus and spleen were taken and weighed, and the organ index was measured. COPD patients have reduced immunity and defense against respiratory pathogenic microorganisms, and thymus and spleen indicators are often used to evaluate host immunity.

6.5 Experimental results

(1)Modeling results

(a) General status of mice during the modeling period

Compared with the control group, the model mice had reduced food intake and severe weight loss. The specific results are shown in Table 1. In addition, the model mice showed symptoms such as wheezing, nodding breathing, deepening and accelerated breathing, abdominal muscle twitching, bunching up and hunched back, etc., and moved independently. decline, reaching a mortality rate of approximately 25%.

Table 1 Body weight growth rate of mice in each group during the modeling period

Note: Compared with the control group, **P<0.01, *P<0.05

(b) Inflammation indicators in mice during the modeling period

Analysis of the results in Table 2 shows that compared with the control group, the leukocytes in the blood of the model mice increased significantly, and the neutrophils and lymphocytes also increased significantly, indicating that a severe inflammatory response occurred in the mice and caused a corresponding immune response.

Table 2 Number of inflammatory cells in each group of mice during the modeling period (10 ⁹ /L)

Note: Compared with the control group, **P<0.01, *P<0.05

(2)Treatment results

(a) Effect of the Tibetan medicine compound of the present invention on the general state of COPD mice

Compared with the control group, the weight growth rate and spontaneous activity of the other groups were significantly reduced; compared with the COPD model group, the Tibetan medicine compound drug group of the present invention had high spontaneous activity activity, good condition, and stable growth rate of weight; model The weight of the mice in the group first increased and then decreased, indicating that the later development of COPD still has a certain impact on weight; the weight of the dexamethasone group showed a downward trend and their activity level was not high. The specific weight results are shown in Table 3.

Table 3 Body weight growth rate of mice in each group after administration

Note: Compared with the control group, **P<0.01, *P<0.05; compared with the COPD group, ##P<0.01, #P<0.05

(b) Effect of the Tibetan medicine compound of the present invention on blood inflammatory cells in COPD mice

Compared with the model group, the white blood cells WBC and lymphocytes Lym in the blood of the Tibetan medicine compound group of the present invention were significantly reduced in a dose-dependent manner, and the neutrophil index showed a dose-dependent decreasing trend. Compared with the model group, the WBC and Lym of the DEX group were significantly reduced, but the Neu index showed a significant increase trend. The specific results are shown in Table 4, which shows that the Tibetan medicine compound extract powder has a good systemic anti-inflammatory effect and is more stable than DEX.

Table 4 Number of blood inflammatory cells in mice in each group after administration (10 ⁹ /L)

Note: Compared with the control group, **P<0.01, *P<0.05; compared with the COPD group, ##P<0.01, #P<0.05

(c) Effect of the Tibetan medicine compound of the present invention on inflammatory cell indicators in BALF of COPD mice

Compared with the model group, the inflammatory leukocyte WBC in the alveolar lavage fluid of the Tibetan medicine compound group of the present invention has a decreasing trend, and the high-concentration drug group has a significant decreasing trend; compared with the model group, the WBC index of the DEX group is significantly lower. Compared with the model, the number of eosinophils in the BALF of the Tibetan medicine compound group and the DEX group was extremely significantly reduced; the number of neutrophils showed a decreasing trend, with no significant difference. The specific results are shown in Table 5, which preliminarily shows that both Tibetan medicine compound drugs and dexamethasone DEX have anti-inflammatory effects on the lungs.

Table 5 Number of inflammatory cells in BALF of mice in each group after administration (10 ⁹ /L)

Note: Compared with the control group, **P<0.01, *P<0.05; compared with the COPD group, ##P<0.01, #P<0.05

(d) The Tibetan medicine compound of the present invention affects IL-6 and TNF-α indicators in the lung tissue of COPD mice

Compared with the model group, the inflammatory factor IL-6 and TNF-α indicators in the Tibetan medicine compound group of the present invention tended to decrease. Among them, the high-concentration drug group showed a significant decrease, and the inflammatory factor index in the DEX group also decreased significantly. Specifically, The results are shown in Table 6. This indicator further illustrates that the Tibetan medicine compound drug of the present invention has obvious anti-inflammatory effect on COPD mice and is dose-dependent, which is equivalent to the anti-inflammatory effect of the positive drug.

Table 6 IL-6 and TNF-α content in lung tissue of mice in each group after administration (pg/mgprot)

Note: Compared with the control group, **P<0.01, *P<0.05; compared with the COPD group, ##P<0.01, #P<0.05

(e) Effects of the Tibetan medicine compound of the present invention on lung function indicators of COPD mice

Compared with the model group, the Tibetan medicine compound group had an increasing trend in forced expiratory volume in 50 ms (FEV50), forced vital capacity (FVC), maximum mid-expiratory flow (MMEF) and maximum expiratory flow (PEF). , among which the high-concentration drug group increased significantly, and the functional residual capacity PRC showed a decreasing trend, among which the high-concentration drug group significantly decreased; compared with the model group, the FEV50, MMEF, PEF and FRC indicators of the dexamethasone DEX group all showed significant improvement. , but the FVC index is worse than that of the model group. The specific results are shown in Table 7. From the above results, it can be seen that LDZK drugs have a dose-dependent prevention and treatment effect on respiratory obstruction in COPD.

Table 7 Lung function indicators of mice in each group after administration

Note: Compared with the control group, **P<0.01, *P<0.05; compared with the COPD group, ##P<0.01, #P<0.05

(f) Effects of the Tibetan medicine compound of the present invention on pathological sections of lung tissue in COPD mice

In order to quantify the pathological changes in lung tissue and compare the results more clearly and directly, a scoring table for the pathological degree of lung tissue can be obtained by reviewing the literature, as shown in Table 8. Please see Figure 1 (×100) and Figure 2 (×200) for the pathological section results of lung tissue.

Compared with the normal group, the alveolar morphology of the model group was abnormal, the structure was destroyed, some alveolar cavities merged with each other to form bullae, and inflammatory cell infiltration was visible in the alveoli; a large number of inflammatory cells were infiltrated into the pulmonary interstitium, and the pulmonary interstitium was significantly thickened; the bronchi The tube wall thickens, the tube lumen becomes narrowed, the epithelial cells on the tube wall become necrotic, and cell debris and mucus appear in the tube lumen. Compared with the model group, the alveolar damage and inflammatory cell infiltration in the DXM group and the low-concentration drug group were slightly improved, but the bronchial damage was not significantly improved; in the high-concentration drug group, most of the alveoli were in normal shape, with some alveolar fusion and inflammatory cells visible. Infiltration and bronchial damage were significantly improved.

Table 8: Lung tissue pathological degree score of mice in each group after administration

(g) Effect of the Tibetan medicine compound of the present invention on the immune organ index of COPD mice

Compared with the normal group, the spleen index and thymus index of the model group and the Tibetan compound drug group were decreased, indicating that COPD modeling caused an imbalance in immune regulation. Compared with the model group, the spleen index and thymus index of the LDZK drug group increased in a dose-dependent manner; the spleen index and thymus index of the dexamethasone group were smaller than those of the model group. The specific results are shown in Table 9. This shows that dexamethasone severely reduces the body's immunity, while LDZK lozenge extract powder enhances the immunity of COPD mice.

Table 9 Immune organ index of mice in each group after administration (mg/g)

Note: Compared with the control group, **P<0.01, *P<0.05; compared with the COPD group, ##P<0.01, #P<0.05

6.6 Summary

(1)About positive drugs

The current main measures to treat COPD are: bronchodilators, including β2 receptor agonists, anticholinergic drugs and theophylline drugs; glucocorticoids; phlegm-reducing drugs, etc. Among them, bronchodilators can very well relieve the symptoms of COPD. However, it has no anti-inflammatory effect and treats the symptoms but not the root cause; therefore, the present invention aims to develop safe and effective anti-inflammatory drugs for COPD. In order to have clear therapeutic indicators to measure the efficacy of the drug, the present invention selects dexamethasone, a glucocorticoid drug already on the market, as a positive control to compare the advantages and disadvantages of the two drugs.

(2) Regarding the feasibility of COPD modeling in the present invention

From the diet, weight growth rate and systemic blood inflammation of the mice during the modeling period, it can be preliminarily determined that this modeling method has caused certain damage to the mice. After the later stage, HE staining of lung tissue, lung function respiratory indicators, blood and BALF The measurement of inflammatory cells, the measurement of inflammatory factors IL-6, TNF-α and the measurement of immune organ index can clarify the pathological process of airway obstruction, emphysema, inflammation and low immunity in mice caused by LPS combined with papain modeling. The symptoms are similar to clinical COPD, which shows that this COPD modeling method is feasible.

(3) Regarding the therapeutic effect of the Tibetan medicine compound in the present invention

From the measurement results of inflammatory cells in blood, bronchoalveolar lavage fluid (BALF) and lung tissue inflammatory factors, it can be seen that the model mice have obvious inflammation from local lung tissue to the whole body. Compared with the model mice, various inflammatory indicators in the Tibetan medicine compound group were significantly improved. From the results of HE staining of lung tissue, respiratory indicators and immune organ index, it can be seen that compared with the model mice, the Tibetan medicine compound drug group has a certain therapeutic effect on respiratory obstruction, emphysema and immunocompromise, and this drug is dose-dependent. treatment effect. In addition, it can be seen from the above results that DEX drugs have a good anti-inflammatory effect, but they can cause a serious decline in immune function, which may be related to the side effects of hormonal drugs.

Example 7

In order to further clarify the anti-inflammatory mechanism of the Tibetan medicine composition, the present invention constructed different in vitro inflammation models to observe the anti-inflammatory protection and regulatory effects of various inflammatory pathway proteins of the drug. Macrophages play an important role in respiratory inflammatory diseases, and the most common inducer of macrophage activation is lipopolysaccharide LPS. LPS-treated RAW 264.7 macrophages have been widely used as a model to study inflammatory responses in vitro. A549 belongs to alveolar type II epithelial cells. LPS or CSE is often used to stimulate A549 to produce an inflammatory response as an in vitro model for lung disease research. Therefore, the present invention uses LPS to induce macrophages RAW264.7 and alveolar type II epithelial cells A549 to construct a lung disease inflammation model.

Autophagy, as a cellular waste removal system, plays a key role in regulating inflammatory responses. The AMPK/mTOR pathway is an important autophagy regulatory pathway. When cells are stimulated by external stimuli, AMPK is activated and undergoes a phosphorylation reaction, which can inhibit the downstream mTOR gene phosphorylation reaction, thereby promoting the occurrence of the autophagy cascade. Activating autophagy can Suppresses the inflammatory response by reducing the secretion of pro-inflammatory cytokines. In addition, multiple studies have shown that oxidative stress inducers such as cigarette smoke and LPS can regulate MAPKs such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK). ways to enhance inflammatory effects. Therefore, the present invention explains the anti-inflammatory mechanism of the compound composition by observing the protein regulatory effects of the AMPK/mTOR pathway and MAPK pathway.

7.1RAW264.7 cell culture and processing

RAW 264.7 cells were purchased from ATCC Cell Bank (USA), cultured and grown in RPMI 1640 medium containing 10% (v/v) fetal calf serum, 100 U/ml penicillin and 100 μg/ml streptomycin, and the incubator conditions were 37°C. ,5%CO2. The MTT method was used to determine the effect of different concentrations of Tibetan medicine compositions on the viability of RAW 264.7 cells; the cells were seeded in a 96-well plate (1×104 cells per well). After the cells adhered to the wall, they were pretreated with appropriate concentrations of drugs. After 24 hours, After LPS (1 μg/ml) stimulated the cells, the supernatant was collected for 24 hours to measure IL-6, IL-1β, TNF-α and other inflammatory factors.

7.2A549 cell culture and processing

A549 cells were from Southeast University and were cultured and grown in high-glucose DMEM medium containing 10% (v/v) fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin in an incubator at 37°C and 5% CO2. The MTT method was used to determine the effects of different concentrations of Tibetan medicine compositions and LPS on A549 cell viability, and to determine the protective effects of drugs on A549 stimulated by LPS at different time points.

7.3Western Blot

The cells were seeded in a 6-well plate (2×105/well), pretreated with Tibetan medicinal composition 24 hours later, and then stimulated with lipopolysaccharide (LPS; Sigma) (1 μg/ml) for 24 hours, and then treated with pre-cooled Wash 3 times with PBS, add pre-cooled RIPA lysis buffer containing protease inhibitors and phosphatase inhibitors, lyse on ice for 20 minutes, centrifuge at 12000r, 4°C for 10 minutes to take the supernatant, and quantify BCA protein to 5 μg/μl. The proteins were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS/PAGE), transferred to PVDF for 1.5 hours, and the membrane was blocked in 5% skim milk prepared with TBST for 2 hours, and the membrane was washed three times for 5 min/ times, incubate with primary antibody (1:1000) at 4°C overnight, and wash the membrane three times, 10 min/time. Incubate with anti-HRPp-coupled rabbit or IgG secondary antibody (1:3000) for 1 hour at room temperature, and wash the membrane three times, 10 minutes each time. Proteins were visualized using a CCD system (Tanon 5200, China) by developing with ECL solution.

7.4 Results

(1) The Tibetan medicine composition of the present invention exerts anti-inflammatory effects by activating the AMPK/mTOR autophagy pathway

As shown in Figure 3, the present invention uses 1 μg/ml LPS to stimulate RAW264.7 to construct an inflammation model, and selects 0.5 and 1 mg/ml Tibetan medicine compositions for intervention. After 24 hours, the levels of inflammatory factors and autophagy protein content indicators are measured. The results showed that LPS treatment significantly increased the secretion of IL-1β, IL-6 and TNF-α in RAW 264.7 cells, and the Tibetan medicine composition could inhibit the expression and release of these inflammatory factors in a dose-dependent manner. Immunoblotting results showed that the drug can significantly increase the phosphorylation of AMPK and reduce the phosphorylation of mTOR, thereby enhancing autophagy and exerting anti-inflammatory effects.

(2) The Tibetan medicine composition of the present invention exerts anti-inflammatory effects by inhibiting the activation of the MAPK pathway

According to the cell viability results in Figure 4, the present invention uses 1 μg/ml LPS to stimulate A549 to construct an inflammation model, and uses 50 and 100 μg/ml Tibetan medicine compositions for intervention, and the protein content index is measured after 24 hours. The results showed that the ERK, JNK, and P38 proteins in the COPD model group were significantly phosphorylated, and the content of downstream inflammatory proteins iNOS and COX2 increased significantly; the drug group had a significant inhibitory effect on the activation state of the MAPK pathway protein and reduced the expression of inflammatory proteins. Exhibits anti-inflammatory effects.

7.5 Discussion and summary

The present invention induces RAW264.7 and A549 cells to produce inflammatory responses through LPS, and both show good anti-inflammatory efficacy after administration and intervention, which is consistent with in vivo research results, and has a regulatory effect on AMPK/MTOR pathway and MAPK pathway proteins. Inflammatory factors such as IL-1β, IL-6 and TNF-α appear in the development process of chronic inflammation in COPD. Excessive inflammatory factors will activate the NF-κB complex, which in turn promotes the expression of more inflammatory genes and amplifies the inflammatory response. Therefore, the present invention selects the release amount of inflammatory factors to measure the anti-inflammatory effect of drugs on macrophages. In addition, inducible NO synthase iNOS is the main enzyme that catalyzes the production of NO under inflammatory conditions, and excess NO may promote the production of cytokines and matrix metalloproteinases, mitochondrial dysfunction and cell apoptosis, thereby accelerating the development of inflammation; Oxygenase-2 (COX-2) promotes the production of prostaglandins (PGs) and plays an important role in chronic inflammation. Therefore, the present invention selects the expression levels of iNOS and COX2 inflammatory proteins to measure the anti-inflammatory protective effect of drugs on alveolar cells. Studies have found that activation of AMPK can improve chronic inflammation and damage in COPD or emphysema by reducing inflammatory responses, slowing down cell aging, inducing autophagy and regulating mitochondrial metabolism; the results of a number of studies on the biological anti-inflammatory activity of medicinal plants have shown It exerts its biological properties by blocking the two main signaling pathways of NF-κB and mitogen-activated protein kinase MAPK, indicating that the MAPK pathway plays a major role in the production of various pro-inflammatory mediators. Therefore, the present invention selects the AMPK/MTOR pathway and the MAPK pathway to illustrate the anti-inflammatory mechanism of the compound composition. The above results show that the composition may regulate autophagy by activating AMPK and thereby inhibiting MTOR phosphorylation, and may affect the expression of inflammatory proteins and the release of inflammatory factors by inhibiting the phosphorylation of the MAPK pathway.

Example 8

In order to ensure the safety of the clinical use of the Tibetan medicine compound of the present invention, this example uses the maximum dosage method to administer the maximum dosage to SD male and female rats in a single dose, and administer the drug extract powder prepared by the method of Example 2 by gavage. , observe the acute toxic reactions after administration, record the toxic reaction time and symptoms, target organs, death time, recovery time, animal weight changes, death and other information to provide information for determining the safety range and toxicity characteristics of the tested drug. in accordance with.

8.1 Animal grouping

In this example, after 14 days of quarantine, the rats were weighed on the last day of the quarantine period, and randomly divided into 4 groups according to body weight, two groups of SD male and female rats respectively, divided into a control group and a drug administration group; there were 12 rats in the control group. , half male and half female; there were 21 animals in the administration group, 10 females and 11 males.

8.2 Conducting toxicity experiments

Based on the dissolution conditions during preparation of the medicinal solution and the difficulty of absorbing the medicinal solution with a No. 16 gavage needle, the maximum dosage concentration of the extract powder of the Tibetan medicine composition of the present invention is 1.5g/ml. Since the maximum single administration volume is 20ml /kg, so the maximum single dose is set at 30g/kg, which is equivalent to 210g of crude drug/kg; the route of administration is set as intragastric administration;

This test adopts the maximum single dose method. The drug group is given a single dose within one day, with a total dose of 30g/kg (292 times the clinically effective dose). The negative control group is given an equal volume of distilled water. After administration, the patients were observed continuously for two weeks, and the results were determined by dissecting the specimens. During the dissection process, one male rat in the medication group was anesthetized and died. No blood or organs were taken, but no lesions were observed in the organs with the naked eye. Blood collection from one female rat in the medication group was unsuccessful, and the organs were removed and observed with the naked eye. There were no lesions in the organs.

8.3 Experimental indicators

(1)Weight testing

The body weight of the animals was measured on the day of administration, and on the 1st, 2nd, 3rd, 5th, 7th, 10th and 14th days after administration.

(2) Blood biochemical indicators

Fourteen days after administration, blood was taken from the abdominal aorta of the rats, and major biochemical indicators in the blood were detected, including AST/ALT (aspartate aminotransferase/alanine aminotransferase), BUN (blood urea nitrogen), and TBI. (total bilirubin) and TP (total protein), which can detect the health status of the liver, kidneys, gallbladder, etc., as well as the nutritional health status of the whole body.

(3) Blood physiological indicators

Fourteen days after administration, blood was taken from the abdominal aorta of the rats, and an automatic hematology analyzer was used to perform statistical analysis on the number of white blood cells, red blood cells, hemoglobin concentration and platelet count in the blood. It can detect inflammation, hematopoietic function, oxygen-carrying capacity and coagulation function in the body. Comprehensive measure of rat health.

(4) Vital organ index

Fourteen days after administration, important organs such as the heart, liver, spleen, lungs, kidneys, and brain were taken and weighed to determine the organ index. During the dissection process, it is observed whether there are obvious organ lesions to determine whether there is a risk of organ damage if a large amount of the Tibetan medicine compound of the present invention is taken.

8.4 Experimental results

(1) Weight growth rate

Female rats experienced significant weight loss on the first day after administration, and began to resume weight gain on the second day. The growth rate on the third day was slower than the weight growth rate of the control group. From the fourth day, the weight data was not significant compared with the control group. Difference; the weight of male rats dropped extremely significantly on the first day after administration, and gradually returned to normal on the second day, with no significant difference from the control group. See Table 10 and Table 11 for specific data. From the above situation, after the Tibetan medicine composition extract powder was administered to female and male rats at the maximum dosage once, there was no abnormality in the body weight growth index.

Table 10 Female rat body weight growth rate

Note: Compared with the control group, ** means P<0.01, * means P<0.05

Table 11 Male rat body weight growth rate

Note: Compared with the control group, ** means P<0.01, * means P<0.05

(2) Blood biochemical indicators

The total protein TP index is related to the nutritional needs and material metabolism capabilities of animals of different ages and genders. Compared with the control group, the total protein TP index of the male rat administration group was significantly lower, but the mean values were not much different. Check the literature. Normal values, and there are no significant differences in other indicators. There was no significant difference in the blood biochemical indicators of female rats. The specific results are shown in Table 12 and Table 13. Judging from the above situation, there were no abnormalities in the blood biochemical indicators of female and male rats after the Tibetan medicine composition extract powder was administered at the maximum dosage once.

Table 12 Biochemical indicators of female rats

Table 13 Biochemical indicators of male rats

Note: Compared with the control group, ** means P<0.01, * means P<0.05

(3) Blood physiological indicators

Compared with the control group, there was no significant difference in routine blood values between the female rat administration group and the male rat administration group, including white blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), and platelets (PLT). The specific results are shown in Table 14 and Table 15. From the above situation, it can be seen that after the Tibetan medicine composition extract powder was administered to female and male rats at the maximum dosage once, there was no abnormality in the blood physiological indicators.

Table 14 Blood routine value indicators of female rats

Table 15 Blood routine value indicators of male rats

(4) Vital organ index

There was no significant difference in the organ index between the female rat administration group and the male rat administration group compared with the control group. The specific results are shown in Table 16 and Table 17. From the above situation, it can be seen that after the Tibetan medicine composition extract powder was administered to female and male rats at the maximum dosage once, there was no abnormality in the organ index indicators.

Table 16 Female rat organ index index (mg/g)

Table 17 Male rat organ index index (mg/g)

8.5 Summary

The present invention evaluates the systemic and organ toxic effects of the Tibetan medicine compound of the present invention based on indicators such as rat weight growth rate, vital organ index, hematology, and blood biochemistry. The results showed that a single large dose of Tibetan medicine composition extract suspension had a short-term and slight slowing effect on the weight gain of male and female rats, and gradually returned to normal after one day; while the heart, liver, spleen, lung, kidney, brain and other organ indexes, blood The blood chemistry and blood biochemistry indicators showed no obvious abnormalities, and no clear tissue damage was found during pathological anatomy. This shows that the maximum single dose of the Tibetan medicine composition of the present invention is 30g/kg, and no obvious toxicity symptoms were observed during the entire test period. This amount is equivalent to 292 times the intended clinical dose.

Claims (9)

1. A Tibetan medicine composition for treating chronic obstructive pneumonia, characterized in that the composition is made of the following raw materials by weight: 120-160 parts of Gentian flower, 50-55 parts of Qinpi, and Rabbit grass. 50-55 parts, nutmeg 19-25 parts, travertine 50-60 parts, slag paste 20-30 parts, licorice 75-85 parts. 2. The Tibetan traditional Chinese medicine composition for treating chronic obstructive pneumonia according to claim 1, characterized in that the composition is made of the following raw materials by weight: 140-160 parts of Gentian flower, 50-50 parts of Qinpi. 55 parts, rabbit ears 50-55 parts, nutmeg 19-23 parts, travertine 50-55 parts, slag paste 26-30 parts, licorice 75-79 parts. 3. The Tibetan medicine composition for treating chronic obstructive pneumonia according to claim 1 or 2, characterized in that the composition is prepared into a Tibetan medicine compound preparation by adding pharmaceutically acceptable auxiliary materials. 4. The Tibetan medicine composition for treating chronic obstructive pneumonia according to claim 3, characterized in that the preparation is one of granules, tablets and capsules. 5. A method for preparing a Tibetan medicinal composition for treating COPD according to claim 1 or 2, characterized in that the method includes the following steps: taking gentian flower, chinensis bark, rabbit ear grass, and nutmeg in proportion, Licorice is soaked in water, boiled or heated under reflux to extract, filter, and the filtrate is concentrated to obtain a concentrated extract, which is then mixed with the slag paste and then alcohol precipitated. The supernatant is concentrated, then combined with travertine, dried, crushed, and sieved. , get extract powder. 6. The preparation method of the Tibetan medicine composition for treating chronic obstructive pulmonary disease according to claim 5, characterized in that the soaking time of adding water is 30-60 minutes. 7. The preparation method of the Tibetan medicine composition for treating chronic obstructive pulmonary disease according to claim 5, characterized in that the decoction or heating and reflux extraction are performed 2-3 times, and each decoction or extraction takes 1-2 hours. The amount of water used for each decoction or extraction is 8-14 times the weight of the medicinal materials. 8. The preparation method of the Tibetan medicinal composition for treating chronic obstructive pulmonary disease according to claim 5, characterized in that the alcohol precipitation involves adding 3-4 times the volume of 95% ethanol, and the alcohol precipitation time is 24 hours. 9. Application of the Tibetan medicine composition according to claim 1 or 2 in the preparation of medicine for the treatment of chronic obstructive pneumonia.