Preparation and application of a Tibetan medicine Mabosong Decoction for Lung Nourishing and its modern formulation

Abstract

This invention discloses a Tibetan medicine formula called Mabosongtang Lung-Nourishing Composition and its preparation method and application in modern formulations, more specifically relating to a preparation method and application of Mabosongtang Lung-Nourishing granules, oral liquid, and capsules. The composition is mainly prepared from the following medicinal materials: Tibetan Lithospermum erythrorhizon, Tibetan Rubia cordifolia, Lithospermum erythrorhizon, Bambusa textilis, Morinda officinalis, Lysimachia christinae, Hippophae rhamnoides, and Glycyrrhiza uralensis. The aqueous extract of this composition, with appropriate excipients, is prepared into granules, oral liquid, and capsules using modern pharmaceutical processes. The three formulations are effective, rationally formulated, advanced in dosage form, and stable in quality. They have the effects of calming the lungs, suppressing blood stasis, and clearing lung heat. They can be used for respiratory symptoms caused by the invasion of pathogenic factors into the lungs, and dysfunction of the lung's vital energy and ascending qi, such as cough, wheezing, shortness of breath, etc. (treatment of lung diseases with the above symptoms).

Description

Technical Field

[0001] This invention belongs to the field of Tibetan medicine, specifically relating to a Tibetan medicine Mabosongtang Feining composition and its modern preparation method and application, and more specifically relating to a Tibetan medicine Mabosongtang Feining granules, oral liquid and capsules preparation method and application. Background Technology

[0002] The basic formula of "Mabosong Decoction" is recorded in various ancient Tibetan medical books. It has a long history of use and rich literature in the Tibetan plateau region of my country, and is one of the main components of the "Mabosong Decoction" series of decoctions. Ancient books such as "Jingzhu Materia Medica" record that it can clear the lungs, reduce inflammation, and stop coughing, and is used to treat lung diseases. "Tibetan Medicine Standards" records that the ingredients are crushed into coarse powder, sieved, mixed, and then decocted in water to make a decoction. The daily dose is 6-9 g, which can clear the lungs, reduce inflammation, and stop coughing, and is used for pulmonary congestion, cough, back pain, hoarseness, dry mouth, etc. However, traditional powders often have a large daily dosage, which is inconvenient to take and has poor patient compliance. To inherit and promote traditional Tibetan medicine, after consulting a large amount of relevant literature, based on Tibetan medical compatibility theory, summarizing the experience of adding or subtracting herbs according to symptoms, and referring to classic prescriptions of Tibetan medical experts throughout history, and after a large number of prescription screenings and years of trials and use, a prescription was finally discovered to have unique efficacy for various lung diseases (acute lung injury, pulmonary interstitial fibrosis, and chronic obstructive pulmonary disease). This prescription was developed by adding five herbs—Bamboo Scutellaria, Horse Root, Winged Sprout, Sea Buckthorn, and Licorice—to the original formula, forming the "Mabosong Decoction Lung-Nourishing Composition." Combined with modern pharmaceutical technology, new products with definite efficacy, reasonable formulation, advanced dosage form, and stable quality, such as "Mabosong Decoction Lung-Nourishing Granules," "Mabosong Decoction Lung-Nourishing Oral Liquid," and "Mabosong Decoction Lung-Nourishing Capsules," were developed. Summary of the Invention

[0003] The purpose of this invention is to provide a Tibetan medicine formula called Mabosongtang Lung-Nourishing Composition and its preparation method. Specifically, it includes Mabosongtang Lung-Nourishing Granules, Mabosongtang Lung-Nourishing Oral Liquid, and Mabosongtang Lung-Nourishing Capsules. This combination and its preparation overcome the drawbacks of traditional decoctions. Years of trials have shown that it is effective in treating respiratory symptoms caused by the invasion of pathogenic factors into the lungs, and functional disorders of the Weiminglong and ascending lung systems, such as cough, wheezing, and shortness of breath caused by lung heat, lung dilation, pulmonary dehydration, and dry cough (as seen in the treatment of lung diseases with the above symptoms).

[0004] To achieve the objectives of this invention, the following implementation scheme is provided.

[0005] In one embodiment, the Tibetan medicine Mabosong Decoction Lung-Nourishing Composition of the present invention is mainly prepared from the following medicinal materials in parts by weight: 10-25 parts of Tibetan Lithospermum erythrorhizon, 10-25 parts of Tibetan Rubia cordifolia, 5-20 parts of Lithospermum erythrorhizon, 5-15 parts of Bambusa textilis, 3-15 parts of Morinda officinalis, 3-13 parts of Lysimachia christinae, 2-11 parts of Hippophae rhamnoides, and 2-8 parts of Glycyrrhiza uralensis.

[0006] In a preferred embodiment, the Tibetan medicine Mabosong Decoction Lung-Nourishing Composition of the present invention is mainly prepared from the following medicinal materials in parts by weight: 10 parts of Tibetan Lithospermum erythrorhizon, 10 parts of Tibetan Rubia cordifolia, 8 parts of Lithospermum erythrorhizon, 5 parts of Bambusa textilis, 4 parts of Morinda officinalis, 3 parts of Lysimachia christinae, 2 parts of Hippophae rhamnoides, and 2 parts of Glycyrrhiza uralensis.

[0007] The Tibetan medicine Mabosong Decoction Lung-Nourishing Composition of the present invention is characterized in that the composition exists in a suitable pharmaceutical preparation form, wherein the preparation form is selected from tablets, capsules, oral liquids, mixtures, lozenges, granules, powders, pills, powders, ointments, elixirs, suspensions, solutions, injections, powder injections, lyophilized powder injections, sprays, spray films, drop pills, drop pills and / or patches, wherein the tablets are selected from sugar-coated tablets, film-coated tablets, enteric-coated tablets or sustained-release tablets; and the capsules are selected from hard capsules, soft capsules or sustained-release capsules.

[0008] More preferably, the Tibetan medicine Mabosong Decoction for Lung Nourishing described above is preferably formulated as granules, oral liquid, or capsules. Each granule sachet weighs 5g and contains no less than 46.0 mg of total quinones, no less than 5.2 mg of isovaleroyl shikonin, and no less than 3.0 mg of alizarin. Each oral liquid bottle contains 20 mL and contains no less than 69.0 mg of total quinones, no less than 7.1 mg of isovaleroyl shikonin, and no less than 4.2 mg of alizarin. Each capsule weighs 0.5g and contains no less than 2.38 mg of total quinones, no less than 0.27 mg of isovaleroyl shikonin, and no less than 0.17 mg of alizarin.

[0009] The present invention also provides a method for preparing Tibetan medicine Mabosongtang Lung-Nourishing Composition or its granules or its oral liquid or its capsules, comprising the following steps:

[0010] 1) Clean, dry, cut into pieces, and grind into coarse powder the roots and rhizomes of the following herbs: gromwell root, madder root and rhizome, horseradish rhizome, whole herb of winged head grass, and licorice root.

[0011] 2) Sea buckthorn fruit, gromwell root, and bamboo shavings are dried and then directly pulverized into coarse powder;

[0012] 3) Weigh the dried medicinal materials into coarse powder according to the formula ratio;

[0013] 4) Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water to extract, the material-to-liquid ratio is 1:10, decoct for 2 hours each time, filter to obtain filtrate, decoct three times, and combine the three extracts.

[0014] 5) Concentrate the filtrate under reduced pressure at a temperature not exceeding 60°C. After concentrating into an extract, freeze-dry it into a powder and store it for later use.

[0015] 6) Take the freeze-dried powder of the above extract and prepare granules by wet granulation: select dextrin and lactose as excipients, the preferred ratio of dextrin to lactose is 1.5:1, and the preferred ratio of excipient to extract is 3:1. After the two are mixed evenly, add 75% ethanol to make a soft mass, sieve and granulate, granulate, dry at 60-65℃, and bag to obtain Mabaosongtang Feining granules.

[0016] 7) Take the freeze-dried powder of the above extract, dissolve it in 8-10 times the amount of distilled water, refrigerate for 24 hours, centrifuge and filter, add 10% xylitol as flavoring agent, 1% sodium benzoate as preservative, and 1% sodium thiosulfate as antioxidant, mix evenly, filter, fill and seal, sterilize to obtain Mabaosongtang Feining oral liquid.

[0017] 8) Take the freeze-dried powder of the above extract, mix it with poloxamer 188 and sodium dodecyl sulfate at a ratio of 1:6:1 to prepare a solid dispersion, and fill it into hard capsules to make Demabaosongtangfeining capsules.

[0018] The following is an explanation of the Tibetan medicine Mabosong Decoction Lung-Nourishing Composition of the present invention:

[0019] Tibetan medicine has made significant contributions to the treatment of lung diseases. As early as the Four Medical Tantras, various symptoms and treatments for lung ailments were recorded. Tibetan medicine considers the lungs, a ministerial organ located high in the chest cavity, to be the repository of the essence of the wind (qi) element among the five elements, and thus hot in nature. When pathogenic factors cause imbalances in the tribal system, the hot, sharp, light, foul, purgative, damp, and greasy properties of tribal system manifest, leading to heat-related symptoms such as sudden and changeable onset, high body temperature, fever, excessive phlegm, and a strong, foul odor in phlegm, sweat, and urine. The lungs' functions of respiration and voice are closely related to the vital energy (vimalung) and ascending energy (rung). The energy of rung, being light and dynamic, becomes an aid in burning away the body's essence when exposed to the heat of tribal system. Therefore, when tribal pathogens invade the lungs, the functions of vimalung and ascending energy become disordered, leading to respiratory symptoms such as coughing, sneezing, hoarseness, wheezing, and shortness of breath. Therefore, Tibetan medicine believes that lung diseases are mainly caused by the disorder of "Chiba and Longxue", and classifies lung diseases as heat syndromes.

[0020] Tibetan medicine has a long history of treating lung heat. Its medications primarily consist of heat-clearing herbs, heat-clearing decoctions, heat-clearing powders, and antipyretic pastes, or purgative formulas to clear heat from the lungs. Judging from the taste and properties of the herbs used, they are mostly sweet, bitter, or astringent. The Mabosong Decoction for Lung-Clearing of this invention, while clearing heat from the lungs, can also stop the spread of blood stasis, thus clearing lung heat and improving symptoms of various lung diseases. The formula includes *Gynostemma pentaphyllum* (Tibetan medicine name: Zhemo), recorded in the 8th-century Tibetan medical classic *Four Tantras*, which is sweet, slightly bitter, and cool in nature. It is described in *Crystal Pearl Materia Medica*, *Tara Materia Medica*, *Blue Glazed Tile*, and *Mirror of Ambrosia Materia Medica* as having the ability to clear heat, cool the blood, nourish the lungs, and treat lung diseases, and is included in the Yunnan Provincial Standards for Traditional Chinese Medicine Decoction Pieces (2005 edition); *Rubia cordifolia* (Tibetan medicine name: Zuo), recorded in *Four Tantras* and *Crystal Pearl Materia Medica* as bitter, astringent, and cold in nature, capable of clearing heat and detoxifying, promoting blood circulation and removing blood stasis, and treating lung heat syndromes, is included in the *Qinghai Provincial Standards for Tibetan Medicine Processing* (2010 edition); and *Gynostemma pentaphyllum* (Tibetan medicine name: Jiaji), mentioned in *Blue Glazed Tile*... Records in medical texts such as *Ganlu Bencao Mingjing* and *Jingzhu Bencao* indicate that it is bitter and astringent, and cool in nature. It is used to clear heat and detoxify, invigorate blood circulation and remove blood stasis, and is primarily used to treat heat-related illnesses in the lungs. It is included in several provincial-level processing standards, such as the 2010 edition of *Qinghai Province Tibetan Medicine Processing Standards*. Tianzhuhuang (Tibetan medicine name: Niujugang) has a history of over 1000 years and is a classic Tibetan medicine for treating pneumonia and various lung diseases. The *Tibetan Medicine Standards* records that it "clears heat and nourishes the lungs, used to treat various lung heat diseases." It is cool in nature and has the effects of clearing heat and detoxifying, resolving phlegm and relieving cough, and calming the nerves. It is also included in the 2020 edition of the *Pharmacopoeia of the People's Republic of China*. Gaoshan horseradish (Tibetan medicine name: Suoluogabao) Included in the 2020 edition of the "Qinghai Province Tibetan Medicine Processing Standards," it is sweet and astringent in taste, and cool in nature. It can clear heat and stop bleeding, and is used for lung heat, turbid heat, and wound bleeding. *Bing Shou Cao* (Tibetan name: Bangziduowo), recorded in the "Four Medical Classics" and "Crystal Pearl Materia Medica," is bitter and cold in nature, with the effects of clearing heat and detoxifying. It is mainly used to treat epidemic diseases, heart disease, and blood disease, and is included in the 2020 edition of the "Pharmacopoeia of the People's Republic of China." *Sea buckthorn* (Tibetan name: Dabu), recorded... Recorded in ancient books such as "Jingzhu Materia Medica", it is sour in taste, cool in nature, sharp and light. It can resolve phlegm, promote digestion and blood circulation. It is mainly used to treat lung diseases, throat diseases, pharyngitis, lung and intestinal tumors, indigestion, etc. It is included in the "Chinese Pharmacopoeia 2020 Edition"; Licorice (Tibetan name: Xiang'er), recorded in ancient books such as "Jingzhu Materia Medica" and "Blue Glass", is sweet in taste and cool in nature. It can clear lung heat, detoxify, and treat wounds, inflammation, heat toxins, and epidemic diseases. It is included in the "Chinese Pharmacopoeia 2020 Edition" and is also a commonly used Chinese medicine.The above eight herbs all have the effects of "clearing heat and detoxifying, promoting blood circulation and removing blood stasis, treating lung heat evil or resisting epidemic diseases". Mabosong Decoction, which is based on "Tibetan purple grass-Tibetan madder-purple grass", is often used as an important component in Tibetan medicine prescriptions for treating lung diseases. Based on this, according to clinical experience and Tibetan medicine compatibility theory, five additional herbs were selected: Tianzhuhuang, alpine horseradish, winged head grass, sea buckthorn and licorice. A total of eight herbs were used to prepare Mabosong Decoction Lung-Nourishing Compound Granules, Oral Liquid and Capsules for treating various lung diseases caused by "Chiba and Longxue" disorders.

[0021] Background of use: The Mabaosong Decoction for Lung Health Combination, Mabaosong Decoction for Lung Health Granules, Mabaosong Decoction for Lung Health Oral Liquid, and Mabaosong Decoction for Lung Health Capsules are used for respiratory symptoms caused by the invasion of pathogenic factors into the lungs and the dysfunction of the Weiminglong and Shangxinglong meridians, such as cough, wheezing, shortness of breath, etc. caused by lung heat, lung dilatation, lung dehydration, and dry cough of the Long type (see the treatment of lung diseases with the above symptoms). It can be used for various lung diseases such as acute lung injury, pulmonary interstitial fibrosis, and chronic obstructive pulmonary disease.

[0022] In one specific embodiment, the preparation method of the Mabaosong Decoction Lung-Nourishing Granules of the present invention includes the following steps:

[0023] 1) Raw material pretreatment: Wash the roots and rhizomes of Tibetan purple gromwell, Tibetan madder, alpine horseradish, whole plant of winged head grass, licorice root, and sea buckthorn fruit with clean water, dry them at 60~65 ℃, cut them into pieces and crush them into coarse powder; also dry purple gromwell root and bamboo yellow at 60~65 ℃ and crush them into coarse powder.

[0024] 2) Formula: Weigh the coarse powder of medicinal materials according to the above standard formula ratio and prepare the mixture.

[0025] 3) Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water, the material-to-liquid ratio is 1:10, decoct for 2 hours each time, filter to obtain filtrate, decoct three times, and combine the three extracts.

[0026] 4) Concentration and freeze-drying: Concentrate the filtrate under reduced pressure at a temperature not exceeding 60°C. After concentrating into an extract, freeze-dry it into a powder and store it under cold for later use.

[0027] 5) Mixing: Take the freeze-dried powder of the above extract and granulate it by wet granulation to prepare granules: select dextrin and lactose as excipients, the preferred ratio of dextrin to lactose is 1.5:1, and the preferred ratio of excipient to extract is 3:1. After the two are mixed evenly, add 75% ethanol to make soft material.

[0028] 6) Granulation: The above soft material is extruded and granulated using a gyratory pellet mill, then sieved and sized.

[0029] 7) Drying: After granulation, dry at 60-65℃, turning the granules frequently during the drying process to prevent clumping. After drying, pass the granules through a 20-mesh sieve and a 60-mesh sieve to remove coarse particles and fine powder. After separating the coarse particles, add fine powder and granulate again using the above method. After drying, mix the granules together and package them accordingly.

[0030] The above method is used to prepare Mabaosong Decoction Lung-Nourishing Granules, 5g / bag, each bag containing not less than 46.0 mg of total quinones, not less than 5.2 mg of isovaleryl shikonin, and not less than 3.0 mg of alizarin.

[0031] The indications for the use of Mabaosong Decoction Lung-Nourishing Granules of this invention are as follows:

[0032] It has the effects of calming the blood, suppressing phlegm, and clearing lung heat. It is used for respiratory symptoms caused by the invasion of pathogenic factors into the lungs and the dysfunction of the lungs' vital energy and ascending phlegm, such as cough, wheezing, shortness of breath, etc. caused by lung heat, lung dilatation, lung dehydration, and dry cough (treatment of lung diseases with the above symptoms).

[0033] Specifications: 5g / bag.

[0034] Dosage and administration: Oral administration. Take one sachet at a time, 1-3 times a day, dissolved in warm water.

[0035] Adverse reactions: Not yet clear.

[0036] This product consists of brown granules with a slightly bitter taste.

[0037] In one specific embodiment, the preparation method of the present invention, Mabaosong Decoction for Lung Nourishing Oral Solution, includes the following steps:

[0038] 1) Raw material pretreatment: Wash the roots and rhizomes of Tibetan purple gromwell, Tibetan madder, alpine horseradish, whole plant of winged head grass, licorice root, and sea buckthorn fruit with clean water, dry them at 60~65 ℃, cut them into pieces and crush them into coarse powder; also dry purple gromwell root and bamboo yellow at 60~65 ℃ and crush them into coarse powder.

[0039] 2) Formula: Weigh the coarse powder of medicinal materials according to the above standard formula ratio and prepare the mixture.

[0040] 3) Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water, the material-to-liquid ratio is 1:10, decoct for 2 hours each time, filter to obtain filtrate, decoct three times, and combine the three extracts.

[0041] 4) Concentration and freeze-drying: Concentrate the filtrate under reduced pressure at a temperature not exceeding 60°C. After concentrating into an extract, freeze-dry it into a powder and store it under cold for later use.

[0042] 5) Dissolving: Take the freeze-dried powder of the above extract, add 10 times the amount of distilled water to dissolve it, refrigerate for 24 hours, centrifuge and filter, and replenish the lost distilled water to 10 times the amount.

[0043] 6) Mixing: Add 10% xylitol as a flavoring agent, 1% sodium benzoate as a preservative, and 1% sodium thiosulfate as an antioxidant. Mix well and filter to obtain the filtrate.

[0044] 7) Dispensing: Take the above filtrate, dispense it, fill and seal it, and sterilize it to obtain the final product.

[0045] The above method is used to prepare Mabaosong Tangfei Ning oral liquid, 20 mL / bottle, each bottle containing not less than 69.0 mg of total quinone, not less than 7.1 mg / bottle of isovaleryl shikonin, and not less than 4.2 mg / bottle of alizarin.

[0046] The indications for the use of the Mabaosong Decoction for Lung Nourishing Oral Solution of this invention are as follows:

[0047] It has the effects of calming the blood, suppressing phlegm, and clearing lung heat. It is used for respiratory symptoms caused by the invasion of pathogenic factors into the lungs and the dysfunction of the lungs' vital energy and ascending phlegm, such as cough, wheezing, and shortness of breath caused by lung heat, lung dilatation, lung dehydration, and dry cough (treatment of lung diseases with the above symptoms).

[0048] Specification: 20 mL / bottle.

[0049] Dosage and administration: Oral administration. One bottle each time, 1-3 times a day.

[0050] Adverse reactions: Not yet clear.

[0051] This product is a brownish-brown liquid with a slightly bitter taste.

[0052] In one specific embodiment, the preparation method of the Mabaosong Decoction for Lung Nourishing Capsules of the present invention includes the following steps:

[0053] 1) Raw material pretreatment: Wash the roots and rhizomes of Tibetan purple gromwell, Tibetan madder, alpine horseradish, whole plant of winged head grass, licorice root, and sea buckthorn fruit with clean water, dry them at 60~65 ℃, cut them into pieces and crush them into coarse powder; also dry purple gromwell root and bamboo yellow at 60~65 ℃ and crush them into coarse powder.

[0054] 2) Formula: Weigh the coarse powder of medicinal materials according to the above standard formula ratio and prepare the mixture.

[0055] 3) Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water, the material-to-liquid ratio is 1:10, decoct for 2 hours each time, filter to obtain filtrate, decoct three times, and combine the three extracts.

[0056] 4) Concentration and freeze-drying: Concentrate the filtrate under reduced pressure at a temperature not exceeding 60°C. After concentrating into an extract, freeze-dry it into a powder and store it under cold for later use.

[0057] 5) Preparation of solid dispersion: Take the freeze-dried powder of the above extract and mix it with poloxamer 188 and sodium dodecyl sulfate at a ratio of 1:6:1.

[0058] 6) Packaging: Prepare a solid dispersion as described above, and fill it into hard capsules to obtain the final product.

[0059] The above method is used to prepare Mabaosong Tangfei Ning capsules, 0.5 g / capsule, each capsule containing not less than 2.38 mg of total quinones, not less than 0.27 mg / capsule of isovaleryl shikonin, and not less than 0.17 mg / capsule of alizarin.

[0060] The indications for the use of the Mabaosong Decoction Lung-Nourishing Capsules of this invention are as follows:

[0061] It has the effects of calming the blood, suppressing phlegm, and clearing lung heat. It is used for respiratory symptoms caused by the invasion of pathogenic factors into the lungs and the dysfunction of the lungs' vital energy and ascending phlegm, such as cough, wheezing, and shortness of breath caused by lung heat, lung dilatation, lung dehydration, and dry cough (treatment of lung diseases with the above symptoms).

[0062] Specification: 0.5 g / capsule.

[0063] Dosage and administration: Oral administration. Take 3 capsules each time, 1 to 3 times a day.

[0064] Adverse reactions: Not yet clear.

[0065] This product is a brownish-brown powder encapsulated.

[0066] The original dosage forms of "Mabosongtang Feining Granules, Mabosongtang Feining Oral Liquid, and Mabosongtang Feining Capsules" were decoctions. However, decoctions have problems such as unstable dosage, inconvenience in taking them, and difficulty in carrying them. Therefore, based on Tibetan medicine theory and using modern pharmaceutical equipment, the inventors have changed the original decoctions into granules, oral liquids, and capsules. They have improved the requirements for particle size, moisture, solubility, and microorganisms in the original prescriptions, and formulated new Tibetan medicine products with controllable dosage, stable process, and feasible production. Attached Figure Description

[0067] Figure 1 Summary Table of Three Batches of Data from the Chinese Experiment of Mabaosong Decoction for Lung Nourishing Granules

[0068] Figure 2 Test results of Mabaosong Decoction for Lung Nourishing Granules

[0069] Figure 3 Table of Content Difference in Mabaosong Decoction and Lung-Nourishing Granules

[0070] Figure 4 Results of content determination of Mabaosong Decoction for Lung Nourishing Granules

[0071] Figure 5 Summary Table of Data from Three Batches of Chinese Experiments on Mabaosong Decoction for Lung Nourishing Oral Liquid

[0072] Figure 6 : Fill volume detection of Mabaosongtang Lung-Nourishing Oral Liquid

[0073] Figure 7 Results of content determination of Ma Baosong Decoction for Lung Nourishing Oral Liquid

[0074] Figure 8 Summary Table of Data from Three Batches of Chinese Experiments on Mabaosong Decoction for Lung Nourishing Capsules

[0075] Figure 9 Table of Content Weight Difference for Mabaosong Decoction Lung-Nourishing Capsules

[0076] Figure 10 Test results of Mabaosong Decoction for Lung Nourishing Capsules

[0077] Figure 11 Content determination results of Mabaosong Decoction for Lung Nourishing Capsules

[0078] Figure 12 Effects of various Tibetan medicine compositions on the wet-to-dry weight ratio of lungs in ALI mice (compared to the blank group, #) P <0.05, ## P <0.01; compared with the model group, * P <0.05,** P <0.01)

[0079] Figure 13 Effects of various Tibetan medicine compositions on MPO activity in the lungs of ALI mice (compared with the blank group, #) P <0.05, ## P <0.01; compared with the model group, * P <0.05,** P <0.01)

[0080] Figure 14 Effects of various Tibetan medicine compositions on lung tissue pathology (H&E staining) in ALI mice

[0081] Figure 15 Effects of various Tibetan medicine compositions on the levels of TNF-α (A), IL-1β (B), IL-6 (C), IL-10 (D) and total protein (E) in BALF of ALI mice (compared with the blank group, #) P <0.05, ## P <0.01; compared with the model group, * P <0.05,** P <0.01)

[0082] Figure 16Effects of various Tibetan medicine compositions on body weight changes in mice with pulmonary fibrosis (compared to the control group, # P <0.05, ## P <0.01; compared with the model group, * P <0.05,** P <0.01)

[0083] Figure 17 Effects of various Tibetan medicine compositions on lung index in mice with pulmonary fibrosis (where, compared with the blank group, # P <0.05, ## P <0.01; compared with the model group, * P <0.05,** P <0.01)

[0084] Figure 18 Effects of various Tibetan medicine compositions on lung tissue pathology (H&E staining) in mice with pulmonary fibrosis

[0085] Figure 19 Effects of various Tibetan medicine combinations on lung tissue pathology (Masson staining) in mice with pulmonary fibrosis

[0086] Figure 20 Effects of various Tibetan medicine compositions on hydroxyproline content in the lungs of mice with pulmonary fibrosis (compared with the blank group, # P <0.05, ## P <0.01; compared with the model group, * P <0.05,** P <0.01)

[0087] Figure 21 Effects of various Tibetan medicine compositions on lung function in COPD mice (comparison with blank control group, #) P <0.05, ## P <0.01, ### P <0.001, #### P <0.0001; compared with the model group, * P <0.05,** P <0.01, *** P <0.001, **** P <0.0001)

[0088] Figure 22 Effects of various Tibetan medicine combinations on lung tissue pathology (H&E staining) in COPD mice

[0089] Figure 23 Effects of various Tibetan medicine compositions on serum TNF-α (A), IL-1β (B), and IL-6 (C) levels in COPD mice (compared to blank control group, #)P <0.05, ## P <0.01, ### P <0.001, #### P <0.0001; compared with the model group, * P <0.05,** P <0.01, *** P <0.001, **** P <0.0001) Detailed Implementation

[0090] Example 1: Preparation of Mabaosong Decoction and Lung-Nourishing Combination

[0091] 1. Prescription composition: 10-25 parts of Tibetan gromwell root, 10-25 parts of Tibetan madder root, 5-20 parts of gromwell root, 5-15 parts of bamboo shavings, 3-15 parts of alpine horseradish, 3-13 parts of winged head grass, 2-11 parts of sea buckthorn, and 2-8 parts of licorice.

[0092] 2. Preparation method:

[0093] 1) Raw material pretreatment: Wash the roots and rhizomes of Lithospermum erythrorhizon, Rubia cordifolia, Moringa oleifera, whole herb of Lysimachia christinae, Glycyrrhiza uralensis, and Hippophae rhamnoides with clean water, dry them at 60-65 ℃, cut them into pieces and grind them into coarse powder; Lithospermum erythrorhizon and Bambusa textilis are also dried at 60-65 ℃ and ground into coarse powder.

[0094] 2) Formula: Weigh the coarse powder of medicinal materials according to the above standard formula ratio, and mix them together to obtain the final product.

[0095] Example 2: Preparation and quality evaluation of Mabaosong Decoction for Lung Nourishing Granules

[0096] 1. Dosage Form Selection

[0097] The original dosage form of "Mabosong Tangfei Ning Granules" was a decoction powder, which required pulverization and decoction under normal pressure. This formula had been used in clinical trials as a decoction for many years with definite efficacy, but it suffered from poor taste, inconvenience in carrying and taking, and easy loss of active ingredients, making it unsuitable for modern medical needs. Therefore, to develop it into a Tibetan medicine preparation that meets modern medical requirements, the original Tibetan medicine formulation process was retained to avoid affecting the efficacy of the active ingredients. Herbs with similar preparation methods to traditional decoctions were used to extract extracts, which were then freeze-dried to obtain freeze-dried powder. This powder was then mixed with a certain proportion of dextrin and lactose to form granules. Granules offer numerous advantages, including ease of administration, rapid dissolution, rapid drug release, relatively simple production process, convenient storage, easy carrying, and accurate dosage. Ultimately, the original dosage form of this formula, a decoction powder, was transformed into granules suitable for modern pharmaceutical needs.

[0098] Experiments have shown that changing this formula to a solid, fast-dissolving granule form makes it convenient to take. This dosage form is significantly more effective, better absorbed, and allows the medicinal effect to be fully realized. Dosage is also easier to control, and the solid dosage form facilitates drug storage.

[0099] 2. Granule Preparation

[0100] 1) Raw material pretreatment: Wash the roots and rhizomes of Tibetan purple gromwell, Tibetan madder, alpine horseradish, whole plant of winged head grass, licorice root, and sea buckthorn fruit with clean water, dry them at 60~65 ℃, cut them into pieces and crush them into coarse powder; also dry purple gromwell root and bamboo yellow at 60~65 ℃ and crush them into coarse powder.

[0101] 2) Formula: Weigh out the coarse powder of the medicinal materials according to the above-mentioned preferred formula ratio (10 parts of Tibetan gromwell, 10 parts of Tibetan madder, 8 parts of gromwell root, 5 parts of bamboo shavings, 4 parts of alpine horseradish, 3 parts of winged head grass, 2 parts of sea buckthorn, and 2 parts of licorice) and prepare the formula.

[0102] 3) Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water, the material-to-liquid ratio is 1:10, decoct for 2 hours each time, filter to obtain filtrate, decoct three times, and combine the three extracts.

[0103] 4) Concentration and freeze-drying: Concentrate the filtrate under reduced pressure at a temperature not exceeding 60°C. After concentrating into an extract, freeze-dry it into a powder and store it under cold for later use.

[0104] 5) Mixing: Take the freeze-dried powder of the above extract and granulate it by wet granulation to prepare granules: select dextrin and lactose as excipients, the preferred ratio of dextrin to lactose is 1.5:1, and the preferred ratio of excipient to extract is 3:1. After the two are mixed evenly, add 75% ethanol to make a soft material.

[0105] 6) Granulation: The above soft material is extruded and granulated using a gyratory pellet mill, then sieved and sized.

[0106] 7) Drying: After granulation, dry at 60-65℃, turning the granules frequently during the drying process to prevent clumping. After drying, pass the granules through a 20-mesh sieve and a 60-mesh sieve to remove coarse particles and fine powder. After separating the coarse particles, add the fine powder and granulate again according to the above method. After drying, mix the granules together and package them (5g / bag).

[0107] 3. Process stability study

[0108] The above prescription dosage was increased tenfold, with 10 kg of medicinal materials used per batch. Three batches of samples were prepared, and granules were produced and packaged according to the above process. The production quality met the national pharmacopoeia standards in key stages such as pulverization, extraction, concentration, freeze-drying, mixing, and granulation. Figure 1 This indicates that the production process has good stability. Each batch contains 2000 bags (5g / bag).

[0109] 4. Quality evaluation of granules

[0110] 1) Particle size: The total percentage of particles that cannot pass through a No. 1 sieve and those that can pass through a No. 5 sieve shall not exceed 15%, in accordance with the provisions for granules in the Pharmacopoeia of the People's Republic of China (0104). Figure 2 ).

[0111] 2) Moisture content: Not exceeding 8.0%, in accordance with the provisions of the Granules (0104) section of the Pharmacopoeia of the People's Republic of China. Figure 2 ).

[0112] 3) Solubility: Perform the test according to the provisions of the Granules (0104) section of the Pharmacopoeia of the People's Republic of China. Take one bag of the test sample, add 200 mL of hot water, stir for 5 minutes, and observe immediately. It should be completely dissolved or only slightly turbid. Figure 2 ).

[0113] 4) Content Weight Variation: The procedure shall be performed according to the provisions under Granules (0104) of the Pharmacopoeia of the People's Republic of China. Take 10 bags of the test sample, remove the packaging, and accurately weigh the contents of each bag. Compare the content of each bag with the labeled content; the content weight variation limit is ±7%. No more than two bags of granules shall exceed the content weight variation limit, and no single bag shall exceed the limit by more than 100%. See [link to relevant documentation]. Figure 3 .

[0114] 5) Microbiological limitations: As specified in the Pharmacopoeia of the People's Republic of China, Granules (0104). Figure 2 ).

[0115] 6) Content determination

[0116] Ten batches of Mabaosong Decoction for Lung Nourishing Granules, prepared according to the above pilot-scale process, were taken. The total quinone content in the granules was determined by ultraviolet spectrophotometry, and the content of isovaleryl shikonin and alizarin in the granules was determined by HPLC. Figure 4 ).

[0117] Figure 4 The results showed that the total quinone content in Mabaosong Tangfei Ning Granules ranged from 53.4 to 61.5 mg / sachet, with an average of 57.6 mg / sachet; the isovaleryl shikonin content ranged from 5.4 to 7.8 mg / sachet, with an average of 6.5 mg / sachet; and the alizarin content ranged from 3.1 to 4.5 mg / sachet, with an average of 3.8 mg / sachet. 80% of the average values ​​were tentatively set as the content limits, meaning that each sachet of this product should contain no less than 46.0 mg of total quinone, no less than 5.2 mg of isovaleryl shikonin, and no less than 3.0 mg of alizarin.

[0118] Example 3: Preparation and quality evaluation of Mabaosong Decoction for Lung Nourishing Oral Solution

[0119] 1. Dosage Form Selection

[0120] The original form of "Mabosong Tangfei Ning Oral Liquid" was a powder, requiring pulverization and decoction under normal pressure. While this formula had been used clinically for many years as a decoction with proven efficacy, its poor taste, inconvenience in carrying and taking it, and the easy loss of active ingredients made it unsuitable for modern medical needs. Therefore, it was researched and developed into a Tibetan medicine preparation that meets modern medical requirements. To avoid affecting the efficacy of the original active ingredients, the original Tibetan medicine formulation process was retained during the modification. Extracts were extracted from medicinal materials similar to those used in traditional decoctions, freeze-dried into freeze-dried powder, dissolved in a specific proportion of distilled water, and flavored with xylitol and sodium benzoate as preservatives. Considering that quinones are the main components of the formula and are easily oxidized, sodium thiosulfate was added as an antioxidant, ultimately producing an oral liquid. This oral liquid offers numerous advantages, including ease of administration, rapid drug release, relatively simple production process, convenient portability, and accurate dosage. Ultimately, the original dosage form of this formula, a decoction or powder, was changed into an oral liquid suitable for modern pharmaceutical needs.

[0121] Experiments have shown that this formula, when converted to an oral liquid, is convenient to take, has significant therapeutic effects, is more easily absorbed, and allows the medicinal effect to be fully exerted. Dosage is also easier to control, and the repackaging and sealing process facilitates drug storage.

[0122] 2. Preparation of oral liquid

[0123] 1) Raw material pretreatment: Wash the roots and rhizomes of Lithospermum erythrorhizon, Rubia cordifolia, Moringa oleifera, whole herb of Lysimachia christinae, Glycyrrhiza uralensis, and Hippophae rhamnoides with clean water, dry them at 60-65 ℃, cut them into pieces and grind them into coarse powder; Lithospermum erythrorhizon and Bambusa textilis are also dried at 60-65 ℃ and ground into coarse powder.

[0124] 2) Formula: Weigh out the coarse powder of the medicinal materials according to the above-mentioned preferred formula ratio (10 parts of Tibetan gromwell, 10 parts of Tibetan madder, 8 parts of gromwell root, 5 parts of bamboo shavings, 4 parts of alpine horseradish, 3 parts of winged head grass, 2 parts of sea buckthorn, and 2 parts of licorice) and prepare the formula.

[0125] 4) Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water, the ratio of material to liquid is 1:10, decoct for 2 hours each time, filter to obtain filtrate, decoct three times, and combine the three extracts.

[0126] 5) Concentration and freeze-drying: Concentrate the filtrate under reduced pressure at a temperature not exceeding 60°C. After concentrating into an extract, freeze-dry it into a powder and store it under cold for later use.

[0127] 6) Dissolving: Take the freeze-dried powder of the above extract, add 10 times the amount of distilled water to dissolve it, refrigerate for 24 hours, centrifuge and filter, and replenish the lost amount of distilled water to 10 times the amount.

[0128] 7) Mixing: Add 10% xylitol as a flavoring agent, 1% sodium benzoate as a preservative, and 1% sodium thiosulfate as an antioxidant. Mix well and filter to obtain the filtrate.

[0129] 8) Dispensing: Take the above filtrate, dispense, fill, and sterilize to obtain the final product.

[0130] 3. Process stability study

[0131] The above prescription dosage was scaled up 10 times, with 10 kg of medicinal materials used per batch. Three batches of samples were prepared, and the oral liquid was produced and packaged according to the above process. The production quality met the national pharmacopoeia standards in key stages such as pulverization, extraction, concentration, freeze-drying, and dissolution. This indicates that the production process has good stability. Each batch contains 1500 bottles (20 mL / bottle). Figure 5 ).

[0132] 4. Quality evaluation of oral liquid

[0133] 1) Relative density test: Performed in accordance with the provisions of General Chapter 0601 of the Pharmacopoeia of the People's Republic of China, with a density range of 1.05 to 1.09.

[0134] 2) pH value test: Performed in accordance with the provisions of General Chapter 0631 of the Pharmacopoeia of the People's Republic of China, with a pH range of 4.3 to 5.6.

[0135] 3) Filling volume inspection: Perform the inspection according to the provisions of General Chapter 0181, Compound Preparations, of the Pharmacopoeia of the People's Republic of China: Take 5 vials of the test sample, pour the contents into standardized graduated cylinders, and inspect at room temperature. Compare the fill volume of each vial with the labeled fill volume. No more than one vial should be less than the labeled fill volume, and the fill volume should not be less than 95% of the labeled fill volume (see...). Figure 6 ).

[0136] 4) Microbial limit testing: The testing shall be conducted in accordance with the provisions of General Chapter 0181, Compound Preparations, of the Pharmacopoeia of the People's Republic of China, and all tests shall comply with the provisions.

[0137] 5) Content determination

[0138] Ten batches of Mabaosong Decoction for Lung Nourishing Oral Solution, prepared according to the above pilot-scale process, were tested. The content of quinone components in the oral solution was determined by ultraviolet spectrophotometry, and the content of isovaleryl shikonin and alizarin in the oral solution was determined by HPLC. Figure 7 ).

[0139] Figure 7The results showed that the total quinone content in Mabaosong Tangfei Ning Oral Solution ranged from 83.2 to 88.4 mg / bottle, with an average of 86.2 mg / bottle; the isovaleryl shikonin content ranged from 7.7 to 10.5 mg / bottle, with an average of 8.9 mg / bottle; and the alizarin content ranged from 4.7 to 5.9 mg / sachet, with an average of 5.3 mg / bottle. 80% of the average values ​​were tentatively set as the content limits, meaning that each bottle of this product should contain no less than 69.0 mg of total quinone, no less than 7.1 mg of isovaleryl shikonin, and no less than 4.2 mg of alizarin.

[0140] Example 4: Preparation and quality evaluation of Mabaosong Decoction for Lung Nourishing Capsules

[0141] 1. Dosage Form Selection

[0142] The original dosage form of "Mabosong Tangfei Ning Capsules" was a decoction powder, which required pulverization and decoction under normal pressure. This formula had been used in clinical trials for many years with definite efficacy, but its poor taste, inconvenience in carrying and taking, and easy loss of active ingredients made it unsuitable for modern medical needs. Therefore, it was researched and developed into a Tibetan medicine preparation that meets modern medical requirements. To avoid affecting the efficacy of the active ingredients in the original preparation, the original Tibetan medicine formulation process was retained during the modification. Extracts were extracted from medicinal materials similar to those used in traditional decoction preparations, and the extracts were freeze-dried into freeze-dried powder. This powder was then mixed with a certain proportion of poloxamer 188 and sodium dodecyl sulfate to form a solid dispersion, which was then encapsulated in hard capsules. Solid dispersions have the advantages of increasing drug solubility and release rate. Encapsulation offers numerous advantages, including ease of administration, masking unpleasant odors, simple production process, convenient storage, easy carrying, and accurate dosage. Ultimately, the original dosage form of this formula, a decoction powder, was transformed into a capsule form suitable for modern pharmaceutical needs.

[0143] Experiments have shown that changing this formula to a solid dispersion capsule makes it convenient to take, significantly improves efficacy, enhances absorption, and allows the medicine to exert its full effect. Dosage is also easier to control, and the solid dosage form facilitates drug storage.

[0144] 2. Capsule Preparation

[0145] 1) Raw material pretreatment: Wash the roots and rhizomes of Tibetan purple gromwell, Tibetan madder, alpine horseradish, whole plant of winged head grass, licorice root, and sea buckthorn fruit with clean water, dry them at 60~65 ℃, cut them into pieces and crush them into coarse powder; also dry purple gromwell root and bamboo yellow at 60~65 ℃ and crush them into coarse powder.

[0146] 2) Formula: Weigh out the coarse powder of the medicinal materials according to the above-mentioned preferred formula ratio (10 parts of Tibetan gromwell, 10 parts of Tibetan madder, 8 parts of gromwell root, 5 parts of bamboo shavings, 4 parts of alpine horseradish, 3 parts of winged head grass, 2 parts of sea buckthorn, and 2 parts of licorice) and prepare the formula.

[0147] 4) Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water, the ratio of material to liquid is 1:10, decoct for 2 hours each time, filter to obtain filtrate, decoct three times, and combine the three extracts.

[0148] 5) Concentration and freeze-drying: Concentrate the filtrate under reduced pressure at a temperature not exceeding 60°C. After concentrating into an extract, freeze-dry it into a powder and store it in a refrigerator for later use.

[0149] 6) Preparation of solid dispersion: Take the freeze-dried powder of the above extract and mix it with poloxamer 188 and sodium dodecyl sulfate at a ratio of 1:6:1.

[0150] 7) Packaging: Prepare a solid dispersion as described above, and fill it into hard capsules to obtain the final product.

[0151] 3. Process stability study

[0152] The above prescription dosage was increased tenfold, with 10 kg of medicinal materials used per batch. Three batches of samples were prepared, and capsules were produced according to the above process. The production quality in key stages such as pulverization, extraction, concentration, freeze-drying, and mixing all met the national pharmacopoeia standards. Figure 8 This indicates that the production process has good stability. Each batch contains 46,000 capsules (0.5g / capsule).

[0153] 4. Quality evaluation of capsules

[0154] 1) Moisture content: Moisture content of traditional Chinese medicine hard capsules should be tested. Take the contents of the test sample and determine the moisture content according to the method for moisture determination (General Rule 0832). The moisture content should not exceed 9.0% and should meet the requirements.

[0155] 2) Content Weight Variation: This shall be performed in accordance with the provisions of General Chapter 0103, Capsules, of the Pharmacopoeia of the People's Republic of China. Take 10 capsules of the test sample, accurately weigh them separately, and pour out the contents (without damaging the capsule shell). Clean the hard capsule shells with a small brush or other suitable tool; then accurately weigh the capsule shells separately again, and calculate the content weight of each capsule and the average content weight. Compare the content weight of each capsule with the average content weight (for capsules with a labeled content weight, the content weight of each capsule should be compared with the labeled content weight). The content weight variation limit is ±10%. No more than 2 capsules may exceed the content weight variation limit, and no single capsule may exceed the limit by more than 100%. Figure 9 ).

[0156] 3) Disintegration time: According to Section 0921 of the General Principles of the Pharmacopoeia of the People's Republic of China (Disintegration Time Test Method), all conditions should meet the requirements. Figure 10 ).

[0157] 4) Microbial limits: The limits shall be implemented in accordance with the provisions of General Chapter 0103, Capsules, of the Pharmacopoeia of the People's Republic of China, and all shall comply with the provisions.

[0158] 5) Content determination

[0159] Ten batches of Mabaosong Decoction for Lung Nourishing were prepared according to the above pilot-scale process. The total quinone content in the granules was determined by ultraviolet spectrophotometry, and the content of isovaleryl shikonin and alizarin in the capsules was determined by HPLC. Figure 11 ).

[0160] Figure 11 The results showed that the total quinone content in Mabaosong Tangfei Ning Capsules ranged from 2.69 to 3.26 mg / capsule, with an average of 2.97 mg / sachet; the isovaleryl shikonin content ranged from 0.29 to 0.38 mg / capsule, with an average of 0.34 mg / capsule; and the alizarin content ranged from 0.17 to 0.25 mg / capsule, with an average of 0.21 mg / capsule. 80% of the average values ​​were tentatively set as the content limits, meaning that each capsule of this product should contain no less than 2.38 mg of total quinone, no less than 0.27 mg / capsule of isovaleryl shikonin, and no less than 0.17 mg / capsule of alizarin.

[0161] Example 5: Acute toxicity evaluation of the Mabaosong decoction and Lung-Nourishing composition

[0162] 1. Preparation of Mabaosong Decoction for Lung Nourishing:

[0163] 1) Raw material pretreatment: Wash the roots and rhizomes of Tibetan purple gromwell, Tibetan madder, alpine horseradish, whole plant of winged head grass, licorice root, and sea buckthorn fruit with clean water, dry them at 60~65 ℃, cut them into pieces and crush them into coarse powder; also dry purple gromwell root and bamboo yellow at 60~65 ℃ and crush them into coarse powder.

[0164] 2) Formula: Weigh the coarse powder of medicinal materials according to the above standard formula ratio and prepare the mixture.

[0165] 3) Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water at a ratio of 1:10, decoct for 2 hours each time, filter to obtain filtrate, decoct three times, and combine the three extracts. 4) Concentration and freeze-drying: Concentrate the filtrate under reduced pressure at a temperature not exceeding 60 ℃, concentrate into an extract, freeze-dry into powder, and store for later use.

[0166] 2. Median lethal dose determination

[0167] Twelve mice were fasted for 12 hours before the experiment was conducted, with four mice in each group (half male and half female). The highest dose group was determined to be greater than 5 g / kg, and the dose was reduced by three times. Three groups were set up (the drug extract was dissolved in warm physiological saline, and high, medium, and low dose groups were prepared, equivalent to 6 g / kg, 2 g / kg, and 0.6 g / kg body weight of raw drug). The preliminary experiment began. If no mice died, the number of mice in the highest dose group was increased to 10 (half male and half female), and the experiment was repeated twice. If no mice died, the experiment was stopped. The median lethal dose (LD50) was greater than 5 g / kg. If mice died, the Kohl's method was followed, and 5-7 experimental groups were set up according to the ratio of total lethal dose to zero lethal dose. The experiment was conducted sequentially until the dose at which all four mice died and the dose at which no mice died were found, and then the formal experiment was conducted.

[0168] Experimental Results: In the preliminary gavage test on the three groups of mice, no abnormalities were observed in their mental state, food and water intake, coat color, or fecal characteristics. Furthermore, when the number of mice in the highest dose group was increased to 10, the results were repeated twice, and the observed indicators were the same as in the preliminary test groups, with no deaths. Therefore, the LD50 of the Mabaosong Decoction Lung-Nourishing Extract was [value missing]. 50 Greater than 5 g / kg (body weight of raw medicinal material).

[0169] 3. Maximum tolerable dose determination

[0170] Twenty mice, half male and half female, adapted to the environment were housed separately and randomly divided into two groups: the Mabaosong Decoction for Lung Health Extract group and the blank control group, with 10 mice in each group. After fasting for 12 hours, the experiment was conducted. The drug extract was dissolved in warm physiological saline to prepare the solution. The test mice were administered the solution by gavage at a dose of 6 g / kg of the original drug every 4 hours for a total of 3 times, until the cumulative gavage dose reached 18 g / kg body weight. The blank control group was administered the same warm physiological saline solution at the same time. Observations were conducted for 4 hours after drug administration. Observations were conducted twice daily for seven consecutive days. The mice's diet, activity, and behavior were recorded for any abnormalities, and any deaths were recorded. If a death occurred, the mouse was promptly dissected to observe any abnormalities in the organs. The maximum tolerated dose of the Mabaosong Decoction for Lung Health Extract in the mice was calculated. After the 7-day observation period, all mice were euthanized, and the organs of the dissected mice were observed for any pathological changes.

[0171] Experimental results: On the same day, the high-dose group drug amount in the median lethal dose test (at the crude drug dose of 6 g / kg) was used to intubate mice 3 times, and they were observed for 7 days and weighed. It was found that there were no abnormalities in the mental state, food intake, water intake, hair color, fecal traits, etc. of the experimental mice in the drug group and the control group, and no deaths occurred. The weight changes of male and female mice were statistically analyzed respectively. The body weights of mice in each group increased significantly, and there was no significant difference in the weight gain of male and female mice between the drug group and the control group (P>0.05). The mice after drug administration were dissected, and no obvious pathological changes were found in each organ through macroscopic observation.

[0172] In summary, when the administration dose of the Mabosong decoction extract reaches the crude drug dose of 6 g / Kg, no obvious toxic reactions occurred in the test mice. It can be seen that the lung-nourishing extract of Mabosong decoction has relatively high safety.

[0173] Example 6 Protective effect of the lung-nourishing composition preparation of Mabosong decoction on the lipopolysaccharide-induced acute lung injury model in mice

[0174] 1 Materials

[0175] 1.1 Animals

[0176] SPF-grade male Kunming (KM) mice, 6-8 weeks old, weighing 18-22 g, 140 in number, were purchased from Chengdu Dashuo Experimental Animal Co., Ltd., with the production license number SCXK (Sichuan) 2020-030. The animals were housed in an SPF-grade animal room with a light cycle of 12 h light / 12 h darkness, an environmental temperature of 20-26 °C, and a relative humidity of 40-70%. They had free access to food and water. This animal experiment protocol has passed the review of the Ethics Review Committee of Sichuan Agricultural University, and the experimental process strictly complied with the Guide for the Care and Use of Laboratory Animals. Before the formal experiment started, all animals were adaptively raised for 7 days.

[0177] 1.2 Test samples

[0178] 1) Preparation of the extract of the original formula group of Mabosong decoction

[0179] Pretreatment of raw materials: The roots and rhizomes of Arnebia euchroma and Rubia cordifolia were washed with clear water, dried at 60-65 °C, cut into pieces and then crushed into coarse powder; the lacca was dried at 60-65 °C and crushed into coarse powder.

[0180] Formula: Weigh the coarse powder of the medicinal materials according to the formula ratio recorded in the above literature (300 g each of Rubia cordifolia and Arnebia euchroma, and 260 g of lacca) and prepare the mixture.

[0181] Extraction: Mix the above-mentioned crude powder of medicinal materials according to the preferred formula ratio (10 parts of Tibetan gromwell root, 10 parts of Tibetan madder root, 8 parts of gromwell root, 5 parts of bamboo shavings, 4 parts of alpine horseradish, 3 parts of winged head grass, 2 parts of sea buckthorn, and 2 parts of licorice root), and then decoct in water to extract. The material-to-liquid ratio is 1:10. Each decoction is 2 hours long, and the filtrate is filtered. The decoction is repeated three times, and the three extracts are combined.

[0182] Concentration and freeze-drying: The filtrate is concentrated under reduced pressure at a temperature not exceeding 60 °C. After being concentrated into an extract, it is freeze-dried into a powder and stored for later use.

[0183] 2) Preparation of the extract from the modified Mabaosong decoction group

[0184] Raw material pretreatment: Wash the roots and rhizomes of Lithospermum erythrorhizon, Rubia cordifolia, horseradish rhizomes, whole plant of Lysimachia christinae, licorice root, and sea buckthorn fruit with clean water, dry them at 60-65 ℃, cut them into pieces and grind them into coarse powder; Lithospermum erythrorhizon and Bambusa textilis are also dried at 60-65 ℃ and ground into coarse powder.

[0185] Formula: Weigh the coarse powder of medicinal materials according to the above standard formulation ratio and prepare the mixture.

[0186] Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water to extract, with a material-to-liquid ratio of 1:10. Each decoction is carried out for 2 hours and filtered to obtain the filtrate. The decoction is carried out three times and the three extracts are combined.

[0187] Concentration and freeze-drying: The filtrate is concentrated under reduced pressure at a temperature not exceeding 60 °C. After being concentrated into an extract, it is freeze-dried into a powder and stored for later use.

[0188] 3) Added preparation group 1: Mabaosong Decoction Lung-Nourishing Granules prepared in Example 2 (batch: 20220601)

[0189] 4) Two groups of flavored preparations: Mabaosong Decoction for Lung Nourishing Oral Solution prepared in Example 3 (batch number: 20220701)

[0190] 5) Three groups of flavored preparations: Mabaosong Decoction Lung-Nourishing Capsules (batch number: 20220801) prepared in Example 4.

[0191] 2. Experimental Methods in this Embodiment

[0192] 2.1 Experimental grouping and drug administration

[0193] All mice were randomly divided into 7 groups of 20 mice each, including a blank control group, a model group, the original Mabaosong Decoction group, a modified Mabaosong Decoction group, modified preparation group 1, modified preparation group 2, and modified preparation group 3. Referring to the clinical use of the Tibetan medicine compound Mabaosong Decoction at a moderate dose of 8g raw drug / day, the equivalent dose for mice was calculated to be 1.04 g / day based on body surface area. The mice were administered the drug at a dose of 0.1 mL / 10g, once daily by gavage to the blank control group (0.9% sodium chloride solution), the model group (0.9% sodium chloride solution), and each test drug group (corresponding to the Tibetan medicine combination solution, 20.8 mg / kg), for 7 consecutive days.

[0194] 2.2 Establishment of an Acute Lung Injury (ALI) Mouse Model

[0195] Thirty minutes after the last administration, animals were anesthetized by inhalation with ether. Mice in the model group and each test drug group were infused with LPS via endotracheal intubation at a dose of 20 mg / kg, while mice in the control group received sterile saline instead. Animals were euthanized by cervical dislocation 24 hours after model establishment. Bronchoalveolar lavage fluid (BALF) and lung tissue were collected for subsequent parameter detection.

[0196] 2.3 Detection Indicators

[0197] 2.3.1 Determination of lung wet / dry weight ratio (W / D)

[0198] The removed lung tissue was rinsed with physiological saline to remove surface blood, then gently patted dry with filter paper, and the wet weight (W) of the lung tissue was obtained. The tissue was then placed in an 80℃ oven until constant weight was achieved, and the dry weight (D) of the tissue was obtained. The W / D value of the lung tissue was calculated.

[0199] 2.3.2 Detection of MPO content in the lungs

[0200] Mouse lung tissue was ground into powder with liquid nitrogen, weighed, and tested according to the MPO kit instructions.

[0201] 2.3.3 Pathological examination of lung tissue

[0202] Fresh mouse lung tissue was cleaned, dried, and cut into 1cm pieces. 3 Tissue blocks of various sizes were fixed in 4% paraformaldehyde. The following steps were performed sequentially: rinsing, gradient dehydration, clearing, paraffin embedding, sectioning, hematoxylin and eosin (H&E) staining, and mounting. Finally, the prepared sections were observed under a microscope.

[0203] 2.3.4 Detection of TNF-α, IIL-1β, L-6, IL-10 and total protein content in BALF

[0204] After BALF collection, the supernatant was collected by centrifugation. The contents of TNF-α, IL-1β, IL-6 and IL-10 were detected by ELISA, and the total protein content was detected by BCA method. The detection was performed according to the kit instructions.

[0205] 2.4 Data Analysis

[0206] The experimental data were analyzed using SPSS 26.0 software. Results are expressed as mean ± standard deviation (±SD). One-way ANOVA was performed on each group using the least significant difference method (LSD). A p-value < 0.05 was considered statistically significant, and a p-value < 0.01 was considered extremely significant. The experimental results were visualized using GraphPad Prism 9 software.

[0207] 3 Experimental Results

[0208] 3.1 Effects of various Tibetan medicine compositions on the wet-to-dry weight ratio of lungs in mice with acute lung injury

[0209] The results are as follows Figure 12 As shown, compared with the blank group, the wet-to-dry weight ratio of the lungs of mice in the model group was significantly increased (P < 0.01); compared with the model group, the wet / dry weight ratio of the lungs of mice in each tested drug group was significantly decreased (P < 0.01). The wet-to-dry weight ratio is an indicator for evaluating the degree of pulmonary edema. This result indicates that all Tibetan medicine compositions can effectively improve the degree of pulmonary edema in ALI mice, with the extract of Jiawei Mabaosong Decoction and its three preparations showing better effects than the original Mabaosong Decoction group.

[0210] 3.2 Effects of various Tibetan medicine compositions on MPO activity in the lungs of mice with acute lung injury

[0211] MPO, an enzyme located in neutrophils, can be used to indirectly reflect neutrophil adhesion and marginalization in lung tissue by measuring its activity. MPO activity detection results are as follows: Figure 13 As shown, compared with the blank group, the MPO enzyme activity in the model group mice was significantly increased (P < 0.01); while compared with the model group, the activity of this enzyme in the lungs of mice in each tested drug group was significantly decreased (P < 0.01). This result indicates that each Tibetan medicine composition can effectively improve MPO activity in the lung tissue of ALI mice, that is, the number of neutrophils in the tissue may be reduced. Among them, the extract of Jiawei Mabaosong Decoction and its three preparations are more effective than the original Mabaosong Decoction group.

[0212] 3.3 Effects of various Tibetan medicine compositions on the pathological characteristics of lung tissue in mice with acute lung injury

[0213] The results are as follows Figure 14As shown: In the control group, the lung tissue structure of mice was normal, with intact alveolar walls, clear alveolar cavities, and no obvious inflammatory cell infiltration in the alveolar spaces. In the model group, the lung tissue of mice showed obvious lesions, with significant thickening of the alveolar septa and alveolar cavities, as well as a large number of inflammatory cell infiltrations. Although the lung tissue of mice in each of the tested drug groups still showed pathological manifestations such as inflammatory cell infiltration and alveolar wall thickening, they all showed a certain degree of improvement compared to the model group. In addition, compared with the original formula of Mabaosong Decoction, the improvement effect of each modified drug group was better. Pathological sections of lung tissue can directly reflect the inflammation status, and this result indicates that each Tibetan medicine combination can effectively alleviate the degree of pulmonary edema and inflammatory cell infiltration in ALI mice.

[0214] 3.4 Effects of various Tibetan medicine compositions on the levels of inflammatory factors TNF-α, IL-1β, IL-6, IL-10 and total protein in bronchoalveolar lavage fluid of mice with acute lung injury.

[0215] The results are as follows Figure 15 As shown in A-15D, compared with the blank group, the levels of various inflammatory factors, including TNF-α, IL-1β, IL-6, and IL-10, in the bronchoalveolar lavage fluid of mice in the model group were significantly increased (P < 0.01). However, compared with the model group, the levels of pro-inflammatory factors TNF-α, IL-1β, and IL-6 in the bronchoalveolar lavage fluid of mice in each of the tested drug groups were significantly decreased (P < 0.01), while the level of IL-10, an anti-inflammatory factor with multiple functions, was significantly increased (P < 0.01). These results indicate that the various Tibetan medicine compositions can improve ALI mice by regulating the secretion of inflammatory factors, with the modified Mabaosong decoction extract and its three preparations showing better effects than the original Mabaosong decoction group.

[0216] Protein concentration in bronchoalveolar lavage fluid is often used to reflect changes in lung tissue permeability and is related to pulmonary microvascular permeability. The total protein content in the bronchoalveolar lavage fluid of each group of mice was measured, and the results are as follows: Figure 15 As shown in Figure E, compared with the blank group, the total protein content in the bronchoalveolar lavage fluid of the model group mice was significantly increased (P < 0.01); and compared with the model group, each tested drug group significantly improved this change. This indicates that the various Tibetan medicine compositions may effectively alleviate the microvascular permeability of the lungs in ALI mice, among which the modified Mabaosong decoction extract and its three preparations were more effective than the original Mabaosong decoction group.

[0217] 4. Conclusion

[0218] The Tibetan medicine compositions provided by the present invention can effectively improve LPS-induced acute lung injury in mice, reduce pulmonary tissue edema, alleviate pulmonary tissue inflammatory infiltration, decrease the permeability of pulmonary microvessels, and regulate the contents of different inflammatory factors in BALF. In addition, by adding flavors to the original formula of Mabao Song Decoction, its effects are better than those of the original Mabao Song Decoction in terms of detection indexes, indicating that the flavored preparation can effectively improve the therapeutic effect of Mabao Song Decoction on acute lung injury.

[0219] Example 7 Protective effect of the lung-soothing composition preparation of Mabao Song Decoction on a bleomycin-induced mouse pulmonary fibrosis model

[0220] In this example, a bleomycin-induced pulmonary fibrosis mouse model was used as the research object to prove the protective effect of the Tibetan medicine compositions provided by the present invention on pulmonary fibrosis.

[0221] 1 Materials

[0222] 1.1 Animals

[0223] SPF-grade male Kunming (KM) mice, 6 - 8 weeks old, weighing 18 - 22 g, 140 in number, were purchased from Chengdu Dashuo Experimental Animal Co., Ltd., with the production license number SCXK (Chuan) 2020 - 030. The animals were housed in an SPF-grade animal room with a light cycle of 12 h light / 12 h darkness, an environmental temperature of 20 - 26 °C, and a relative humidity of 40 - 70%, and were allowed free access to food and water. The animal experiment protocol has passed the review of the Ethics Review Committee of Sichuan Agricultural University, and the experimental process strictly complied with the Guide for the Care and Use of Laboratory Animals. Before the formal experiment began, all animals were adaptively raised for 7 days.

[0224] 1.2 Sample preparation

[0225] 1) Preparation of the original extract of Mabao Song Decoction

[0226] Raw material pretreatment: The roots of Arnebia euchroma, the roots and rhizomes of Rubia cordifolia were washed with clear water, dried at 60 - 65 °C, cut into pieces and then crushed into coarse powder; Gummi Arabicum was dried at 60 - 65 °C and crushed into coarse powder.

[0227] Formula: According to the formula ratio recorded in the above literature (300 g each of Rubia cordifolia and Arnebia euchroma, 260 g of Gummi Arabicum), the coarse powder of the medicinal materials was weighed and prepared.

[0228] Extraction: After mixing the above-mentioned coarse powder of medicinal materials according to the formula ratio, water decoction extraction was carried out with a material-liquid ratio of 1:10. Each decoction was for 2 h, and the filtrate was obtained by filtration. Decoction was carried out three times, and the three extraction liquids were combined.

[0229] Concentration and freeze-drying: The filtrate is concentrated under reduced pressure at a temperature not exceeding 60 °C. After being concentrated into an extract, it is freeze-dried into a powder and stored for later use.

[0230] 2) Preparation of Jiawei Mabaosong Decoction Extract

[0231] Raw material pretreatment: Wash the roots and rhizomes of Lithospermum erythrorhizon, Rubia cordifolia, horseradish rhizomes, whole plant of Lysimachia christinae, licorice root, and sea buckthorn fruit with clean water, dry them at 60-65 ℃, cut them into pieces and grind them into coarse powder; Lithospermum erythrorhizon and Bambusa textilis are also dried at 60-65 ℃ and ground into coarse powder.

[0232] Formula: Weigh the coarse powder of medicinal materials according to the above standard formulation ratio and prepare the mixture.

[0233] Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water to extract, with a material-to-liquid ratio of 1:10. Each decoction is carried out for 2 hours and filtered to obtain the filtrate. The decoction is carried out three times and the three extracts are combined.

[0234] Concentration and freeze-drying: The filtrate is concentrated under reduced pressure at a temperature not exceeding 60 °C. After being concentrated into an extract, it is freeze-dried into a powder and stored for later use.

[0235] 3) Added preparation group 1: Mabaosong Decoction Lung-Nourishing Granules prepared in Example 2 (batch: 20220601)

[0236] 4) Two groups of flavored preparations: Mabaosong Decoction for Lung Nourishing Oral Solution prepared in Example 3 (batch number: 20220701)

[0237] 5) Three groups of flavored preparations: Mabaosong Decoction Lung-Nourishing Capsules (batch number: 20220801) prepared in Example 4.

[0238] 2. Experimental Methods in this Embodiment

[0239] 140 SPF-grade male ICR mice, 6-8 weeks old and weighing 18-22 g, were randomly divided into 7 groups. The grouping and administration methods were the same as in Example 6. After acclimatization, all groups except the control group were instilled with bleomycin hydrochloride (Hanhui Pharmaceutical Co., Ltd.) via endotracheal infusion at a dose of 3 U / kg to establish the model. The control group was treated with sterile saline. This was designated as day 0 of the modeling process. From day 1, each test drug group was administered the corresponding Tibetan medicine composition solution by gavage, while the control and model groups were treated with saline. Administration was once daily for 21 consecutive days. From day 0 of the modeling process, mouse weight was recorded every 7 days to observe changes in weight during the experiment. One hour after the last administration, the mice were euthanized. The intact lungs were harvested, the surface blood was washed away with pre-cooled saline, the lungs were gently patted dry with filter paper, and then weighed. The lung index was calculated as: lung weight (mg) / mouse body weight (g). After weighing, the left lung was fixed in 4% paraformaldehyde, and paraffin sections were prepared. The histopathology of the mouse lung tissue was evaluated by H&E and Masson staining, respectively. Approximately 50 g of tissue was accurately weighed from the right lung, and the hydroxyproline (HYP) content in the lung was detected using an alkaline hydrolysis kit (Nanjing Jiancheng Technology Co., Ltd.). The data analysis methods were the same as in Example 6.

[0240] 3. Experimental Results of This Example

[0241] 3.1 Effects of various Tibetan medicine compositions on body weight changes in mice with pulmonary fibrosis

[0242] The results are as follows Figure 16 As shown: On day 0 of modeling, there was no significant difference in the initial body weight of all mice. After modeling, it was obvious that the body weight of the model group mice increased slowly, and on days 7, 14, and 21, their body weight decreased significantly compared with the control group mice (P < 0.01); while compared with the model group, the mice in each test drug group showed significant improvement in this body weight change (P < 0.05).

[0243] 3.2 Effects of various Tibetan medicine compositions on lung index in mice with pulmonary fibrosis

[0244] The results are as follows Figure 17 As shown, compared with the blank group, the lung index of mice in the model group was significantly increased (P<0.01); while compared with the model group, the lung index of mice in each test drug group was significantly decreased (P<0.05).

[0245] 3.3 Effects of various Tibetan medicine compositions on the pathological characteristics of lung tissue in mice with pulmonary fibrosis:

[0246] H&E staining results of lung tissue from pulmonary fibrosis mice are as follows: Figure 18As shown: In the control group, the lung tissue of mice was normal, the alveolar structure was intact, and there was no obvious inflammatory cell infiltration in the alveolar spaces; in the model group, the lung tissue of mice showed obvious lesions, including significant destruction of the lung tissue structure, disappearance of alveolar structure, thickening of the pulmonary septa, a large number of inflammatory cell infiltrations, and an increased number of fibroblasts; while in the lung tissue of mice in each of the test drug groups, the above pathological changes were milder than those in the model group. These results indicate that the Tibetan medicine compositions can effectively improve lung inflammation in mice with pulmonary fibrosis.

[0247] Masson staining, a classic method for staining collagen fibers, is often used to observe the degree of collagen fiber deposition in the lungs of pulmonary fibrosis models. The results of Masson staining of lung tissue from pulmonary fibrosis mice are shown below. Figure 19 As shown: In the blank control group, the lung tissue structure of mice was normal, with a small amount of collagen deposition observed around the blood vessel walls or bronchial walls, but no obvious collagen deposition was observed in other areas; in the model group, the lung tissue structure of mice showed obvious damage, with a large number of blue collagen fibers deposited diffusely around the alveolar walls, alveolar septa, bronchial walls, and blood vessel walls; while in the lung tissue of mice in each of the test drug groups, collagen fiber deposition was significantly reduced compared to the model group. This result indicates that the Tibetan medicine compositions can effectively improve the degree of pulmonary fibrosis in mice with pulmonary fibrosis.

[0248] 3.4 Effects of various Tibetan medicine compositions on the hydroxyproline content in lung tissue of mice with pulmonary fibrosis

[0249] HYP, the most abundant imino acid in collagen, is often used as an important indicator of the degree of connective tissue deposition in the lungs. The results of HYP content detection in the lungs of various groups of mice are as follows: Figure 20 As shown, compared with the control group, the HYP content in the lungs of mice in the model group was significantly increased (P < 0.01); while the HYP content in the lungs of mice in each of the test drug groups was significantly decreased compared with the control group (P < 0.01). This result indicates that the Tibetan medicine compositions can effectively reduce collagen deposition in the lungs of mice with pulmonary fibrosis.

[0250] 3. Conclusion

[0251] The Tibetan medicine composition provided by this invention has a certain protective effect against bleomycin-induced pulmonary fibrosis in mice, improves the pathological changes in lung tissue of mice with pulmonary fibrosis, reduces inflammatory cell infiltration and collagen deposition in the lungs, and alleviates the degree of pulmonary fibrosis. Furthermore, the results show that the modified Mabaosong Decoction groups have better therapeutic effects on pulmonary fibrosis, indicating that the modified preparations can effectively improve the therapeutic effect of the original Mabaosong Decoction on pulmonary fibrosis.

[0252] Example 8: The effect of the Mabaosong decoction and lung-nourishing composition on improving cigarette smoke-induced COPD in mice.

[0253] 1 Materials

[0254] 1.1 Animals

[0255] SPF-grade male Kunming (KM) mice, 6 - 8 weeks old, weighing 18 - 22 g, 140 in number, were purchased from Chengdu Dashuo Laboratory Animal Co., Ltd., with the production license number SCXK (Sichuan) 2020 - 030. The animals were housed in an SPF-grade animal room with a light cycle of 12 h light / 12 h darkness, an environmental temperature of 20 - 26 °C, and a relative humidity of 40 - 70%. They had free access to food and water. This animal experiment protocol has passed the review of the Ethics Committee of Sichuan Agricultural University, and the experimental process strictly adhered to the Guide for the Care and Use of Laboratory Animals. Before the formal experiment began, all animals were adaptively housed for 7 days.

[0256] 1.2 Test Samples

[0257] 1) Preparation of the extract of the original formula group of Mabao Song Decoction

[0258] Raw material pretreatment: The roots of Arnebia euchroma, the roots and rhizomes of Rubia tibetica were washed with clean water, dried at 60 - 65 °C, cut into pieces and then crushed into coarse powder; the galls of Laccifer lacca were dried at 60 - 65 °C and crushed into coarse powder.

[0259] Formula: Weigh the coarse powder of the medicinal materials according to the formula ratio recorded in the above literature (300 g each of Rubia cordifolia and Arnebia euchroma, 260 g of the galls of Laccifer lacca) and make the preparation.

[0260] Extraction: After mixing the above-mentioned coarse powder of the medicinal materials according to the formula ratio, extract by boiling in water. The ratio of material to liquid is 1:10. Each boiling lasts for 2 h, and the filtrate is obtained by filtration. Boil three times and combine the extraction solutions of the three times.

[0261] Concentration and lyophilization: Concentrate the filtrate under reduced pressure, with the concentration temperature not exceeding 60 °C. After concentrating into an extract, lyophilize it into powder and store it in the refrigerator for later use.

[0262] 2) Preparation of the extract of the modified Mabao Song Decoction group

[0263] Raw material pretreatment: The roots of Arnebia euchroma, the roots and rhizomes of Rubia tibetica, the rhizomes of Lepidium sativum, the whole herb of Pterocephalus hookeri, the roots of Glycyrrhiza uralensis, and the fruits of Hippophae rhamnoides were washed with clean water, dried at 60 - 65 °C, cut into pieces and then crushed into coarse powder; the galls of Laccifer lacca and the concretion silicea Bambusae were also dried at 60 - 65 °C and crushed into coarse powder.

[0264] Formula: Weigh out the coarse powder of the medicinal materials according to the above-mentioned preferred formula ratio (10 parts of Tibetan gromwell, 10 parts of Tibetan madder, 8 parts of gromwell root, 5 parts of bamboo shavings, 4 parts of alpine horseradish, 3 parts of winged head grass, 2 parts of sea buckthorn, and 2 parts of licorice) and prepare the formula.

[0265] Extraction: Mix the above-mentioned crude powder of medicinal materials according to the formula ratio, decoct with water to extract, with a material-to-liquid ratio of 1:10. Each decoction is carried out for 2 hours and filtered to obtain the filtrate. The decoction is carried out three times and the three extracts are combined.

[0266] Concentration and freeze-drying: The filtrate is concentrated under reduced pressure at a temperature not exceeding 60 °C. After being concentrated into an extract, it is freeze-dried into a powder and stored for later use.

[0267] 3) Added preparation group 1: Mabaosong Decoction Lung-Nourishing Granules prepared in Example 2 (batch: 20220601)

[0268] 4) Two groups of flavored preparations: Mabaosong Decoction for Lung Nourishing Oral Solution prepared in Example 3 (batch number: 20220701)

[0269] 5) Three groups of flavored preparations: Mabaosong Decoction Lung-Nourishing Capsules (batch number: 20220801) prepared in Example 4.

[0270] 2 Experimental Methods

[0271] 2.1 Experimental grouping and drug administration

[0272] 140 SPF-grade male ICR mice, 6-8 weeks old and weighing 18-22 g, were randomly divided into 7 groups. The grouping and administration methods were the same as in Example 6. From week 9 to week 12 after modeling, the mice were administered the drug by gavage 0.5 hours before the start of modeling each afternoon, for a total of 4 weeks.

[0273] 2.2 Establishment of COPD mouse model:

[0274] Mice in the model group and each drug-treated group underwent a 12-week cigarette smoke exposure model. Each exposure lasted 40 minutes, twice daily, 6 days a week, for a total of 12 weeks. The exposure time was at least twice a day, each exposure lasting at least 40 minutes, with an interval of at least 4 hours between exposures. Mice in the control group were exposed to normal air. After the 12-week experiment, various lung function indicators of mice in each group were measured using a small animal pulmonary function analyzer. Mice were sacrificed, and serum and lung tissue were collected for subsequent indicator testing.

[0275] 2.3 Detection Indicators

[0276] 2.3.1 Pulmonary function test:

[0277] Mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital solution (50 mg / kg). After fixing the mice in a supine position, the skin on their necks was cut open to expose the trachea. A small incision was made in the middle of the trachea, and the trachea was inserted. The incision was fixed with surgical sutures. The mice were then placed in the pulmonary function instrument's cage, and the trachea was connected to the instrument. The following pulmonary function indicators were measured: functional residual capacity (FRC), static compliance (Cchord), and airway resistance (RI). Under anesthesia, pulmonary function indicators were measured three times consecutively, and the average value was calculated.

[0278] 2.3.2 Pathological examination of lung tissue:

[0279] Fresh mouse lung tissue was cleaned, dried, and cut into 1cm pieces. 3 Tissue blocks of various sizes were fixed in 4% paraformaldehyde. The following steps were performed sequentially: rinsing, gradient dehydration, clearing, paraffin embedding, sectioning, hematoxylin and eosin (H&E) staining, and mounting. Finally, the prepared sections were observed under a microscope.

[0280] 2.3.3 Detection of TNF-α, II-1β, and L-6 in serum:

[0281] The levels of TNF-α, IL-1β, and IL-6 in the serum of mice in each group were detected by ELISA, according to the kit instructions.

[0282] 2.4 Data Analysis

[0283] The experimental data were analyzed using SPSS 26.0 software. Results are expressed as mean ± standard deviation (±SD). One-way ANOVA was performed on each group using the least significant difference method (LSD). P <0.05 indicates a significant difference. P <0.01 indicates a highly significant difference, all of which are statistically significant. Experimental results were visualized using GraphPad Prism 9 software.

[0284] 3 Experimental Results

[0285] 3.1 Effects of various Tibetan medicine compositions on lung function in COPD mice

[0286] The results are as follows Figure 21As shown in AC, compared with the blank group, the FRC, Cchord, and RI of mice in the model group were significantly increased; compared with the model group, the FRC, Cchord, and RI of mice in each test drug group were significantly decreased, which was statistically significant. This result indicates that each Tibetan medicine composition can effectively improve lung function in COPD mice.

[0287] 3.2 Effects of various Tibetan medicine compositions on the pathological characteristics of lung tissue in mice with acute lung injury

[0288] The results are as follows Figure 22 As shown: In the blank group, the bronchial mucosal epithelial structure of the lung tissue of mice was intact, the cilia were neatly arranged, no glandular hyperplasia was observed, the tracheal wall was smooth, no thickening of pulmonary vascular smooth muscle was observed, no inflammatory cell infiltration was observed around the bronchi and blood vessels, and no obvious rupture or fusion of alveolar septa was observed. In the COPD group, the bronchial wall of the lung tissue of mice was thickened, with obvious destruction of lung tissue structure, shedding of bronchial mucosal epithelium, thickening of bronchial smooth muscle, a large number of inflammatory cell infiltrations in the pulmonary interstitium, glandular hyperplasia, obvious alveolar dilation, some alveolar septal rupture, irregular enlargement and fusion of alveoli to form bullae, and thickening of the vessel walls of some small and medium-sized blood vessels. The lung tissue structure of each treatment group was improved and repaired to a certain extent. It was observed that the peribronchial inflammatory cell infiltration was improved compared with the COPD group, the alveolar dilation and rupture were reduced, and the bronchial wall was thinned. The results indicate that each Tibetan medicine composition can effectively alleviate inflammatory cell infiltration, emphysema and bronchial structural destruction in the lung tissue of COPD mice.

[0289] 3.3 Effects of various Tibetan medicine compositions on serum inflammatory factors TNF-α, IL-1β, and IL-6 in COPD mice

[0290] The results are as follows Figure 23 As shown in AC, compared with the blank group, the serum levels of inflammatory factors TNF-α, IL-1β, and IL-6 in the model group mice were significantly increased; while compared with the model group, the serum levels of pro-inflammatory factors TNF-α, IL-1β, and IL-6 in the test drug groups were significantly decreased. This result indicates that the Tibetan medicine compositions can improve COPD in mice by regulating the secretion of inflammatory factors.

[0291] 4. Conclusion

[0292] The Tibetan medicine compositions provided by this invention can effectively improve cigarette smoke-induced COPD in mice, improve lung function, reduce emphysema, decrease inflammatory infiltration in lung tissue, and regulate the levels of different inflammatory factors in serum. Furthermore, by adding ingredients to the original formula of Mabaosong Decoction, the modified formula showed better results than the original formula in terms of detection indicators, indicating that the modified preparation can effectively enhance the therapeutic effect of Mabaosong Decoction on COPD.

Claims

1. The use of Tibetan medicine Mabao Songtong Feining composition in the preparation of a drug for treating pulmonary fibrosis, chronic obstructive pulmonary disease; characterized in that The composition is prepared from the following weight parts of medicinal materials: Arnebia euchroma 10-25 parts, Rubia yunnanensis 10-25 parts, Radix Baphlami 5-20 parts, Radix Curculiginis 5-15 parts, Eutrema japonicum 3-15 parts, Pterocephalus tottus 3-13 parts, Hippophae rhamnoides 2-11 parts, and Glycyrrhiza uralensis 2-8 parts; the composition preparation comprises the following steps: 1) washing, drying, cutting, and then crushing into coarse powder for Arnebia euchroma roots, Rubia yunnanensis roots and rhizomes, Eutrema japonicum rhizomes, Pterocephalus tottus whole plants, and Glycyrrhiza uralensis roots; 2) directly crushing into coarse powder for Hippophae rhamnoides fruits, Radix Baphlami, and Radix Curculiginis; 3) weighing the dried medicinal material coarse powder according to the prescription proportion; 4) mixing the above medicinal material coarse powder according to the prescription proportion, then water decocting and extracting, with a material-liquid ratio of 1:10, decocting for 2 hours each time, filtering to obtain a filtrate, decocting for three times, and combining the three times of extraction liquid; 5) reducing pressure and concentrating the filtrate, with a concentration temperature of no more than 60 DEG C, then freeze-drying the extract into a powder, and storing in cold storage for later use; 6) taking the above extract freeze-dried powder, and preparing granules by wet granulation: selecting dextrin and lactose as auxiliary materials, preferably with a dextrin and lactose ratio of 1.5:1, and preferably with an auxiliary material amount and extract ratio of 3:1, mixing them uniformly, then adding 75% ethanol to prepare soft material, sieving and granulating, whole granulating, drying at 60-65 DEG C, and then bagging to obtain the Mahuosong Tang Feining granules; 7) taking the above extract freeze-dried powder, dissolving in 8-10 times distilled water, storing in cold storage for 24 hours, then centrifuging and filtering, adding 10% xylitol as a flavoring agent, 1% sodium benzoate as a preservative, and 1% sodium thiosulfate as an antioxidant, mixing uniformly, filtering, filling and sealing, sterilizing, and then obtaining the Mahuosong Tang Feining oral liquid; 8) taking the above extract freeze-dried powder, mixing with poloxamer 188 and sodium dodecyl sulfate according to a ratio of 1:6:1 to prepare a solid dispersion, and then loading into hard capsules to obtain the Mahuosong Tang Feining capsules.

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