Application of yin-nourishing and lung-clearing composition in preparation of medicine for treating pulmonary fibrosis

Abstract

The invention belongs to the technical field of traditional Chinese medicine, and relates to application of a yin-nourishing and lung-clearing composition in preparation of medicine for preventing and / or treating pulmonary fibrosis, and the yin-nourishing and lung-clearing composition is prepared from the following raw materials: rehmannia, radix scrophulariae, radix ophiopogonis, moutan bark, bulbus fritillariae cirrhosae, radix paeoniae alba, mint and liquorice. The invention provides the application of the yin-nourishing and lung-clearing composition in preparing the medicine for preventing and / or treating the pulmonary fibrosis, bleomycin is injected into the trachea to construct a pulmonary fibrosis mouse model, and the effect of the yin-nourishing and lung-clearing composition on treating the pulmonary fibrosis is determined from the three aspects of the lung coefficient, the pathological change and the hydroxyproline content of a model animal. Basic data support is provided for increasing clinical application of the composition for nourishing yin and clearing away lung-heat.

Description

Application of Yin-nourishing and Lung-clearing Composition in the Preparation of Drugs for Treating Pulmonary Fibrosis Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology and relates to the application of a Yin-nourishing and lung-clearing composition in the preparation of drugs for the prevention and / or treatment of pulmonary fibrosis. Background Technology

[0002] Pulmonary fibrosis is a common chronic, interstitial lung disease among respiratory system diseases. Its main clinical manifestations include progressive dyspnea, chest tightness, cough, and crackles at the bases of both lungs, which may be accompanied by clubbing of the fingers. As the disease progresses, it can lead to death from respiratory failure. In recent years, the incidence and prevalence of pulmonary fibrosis have been increasing annually, with a median survival of 2-3 years after diagnosis. Treatment mainly includes drug therapy and non-drug therapy. Drug therapy mainly includes pirfenidone, nintedanib, glucocorticoids, and immunosuppressants. These drugs can relieve symptoms but cannot slow the progression of the disease, and long-term use of these drugs can easily cause adverse reactions. Non-drug therapy includes oxygen therapy, pulmonary rehabilitation, and lung transplantation. Lung transplantation can improve clinical symptoms and prolong patient survival time and is the only curative treatment. However, lung resources are limited, the surgery is expensive, and post-transplant complications such as heart failure, gastroesophageal reflux disease, diabetes, and obesity must be considered. Therefore, new treatment methods are needed.

[0003] In ancient Chinese medicine, there was no specific disease name for pulmonary fibrosis. Based on the patient's clinical manifestations, physicians categorized it under the traditional Chinese medicine terms "lung obstruction" or "lung atrophy." In terms of treatment, based on its pathogenesis, traditional Chinese medicine often focuses on tonifying qi and nourishing yin, promoting blood circulation and removing blood stasis, and eliminating phlegm and unblocking the meridians.

[0004] This Yin-nourishing and Lung-clearing composition has the effects of nourishing Yin and moistening dryness, clearing the lungs and relieving sore throat. It is used for Yin deficiency and lung dryness, dry and sore throat, dry cough with little phlegm. It is composed of Rehmannia glutinosa, Scrophularia ningpoensis, Ophiopogon japonicus, Paeonia suffruticosa, Fritillaria cirrhosa, Paeonia lactiflora, Mentha haplocalyx, and Glycyrrhiza uralensis. Rehmannia glutinosa nourishes Yin and clears heat; Scrophularia ningpoensis clears heat and nourishes Yin, clears deficiency heat and detoxifies; Ophiopogon japonicus clears the lungs and nourishes Yin; Paeonia lactiflora nourishes blood and benefits Yin; Paeonia suffruticosa cools the blood and detoxifies, invigorates blood and removes blood stasis; Fritillaria cirrhosa clears heat and moistens the lungs, resolves phlegm and relieves cough; Mentha haplocalyx promotes lung function and relieves sore throat, moistens the throat and relieves itching; Glycyrrhiza uralensis is sweet and moistening, clears heat and detoxifies, and harmonizes the other herbs. It has the functions of nourishing Yin and clearing the lungs, clearing heat and relieving sore throat, resolving phlegm, invigorating blood and removing blood stasis, and is used for Yin deficiency and lung dryness, dry and sore throat, dry cough with little phlegm, or blood in phlegm. In recent years, extensive clinical trials have shown that this drug is also commonly used to treat upper respiratory tract infections, acute and chronic bronchitis, bronchiectasis, diphtheria, pharyngitis, tonsillitis, oral ulcers, and other diseases, especially those with symptoms of lung dryness and yin deficiency. Summary of the Invention

[0005] The main objective of this invention is to provide the application of the Yin-nourishing and lung-clearing composition in the preparation of drugs for the prevention and / or treatment of pulmonary fibrosis.

[0006] This invention provides the application of a Yin-nourishing and Lung-clearing composition in the preparation of drugs for the prevention and / or treatment of pulmonary fibrosis. The raw materials of the Yin-nourishing and Lung-clearing composition include: Rehmannia glutinosa, Scrophularia ningpoensis, Ophiopogon japonicus, Paeonia suffruticosa, Fritillaria cirrhosa, Paeonia lactiflora, Mentha haplocalyx, and Glycyrrhiza uralensis.

[0007] The Yin-nourishing and Lung-clearing composition is a pure traditional Chinese medicine formulation with high safety, simple formula, low cost, and no side effects. This invention established a mouse model of pulmonary fibrosis through intratracheal injection of bleomycin, and clarified the therapeutic effect of the Yin-nourishing and Lung-clearing composition on pulmonary fibrosis by analyzing the lung coefficient, pathological changes, and hydroxyproline content in the model animals.

[0008] Preferably, the raw materials of the Yin-nourishing and lung-clearing composition in the application include the following components in parts by weight: Rehmannia glutinosa: 5-10 parts; Ophiopogon japonicus: 5-8 parts; Scrophularia ningpoensis: 3-7 parts; Fritillaria cirrhosa: 3-5 parts; Paeonia lactiflora: 5-7 parts; Paeonia suffruticosa: 3-5 parts; Mentha haplocalyx: 3-5 parts; Glycyrrhiza uralensis: 2-5 parts.

[0009] More preferably, the raw materials of the Yin-nourishing and lung-clearing composition in the application include the following components in parts by weight: Rehmannia glutinosa: 5 parts; Ophiopogon japonicus: 5 parts; Scrophularia ningpoensis: 3 parts; Fritillaria cirrhosa: 3 parts; Paeonia lactiflora: 5 parts; Paeonia suffruticosa: 3 parts; Mentha haplocalyx: 3 parts; Glycyrrhiza uralensis: 2 parts.

[0010] Furthermore, the Yin-nourishing and lung-clearing composition also contains pharmaceutically acceptable excipients, such as sucrose.

[0011] Furthermore, the dosage form of the Yin-nourishing and lung-clearing composition is a paste, capsule, granule, tablet, pill, or oral liquid.

[0012] Preferably, the Yin-nourishing and lung-clearing composition is a Yin-nourishing and lung-clearing paste.

[0013] Furthermore, the Yin-nourishing and Lung-clearing composition is prepared from the water extract, alcohol extract, or water-extracted alcohol precipitate of the raw materials of the Yin-nourishing and Lung-clearing composition.

[0014] Furthermore, the pulmonary fibrosis is idiopathic pulmonary fibrosis or secondary pulmonary fibrosis.

[0015] Furthermore, the Yin-nourishing and lung-clearing composition is used to inhibit or reduce the content of hydroxyproline in lung tissue.

[0016] Furthermore, the Yin-nourishing and lung-clearing composition is used to reduce the lung coefficient caused by pulmonary fibrosis or to inhibit the proliferation and activation of pulmonary fibroblasts.

[0017] Furthermore, the Yin-nourishing and lung-clearing composition is used to improve pathological changes in lung tissue, including one or more of the following: reduced alveolar inflammation, mild widening of alveolar septa, clearer lung tissue structure, reduced content of blue collagen fibers, or reduced degree of pulmonary fibrosis.

[0018] Furthermore, in the application, the Yin-nourishing and Lung-clearing composition is a Yin-nourishing and Lung-clearing paste, with a human dosage of 9 ml / kg.

[0019] In the mouse model of pulmonary fibrosis constructed in this invention, there were high-dose group (18 ml / kg), medium-dose group (9 ml / kg), and low-dose group (4.5 ml / kg). These doses are the daily dosage for mice, which are 2 times, 1 times, and 0.5 times the human dosage, respectively.

[0020] Compared with the prior art, the present invention has the following significant beneficial effects: The present invention provides the application of the Yin-nourishing and Lung-clearing composition in the preparation of drugs for treating pulmonary fibrosis. A mouse model of pulmonary fibrosis is constructed by intratracheal injection of bleomycin. The lung coefficient, pathological changes and hydroxyproline content of the model animals are used to clarify that the Yin-nourishing and Lung-clearing composition has the effect of treating pulmonary fibrosis, providing basic data support for increasing the clinical application of the Yin-nourishing and Lung-clearing composition. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0022] Figure 1 is an HE staining (200×) image of lung tissue in an embodiment of the present invention; A: blank group; B: model group; C: pirfenidone tablet group; D: high-dose Yangyin Qingfei ointment group; E: medium-dose Yangyin Qingfei ointment group; F: low-dose Yangyin Qingfei ointment group; Figure 2 is a Masson staining (200×) image of lung tissue in an embodiment of the present invention; A: blank group; B: model group; C: pirfenidone tablet group; D: high-dose Yangyin Qingfei ointment group; E: medium-dose Yangyin Qingfei ointment group; F: low-dose Yangyin Qingfei ointment group. Detailed Implementation

[0023] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0024] Unless otherwise specified in the embodiments of the present invention, the techniques or conditions described in the literature in this field or the product instructions shall be followed; if the manufacturers of the reagents or instruments used are not specified, they are all conventional products that can be purchased through legitimate channels.

[0025] Example: The Yin-nourishing and Lung-clearing composition used in this example is Yin-nourishing and Lung-clearing Paste, which is produced by Beijing Tongrentang Technology Development Co., Ltd. The raw materials include the following components by weight: Rehmannia glutinosa: 5 parts; Ophiopogon japonicus: 5 parts; Scrophularia ningpoensis: 3 parts; Fritillaria cirrhosa: 3 parts; Paeonia lactiflora: 5 parts; Paeonia suffruticosa: 3 parts; Mentha haplocalyx: 3 parts; Glycyrrhiza uralensis: 2 parts.

[0026] The efficacy of the Yin-nourishing and Lung-clearing ointment was verified as follows: I. Model establishment: ICR mice weighing 18-20g were acclimatized for 5 days. Twelve mice were randomly selected as the blank group, and the remaining mice were used to prepare a pulmonary fibrosis mouse model by intratracheal injection of bleomycin.

[0027] The method is as follows: Prepare a 2.5 mg / ml bleomycin hydrochloride (BLM) solution. Calculate the required BLM dosage (0.5 mg / 100g) based on the mouse's weight. Use the calculated dosage; the modeling dose is 5 mg / kg, i.e., 20 μL of bleomycin hydrochloride per 10g animal. Anesthetize the animal using a small animal gas anesthesia device. Place the animal on a rodent board, pull out its tongue, and insert an indwelling intravenous catheter into the trachea. Place cotton at the needle hole; even movement of the cotton indicates successful insertion. Draw 150 μL of air into the syringe, then inhale the corresponding volume of bleomycin based on the animal's weight, slowly injecting it into the trachea. Remove the needle core and shake the mouse vertically left and right to ensure even distribution of the drug in the lungs. Immediately after injection, stand the mouse upright and rotate it left and right to ensure even distribution of the drug in the lungs. After the animal regains consciousness, raise it as usual.

[0028] II. Grouping and Drug Administration: Mice surviving 24 hours after surgery were randomly divided into a model group, a pirfenidone tablet group (270 mg / kg), a high-dose Yangyin Qingfei ointment group (18 ml / kg), a medium-dose Yangyin Qingfei ointment group (9 ml / kg), and a low-dose Yangyin Qingfei ointment group (4.5 ml / kg). All groups were administered the drug by gavage at the same time, with a volume of 20 ml / kg. The blank group and the model group were given the same volume of deionized water.

[0029] III. Pathological and Histological Examination (1) Measurement of Lung Coefficient in Mice When collecting samples, first weigh the mouse body weight, anesthetize the mouse, quickly open the chest cavity, expose the lungs, carefully remove the lung lobes, and use filter paper to absorb the residual blood, weigh the whole lung wet weight, and calculate the lung coefficient. Lung coefficient = whole lung wet weight / body weight × 100%.

[0030] (2) Pathological observation of lung tissue: Dissect the thoracic cavity, cut out the right lung tissue, rinse with pre-cooled 4°C physiological saline, absorb moisture with filter paper, fix the right lung of the mouse in 4% paraformaldehyde solution, dehydrate and embed routinely, and perform HE and Masson staining after sectioning to evaluate inflammation, pathological changes and blue collagen status.

[0031] IV. Detection of Hydroxyproline (HYP) Content, a Specific Indicator of Pulmonary Fibrosis: 0.8–1 g of lung tissue from each group of mice was homogenized with physiological saline at a ratio of 1:9. The homogenate was centrifuged at 3000 r / min for 10 min at 4℃, and the supernatant was collected. The HYP content in the lung tissue homogenate was determined using a HYP ELISA kit, and the OD value was measured using an enzyme-linked immunosorbent assay (ELISA) reader.

[0032] 1. The results of lung coefficient measurement in mice showed that, compared with the blank group, the lung coefficient of the model group was significantly increased (p < 0.01). Compared with the model group, the lung coefficients of the pirfenidone group, the high-dose Yangyin Qingfei ointment group, and the low-dose Yangyin Qingfei ointment group were decreased (p < 0.05). The lung coefficient of the medium-dose Yangyin Qingfei ointment group showed a decreasing trend, but there was no statistical difference (p > 0.05). As shown in Table 1, this indicates that the Yangyin Qingfei ointment of the present invention has the effect of reducing the severity of pulmonary fibrosis.

[0033] Table 1. Effects of Yin-nourishing and Lung-clearing Paste on Lung Coefficient in Mice with Pulmonary Fibrosis ( )

[0034] Compared with the blank group: ##P<0.01; Compared with the model group: *P<0.05.

[0035] 2. Pathological changes in lung tissue and microscopic observation under Masson staining: In the blank group, the lung tissue structure was clear, with intact and clearly defined alveolar walls and epithelial structures. A small amount of inflammatory cell infiltration was observed around the bronchi, and the lungs contained little or no blue-stained collagen fibers. In the model group, the lung tissue structure was unclear, with significantly widened alveolar septa, alveolar structure destruction, alveolar collapse, extensive inflammatory cell infiltration around the bronchi, and focal or diffuse blue-stained collagen fibers within the lungs. Compared with the model group, the low-dose group of Yangyin Qingfei ointment showed clearer lung tissue structure, slightly widened alveolar septa, fibrosis, and a small or focal distribution of blue-stained collagen fibers within the lungs (see Figures 1 and 2). This indicates that the Yangyin Qingfei ointment of this invention can reduce inflammatory cell infiltration, slightly widen alveolar septa, improve lung tissue structure, reduce alveolar structure destruction, decrease fibrosis, and reduce the distribution of blue collagen fibers, thus alleviating pulmonary fibrosis and protecting the pathological tissue of the lungs.

[0036] 3. The results of hydroxyproline (HYP) content detection in lung tissue showed that, compared with the blank group, the HYP content in the model group was significantly increased (p<0.01). Compared with the model group, the HYP content in the pirfenidone group, and the high, medium and low dose groups of Yangyin Qingfei ointment were significantly decreased (p<0.01). The results are shown in Table 2. This indicates that Yangyin Qingfei ointment of the present invention can reduce the content of the biomarker HYP in lung tissue, and that Yangyin Qingfei ointment can reduce the collagen deposition content in model mice, thereby reducing the severity of pulmonary fibrosis. This indicates that Yangyin Qingfei ointment has a therapeutic effect on pulmonary fibrosis.

[0037] Table 2. Effects of Yin-nourishing and Lung-clearing Paste on Lung Coefficient in Mice with Pulmonary Fibrosis ( )

[0038] Compared with the blank group: ## P<0.01; Compared with the model group: ** P<0.01.

[0039] This invention utilizes tracheal injection of bleomycin to replicate a mouse model of pulmonary fibrosis. The pharmacodynamic effects of Yangyin Qingfei ointment in treating pulmonary fibrosis were verified from three aspects: lung coefficient, pathological changes, and hydroxyproline content in the model animals. The results were reliable.

[0040] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. The use of the Yin-nourishing and Lung-clearing composition in the prevention and / or preparation of drugs for treating pulmonary fibrosis, wherein the raw materials of the Yin-nourishing and Lung-clearing composition include: Rehmannia glutinosa, Scrophularia ningpoensis, Ophiopogon japonicus, Paeonia suffruticosa, Fritillaria cirrhosa, Paeonia lactiflora, Mentha haplocalyx, Glycyrrhiza uralensis.

2. The application according to claim 1, characterized in that, The raw materials of the Yin-nourishing and lung-clearing composition include the following components in parts by weight: Rehmannia glutinosa: 5-10 parts; Ophiopogon japonicus: 5-8 parts; Scrophularia ningpoensis: 3-7 parts; Fritillaria cirrhosa: 3-5 parts; Paeonia lactiflora: 5-7 parts; Paeonia suffruticosa: 3-5 parts; Mentha haplocalyx: 3-5 parts; Glycyrrhiza uralensis: 2-5 parts.

3. The application according to claim 2, characterized in that, The raw materials of the Yin-nourishing and lung-clearing composition include the following components in parts by weight: Rehmannia glutinosa: 5 parts; Ophiopogon japonicus: 5 parts; Scrophularia ningpoensis: 3 parts; Fritillaria cirrhosa: 3 parts; Paeonia lactiflora: 5 parts; Paeonia suffruticosa: 3 parts; Mentha haplocalyx: 3 parts; Glycyrrhiza uralensis: 2 parts.

4. The application according to any one of claims 1-3, characterized in that, The dosage form of the Yin-nourishing and lung-clearing composition is ointment, capsule, granule, tablet, pill or oral liquid.

5. The application according to claim 4, characterized in that, The Yin-nourishing and Lung-clearing composition is a Yin-nourishing and Lung-clearing paste.

6. The application according to any one of claims 1-3, characterized in that, The Yin-nourishing and Lung-clearing composition is prepared from the water extract, alcohol extract, or water-extracted alcohol precipitate of the raw materials of the Yin-nourishing and Lung-clearing composition.

7. The application according to any one of claims 1-3 or 5, characterized in that, The pulmonary fibrosis is either idiopathic pulmonary fibrosis or secondary pulmonary fibrosis.

8. The application according to any one of claims 1-3 or 5, characterized in that, The Yin-nourishing and lung-clearing composition is used to inhibit or reduce the content of hydroxyproline in lung tissue.

9. The application according to any one of claims 1-3 or 5, characterized in that, The Yin-nourishing and lung-clearing composition is used to reduce the lung coefficient caused by pulmonary fibrosis or to inhibit the proliferation and activation of pulmonary fibroblasts.

10. The application according to any one of claims 1-3 or 5, characterized in that, The Yin-nourishing and lung-clearing composition is used to improve pathological changes in lung tissue, including one or more of the following: reduced alveolar inflammation, mild widening of alveolar septa, clearer lung tissue structure, reduced content of blue collagen fibers, or reduced degree of pulmonary fibrosis.

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