A Chinese medicine composition, preparation and application thereof

Abstract

The present invention relates to the field of traditional Chinese medicine, and more specifically to a traditional Chinese medicine composition for treating acute lung injury. The composition is made from the following raw materials in parts by weight: 2-10 parts of roasted licorice root, 3-10 parts of ginger, 3-10 parts of ginseng, 10-40 parts of raw rehmannia root, 3-10 parts of cassia twig, 3-10 parts of donkey-hide gelatin, 5-20 parts of ophiopogon japonicus, 5-20 parts of hemp seed, and 5-30 parts of jujube. The present invention also provides a traditional Chinese medicine compound preparation for treating acute lung injury and its use.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and in particular relates to a traditional Chinese medicine composition for treating acute lung injury, and a preparation and application thereof. Background Art

[0002] Acute lung injury (ALI) is a severe inflammatory response in the lungs, usually triggered by severe infection, trauma or other serious diseases.

[0003] Currently, there are no specific treatments for ALI itself. Most treatment strategies focus on supportive care, such as oxygen therapy, mechanical ventilation, fluid management, and elimination of underlying causes. Traditional Chinese medicine offers significant advantages in improving clinical symptoms, slowing pathological progression, and improving quality of life in patients with ALI. In traditional Chinese medicine, ALI falls under the categories of "asthma and collapse," "chest congestion," and "sudden asthma." Its pathogenesis is the gradual depletion of vital energy by pathogenic factors, leading to a deficiency of vital energy and a consequent weakening of vital energy. This ultimately leads to a deficiency of both Qi, blood, and Yin and Yang, ultimately depleting the body.

[0004] Based on prescriptions from classic medical texts and long-term practical experience, traditional Chinese medicine compositions are formulated by selecting Chinese medicinal herbs with specific pharmacological effects. A key approach to the prevention and treatment of ALI is the integrated treatment of traditional Chinese medicine (TCM) and modern medicine, guided by TCM syndrome differentiation theory. Chinese medicinal herbs are readily available and affordable, making them highly suitable for public access. Therefore, research on TCM formulas that can prevent and treat ALI is of great practical significance. Summary of the Invention

[0005] The purpose of the present invention is to provide a traditional Chinese medicine composition for treating acute lung injury, and its preparation and application.

[0006] The first aspect of the present invention provides a traditional Chinese medicine composition for treating acute lung injury, which is prepared from the following raw materials in parts by weight: 2-10 parts of roasted licorice, 3-10 parts of ginger, 3-10 parts of ginseng, 10-40 parts of raw rehmannia, 3-10 parts of cassia twig, 3-10 parts of donkey-hide gelatin, 5-20 parts of ophiopogon, 5-20 parts of hemp seed, and 5-30 parts of jujube.

[0007] In a preferred example, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 3-9 parts of roasted licorice, 4-8 parts of ginger, 3-8 parts of ginseng, 10-35 parts of raw rehmannia, 4-9 parts of cinnamon twig, 4-9 parts of donkey-hide gelatin, 6-12 parts of ophiopogon, 6-15 parts of hemp seed, and 6-15 parts of jujube.

[0008] In a preferred example, the Chinese medicine composition is made of the following raw materials in parts by weight: 2 parts of roasted licorice, 3 parts of ginger, 3 parts of ginseng, 10 parts of raw rehmannia, 3 parts of cinnamon twig, 3 parts of donkey-hide gelatin, 5 parts of ophiopogon, 5 parts of hemp seed, and 5 parts of jujube.

[0009] In another preferred example, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 4 parts of roasted licorice, 4 parts of ginger, 4 parts of ginseng, 15 parts of raw rehmannia, 4 parts of cinnamon twig, 4 parts of donkey-hide gelatin, 7 parts of ophiopogon, 7 parts of hemp seed, and 10 parts of jujube.

[0010] In another preferred example, the Chinese medicine composition is made of the following raw materials in parts by weight: 7 parts of roasted licorice, 7 parts of ginger, 7 parts of ginseng, 27 parts of raw rehmannia, 7 parts of cinnamon twig, 7 parts of donkey-hide gelatin, 14 parts of ophiopogon, 14 parts of hemp seed, and 20 parts of jujube.

[0011] In another preferred example, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 8 parts of roasted licorice, 6 parts of ginger, 4 parts of ginseng, 15 parts of raw rehmannia, 6 parts of cinnamon twig, 4 parts of donkey-hide gelatin, 12 parts of ophiopogon, 15 parts of hemp seed, and 12 parts of jujube.

[0012] In another preferred example, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 10 parts of roasted licorice, 10 parts of ginger, 10 parts of ginseng, 40 parts of raw rehmannia, 10 parts of cinnamon twig, 10 parts of donkey-hide gelatin, 20 parts of ophiopogon, 20 parts of hemp seed, and 30 parts of jujube.

[0013] In another preferred example, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 8 parts of roasted licorice, 6 parts of ginger, 4 parts of ginseng, 32 parts of raw rehmannia, 6 parts of cinnamon twig, 4 parts of donkey-hide gelatin, 16 parts of ophiopogon, 16 parts of hemp seed, and 12 parts of jujube.

[0014] The second aspect of the present invention is to provide a Chinese medicine compound preparation for treating acute lung injury, wherein the Chinese medicine compound preparation is a decoction, granules, powder, capsule, tablet or oral liquid with the above-mentioned Chinese medicine composition as the active ingredient.

[0015] The present invention also provides use of the above-mentioned traditional Chinese medicine composition in preparing a medicine for treating acute lung injury.

[0016] The details of various aspects of the present invention will be described in detail in the following sections. The features, objectives and advantages of the present invention will become more apparent through the description below and the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The Chinese medicine composition alleviates the pathological damage of lung tissue in ALI mice

[0018] Figure 2 This shows that the Chinese medicine combination has an effect on the pathological score of lung injury in ALI mice

[0019] Figure 3 The results showed that the Chinese herbal medicine composition reduced the mRNA expression of inflammatory factors IL-1β, IL-6, TNF-α and hypoxia-inducible factor Hif-1α in the lung tissue of ALI mice.

[0020] Figure 4 The results showed that the Chinese herbal medicine composition reduced the protein expression of serum inflammatory factors IL-1β, IL-6, IL-17, and TNF-α in ALI mice. DETAILED DESCRIPTION

[0021] The Chinese medicinal materials used by the inventors in preparing the pharmaceutical composition of the present invention are from the following sources: Radix Glycyrrhizae (Roasted Licorice), a processed product of licorice root, is derived from the dried roots and rhizomes of Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat., or Glycyrrhiza glabra L. (Leguminosae); Ginger, is the fresh rhizome of Zingiber officinale Rosc. (Zingiberaceae); Ginseng, is the dried roots and rhizomes of Panax ginseng CA mel. (Araliaceae); Rehmannia glutinosa, is the fresh or dried tuberous root of Rehmannia glutinosa Libosch. (Scrophulariaceae); Cinnamon Twig, is the dried young branches of Cinnamomum cassia Presl. (Lauraceae); and Donkey-hide Gelatin, is a solid gelatin made by decocting and concentrating the dried or fresh bark of Equus asinus L. (Equus asinus L.). Ophiopogon japonicus (Lf) Ker-Gawl., a plant of the Liliaceae family. Hemp seed (Hemp seed) is the dried mature fruit of Cannabis sativa L., a plant of the Moraceae family. Jujube (Ziziphus jujuba Mill.), a plant of the Rhamnaceae family. Based on years of clinical experience, the present inventors believe that both the six exogenous pathogens and the seven internal emotions can contribute to this disease, but that insufficient vital energy and a loss of both qi, blood, yin, and yang permeate the entire condition. In Traditional Chinese Medicine, ALI falls under the categories of "asthma and collapse," "chest congestion," and "sudden asthma." Its pathogenesis is that pathogenic factors gradually damage the body's vital energy, leading to insufficient vital energy and a deficiency of vital energy, resulting in a weakening of both qi, blood, yin, and yang, ultimately overwhelming the body. According to Traditional Chinese Medicine theory, this prescription has the effects of invigorating qi, nourishing yin, and promoting yang, primarily focusing on strengthening vital energy, in line with the Yellow Emperor's Classic of Internal Medicine's statement that "when vital energy is present within, pathogenic factors cannot prevail." The present Chinese medicine composition contains nine herbs. Roasted licorice is the main ingredient, tonifying the middle and replenishing qi. Ginseng and jujube nourish the heart and spleen, while combined with roasted licorice root, they contribute to the production of qi and blood, replenishing lung qi and moistening the lungs to relieve coughs. Donkey-hide gelatin, hemp seed, and Ophiopogon japonicus nourish yin and blood, and donkey-hide gelatin and Ophiopogon japonicus are also known to nourish lung yin and moisten dry lungs. Donkey-hide gelatin, hemp seed, Ophiopogon japonicus, ginseng, and jujube serve as assistant herbs. Cinnamon twig and ginger warm yang and invigorate qi, promoting the flow of qi and blood, ensuring a continuous flow of qi and blood, and making rich flavors nourishing without being greasy, serving as adjuvant herbs. Rehmannia root and hemp seed, combined with donkey-hide gelatin and Ophiopogon japonicus, embody the concept of "metal and water complementing each other." Therefore, these herbs, when combined, achieve the goal of "invigorating lung qi, nourishing lung yin, and promoting lung yang."

[0022] The Chinese medicine composition of the present invention can be prepared according to conventional methods in the art, for example: all medicinal materials are placed in water and soaked for 0.5 to 2 hours, all the medicinal materials are boiled together over high heat and then simmered over low heat for half an hour; after removing the residue and taking the juice, the same method is continued, and the two decoctions are mixed; during the preparation process, the amount of water added is controlled so that the water surface exceeds the surface of the medicinal materials by 2 to 3 cm.

[0023] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. Any methods and materials similar or equivalent to those described herein can be applied to the present invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally performed under conventional conditions or as recommended by the manufacturer.

[0024] The above-mentioned features of the present invention or the features described in the embodiments may be combined in any combination. All features disclosed in this patent specification may be used in any combination, and each feature disclosed in the specification may be replaced by any alternative feature that can provide the same, equal, or similar purpose. Therefore, unless otherwise specified, the features disclosed are only general examples of equal or similar features.

[0025] The raw materials used in this invention, such as licorice root, ginger, ginseng, raw rehmannia, cinnamon twig, donkey-hide gelatin, ophiopogon japonicus, hemp seed, and jujube, all comply with the relevant provisions under the main text of the Chinese Pharmacopoeia 2020 edition, Part I. Each medicinal material has been processed by cleaning, cutting, processing, and crushing. Before feeding, it has been identified that the actual medicinal material is consistent with the name and the quality meets the national pharmacopoeia standards (the specific medicinal material identification method is implemented in accordance with the pharmacopoeia standards).

[0026] The traditional Chinese medicine composition of the present invention has the effects of invigorating qi, nourishing yin and promoting yang, can effectively improve the symptoms of lung injury, prevent the aggravation of lung injury, and can prevent and treat acute lung injury. It has definite efficacy, few side effects, and low cost. It is especially suitable for use during the epidemic season of respiratory viruses and has promotion and application value.

[0027] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, all percentages, ratios, proportions, or parts are by weight.

[0028] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0029] The above-mentioned features of the present invention or the features described in the embodiments may be combined in any combination. All features disclosed in this patent specification may be used in any combination, and each feature disclosed in the specification may be replaced by any alternative feature that can provide the same, equal, or similar purpose. Therefore, unless otherwise specified, the features disclosed are only general examples of equal or similar features.

[0030] Example 1: Preparation of Chinese medicine composition

[0031] Weigh 2g of roasted licorice root, 3g of ginger, 3g of ginseng, 10g of raw rehmannia, 3g of cinnamon twig, 3g of donkey-hide gelatin, 5g of ophiopogon, 5g of hemp seed, and 5g of jujube, and soak all the medicinal materials in water for 0.5 hour. Boil all the medicinal materials together over high heat, then simmer over low heat for half an hour; remove the residue and take the juice, then continue this process, mixing the two decoctions; during the preparation process, control the amount of water added so that the water surface exceeds the surface of the medicinal materials by 2-3 cm.

[0032] Example 2: Preparation of Chinese medicine composition

[0033] Weigh 4g of roasted licorice root, 4g of ginger, 4g of ginseng, 15g of raw rehmannia root, 4g of cinnamon twig, 4g of donkey-hide gelatin, 7g of ophiopogon japonicus, 7g of hemp seed, and 10g of jujube, and soak all the medicinal materials in water for 0.5 hour. Boil all the medicinal materials together over high heat, then simmer over low heat for half an hour; remove the residue and take the juice, then continue this process, mixing the two decoctions; during the preparation process, control the amount of water added so that the water surface exceeds the surface of the medicinal materials by 2-3 cm.

[0034] Example 3: Preparation of Chinese medicine composition

[0035] Weigh 7g of roasted licorice root, 7g of ginger, 7g of ginseng, 27g of raw rehmannia root, 7g of cinnamon twig, 7g of donkey-hide gelatin, 14g of ophiopogon japonicus, 14g of hemp seed, and 20g of jujube, and soak all the medicinal materials in water for 1 hour. Boil all the medicinal materials together over high heat, then simmer over low heat for half an hour; remove the residue and take the juice, then continue this method, and mix the two decoctions; during the preparation process, control the amount of water added so that the water surface exceeds the surface of the medicinal materials by 2-3 cm.

[0036] Example 4: Preparation of Chinese medicine composition

[0037] Weigh 8g of roasted licorice root, 6g of ginger, 4g of ginseng, 15g of raw rehmannia, 6g of cinnamon twig, 4g of donkey-hide gelatin, 12g of ophiopogon japonicus, 15g of hemp seed, and 12g of jujube, and soak all the medicinal materials in water for 1 hour. Boil all the medicinal materials together over high heat, then simmer over low heat for half an hour. Remove the residue and take the juice, then continue this process, mixing the two decoctions. During the preparation process, control the amount of water added so that the water surface is 2-3 cm above the surface of the medicinal materials.

[0038] Example 5: Preparation of Chinese medicine composition

[0039] Weigh 10g of roasted licorice root, 10g of ginger, 10g of ginseng, 40g of raw rehmannia, 10g of cinnamon twig, 10g of donkey-hide gelatin, 20g of ophiopogon japonicus, 20g of hemp seed, and 30g of jujube, and soak all the medicinal materials in water for 1 hour. Boil all the medicinal materials together over high heat, then simmer over low heat for half an hour; remove the residue and take the juice, then continue this method, and mix the two decoctions; during the preparation process, control the amount of water added so that the water surface is 2-3 cm above the surface of the medicinal materials.

[0040] Example 6: Preparation of Chinese medicine composition

[0041] Weigh 8g of roasted licorice root, 6g of ginger, 4g of ginseng, 32g of raw rehmannia, 6g of cinnamon twig, 4g of donkey-hide gelatin, 16g of ophiopogon japonicus, 16g of hemp seed, and 12g of jujube, and soak all the medicinal materials in water for 1.5 hours. Boil all the medicinal materials together over high heat, then simmer over low heat for half an hour; remove the residue and take the juice, then continue this process, mixing the two decoctions; during the preparation process, control the amount of water added so that the water surface exceeds the surface of the medicinal materials by 2-3 cm.

[0042] Example 7: Pharmaceutical research on Chinese medicine composition

[0043] 7.1 Animals:

[0044] Sixty healthy male C57BL / 6 mice, SPF grade, weighing (20-22) g, were obtained from Shanghai SLACK Laboratory Animal Co., Ltd. (certificate number: 20220004035589). They were fed with a special standard pellet feed and water. Animal ethics review number: PZSHUTCM2310260004. The experiment was conducted in the Experimental Animal Center of Shanghai University of Traditional Chinese Medicine (SYCK (Shanghai) 2020-0009).

[0045] 7.2 Main equipment and instruments:

[0046] A 3K15 low-temperature refrigerated centrifuge (Sigma, USA); a 680 microplate reader (Bio-Rad, USA); a real-time fluorescence quantitative PCR instrument (QuantSTudio3, USA); a PCR instrument (Mastercycler nexus, Germany); an SDS-PAGE electrophoresis apparatus (MINIProtean2 Bio-Rad, USA); and a pathology section machine (Shanghai Leica Instrument Co., Ltd., China).

[0047] 7.3 Drugs and Reagents:

[0048] Dexamethasone tablets, Tianjin Xinyi Jinjin Pharmaceutical Co., Ltd., batch number 501220321; lipopolysaccharide, Sigma-Aldrich, batch number 0000153963; IL-6 ELISA kit (Nanjing Boyan Biotechnology, catalog number: BY-EM220188); IL-1β ELISA kit (Nanjing Boyan Biotechnology, catalog number: BY-EM220174); TNF-α ELISA kit (Nanjing Boyan Biotechnology, catalog number: BY-EM220852); IL-17 ELISA kit (Nanjing Boyan Biotechnology, catalog number: BY-EM220852); SA kit (Nanjing Boyan Biotechnology, catalog number: BY-EM220170); Hif-1α ELISA kit (Yuanxin Biotechnology, catalog number: YX-E20354); RNA extraction kit (B0004D, EZ Bioscience, Suzhou, China); reverse transcription kit (TakaRa, China, catalog number: RR036A); qPCR kit (Novozymes, China, catalog number: Q711-02); primer sequences (Sangon Biotechnology Co., Ltd., China).

[0049] The Chinese medicine composition prepared according to Example 4 was reheated and concentrated to 38.4 ml, i.e., a concentration of 2.132 g / ml, and then cooled and stored at 4°C for later use.

[0050] 7.4 Statistical methods:

[0051] All data were statistically analyzed using SPSS 29.0 software. All experiments were repeated 3 times or more, and the data were After the homogeneity of variance test, one-way analysis of variance was performed for data from more than two groups, and the independent sample t-test was used to test the differences between two groups. P < 0.05 was considered statistically significant.

[0052] 7.5 Observation of general condition of mice:

[0053] Experimental methods:

[0054] Healthy male SPF-grade C57BL / 6 mice were randomly divided into a normal group, a model group, a Chinese herbal composition (high, medium, and low doses), and a dexamethasone group; 10 mice per group. They weighed (20-22) g and were housed in an SPF-grade barrier environment with a 12-h light-dark cycle, (24±2)°C, and a relative humidity of (55±5)%. They had free access to food and water. After one week of adaptive feeding, the Chinese herbal composition (high, medium, and low doses) groups were given 0.2 ml of the corresponding concentration of Chinese herbal composition (high dose 21.32 g / kg, medium dose 10.66 g / kg, low dose 5.33 mg / kg) once daily. The dexamethasone group was given 0.2 ml of dexamethasone (5 mg / kg) by oral gavage. The normal and model groups were given 0.2 ml of normal saline by oral gavage once daily. On the 7th day after gavage, the mice were anesthetized and fixed in a supine position on the operating table. The throat and glottis were exposed. Under bright field conditions, a soft intravenous cannula was slowly inserted into the airway, and 50ul of LPS was dripped at a dose of 5mg / kg. Mice in the model group, the Chinese medicine composition (high, medium, and low dose) group, and the dexamethasone group were all given LPS tracheal instillation to establish the model. The normal group was given 50ul of normal saline tracheal instillation. During the experiment, the mental state, activity, fur gloss, and mortality of all mice were observed. 24 hours after modeling, the mice were processed, and blood was collected to separate serum for serological testing. The right lobe of the lung was fixed in 4% paraformaldehyde, dehydrated in gradient ethanol, embedded in paraffin, and sliced for lung tissue pathological morphology. The remaining lungs were quickly frozen in liquid nitrogen and stored in a -80°C refrigerator for later use.

[0055] Experimental results:

[0056] Three mice in the model group died during the experiment. Following intratracheal instillation of LPS, the model group experienced transient dyspnea and gradually developed symptoms of Qi and Yin deficiency, including curling up, lethargy, sluggish reactions, a marked decrease in activity, decreased food and water intake, dry and dull fur, hair loss, and hard stools. Compared with the model group, the TCM combination group, and the dexamethasone group, the mice in the normal group displayed a healthy spirit, were active, responsive, ate normally, and had smooth fur. Following oral administration, the mice in the TCM combination and dexamethasone groups showed minimal response to the modeling drug, LPS, and all of these Qi and Yin deficiency symptoms were alleviated to some extent.

[0057] 7.6 Observation on the inhibitory effect of Chinese herbal medicine composition on dyspnea in mice

[0058] Experimental methods:

[0059] Two experimental technicians observed the symptoms of dyspnea in mice and heard obvious dry or wet rales or sputum sounds without using a stethoscope. The onset time was recorded and the average of the two records was taken.

[0060] Experimental results:

[0061] Table 1 demonstrates the inhibitory effect of the Chinese herbal composition on dyspnea in mice. All mice in the model group experienced dyspnea. Compared with the model group, the duration of dyspnea in the high-, medium-, and low-dose groups of the Chinese herbal composition was significantly reduced (P < 0.01). However, the duration of dyspnea in the high-dose group was not significantly different from that in the dexamethasone group (P > 0.05). This suggests that the Chinese herbal composition alleviated dyspnea symptoms, with the degree of relief in the high-dose group being similar to that observed with the Western medicine dexamethasone.

[0062] Table 1 Comparison of the onset time of dyspnea in mice in each group

[0063]

[0064] Note: Compared with the model group, * indicates: P < 0.05, ** indicates: P < 0.01; compared with the dexamethasone group: △ indicates: P < 0.05, △△ indicates: P < 0.01

[0065] 7.7 Observation of mouse lung tissue pathology

[0066] Experimental methods:

[0067] Fresh lung tissue from mice was fixed in 4% paraformaldehyde. Paraffin sections were dehydrated as usual using the following procedure: 75% ethanol for 2 hours → 85% ethanol for 2 hours → 90% ethanol for 2 hours → 100% ethanol for 45 minutes → 100% ethanol for 45 minutes → ethanol + xylene (1:1) for 10 minutes → xylene for 8 minutes → xylene for 8 minutes → paraffin for 1 hour → paraffin for 1 hour → paraffin for 1 hour. Paraffin embedding was performed as usual. The paraffin block was placed in a -20°C freezer and allowed to cool. The paraffin block was then clamped to the paraffin section holder and cut into 4-6 microns. The sections were placed on the surface of the microtome (water temperature 40°C). The tissue sections were removed with a glass slide and baked in a 60°C oven for 1 hour before use. Paraffin sections were dewaxed to water using the following procedure: xylene I for 20 minutes → xylene II for 20 minutes → anhydrous ethanol I for 5 minutes → anhydrous ethanol II for 5 minutes → 75% alcohol for 5 minutes in a gradient dewaxing process, and then washed with water. Stain sections in hematoxylin for 3-5 minutes. Rinse with tap water to remove excess stain. Incubate with 1% hydrochloric acid for several seconds. Rinse with tap water, then with 0.6%-0.7% ammonia solution to blue the sections. Rinse with running water for several seconds. Dehydrate paraffin sections in a gradient of alcohol (85% ethanol → 95% ethanol) and stain in eosin solution for 5 minutes. Transparent paraffin sections by sequentially placing sections in anhydrous ethanol I for 5 minutes, anhydrous ethanol II for 5 minutes, anhydrous ethanol III for 5 minutes, n-butanol for 5 minutes, xylene I for 5 minutes, and xylene II for 5 minutes. Remove sections from the xylene, allow to dry briefly, and mount with neutral gum. Observe sections under a light microscope and preserve images.

[0068] Experimental results:

[0069] HE staining observation of mouse lung tissue

[0070] Figure 1 The effect of the Chinese medicine composition on the pathological morphology of lung tissue in ALI mice was reflected; Figure 1 The normal group showed clear alveolar structure, uniform alveolar size, and intact airway epithelial structure, with no obvious pathological changes. In the model group, the lung tissue was disorganized, with numerous red blood cells visible in the alveolar cavity and interstitium, and flake-like blood foci visible to the naked eye. The alveolar walls were significantly thickened, with hyaline membranes forming on the alveolar walls. Some bronchial epithelial cells were sloughed, and there was infiltration of neutrophils and lymphocytes. These pathological changes were alleviated in all groups treated with the Chinese medicine composition.

[0071] 7.8 Pathological scoring of mouse lung tissue injury

[0072] Experimental methods:

[0073] Lung tissue sections were scored on a scale of 0 to 4 based on the degree of localized pulmonary necrosis, neutrophil-erythrocyte infiltration, alveolar septal thickening, alveolar luminal enlargement, and hyaline membrane formation: 0 for no lesion, 1 for lesion covering <25%, 2 for lesion covering 25% to 50%, 3 for lesion covering 51% to 75%, and 4 for lesion covering >75%. Ten high-power fields were scored for each section.

[0074] Experimental results:

[0075] Figure 2 The results show that the Chinese medicine combination can improve the pathological score of lung injury in ALI mice; in the figure, compared with the normal group, #: P < 0.05, ##: P < 0.01; compared with the model group, *: P < 0.05, **: P < 0.01; compared with the dexamethasone group: △: P < 0.05, △△: P < 0.01; Figure 2 As can be seen, compared with the normal group, the lung injury pathology scores of mice in the model group were significantly increased (P < 0.01). Compared with the model group, the lung injury scores of mice in the high, medium, and low dose groups of the Chinese medicine composition were significantly reduced (P < 0.01), and the lung injury scores of mice in the high dose group were not significantly different from those in the dexamethasone group (P > 0.05). This indicates that the Chinese medicine composition alleviated the lung pathology of mice, and the degree of reduction in the high dose group was similar to that of the Western medicine dexamethasone.

[0076] 7.9 Determination of mRNA Expression of Inflammatory Factors IL-1β, IL-6, TNF-α, and Hypoxia-Inducible Factor Hif-1α in Mouse Lung Tissue

[0077] Experimental methods:

[0078] RNA was extracted from fresh lung tissue using an RNA extraction kit and then reverse transcribed into cDNA. The primer sequences are shown in Table 2. TM3 lines were subjected to real-time quantitative PCR. The data were analyzed by relative quantitative method, with GAPDH as internal reference, 2 -△△Ct The relative expression of mRNA was determined.

[0079] Table 2 Primer sequences

[0080]

[0081] Experimental results:

[0082] Figure 3 The results showed that the Chinese medicine composition can effectively inhibit the mRNA expression of inflammatory factors IL-1β, IL-6, TNF-α and hypoxia-inducible factor Hif-1α in lung tissue of ALI mice; in the figure, compared with the normal group, #: P < 0.05, ##: P < 0.01; compared with the model group, *: P < 0.05, **: P < 0.01; compared with the dexamethasone group: △: P < 0.05, △△: P < 0.01; compared with the dexamethasone group, △: P < 0.05, △△: P < 0.01; Figure 3 It can be seen that compared with the normal group, the expression of IL-1β, IL-6, TNF-α, and Hif-1α mRNA in the lung tissue of mice in the model group was significantly increased (P<0.05). Compared with the model group, the expression of IL-1β, IL-6, TNF-α, and Hif-1α mRNA in the lung tissue of mice in the high, medium, and low dose groups of the Chinese medicine composition was significantly reduced (P<0.05, P<0.01). There was no significant difference in the expression of IL-6 and TNF-α mRNA in the high dose group and the dexamethasone group (P>0.05). This shows that the Chinese medicine composition significantly reduced the gene expression of inflammatory factors and hypoxia-inducible factors in the lung tissue of mice, and some indicators were similar to those of the Western medicine dexamethasone.

[0083] 7.10 Determination of protein expression of inflammatory factors IL-1β, IL-6, IL-17, and TNF-α in mouse serum

[0084] Experimental methods:

[0085] Mouse serum was collected and assayed for IL-1β, IL-6, IL-17, and TNF-α levels according to the ELISA kit instructions. Standards and samples were added to pre-coated enzyme plates. The respective biotinylated antigens were added and incubated at 37°C for 30 minutes. After adding the developer and terminator, the OD value of each well was measured using a microplate reader at 450 nm. A linear regression equation was established between the standard concentration and its OD value to calculate the sample concentration.

[0086] Experimental results:

[0087] Figure 4The results show that the Chinese medicine composition can improve the protein expression of IL-1β, IL-6, IL-17 and TNF-α in the serum of ALI mice; in the figure, compared with the normal group, #: P < 0.05, ##: P < 0.01; compared with the model group, *: P < 0.05, **: P < 0.01; compared with the dexamethasone group: △: P < 0.05, △△: P < 0.01; Figure 4 It can be seen that compared with the normal group, the expression of IL-1β, IL-6, IL-17, and TNF-α proteins in the serum of mice in the model group was significantly increased (P<0.01). Compared with the model group, the expression of IL-1β protein in the serum of mice in the high, medium, and low dose groups of the Chinese medicine composition was significantly reduced (P<0.05, P<0.01). The expression of IL-1β protein in the serum of mice in the high, medium, and low dose groups of the Chinese medicine composition was significantly reduced (P<0.05, P<0.01), and the expression of IL-6, IL-17, and TNF-α proteins in the serum of mice in the high and medium dose groups of the Chinese medicine composition was significantly reduced (P<0.05, P<0.01). Compared with the dexamethasone group, some indicators had no significant differences with the dexamethasone group (P>0.05). It shows that the Chinese medicine composition reduces the level of inflammatory factors in the serum of mice, and the degree of reduction of some indicators is similar to that of the Western medicine dexamethasone. Example 8 Comparative study of Chinese medicine composition and literature prescriptions

[0088] Experimental animals:

[0089] Thirty-two healthy male C57BL / 6 mice, SPF grade, weighing (20-22) g, were obtained from Shanghai SLACK Laboratory Animal Co., Ltd. (certificate number: 20220004035589). They were fed with a special standard pellet feed and water. Animal ethics review number: PZSHUTCM2310260004. The experiment was conducted in the Experimental Animal Center of Shanghai University of Traditional Chinese Medicine (SYCK (Shanghai) 2020-0009).

[0090] Experimental drugs:

[0091] The Chinese medicine composition was prepared according to Example 4

[0092] Literature:

[0093] 30g of roasted licorice root, 9g of dried ginger, 15g of ginseng, 15g of cooked rehmannia root, 15g of cinnamon twig, 9g of donkey-hide gelatin (melted), 15g of ophiopogon japonicus, 9g of hemp seed, 9g of jujube, 15g of northern adenophora root, 12g of leech, and 12g of arisaema. Decoction in cold water until 300ml of juice is collected. References (Zhang Yangyang, Sun Cong, Yang Xingmei, et al. Study on the efficacy of modified roasted licorice decoction combined with enalapril in the treatment of chronic obstructive pulmonary disease and heart failure [J]. Shaanxi Journal of Traditional Chinese Medicine, 2021, 42(09): 1219-1222.).

[0094] Experimental methods:

[0095] Mice were randomly divided into a normal group, a model group, a Chinese herbal medicine combination group, and a literature prescription group. Each group had 8 mice. After one week of adaptive feeding, the Chinese herbal medicine combination and literature prescription groups were gavaged once daily with the Chinese herbal medicine combination and literature prescription, respectively. The normal and model groups were gavaged with an equal volume of normal saline once daily. On the seventh day of gavage, mice were anesthetized and fixed in a supine position on an operating table. The pharynx and glottis were exposed. Under bright field vision, a soft intravenous cannula was slowly inserted into the airway, and 50 μl of LPS was dripped at a dose of 5 mg / kg. LPS was administered intratracheally to mice in the model, Chinese herbal medicine combination, and literature prescription groups to establish the model. The normal group received 50 μl of normal saline. All mice were observed for dyspnea during the experiment, and the onset time was recorded. Twenty-four hours after modeling, mice were treated, and blood was collected and serum was separated for serological analysis. Interleukin-1β, IL-6, and Hif-1α levels were measured according to the ELISA kit instructions.

[0096] Statistical methods:

[0097] All data were statistically analyzed using SPSS 29.0 software. After the homogeneity of variance test, one-way analysis of variance was used for data from more than two groups. P < 0.05 was considered statistically significant.

[0098] Experimental results:

[0099] Table 3 demonstrates that the TCM composition is superior to the literature-based prescription in suppressing dyspnea in mice; all mice in the model group experienced dyspnea. As shown in Table 3, compared with the model group, the TCM composition group experienced significantly shorter dyspnea duration (P < 0.01) and a superior suppressive effect on dyspnea compared to the literature-based prescription group (P < 0.01).

[0100] Table 3 Comparison of respiratory distress onset time in mice of each group

[0101]

[0102] Note: Compared with the model group, * indicates: P < 0.05, ** indicates: P < 0.01; compared with the literature: △ indicates: P < 0.05, △△ indicates: P < 0.01

[0103] Table 4 shows the effects of the Chinese herbal composition on inflammatory factors and hypoxia-inducible factors in mice. As shown in Table 4, compared with the normal group, the expression of inflammatory factors IL-1β, IL-6, and hypoxia-inducible factor Hif-1α in the model group was significantly increased (P < 0.01). The Chinese herbal composition group had a superior inhibitory effect on the expression of inflammatory factors and hypoxia-inducible factors in mice compared with the reference group (P < 0.01).

[0104] Table 4 Levels of inflammatory factors and hypoxia-inducible factors in mouse serum ( pg / ml)

[0105]

[0106] Note: Compared with the normal group, # indicates: P < 0.05, ## indicates: P < 0.01; compared with the model group, * indicates: P < 0.05, ** indicates: P < 0.01; compared with the literature: △ indicates: P < 0.05, △△ indicates: P < 0.01

[0107] Example 9 Comparative Study of Chinese Medicine Composition and Separated Prescriptions

[0108] Experimental animals:

[0109] 96 healthy male C57BL / 6 mice, SPF grade, weighing (20-22) g, were obtained from Shanghai SLACK Laboratory Animal Co., Ltd., with a certificate number of 20220004035589. They were fed with a dedicated standard pellet feed and water. Animal ethics review number: PZSHUTCM2310260004. The experiment was conducted in the Experimental Animal Center of Shanghai University of Traditional Chinese Medicine (SYCK (Shanghai) 2020-0009). Experimental drugs:

[0110] The Chinese medicine composition was prepared according to Example 4

[0111] The ingredients of the split prescription 1 are: 6g ginger, 4g ginseng, 15g raw rehmannia, 6g cinnamon twig, 4g donkey-hide gelatin, 12g ophiopogon, 15g hemp seed, and 12g jujube. The ingredients of the split prescription 2 are: 8g roasted licorice root, 4g ginseng, 15g raw rehmannia, 6g cinnamon twig, 4g donkey-hide gelatin, 12g ophiopogon, 15g hemp seed, and 12g jujube. The ingredients of the split prescription 3 are: 8g roasted licorice root, 6g ginger, 15g raw rehmannia, 6g cinnamon twig, 4g donkey-hide gelatin, 12g ophiopogon, 15g hemp seed, and 12g jujube.

[0112] The composition of prescription 4 is: 8g of roasted licorice root, 6g of ginger, 4g of ginseng, 6g of cinnamon twig, 4g of donkey-hide gelatin, 12g of ophiopogon, 15g of hemp seed, 12g of jujube, the composition of prescription 5 is: 8g of roasted licorice root, 6g of ginger, 4g of ginseng, 15g of raw earth, 4g of donkey-hide gelatin, 12g of ophiopogon, 15g of hemp seed, 12g of jujube, the composition of prescription 6 is: 8g of roasted licorice root, 6g of ginger, 4g of ginseng, 15g of raw earth, 15g of cinnamon twig, 12g of ophiopogon, 12g of hemp seed, 15g of jujube g, Decomposed prescription 7: 8g of roasted licorice root, 6g of ginger, 4g of ginseng, 15g of raw earth, 6g of cinnamon twig, 4g of donkey-hide gelatin, 15g of hemp seed, 12g of jujube, Decomposed prescription 8: 8g of roasted licorice root, 6g of ginger, 4g of ginseng, 15g of raw earth, 6g of cinnamon twig, 4g of donkey-hide gelatin, 12g of ophiopogon japonicus, 12g of jujube, Decomposed prescription 9: 8g of roasted licorice root, 6g of ginger, 4g of ginseng, 15g of raw earth, 6g of cinnamon twig, 4g of donkey-hide gelatin, 12g of ophiopogon japonicus, 15g of hemp seed.

[0113] Preparation method of the split prescription medicine: prepare it according to the method of Example 4.

[0114] Experimental methods:

[0115] Mice were randomly divided into a normal group, a model group, a Chinese herbal combination group, and a separate prescription group (Separate Prescription 1, Separate Prescription 2, Separate Prescription 3, Separate Prescription 4, Separate Prescription 5, Separate Prescription 6, Separate Prescription 7, Separate Prescription 8, and Separate Prescription 9). Each group had 8 mice. After one week of adaptive feeding, the Chinese herbal combination and separate prescription groups were administered the Chinese herbal combination and separate prescription drugs, respectively, once daily. The normal and model groups were gavaged with an equal volume of normal saline once daily. On the seventh day of gavage, mice were anesthetized and fixed in a supine position on an operating table. The pharynx and glottis were exposed. Under bright field vision, a soft intravenous cannula was slowly inserted into the airway, and 50 μl of LPS was dripped at a dose of 5 mg / kg. LPS was administered intratracheally to establish the model in the model, Chinese herbal combination, and separate prescription groups. The normal group was administered 50 μl of normal saline. All mice were observed for dyspnea during the experiment, and the onset time was recorded. Twenty-four hours after modeling, mice were treated, and blood was collected and serum was isolated for serological testing.

[0116] Statistical methods:

[0117] All data were statistically analyzed using SPSS 29.0 software. After the homogeneity of variance test, one-way analysis of variance was used for data from more than two groups. P < 0.05 was considered statistically significant.

[0118] Experimental results:

[0119] Table 5 demonstrates that the TCM combination significantly inhibited dyspnea symptoms, inflammatory factors, and hypoxia-inducible factor (HIF) in mice compared to the individual components of the combined formula. All mice in the model group developed dyspnea. Table 5 shows that compared with the model group, the TCM combination and the individual components of the combined formula significantly reduced dyspnea duration (P < 0.01). However, the therapeutic effects of any of the individual components that omitted a single herb were significantly inferior to those of the TCM combination, with statistically significant differences (P < 0.01). Compared with the normal group, serum levels of the inflammatory factor IL-6 and the hypoxia-inducible factor Hif-1α were significantly elevated in the model group (P < 0.01). Compared with the model group, serum levels of the inflammatory factors IL-6 and Hif-1α were significantly decreased in the TCM combination and the individual components of the combined formula (P < 0.01). However, the therapeutic effects of any of the individual components that omitted a single herb were significantly inferior to those of the TCM combination, with statistically significant differences (P < 0.01).

[0120] Table 5 Comparison of dyspnea onset time, inflammatory factors and hypoxia-inducible factors in mice of each group

[0121]

[0122] Note: Compared with the normal group, # means: P < 0.05, ## means: P < 0.01; compared with the model group, * means: P < 0.05, ** means: P < 0.01;

[0123] Example 10 Comparative Study of Treatment with Chinese Medicine Composition and Conventional Western Medicine Treatment

[0124] Research subjects

[0125] The patient went to the outpatient clinic of Traditional Chinese Medicine Internal Medicine of Shuguang Hospital and was diagnosed with acute lung injury. He was in the recovery period and was diagnosed with both Qi and Yin deficiency by Traditional Chinese Medicine.

[0126] Western medicine diagnostic criteria:

[0127] The diagnostic criteria for acute lung injury (ALI) follow the 2012 Berlin Definition (Acute Respiratory Distress Syndrome: The Berlin Definition): pulmonary imaging changes, PaO2 / FiO2 ≤ 300 mmHg, and pulmonary edema caused by heart failure or fluid overload excluded. Onset must be within the past month.

[0128] TCM diagnostic criteria:

[0129] Refer to the chapter "Asthma" in the "Tenth Edition of the National TCM Colleges and Universities Planning Textbook for Internal Medicine". The main clinical manifestations are shortness of breath when moving, dyspnea, general fatigue, opening the mouth and raising the shoulders, twitching of the nasal wings, inability to lie flat, fatigue, spontaneous sweating or night sweats, low-grade fever, hot hands and feet, poor appetite, dry mouth and tongue, and red tongue with little coating. Inclusion criteria: (1) 18 years old ≤ ≥ 80 years old; (2) meet the above-mentioned Western medicine and TCM diagnostic criteria; (3) the patient and family members agree and are willing to sign the informed consent form.

[0130] Exclusion criteria:

[0131] (1) Patients who are not in the recovery stage or are in critical condition. (2) Patients with other serious systemic diseases, heart failure, severe liver and kidney dysfunction, etc. (3) Patients with abnormal consciousness or mental illness. (4) Patients with allergic constitution.

[0132] Research Methodology:

[0133] Patients in the recovery stage of acute lung injury and with both qi and yin deficiency according to TCM syndrome differentiation were randomly divided into a Chinese medicine group and a control group, with 30 cases in each group. Conventional Western medicine treatment of acute lung injury was based on the "Guidelines for the Diagnosis and Treatment of Acute Lung Injury / Acute Respiratory Distress Syndrome (2006)". The control group was given conventional Western medicine treatment (including oxygen therapy, antibiotics, glucocorticoids, fluid management, etc.). The Chinese medicine group was treated with a Chinese medicine composition (8g of roasted liquorice, 6g of ginger, 4g of ginseng, 15g of raw earth, 6g of cassia twig, 4g of donkey-hide gelatin, 12g of ophiopogon, 15g of hemp seed, and 12g of jujube) on the basis of conventional Western medicine treatment. The preparation method was the same as in Example 4. One dose was taken daily, orally after meals, and 7 days was one course of treatment. The efficacy was observed after 7 days of treatment.

[0134] Efficacy criteria:

[0135] Patient scores before and after treatment were recorded using the Traditional Chinese Medicine (TCM) syndrome scoring scale, as described in the "Clinical Disease Diagnosis Based on Cure and Improvement Criteria" and the "Guiding Principles for Clinical Research of New Traditional Chinese Medicines." The formula for calculation is: efficacy rate = (total score before treatment - total score after treatment) / total score before treatment × 100. Cured: Patients experience no significant shortness of breath or fatigue on movement, with an oxygenation index > 300 mHg; TCM syndrome score reduction of ≥ 90%; Markedly effective: Patients experience only mild shortness of breath and no significant fatigue; TCM syndrome score reduction of 60% or less, < 90%; Improved: Patients experience improved shortness of breath on movement, but some symptoms persist; TCM syndrome score reduction of 30% or less, < 60%; Ineffective: Patients experience similar shortness of breath and wheezing, with minimal improvement, but with frequent interruptions or noticeable gasps when speaking, and still experience symptoms such as mental fatigue, general fatigue, dry mouth, spontaneous sweats, and night sweats; TCM syndrome score reduction of < 30%. The total efficacy rate is calculated as: cure rate + markedly effective rate + improvement rate.

[0136] Observation of inflammation indicators:

[0137] The neutrophil ratio (NEUT%) and C-reactive protein (CRP) in the blood of the two groups of patients were detected on the 7th day of treatment. Safety monitoring:

[0138] Observe the patient for adverse reactions such as allergic reactions, diarrhea, vomiting, and dizziness.

[0139] Statistical analysis:

[0140] All data were processed using SPSS 29.0 statistical software. After the homogeneity of variance test, the independent sample t test was used to compare the groups before and after treatment, and the one-way analysis of variance was used to compare within the same group. The count data were expressed as the number of cases (%), and the categorical count data were expressed using Pearson χ 2 P < 0.05 was considered statistically significant.

[0141] Research Results:

[0142] The general information of the patients was collected, and there was no significant difference in age, gender, and smoking history. The two groups of patients were comparable. Table 6 shows the efficacy of the Chinese medicine composition on acute lung injury. The total effective rate of the Chinese medicine group can reach 96.7%, and the efficacy of the Chinese medicine group is better than that of the control group. The difference is statistically significant (P<0.01). It shows that the Chinese medicine composition significantly relieves the symptoms of patients. The total effective rate of combined treatment with the Chinese medicine composition is better than that of conventional Western medicine treatment alone. Table 7 shows the effect of the Chinese medicine composition on inflammatory indicators of patients with acute lung injury. Compared with the control group, the neutrophil ratio and C-reactive protein of patients in the Chinese medicine group were significantly reduced (P<0.01). It shows that the Chinese medicine composition can reduce inflammation in patients. Through safety monitoring, no adverse reactions occurred in patients in the Chinese medicine group. It shows that the Chinese medicine composition has good safety, definite clinical efficacy, and important clinical value for acute lung injury.

[0143] Table 6 Comparison of the efficacy of Chinese medicine compositions on acute lung injury

[0144]

[0145] Table 7 Comparison of inflammatory indicators between the two groups of patients

[0146]

[0147] Note: Compared with before treatment, # means P < 0.05, ## means P < 0.01; compared with the control group after treatment, * means P < 0.05, ** means P < 0.01;

[0148] The various aspects of the present invention have been described above. However, it should be understood that, without departing from the spirit of the present invention, those skilled in the art may make equivalent changes and modifications thereto, which also fall within the scope of the appended claims.

Claims

1. Use of a traditional Chinese medicine composition in the preparation of a medicine for treating acute lung injury in the recovery period, wherein the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 8 parts of roasted licorice, 6 parts of ginger, 4 parts of ginseng, 15 parts of raw rehmannia, 6 parts of cassia twig, 4 parts of donkey-hide gelatin, 12 parts of ophiopogon, 15 parts of hemp seed, and 12 parts of jujube.

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