Application of traditional Chinese medicine composition in preparation of medicine for preventing and / or treating lung injury caused by low pressure and low oxygen

Abstract

The invention discloses application of a traditional Chinese medicine composition in preparation of a medicine for preventing and / or treating lung injury caused by low pressure and low oxygen, and belongs to the technical field of traditional Chinese medicines. The traditional Chinese medicine composition is prepared from the following components in parts by weight: 2 parts of rhododendron anthopogonoide, 1 part of cardamom and 1 part of galangal. The traditional Chinese medicine composition not only has the functions of soothing the liver, invigorating the stomach, strengthening the body resistance, but also has good effects of improving lung functions and preventing lung injury. From the Tibetan medicine prescription theory, the rhododendron anthopogonoide is a monarch drug and has the effects of inducing diuresis to alleviate edema, clearing away lung heat and improving bronchitis; the cardamom is a ministerial drug and has the effects of dispelling cold, warming kidney and astringing pulse orifices; the galangal is an adjuvant drug and has the effects of warming the stomach and improving the functions of the spleen and the stomach. Through reasonable compatibility of the three medicinal materials, the effects of improving the stomach function, promoting digestion and absorption, improving immunity, clearing away the lung-heat, diminishing swelling, clearing heat and resisting inflammation can be achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and in particular to the use of a traditional Chinese medicine composition in preparing a medicine for preventing and / or treating lung injury caused by hypobaric hypoxia. Background Art

[0002] Low-pressure and low-oxygen environments mainly exist in high-altitude areas. They are characterized by low air pressure and low oxygen partial pressure. This environment has a significant impact on the physiological functions of humans and animals, especially on lung tissue. In high-altitude areas, as the altitude increases, the air becomes thinner and the oxygen content decreases. The amount of oxygen inhaled by the human body decreases, leading to hypoxia of tissues and organs, which in turn triggers a series of physiological and pathological changes. In high-altitude areas, about 25% of people will experience acute mountain reaction, and severe cases may develop into high-altitude pulmonary edema, which may even be life-threatening. High-altitude pulmonary edema is a severe type of acute mountain sickness, and its pathogenesis is mainly related to pulmonary vasoconstriction, pulmonary hypertension, increased pulmonary capillary permeability, and alveolar fluid exudation. In addition, long-term exposure to low-pressure and low-oxygen environments may also lead to chronic mountain sickness, such as high-altitude polycythemia and high-altitude heart disease.

[0003] Lung injury caused by hypobaric hypoxia seriously affects the work efficiency of workers in plateau areas and reduces the life experience of travelers. Currently, there is a lack of drugs that can be used to prevent plateau hypoxic injury and there are deficiencies. Summary of the invention

[0004] The purpose of the present invention is to provide a Chinese medicine composition for use in the preparation of a drug for preventing and / or treating lung injury caused by hypobaric hypoxia, so as to solve the problems existing in the above-mentioned prior art.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] One of the technical solutions of the present invention is the use of a traditional Chinese medicine composition in the preparation of a drug for preventing and / or treating lung injury caused by hypobaric hypoxia, wherein the traditional Chinese medicine composition is made of the following components in parts by weight: 2 parts of Rhododendron australis, 1 part of Cardamom and 1 part of Alpinia galanga.

[0007] Based on the above technical solution, the present invention has the following technical effects:

[0008] The prescription of the present invention not only has the functions of soothing the liver and invigorating the stomach and strengthening the body, but also has a good effect of improving lung function and preventing lung damage. From the perspective of Tibetan medicine prescription theory, Rhododendron anthopogonoides Maxim is the main medicine, which has the effects of promoting diuresis and reducing swelling, clearing lung heat, and improving bronchitis; Elettaria cardamomum (L.) Maton is the secondary medicine, which has the effects of dispelling cold, warming the kidney, and astringing the meridians; Alpinia officinarum Hance is the adjuvant medicine, which has the effects of warming the stomach and improving the spleen and stomach functions. Through the reasonable compatibility of the above three medicinal materials, it can play the role of improving stomach function, promoting digestion and absorption, enhancing immunity, clearing the lungs and reducing swelling, and clearing heat and anti-inflammation. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0010] Figure 1 The effect of this formula (SWDJ) on the lung function of hypobaric hypoxic mice. Among them, A, respiratory rate; B, minute ventilation; C, cumulative air volume; D, maximum inspiratory flow rate; E, maximum expiratory flow rate; F, 50% maximum expiratory flow rate; G, end-expiratory pause; H, end-inspiratory pause; I, relaxation time; J, tidal volume; K, inspiration-expiration time ratio. n = 8, compared with the normal group ## P<0.01; compared with the model group ** P<0.01, * P<0.05.

[0011] Figure 2 The effect of this formula on the hematological indexes of hypobaric and hypoxic mice. Among them, A, red blood cell count; B, hematocrit; C, hemoglobin; D, mean corpuscular volume; E, white blood cell count; F, neutrophil count. n = 12, compared with the normal group ## P<0.01; compared with the model group ** P<0.01, * P<0.05.

[0012] Figure 3 The effect of the extract of this formula (SWDJ) on the pathological changes of lung tissue in hypobaric and hypoxic mice. A, lung tissue morphology; B, lung water content LWC; C, lung tissue H&E staining; D, tissue Tunel staining. n = 8, compared with the normal group ##P<0.01; compared with the model group * P<0.05.

[0013] Figure 4 The effect of this formula (SWDJ) on the expression of HIF-1α and AQP1 in the lung tissue of hypobaric hypoxic mice. A, representative image of HIF-1α immunohistochemical staining in lung tissue; B, representative image of AQP1 immunohistochemical staining in lung tissue; C, quantitative analysis of HIF-1α positive area; D, quantitative analysis of AQP1 positive area. n = 3, compared with the normal group ## P<0.01; compared with the model group ** P<0.01, * P<0.05. DETAILED DESCRIPTION

[0014] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0015] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0016] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0017] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0018] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0019] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.

[0020] An embodiment of the present invention provides an application of a traditional Chinese medicine composition in the preparation of a drug for preventing and / or treating lung injury caused by hypobaric hypoxia. The traditional Chinese medicine composition is made of the following components in parts by weight: 2 parts of Rhododendron australis, 1 part of Cardamom and 1 part of Alpinia galanga.

[0021] In some specific embodiments, the method for preparing the Chinese medicine composition comprises the following steps:

[0022] (1) extracting Rhododendron australis, Elaeagnus cardamomum and Alpinia officinalis with water, and concentrating under reduced pressure to obtain a water extract;

[0023] (2) The water extract is subjected to alcohol precipitation to obtain the Chinese medicine composition.

[0024] In some specific embodiments, the water extraction conditions are: the medicinal material is added with 15 mL / g of water and boiled for extraction 3 times, and each extraction time is 1 hour.

[0025] In some specific embodiments, the method of vacuum concentration is: vacuum concentration until the amount of crude drug contained in each milliliter of drug solution is 0.5 g / mL.

[0026] In some specific embodiments, the alcohol precipitation conditions are: the alcohol content of the drug solution is 80%, and the alcohol precipitation time is 18 hours.

[0027] In some specific embodiments, the application includes at least one of the following applications:

[0028] (1) Use of a Chinese medicine composition in the preparation of a drug for preventing and / or treating pulmonary dysfunction caused by hypobaric hypoxia;

[0029] (2) Use of Chinese medicine compositions in the preparation of blood regulation drugs;

[0030] (3) Use of a Chinese medicine composition in the preparation of a drug for reducing lung water content;

[0031] (4) Use of the Chinese medicine composition in the preparation of a drug for inhibiting the expression of HIF-1a and AQP1 in lung tissue.

[0032] In some specific embodiments, the medicament further comprises a pharmaceutically acceptable carrier.

[0033] In some specific embodiments, the pharmaceutically acceptable carrier is suitable for powders, granules, tablets, pills, capsules, oral solutions or pastes.

[0034] This prescription is composed of three medicinal materials: Rhododendron anthopogonoides Maxim, Elettariacardamomum(L.)Maton, and Alpinia officinarum Hance.

[0035] According to Tibetan medicine theory, this formula not only has the functions of soothing the liver and strengthening the stomach and strengthening the body, but also has a good effect of improving lung function and preventing lung damage. From the perspective of Tibetan medicine prescription theory, Rhododendronanthopogonoides Maxim is the main medicine, which has the effects of diuresis and swelling, clearing lung heat, and improving bronchitis; Elettaria cardamomum (L.) Maton is the ministerial medicine, which has the effects of dispelling cold, warming the kidney, and astringing the meridians; Alpinia officinarum Hance is the adjuvant medicine, which has the effects of warming the stomach and improving spleen and stomach function. Through the reasonable combination of the above three medicinal materials, it can improve stomach function, promote digestion and absorption, enhance immunity, clear the lungs and swelling, and clear heat and anti-inflammation.

[0036] Example 1

[0037] (I) Screening of the best formula ratio

[0038] The present invention is based on the research method of everted intestinal sac method and combines the traditional Tibetan medicine clinical medication experience and measurement and verification. 50 ) was used as an indicator to screen the optimal ratio and extraction method of Rhododendron australis, Cardamom and Alpinia galanga. The results are shown in Table 1. The extract obtained by water extraction and alcohol precipitation of the three herbs of Rhododendron australis, Cardamom and Alpinia galanga in a weight ratio of 2:1:1 has the best efficacy (EC 50 The lowest was 0.3515), which was significantly different from the extracts in other ratio groups (P<0.01).

[0039] Table 1 The median effective dose of the drugs in this prescription

[0040]

[0041] (II) Screening of the best extraction process

[0042] 1. Water extraction process screening

[0043] Using L9(3 4) table for orthogonal experimental design, with extract yield and hyperoside content as evaluation indicators, water addition (A, mL / g), extraction time (B, h), and extraction times (C, times) as factors, and comprehensive scoring method for evaluation, with weight coefficients of 50% and 50% respectively, and comprehensive score = extract yield / maximum extract yield × 50% × 100 + hyperoside content / maximum hyperoside content × 50% × 100. According to the prescription weight ratio of Rhododendron australis: Cardamom: Alpinia officinalis (2:1:1), 9 portions of medicinal material mixture were weighed, each portion was 20g, and the optimal level of process combination was selected. The factor level design is shown in Table 2. From the experimental results shown in Table 3, it can be seen that the influence of each factor is C>A>B, that is, the number of extractions has the greatest influence on the extraction effect, followed by the water addition, and finally the extraction time. The best extraction process obtained by the orthogonal experiment is A2B1C3, that is, adding 15 times the amount of water and boiling for 3 times, each time for 1h. The optimal extraction process obtained by orthogonal experiment was used as the condition to verify the optimal process, that is, 15 times the amount of water was added to extract 3 times, each time for 1 hour. According to the prescription compatibility ratio, a total of 20g of medicinal materials were weighed, and three portions of medicinal materials were weighed in parallel, and 15 times the amount of water was added to extract 3 times, each time for 1 hour. All extracts were combined, filtered, and concentrated to a thick paste under reduced pressure. After drying in a vacuum drying oven at 60°C, the dry paste yield was calculated. The experimental results are shown in Table 4. The average extract yield was 23.69% (RSD = 1.32%, n = 3) and the average hyperoside content was 7.296mg / g (RSD = 1.02%, n = 3).

[0044] Table 2 Water extraction factor level table

[0045]

[0046] Table 3 Results of orthogonal test of water extraction process

[0047]

[0048]

[0049] Note: The standard curve used for content determination is Y=29.854X-0.1288R 2 =0.9999.

[0050] Table 4 Optimal water extraction process verification test

[0051]

[0052] Note: The standard curve used for content determination is Y=30.999X-0.1007R 2 =0.9999.

[0053] 2 Alcohol precipitation process screening

[0054] After the preliminary study of the above water extraction process route and process optimization test, the optimal water extraction process conditions were obtained, and the extraction process (adding 15 times the amount of water for extraction 3 times, each time for 1 hour) has been determined. However, the water extract usually contains a large amount of impurities such as resin colloid, starch, protein, etc. These components may cause precipitation, rancidity, etc. during the storage of the liquid medicine. Therefore, it is very necessary to purify the water extract not only to enrich the effective ingredients, but also to ensure the quality and stability of the liquid medicine.

[0055] On the basis of previous studies, the amount of crude drug per milliliter of liquid medicine (A, g / mL), alcohol content (B, %), and alcohol precipitation time (C, h) were used as the investigation factors, and the extract yield and hyperoside content were used as the optimization indicators of alcohol precipitation conditions. 4 ) table for orthogonal experimental design, and the comprehensive scoring method was used for evaluation, with the weight coefficients of 50% and 50% respectively, and the comprehensive score = extract yield / maximum extract yield × 50% × 100 + hyperoside content / maximum hyperoside content × 50% × 100. According to the prescription ratio of Rhododendron australis: Cardamom: Alpinia officinalis (2:1:1), 9 portions of medicinal material mixture were weighed, each 200g, and the optimal level of process combination was selected. The factor level design is shown in Table 5. From the experimental results shown in Table 6, it can be seen that the influence of the three factors is A>B>C, that is, the amount of crude drug contained in each milliliter of liquid has the greatest influence on the alcohol precipitation effect, followed by the alcohol content, and finally the alcohol precipitation time. The optimal alcohol precipitation process obtained by the orthogonal experiment is A1B3C2, that is, the amount of crude drug contained in each milliliter of liquid is 0.5 (g / mL), the alcohol content of the liquid is 80% (volume fraction), and the alcohol precipitation time is 18 hours.

[0056] Table 5 Factor level table

[0057]

[0058] Table 6 Orthogonal test results of alcohol precipitation process

[0059]

[0060] Note: The standard curve used for content determination is Y=29.43X+0.2623R 2 =0.9999.

[0061] Example 2

[0062] The effect of this prescription on preventing lung injury caused by hypobaric hypoxia

[0063] 60 SPF C57BL / 6 male mice, weighing 20g±2g, were housed in the SPF animal barrier of the Experimental Animal Center of South-Central University for Nationalities. The experiment officially began after the mice were adapted to feeding for one week. The mice were randomly divided into 5 groups, 12 in each group: normal control group (Control), hypobaric hypoxia model group (HHM), acetazolamide administration group (ACTZ, 0.0758g / kg), low-dose group of this group (SWDJ-L, 2.275g / kg) and high-dose group of this group (SWDJ-H, 11.375g / kg), pre-administered for 14 days.

[0064] One hour after intragastric administration on the 14th day, except for the blank control group, the other groups were placed in a low-pressure simulation cabin for experimental animals to simulate plateau environment exposure for 24 hours, with food and drinking water guaranteed, and ascended to a simulated altitude of 6000m at a speed of 10m / s. After the model was completed, respiratory function testing, blood cell analysis, lung tissue water content determination, lung tissue hematoxylin-eosin staining, lung tissue TUNEL staining, immunohistochemical analysis and other experimental methods were used to comprehensively evaluate the effect of this formula in preventing lung injury caused by low pressure and hypoxia.

[0065] (I) Effect of this prescription on the lung function of low-pressure and low-oxygen mice

[0066] Pulmonary function indicators can determine the status of airway patency, respiratory muscle strength, lung tissue elasticity, ventilation function, storage capacity, etc. when lung diseases are disordered. They are sensitive indicators reflecting the degree of lung damage. Therefore, this experiment used a small animal whole-body non-invasive pulmonary function instrument to detect the basic lung function data of high-altitude hypoxic mice. There were 8 samples in each group. The detection indicators included mouse respiratory rate (F), minute ventilation (Mv), cumulative air volume (Av), maximum inspiratory flow rate (PIF), maximum expiratory flow rate (PEF), 50% maximum expiratory flow rate (EF50), end-expiratory pause (EEP), end-inspiratory pause (EIP), relaxation time (TR), tidal volume (VT), inspiration-expiration time ratio (Ti / Te), etc. Figure 1 As shown, compared with the normal group, the lung function test of mice with hypobaric hypoxia-induced lung injury showed an increase in f, MV, AV, PIF, PEF and EF50% ( Figure 1 moderate AF), decreased EEP, EIP, and RT ( Figure 1 GI), while this formula reversed the pulmonary dysfunction, but there was no significant difference in VT and Ti / Te ( Figure 1 (J, K).

[0067] (II) The regulatory effect of this formula on the hematological parameters of low-pressure and low-oxygen mice

[0068] The results are as follows Figure 2As shown in the figure, compared with the normal group, the concentrations of white blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), and neutrophil granulocytes (NEUT) in the hypobaric hypoxia model group were significantly increased, and the hematocrit (HCT) was significantly increased, however, the mean corpuscular volume (MCV) level did not change significantly. The treatment of this formula (SWDJ) significantly reduced the HCT of hypoxic mice and the concentrations of RBC, HGB, WBC, and NEUT in the blood, among which the high-dose group of this formula (SWDJ) had the most obvious effect, which was basically equivalent to acetazolamide.

[0069] (III) Protective effect of this formula on lung tissue of hypobaric and hypoxic mice

[0070] The results showed that compared with the normal control group, the lung tissue of the hypobaric hypoxic model group showed obvious black spots, thickened alveolar septa, collapsed or even disappeared alveolar cavities, and compensatory expansion of some alveolar cavities. Plasma components leaked from blood vessels and accumulated in the alveoli. A large number of inflammatory cells infiltrated, mainly neutrophils, and a small number of lung epithelial cells necrotized and fell off. TUNE staining results showed that hypoxia caused a large number of alveolar epithelial cells to apoptosis. Both ACTZ and the extract of this formula (SWDJ) significantly improved the pathological changes of hypobaric hypoxic lung tissue and reduced the apoptosis of lung epithelial cells ( Figure 3 In addition, compared with the normal control group, the lung water content in the hypobaric hypoxia model group was significantly increased, while the positive control group ACTZ and the extract of this formula (SWDJ) significantly reduced the lung water content, among which the high dose had the most obvious effect ( Figure 3 (B).

[0071] (IV) Effect of this prescription on the expression of HIF-1α and AQP1 in lung tissue of hypobaric and hypoxic mice

[0072] The study found that compared with the normal control group, the expression of hypoxia-inducible factor HIF-1a in lung tissue of the hypobaric hypoxia model group was significantly increased ( Figure 4 AQP1 expression, which reflects the degree of pulmonary edema, increased significantly ( Figure 4 B, D), while the high dose of this formula (SWDJ) and ACTZ could inhibit the expression of HIF-1a and AQP1 in lung tissues to varying degrees.

[0073] (V) Conclusion

[0074] The present invention uses multiple indicators to prove the preventive effect of this prescription (SWDJ) on lung injury in mice caused by hypobaric hypoxia. The hypoxia and cold conditions of the plateau environment have a significant effect on lung function, leading to dyspnea and increased respiratory rate. In individuals rising from the plain to the plateau, hypoxia leads to an increase in respiratory rate and depth, which leads to an accelerated F value in the hypobaric hypoxic model group (HHM). In addition, the minute ventilation (MV) increased significantly, while the respiratory time ratio (Ti / Te) decreased significantly, which may be related to enhanced respiratory drive. The results of this study showed that the water content of the lung tissue in the hypobaric hypoxic model group (HHM) increased, and the expression of the water channel protein AQP1 increased, both indicating that pulmonary edema occurred in the model group, and this prescription (SWDJ) can significantly reduce the water content of the lung tissue and the expression of AQP1, and has a protective effect. In addition, the treatment of this prescription (SWDJ) effectively inhibited the apoptosis of lung epithelial cells in the lung tissue induced by hypobaric hypoxia and pathological damage to the lung tissue. Based on the above research results, it is concluded that (SWDJ) has a protective effect on low-pressure and hypoxia-induced lung injury in mice.

[0075] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation methods of the present invention. For ordinary technical users in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. Use of a Chinese medicine composition in the preparation of a drug for preventing and / or treating lung injury caused by hypobaric hypoxia, characterized in that: The traditional Chinese medicine composition is prepared from the following components in parts by weight: 2 parts of Rhododendron australis, 1 part of Cardamomum villosum and 1 part of Alpinia officinalis.

2. The use according to claim 1, characterized in that: The preparation method of the Chinese medicine composition comprises the following steps: (1) extracting Rhododendron australis, Elaeagnus cardamomum and Alpinia officinalis with water, and concentrating under reduced pressure to obtain a water extract; (2) The water extract is subjected to alcohol precipitation to obtain the Chinese medicine composition.

3. The use according to claim 2, characterized in that: The water extraction conditions are as follows: the medicinal material is added with 15 mL / g of water and boiled for extraction 3 times, with each extraction time being 1 hour.

4. The use according to claim 2, characterized in that: The method of vacuum concentration is: vacuum concentration until the amount of crude drug contained in each milliliter of drug solution is 0.5 g / mL.

5. The use according to claim 2, characterized in that: The alcohol precipitation conditions are as follows: the alcohol content of the drug solution is 80% and the alcohol precipitation time is 18 hours.

6. The use according to claim 1, characterized in that: The application includes at least one of the following applications: (1) Use of a Chinese medicine composition in the preparation of a drug for preventing and / or treating pulmonary dysfunction caused by hypobaric hypoxia; (2) Use of Chinese medicine compositions in the preparation of blood regulation drugs; (3) Use of a Chinese medicine composition in the preparation of a drug for reducing lung water content; (4) Use of the Chinese medicine composition in the preparation of a drug for inhibiting the expression of HIF-1a and AQP1 in lung tissue.

7. The use according to claim 1, characterized in that: The drug also includes a pharmaceutically acceptable carrier.

8. The use according to claim 7, characterized in that: The pharmaceutically acceptable carrier is suitable for powders, granules, tablets, pills, capsules, oral liquids or pastes.

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